{"gene":"TNNI3","run_date":"2026-04-28T21:42:59","timeline":{"discoveries":[{"year":1996,"finding":"The human cardiac troponin I gene (TNNI3) comprises eight exons within 6.2 kb of genomic DNA. Transfection experiments showed that 2300 bp of 5'-flanking sequence drives expression in cardiac myocytes and skeletal muscle cells but not fibroblasts, and several conserved putative cis-acting elements were identified in the proximal promoter, indicating that additional sequences are required to confer cardiac specificity.","method":"Genomic cloning, sequencing, transfection reporter assays in cardiac myocytes and fibroblasts","journal":"Genomics","confidence":"Medium","confidence_rationale":"Tier 2 — direct functional transfection assay; single lab study","pmids":["8661099"],"is_preprint":false},{"year":1997,"finding":"Mutations in TNNI3 (cardiac troponin I) were identified as a cause of hypertrophic cardiomyopathy, establishing cTnI as an HCM disease gene. Key mutations included Arg145Gly (linked by family analysis) and Lys206Gln (de novo), demonstrating that the inhibitory subunit of the troponin complex is essential for normal cardiac function.","method":"Candidate gene sequencing in 184 unrelated HCM patients, family linkage analysis, identification of de novo mutations","journal":"Nature genetics","confidence":"High","confidence_rationale":"Tier 2 — genetic epistasis/linkage in large cohort, replicated across multiple independent labs subsequently","pmids":["9241277"],"is_preprint":false},{"year":1998,"finding":"The crystal structure of troponin C in complex with the N-terminal fragment of troponin I (TnI residues 1–47) at 2.3 Å resolution revealed that TnC adopts a compact globular conformation (vs. elongated dumbbell in free TnC) with the central helix bent 90°. The TnI1–47 alpha-helix spans the surface of TnC, stabilizing the compact conformation via contacts with both N- and C-terminal lobes, with the amphiphilic C-terminus of TnI1–47 anchored in the hydrophobic pocket of the TnC C-lobe.","method":"X-ray crystallography (2.3 Å), single isomorphous replacement + MAD phasing","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1 — crystal structure at atomic resolution","pmids":["9560191"],"is_preprint":false},{"year":1999,"finding":"NMR structure of the cardiac TnC N-terminal regulatory domain (cNTnC) in complex with Ca2+ and the cTnI switch peptide (residues 147–163) showed that cTnI147–163 binding induces an 'open' conformation of cNTnC (similar to Ca2+-saturated skeletal TnC), even though Ca2+ alone does not open cNTnC. The cTnI peptide adopts an alpha-helical conformation at residues 150–157 bound in the hydrophobic pocket of cNTnC, establishing the molecular basis for Ca2+-dependent activation of cardiac muscle.","method":"Multinuclear multidimensional NMR spectroscopy, solution structure determination","journal":"Biochemistry","confidence":"High","confidence_rationale":"Tier 1 — NMR structure with functional validation of binding mechanism","pmids":["10387074"],"is_preprint":false},{"year":2003,"finding":"The crystal structure of the core domain of human cardiac troponin (TnC + TnI + TnT) in the Ca2+-saturated form revealed that the core domain is divided into structurally distinct subdomains connected by flexible linkers. An alpha-helical coiled-coil between TnT and TnI forms the rigid IT arm (~80 Å), which bridges tropomyosin-anchoring regions. Ca2+ binding to the regulatory site of TnC removes the C-terminal portion of TnI from actin, altering mobility/flexibility of troponin and tropomyosin on the actin filament.","method":"X-ray crystallography (crystal structures at 46 kDa and 52 kDa core domain constructs)","journal":"Nature","confidence":"High","confidence_rationale":"Tier 1 — landmark crystal structure of ternary troponin complex","pmids":["12840750"],"is_preprint":false},{"year":2003,"finding":"TNNI3 mutations were identified as causes of idiopathic restrictive cardiomyopathy (RCM). In a large family with both RCM and HCM, linkage to TNNI3 was established (lod score 4.8) and a novel missense mutation identified. Screening nine additional unrelated RCM patients found TNNI3 mutations in six, including two de novo mutations in young patients. All mutations mapped to conserved, functionally important domains, establishing TNNI3 as an RCM susceptibility gene.","method":"Linkage analysis, candidate gene mutation screening, family co-segregation studies","journal":"The Journal of clinical investigation","confidence":"High","confidence_rationale":"Tier 2 — linkage analysis plus multi-family replication; established new disease association","pmids":["12531876"],"is_preprint":false},{"year":2004,"finding":"Muscle-specific RING finger protein 1 (MuRF1) was identified as a bona fide ubiquitin E3 ligase that ubiquitylates and degrades cardiac troponin I (cTnI). Yeast two-hybrid screening identified cTnI as a MuRF1 interactor; co-IP confirmed the association in vitro and in cardiomyocytes. MuRF1 reduced steady-state cTnI levels via RING finger-dependent ubiquitin ligase activity, accumulation of poly-ubiquitylated cTnI intermediates, and proteasome-dependent degradation. MuRF1 overexpression in cardiomyocytes reduced contractility indices.","method":"Yeast two-hybrid screen, Co-immunoprecipitation, in vitro ubiquitylation assay, cardiomyocyte overexpression with contractility readout","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1-2 — in vitro reconstitution of ubiquitylation + multiple cellular assays + functional outcome","pmids":["15601779"],"is_preprint":false},{"year":2004,"finding":"PKC phosphorylation of cTnI at Ser43 and Ser45 modulates cardiac contractility in vivo. Transgenic mice with Ala substitutions at these sites (preventing PKC phosphorylation) showed 30% greater +dP/dt and 18% greater −dP/dt at baseline compared with wild-type, and had negligible inotropic response to isoproterenol or phenylephrine/propranolol. Mutation of PKC sites was also associated with enhanced PKA-dependent phosphorylation of cTnI, demonstrating interdependence of phosphorylation sites.","method":"Transgenic mouse model (Ala substitutions at S43/S45), in situ hemodynamics, back-phosphorylation assays, cAMP measurements","journal":"American journal of physiology. Heart and circulatory physiology","confidence":"High","confidence_rationale":"Tier 2 — in vivo genetic loss-of-function with defined hemodynamic phenotype and biochemical follow-up","pmids":["14726296"],"is_preprint":false},{"year":2006,"finding":"DSCR1L2 (Down Syndrome Critical Region gene 1-like 2) protein interacts directly with cardiac troponin I (TNNI3). Interaction was identified by yeast two-hybrid screening of a human heart cDNA library and confirmed by yeast co-transformation and GST fusion protein pull-down assays. A novel DSCR1L2 splice isoform lacking two central exons was identified; both isoforms interact with TNNI3, and exon 2 of DSCR1L2 is critical for binding, suggesting a possible role for this calcineurin-pathway family member in cardiac contraction.","method":"Yeast two-hybrid screening (human heart cDNA library), yeast co-transformation, GST pulldown assay, quantitative RT-PCR","journal":"Gene","confidence":"Medium","confidence_rationale":"Tier 3 — confirmed by two orthogonal binding assays (yeast two-hybrid + GST pulldown) but no functional consequence established","pmids":["16516408"],"is_preprint":false},{"year":2007,"finding":"A novel one-nucleotide frameshift deletion in TNNI3 (exon 7) causing premature stop codon and truncation of the C-terminal portion of cTnI was found to cause restrictive cardiomyopathy. Western blot analysis of myocardial tissue showed approximately 50% reduction in total troponin I content, indicating haploinsufficiency of cTnI protein as a mechanism for RCM.","method":"Candidate gene sequencing, Western blot of myocardial tissue","journal":"International journal of cardiology","confidence":"Medium","confidence_rationale":"Tier 2-3 — Western blot directly quantified protein reduction in patient tissue; single case","pmids":["18006163"],"is_preprint":false},{"year":2007,"finding":"MuRF1 with the E2 enzyme UbcH5 synthesizes mixed forked ubiquitin chains on troponin I containing all seven possible linkages (predominantly Lys48, Lys63, Lys11), and troponin I linked to these forked chains was degraded poorly by purified 26S proteasomes compared to troponin I linked to homogeneous Lys48 or Lys63 chains. With UbcH13/Uev1a, MuRF1 attaches Lys63 chains, and with UbcH1 it attaches Lys48 chains, both of which support efficient degradation, establishing that chain topology determines proteasomal degradation efficiency of cTnI.","method":"In vitro ubiquitylation assay with purified components, mass spectrometry of Ub chain linkages, 26S proteasome degradation assay","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 — fully reconstituted in vitro system with MS characterization and degradation assays","pmids":["17426036"],"is_preprint":false},{"year":2008,"finding":"Expression of the HCM-associated cTnI-R145G mutation in adult rat cardiomyocytes significantly decreased baseline sarcomere shortening. Upon β-adrenergic stimulation with isoproterenol, rates of shortening and re-lengthening were depressed in cTnI-R145G-expressing cells but comparable to controls when using the β2-selective blocker ICI118,551, indicating that the R145G mutation alters β-adrenergic receptor subtype-dependent modulation of contractility.","method":"Adenovirus-mediated expression of cTnI-R145G in adult rat cardiomyocytes, sarcomere shortening measurements, pharmacological β-adrenergic dissection","journal":"Pflugers Archiv : European journal of physiology","confidence":"Medium","confidence_rationale":"Tier 2 — direct expression of mutant protein in primary cardiomyocytes with functional readout","pmids":["18548271"],"is_preprint":false},{"year":2010,"finding":"PAK3-mediated phosphorylation of cTnI at Ser151 (studied via S151E phosphomimic) increases Ca2+ sensitivity of myofilament by shortening inter-site distances between cTnC and cTnI (monitored by FRET in reconstituted thin filaments) and reducing kinetic rates of Ca2+-dissociation-induced structural changes in the regulatory region of cTnI. The structural effects mimic those of strong actomyosin crossbridges.","method":"Steady-state and time-resolved FRET, stopped-flow kinetics in reconstituted thin filaments containing cTnI-S151E","journal":"Journal of molecular biology","confidence":"High","confidence_rationale":"Tier 1 — in vitro reconstitution with multiple spectroscopic methods","pmids":["20540949"],"is_preprint":false},{"year":2013,"finding":"Phosphorylation of cTnI at serines 23/24 by PKA is a key regulator of length dependence of force generation (Frank-Starling mechanism) in cardiac myocytes. Exchange of unphosphorylated recombinant cTn into permeabilized rat cardiac myocytes or slow-twitch skeletal muscle fibers produced a shallow length-tension relationship; PKA treatment or use of cTn with S23/S24 mutated to Asp (phosphomimetic) restored a steep length-tension relationship.","method":"Troponin exchange into permeabilized cardiac myocytes and skeletal muscle fibers, PKA treatment, phosphomimetic mutation (S23D/S24D), force-length measurements","journal":"The Journal of physiology","confidence":"High","confidence_rationale":"Tier 1-2 — reconstitution approach in multiple muscle preparations with genetic and pharmacological validation","pmids":["23836688"],"is_preprint":false},{"year":2014,"finding":"The Ca2+-regulatory function of the cTnI inhibitory peptide (Ip) region requires cooperation with the C-terminus of cTnT. In vitro motility assays showed that cTnI-R145G (within the Ip) alone impaired switching off of actomyosin at low Ca2+; cTnT-R278C partially rescued high-Ca2+ defects of cTnI-R145G but exacerbated loss of inhibitory function at low Ca2+. Bioinformatics analysis revealed evolutionary and structural relatedness between the affected regions of cTnI and cTnT.","method":"In vitro motility assay with reconstituted troponin complexes (single and double mutants), varying Ca2+ concentration, temperature, and HMM density","journal":"Archives of biochemistry and biophysics","confidence":"Medium","confidence_rationale":"Tier 2 — in vitro reconstitution with functional readout; single lab","pmids":["24418317"],"is_preprint":false},{"year":2015,"finding":"Molecular dynamics simulations of the cTnI-R145G HCM mutation revealed that R145G does not alter overall cTn dynamics but stabilizes Ca2+-coordinating interactions in cTnC. The R145G mutation blunted intramolecular interactions between the cTnI N-terminal extension (residues 1–39) and the inhibitory peptide that are normally induced by PKA phosphorylation of S23/S24 (β-adrenergic signaling), providing a molecular explanation for why R145G reduces β-adrenergic modulation of cTn.","method":"Triplicate 150 ns molecular dynamics simulations of cTnC(1–161)–cTnI(1–172)–cTnT(236–285) complexes with R145G, R145G/S23D/S24D, and R145G/pS23/pS24","journal":"Biophysical journal","confidence":"Medium","confidence_rationale":"Tier 4 — computational simulation, but replicated triplicates with explicit comparison to phosphomimetic states","pmids":["25606687"],"is_preprint":false},{"year":2015,"finding":"TNNI3 encodes the inhibitory subunit of the troponin complex and has evolved from TNNI1 and TNNI2 (slow and fast skeletal isoforms) through vertebrate evolution. cTnI has a unique N-terminal cardiac-specific extension (residues 1–32) that is phosphorylated by PKA at S23/S24 and by PKC at S43/S45, and contains an inhibitory peptide region and a switch peptide that interact with cTnC. Developmental regulation switches expression from slow skeletal TNNI1 to TNNI3 postnatally in cardiac muscle.","method":"Comparative genomic and phylogenetic analysis, review of biochemical and PTM literature","journal":"Gene","confidence":"Medium","confidence_rationale":"Tier 3-4 — review synthesizing published experimental data; supported by extensive primary literature","pmids":["26526134"],"is_preprint":false},{"year":2016,"finding":"In cardiomyocytes from cTnI-G203S HCM mice, L-type Ca2+ channel (ICaL) inactivation was significantly faster. Activation of ICaL caused a greater increase in mitochondrial membrane potential and metabolic activity in cTnI-G203S myocytes compared to wild-type. This hypermetabolic response resulted from impaired communication between ICaL and F-actin, involving reduced actin-myosin dynamics and block of the mitochondrial voltage-dependent anion channel. L-type Ca2+ channel antagonists normalized mitochondrial membrane potential.","method":"Patch-clamp electrophysiology, mitochondrial membrane potential measurements, metabolic activity assays, pharmacological manipulation (nisoldipine, diltiazem) in isolated cardiomyocytes from transgenic cTnI-G203S mice","journal":"The Journal of physiology","confidence":"Medium","confidence_rationale":"Tier 2 — multiple electrophysiological and metabolic measurements in disease model with pharmacological rescue","pmids":["27062056"],"is_preprint":false},{"year":2016,"finding":"Estrogen treatment of neonatal mouse hearts increased HDAC activity and HDAC5 binding at the cTnI promoter, reduced histone H3K9 acetylation at the cTnI promoter, and decreased cTnI expression at both mRNA and protein levels. SAHA (an HDAC inhibitor) rescued cTnI expression by inhibiting HDAC5 binding and restoring H3K9 acetylation at the cTnI promoter, indicating that HDAC5-mediated histone deacetylation represses TNNI3 transcription.","method":"Chromatin immunoprecipitation (ChIP), colorimetric HDAC/HAT activity assays, RT-PCR, Western blot, pharmacological HDAC inhibition in neonatal mouse hearts","journal":"Journal of cellular biochemistry","confidence":"Medium","confidence_rationale":"Tier 2 — ChIP with functional pharmacological rescue; single lab","pmids":["27379430"],"is_preprint":false},{"year":2017,"finding":"EGCG (epigallocatechin-3-gallate) improved cardiac diastolic function in aged mice by reversing age-related low cTnI expression. Mechanistically, EGCG inhibited HDAC1 and HDAC3 expression and reduced HDAC1 binding to the cTnI proximal promoter, increased H3K9 acetylation at the promoter, and enhanced binding of transcription factors GATA4 and Mef2c to the cTnI promoter.","method":"ChIP assay, Western blot, echocardiography, HDAC1/HDAC3 expression analysis, transcription factor binding assay in aged mice treated with EGCG","journal":"Journal of cellular and molecular medicine","confidence":"Medium","confidence_rationale":"Tier 2 — ChIP and multiple molecular assays with in vivo functional readout; single lab","pmids":["28382690"],"is_preprint":false},{"year":2019,"finding":"cTnI is modified by arginine methylation in human myocardium. Mass spectrometry identified methylation sites at R74/R79 and R146/R148 in human cardiac samples. PRMT1 methylated an extended cTnI inhibitory peptide at R146 and R148 in vitro. HCM-associated mutations at R145 hampered R146/R148 methylation by PRMT1 in vitro. Methylation-deficient R146A/R148A cTnI expressed in H9c2 cells induced a 32% increase in cell size (hypertrophy), and Western blot showed reduced arginine methylation levels in HCM vs. DCM biopsies and in a rat cardiac hypertrophy model.","method":"Western blot with anti-methylarginine antibody, mass spectrometry, in vitro PRMT1 methylation assay, cell size measurement in H9c2 cells transfected with methylation-deficient mutant","journal":"International journal of cardiology","confidence":"Medium","confidence_rationale":"Tier 2 — mass spectrometry identification + in vitro enzyme assay + cell-based functional consequence; single lab","pmids":["30772011"],"is_preprint":false},{"year":2019,"finding":"In restrictive cardiomyopathy mice carrying the cTnI-193His mutation, PDE4d expression was epigenetically downregulated. cTnI was detected in the nucleus of cardiomyocytes by immunofluorescence and Western blot. Co-immunoprecipitation demonstrated direct interaction between cTnI and SMYD1 (histone methyltransferase) and between cTnI and HDAC1. Overexpression of mutated cTnI in cultured cardiomyocytes reduced PDE4d expression, suggesting nuclear cTnI modulates histone modifications and gene expression via interaction with chromatin-modifying enzymes.","method":"Immunofluorescence, Western blot of nuclear fractions, Co-IP, ChIP, cardiomyocyte overexpression studies","journal":"Science China. Life sciences","confidence":"Medium","confidence_rationale":"Tier 2-3 — multiple methods (nuclear fractionation, Co-IP, ChIP) but findings require independent replication","pmids":["30900165"],"is_preprint":false},{"year":2019,"finding":"Panel sequencing of 80 pediatric cardiomyopathy patients identified 5 pathogenic TNNI3 variants; one patient carried a homozygous truncating variant (c.24+2T>A). Protein and transcript analysis on heart biopsies from this individual showed complete absence of TNNI3 protein and compensatory upregulation of the fetal slow skeletal isoform TNNI1, demonstrating that total loss of cTnI triggers a developmental isoform switch.","method":"Next-generation sequencing panel, Western blot and transcript analysis of endomyocardial biopsy","journal":"Clinical genetics","confidence":"Medium","confidence_rationale":"Tier 2 — direct protein analysis of patient cardiac tissue demonstrating isoform compensation","pmids":["31568572"],"is_preprint":false},{"year":2020,"finding":"RCM-associated cTnI mutations R170G and R170W (in the regulatory C-terminus) strongly enhanced Ca2+ sensitivity in skinned cardiac fibers. Both mutants increased affinity of the troponin complex for tropomyosin; R170G strengthened and R170W weakened actin binding. Electron microscopy of reconstituted thin filaments showed wavy, broken filaments with R170G/W. MyBP-C N-terminal fragment (cMyBPC-C0C2) that normally reduces troponin binding to actin and increases cooperativity of thin filament activation had no effect in the presence of R170G/W cTn, and microscale thermophoresis identified direct cMyBPC-C0C2 binding to cTnI and cTnT (but not cTnC).","method":"Skinned fiber Ca2+-sensitivity measurements, electron microscopy of reconstituted thin filaments, co-sedimentation assays, microscale thermophoresis","journal":"PloS one","confidence":"High","confidence_rationale":"Tier 1-2 — multiple orthogonal methods (reconstitution, EM, MST, fiber mechanics) in one study","pmids":["32182250"],"is_preprint":false},{"year":2020,"finding":"miR-449 regulates cTnI expression in cardiomyocytes by targeting HDAC1. miR-449 sponges HDAC1, relieving HDAC1-mediated deacetylation of H3K4 and H3K9 at the cTnI promoter GATA element, which in turn recruits transcription factor GATA4 to increase TNNI3 transcription. In vivo, miR-449 agomiR intervention in aged mice with low cTnI expression restored cTnI levels and improved cardiac function.","method":"Dual-luciferase reporter assay (miR-449/HDAC1 binding site), biochemical analysis, in vivo miR-449 agomiR injection in aged mice, echocardiography","journal":"European review for medical and pharmacological sciences","confidence":"Medium","confidence_rationale":"Tier 2 — reporter assay + in vivo rescue with defined molecular mechanism; single lab","pmids":["33378032"],"is_preprint":false},{"year":2021,"finding":"De novo cTnI-D127Y mutation (infantile restrictive cardiomyopathy) increased Ca2+ sensitivity and reduced maximum force in skinned cardiomyocytes. In reconstituted thin filaments, cTnI-D127Y showed increased actin/tropomyosin binding affinity and compromised structural integrity (EM). Levosimendan and EGCG (troponin-targeting agents) partially stabilized thin filament structure and improved contractile parameters in vitro for both cTnI-D127Y and the companion cTnC-G34S mutation.","method":"Skinned fiber mechanics, reconstituted thin filament assays, electron microscopy, protein interaction binding assays, pharmacological intervention (levosimendan, EGCg)","journal":"International journal of molecular sciences","confidence":"Medium","confidence_rationale":"Tier 1-2 — reconstitution plus EM plus pharmacological rescue; single lab","pmids":["34502534"],"is_preprint":false},{"year":2024,"finding":"iPSC-derived cardiomyocytes and engineered heart tissues (EHT) from a patient carrying TNNI3 R170W showed prolonged tau (relaxation time constant) in Ca2+ kinetics and an increased ratio of relaxation force to contractile force. CRISPR/Cas9 isogenic correction of R170W reversed these defects. Overexpression of wild-type TNNI3 in R170W-iPSC-CMs and EHTs also rescued impaired relaxation, demonstrating that cTnI levels and function directly determine diastolic dysfunction in RCM.","method":"iPSC generation, CRISPR/Cas9 isogenic correction, EHT formation, Ca2+ kinetics imaging, force measurements, TNNI3 overexpression rescue","journal":"Development, growth & differentiation","confidence":"High","confidence_rationale":"Tier 2 — isogenic correction + overexpression rescue in patient-derived model with functional readout","pmids":["38193576"],"is_preprint":false},{"year":2024,"finding":"Patient-specific iPSC-derived cardiomyocytes heterozygous and homozygous for TNNI3 R170W showed impaired diastolic function (cell motion analysis) compared to CRISPR-corrected isogenic lines and three independent healthy controls. Intracellular Ca2+ oscillation and troponin I immunocytochemistry were not significantly altered, and myofibril/mitochondrial ultrastructure appeared intact by electron microscopy, but RNA-seq revealed altered pathways in cardiac muscle development/contraction, extracellular matrix-receptor interaction, and TGF-β signaling.","method":"iPSC-CM differentiation, CRISPR/Cas9 isogenic lines, cell motion analysis, Ca2+ imaging, immunocytochemistry, electron microscopy, RNA sequencing","journal":"Journal of the American Heart Association","confidence":"Medium","confidence_rationale":"Tier 2 — isogenic comparison with multiple readouts; transcriptomic data are associative","pmids":["38497452"],"is_preprint":false}],"current_model":"TNNI3-encoded cardiac troponin I (cTnI) is the inhibitory subunit of the sarcomeric troponin complex, whose switch peptide (residues 147–163) binds the N-terminal regulatory domain of cTnC in a Ca2+-dependent manner (inducing the open cTnC conformation) while its inhibitory peptide (residues ~128–147) normally restrains actin-tropomyosin; the cardiac-specific N-terminal extension is phosphorylated by PKA at S23/S24 (increasing length-dependent Ca2+ sensitivity and Frank-Starling response) and by PKC at S43/S45 (reducing contractility), PAK3 at S151 (increasing Ca2+ sensitivity via altered cTnC–cTnI distances), and by PRMT1 at R146/R148 (arginine methylation that modulates inhibitory/switch peptide function); cTnI is degraded by the E3 ubiquitin ligase MuRF1 in a RING finger- and UbcH5-dependent manner through mixed-linkage polyubiquitin chains; a fraction of cTnI localizes to the cardiomyocyte nucleus where it interacts with HDAC1 and SMYD1 to epigenetically regulate target genes including PDE4d; and TNNI3 transcription is regulated by HDAC1/HDAC3/HDAC5-mediated histone deacetylation at its promoter, counteracted by GATA4 and MEF2C binding, with mutations throughout the gene causing HCM, RCM, or DCM depending on their functional impact on Ca2+ sensitivity and thin filament structure."},"narrative":{"teleology":[{"year":1996,"claim":"Defining the genomic organization and cardiac-enriched promoter of TNNI3 established a framework for understanding its transcriptional regulation, revealing that 2.3 kb of 5′-flanking sequence drives expression in cardiac but not non-muscle cells, though full cardiac specificity required additional elements.","evidence":"Genomic cloning, sequencing, and transfection reporter assays in cardiac myocytes and fibroblasts","pmids":["8661099"],"confidence":"Medium","gaps":["Minimal regulatory elements sufficient for cardiac specificity not mapped","No in vivo transgenic validation of promoter constructs"]},{"year":1997,"claim":"Identification of TNNI3 mutations in HCM families established cTnI as a disease gene and demonstrated that the inhibitory subunit of the troponin complex is essential for normal cardiac function, opening the mechanistic question of how specific residue changes alter contractility.","evidence":"Candidate gene sequencing in 184 unrelated HCM patients with family linkage analysis","pmids":["9241277"],"confidence":"High","gaps":["Molecular mechanism by which individual mutations (e.g., R145G) alter troponin function not yet determined","No functional reconstitution at this stage"]},{"year":1998,"claim":"The first crystal structure of a TnC–TnI fragment complex revealed that TnI binding converts TnC from an elongated to a compact globular conformation, providing the first atomic-level view of how the inhibitory subunit contacts both lobes of TnC.","evidence":"X-ray crystallography at 2.3 Å resolution with MAD phasing","pmids":["9560191"],"confidence":"High","gaps":["Only N-terminal TnI fragment (1–47) resolved; inhibitory and switch peptide regions not visualized","No Ca²⁺-dependent conformational changes captured"]},{"year":1999,"claim":"NMR resolution of the cTnI switch peptide (147–163) bound to the Ca²⁺-loaded cTnC N-terminal domain revealed that cTnI binding—not Ca²⁺ alone—induces the open conformation of cNTnC, establishing the molecular basis for Ca²⁺-dependent activation of cardiac muscle.","evidence":"Multinuclear multidimensional NMR spectroscopy and solution structure determination","pmids":["10387074"],"confidence":"High","gaps":["Isolated peptide–domain complex; behavior in the context of the full troponin–tropomyosin–actin filament not addressed","Inhibitory peptide conformation not resolved simultaneously"]},{"year":2003,"claim":"Two landmark advances: the crystal structure of the ternary cardiac troponin core domain revealed the IT arm coiled-coil architecture and Ca²⁺-dependent release of cTnI from actin, while genetic linkage and multi-family screening established TNNI3 as a cause of restrictive cardiomyopathy in addition to HCM.","evidence":"X-ray crystallography of ternary troponin core domain (Nature); linkage analysis (LOD 4.8) plus mutation screening in RCM families (JCI)","pmids":["12840750","12531876"],"confidence":"High","gaps":["Full-length troponin on the thin filament not structurally resolved","Mechanism by which the same gene causes HCM vs. RCM not clarified at structural level"]},{"year":2004,"claim":"Parallel discoveries showed that cTnI is a substrate of the E3 ubiquitin ligase MuRF1 (linking proteasomal degradation to contractile protein turnover) and that PKC phosphorylation at S43/S45 attenuates contractility in vivo, with cross-talk between PKC and PKA phosphorylation sites.","evidence":"Yeast two-hybrid, co-IP, in vitro ubiquitylation, and cardiomyocyte contractility assays for MuRF1; transgenic S43A/S45A mice with in situ hemodynamics for PKC sites","pmids":["15601779","14726296"],"confidence":"High","gaps":["Physiological signals controlling MuRF1-mediated cTnI turnover rates in vivo unclear","PKC isoform specificity for S43/S45 phosphorylation in intact heart not defined"]},{"year":2007,"claim":"Reconstitution of MuRF1-mediated ubiquitylation with defined E2 enzymes revealed that ubiquitin chain topology (mixed forked chains via UbcH5 vs. homogeneous K48 or K63 chains) determines cTnI degradation efficiency by the 26S proteasome, adding a regulatory layer to sarcomeric protein quality control.","evidence":"In vitro ubiquitylation with purified components, mass spectrometry of chain linkages, 26S proteasome degradation assays","pmids":["17426036"],"confidence":"High","gaps":["In vivo chain type preference on endogenous cTnI not confirmed","Whether deubiquitinases selectively edit specific chain types on cTnI is unknown"]},{"year":2010,"claim":"PAK3-mediated phosphorylation of cTnI at S151 was shown to increase myofilament Ca²⁺ sensitivity by shortening inter-site cTnC–cTnI distances and slowing Ca²⁺-dissociation-induced structural transitions, mimicking the effect of strong crossbridges and identifying a third kinase-specific regulatory input on cTnI.","evidence":"Steady-state and time-resolved FRET, stopped-flow kinetics in reconstituted thin filaments with S151E phosphomimetic","pmids":["20540949"],"confidence":"High","gaps":["In vivo contribution of PAK3 to cTnI phosphorylation at S151 under physiological stimuli not quantified","Interplay between S151 phosphorylation and S23/S24 or S43/S45 not tested"]},{"year":2013,"claim":"PKA phosphorylation of cTnI at S23/S24 was established as the molecular switch for length-dependent Ca²⁺ sensitivity (Frank–Starling mechanism), as unphosphorylated troponin exchange abolished steep length–tension relationships that were restored by PKA treatment or phosphomimetic substitution.","evidence":"Troponin exchange in permeabilized cardiac myocytes and skeletal fibers, PKA treatment, S23D/S24D phosphomimetics, force-length measurements","pmids":["23836688"],"confidence":"High","gaps":["Structural mechanism by which N-terminal extension phosphorylation transmits length sensing to the regulatory domain not resolved","Contribution of titin compliance to the observed effects not dissected"]},{"year":2015,"claim":"MD simulations of the R145G HCM mutation provided a molecular explanation for its blunted β-adrenergic response: R145G stabilizes Ca²⁺ coordination in cTnC and prevents PKA-phosphorylation-induced intramolecular contact between the N-terminal extension and the inhibitory peptide, consistent with experimental observation of impaired β-adrenergic modulation in R145G cardiomyocytes.","evidence":"Triplicate 150 ns molecular dynamics simulations of troponin subcomplexes with R145G and phosphomimetic variants; complemented by earlier cardiomyocyte sarcomere shortening data","pmids":["25606687","18548271"],"confidence":"Medium","gaps":["Computational prediction; direct experimental validation of altered intramolecular contacts in full-length cTnI-R145G lacking","Simulations limited to 150 ns timescale"]},{"year":2016,"claim":"Epigenetic regulation of TNNI3 transcription was uncovered: HDAC5 binds the cTnI promoter and deacetylates H3K9, repressing transcription—an effect reversed by HDAC inhibitor SAHA—adding a transcriptional regulatory layer to cTnI biology.","evidence":"ChIP, HDAC activity assays, RT-PCR, Western blot, and HDAC inhibitor treatment in neonatal mouse hearts","pmids":["27379430"],"confidence":"Medium","gaps":["Physiological contexts in which HDAC5 is recruited to the TNNI3 promoter beyond estrogen exposure unclear","Whether HDAC5 directly binds DNA or is recruited via a transcription factor complex not determined"]},{"year":2017,"claim":"HDAC1 and HDAC3 were further implicated in TNNI3 promoter repression, with GATA4 and MEF2C binding shown to activate transcription when HDAC-mediated deacetylation is relieved, defining a complete activator–repressor switch at the TNNI3 locus.","evidence":"ChIP, Western blot, echocardiography, and EGCG treatment in aged mice","pmids":["28382690"],"confidence":"Medium","gaps":["Direct HDAC1/3 recruitment mechanism not resolved","Whether age-related cTnI decline is a cause or consequence of cardiac dysfunction not established"]},{"year":2019,"claim":"Three concurrent advances expanded cTnI biology: (1) arginine methylation at R146/R148 by PRMT1 was discovered in human myocardium, with HCM mutations at R145 impairing this modification and methylation-deficient cTnI inducing hypertrophy; (2) nuclear cTnI was shown to interact with SMYD1 and HDAC1, epigenetically regulating PDE4d expression; (3) complete loss of cTnI protein in a patient with a homozygous truncating variant triggered compensatory re-expression of fetal TNNI1.","evidence":"Mass spectrometry, in vitro PRMT1 assays, H9c2 cell transfection (PMID:30772011); nuclear fractionation, co-IP, ChIP in RCM mice (PMID:30900165); Western blot/transcript analysis of endomyocardial biopsy (PMID:31568572)","pmids":["30772011","30900165","31568572"],"confidence":"Medium","gaps":["Nuclear cTnI findings require independent replication and quantification of nuclear vs. sarcomeric pools","Functional consequence of R146/R148 methylation on thin filament Ca²⁺ regulation not tested in reconstituted systems","Whether TNNI1 compensation is sufficient for long-term cardiac function unknown"]},{"year":2020,"claim":"Reconstitution and EM studies of RCM-associated R170G/W mutations showed increased tropomyosin affinity, disrupted thin filament structural integrity, and loss of cMyBP-C regulatory modulation—demonstrating that a single cTnI residue change can simultaneously affect multiple protein–protein interfaces within the thin filament.","evidence":"Skinned fiber mechanics, reconstituted thin filament EM, co-sedimentation, microscale thermophoresis","pmids":["32182250"],"confidence":"High","gaps":["Whether loss of MyBP-C modulation is a general feature of RCM mutations or specific to R170 unclear","In vivo consequence of altered thin filament integrity not assessed in animal model"]},{"year":2024,"claim":"Patient-derived iPSC-cardiomyocyte and engineered heart tissue models of TNNI3 R170W RCM demonstrated that the mutation directly causes diastolic dysfunction (prolonged relaxation), which was reversed by CRISPR isogenic correction and wild-type TNNI3 overexpression, establishing a causal link from genotype to diastolic phenotype and validating gene augmentation as a potential therapeutic strategy.","evidence":"iPSC-CM generation, CRISPR/Cas9 isogenic correction, EHT formation, Ca²⁺ kinetics imaging, force measurements, TNNI3 overexpression rescue","pmids":["38193576","38497452"],"confidence":"High","gaps":["Long-term in vivo efficacy of TNNI3 gene augmentation not tested","Whether TNNI3 overexpression causes dominant-negative effects at supraphysiological levels not evaluated","Transcriptomic changes (TGF-β, ECM pathways) observed in mutant cells await mechanistic dissection"]},{"year":null,"claim":"Key unresolved questions include: the structural basis for how individual TNNI3 mutations differentially cause HCM, RCM, or DCM; the physiological significance and regulation of nuclear cTnI; the in vivo dynamics and chain-type specificity of MuRF1-mediated cTnI turnover; and whether gene therapy or allele-specific silencing can correct TNNI3-associated cardiomyopathies.","evidence":"","pmids":[],"confidence":"Low","gaps":["No high-resolution cryo-EM structure of the full thin filament with disease-mutant troponin","Nuclear cTnI interactome and gene regulatory targets not comprehensively mapped","No clinical gene therapy trials for TNNI3 cardiomyopathies"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[3,4,12,13,14]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[4,14,23]},{"term_id":"GO:0005198","term_label":"structural molecule activity","supporting_discovery_ids":[4,23]}],"localization":[{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[4,6,23]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[21]}],"pathway":[{"term_id":"R-HSA-397014","term_label":"Muscle contraction","supporting_discovery_ids":[3,4,7,12,13,14,23,26]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[7,13,17]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[6,10,20]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[1,5,9,11,25,26]}],"complexes":["cardiac troponin complex (cTnC–cTnI–cTnT)","thin filament (troponin–tropomyosin–actin)"],"partners":["TNNC1","TNNT2","TRIM63","HDAC1","SMYD1","PRMT1","MYBPC3","RCAN3"],"other_free_text":[]},"mechanistic_narrative":"TNNI3 encodes cardiac troponin I (cTnI), the inhibitory subunit of the sarcomeric troponin complex that governs Ca²⁺-dependent regulation of cardiac muscle contraction. The cTnI switch peptide (residues 147–163) binds the N-terminal regulatory domain of troponin C in a Ca²⁺-dependent manner to induce its open conformation, while the inhibitory peptide (~residues 128–147) restrains actin–tropomyosin at low Ca²⁺; these interactions, resolved by crystal and NMR structures of the ternary troponin complex, are modulated by PKA phosphorylation at S23/S24 (governing length-dependent activation and the Frank–Starling mechanism), PKC phosphorylation at S43/S45 (reducing contractility), PAK3 phosphorylation at S151 (increasing Ca²⁺ sensitivity), and PRMT1-mediated arginine methylation at R146/R148 [PMID:10387074, PMID:12840750, PMID:23836688, PMID:14726296, PMID:20540949, PMID:30772011]. cTnI is targeted for proteasomal degradation by the E3 ubiquitin ligase MuRF1, which assembles mixed-linkage polyubiquitin chains via UbcH5 whose topology determines degradation efficiency [PMID:15601779, PMID:17426036]. Mutations throughout TNNI3 cause hypertrophic, restrictive, or dilated cardiomyopathy by altering Ca²⁺ sensitivity, thin-filament integrity, and diastolic relaxation, as demonstrated by family linkage studies, reconstituted fiber mechanics, and CRISPR-corrected patient iPSC-cardiomyocyte models [PMID:9241277, PMID:12531876, PMID:32182250, PMID:38193576]."},"prefetch_data":{"uniprot":{"accession":"P19429","full_name":"Troponin I, cardiac muscle","aliases":["Cardiac troponin I"],"length_aa":210,"mass_kda":24.0,"function":"Troponin I is the inhibitory subunit of troponin, the thin filament regulatory complex which confers calcium-sensitivity to striated muscle actomyosin ATPase activity","subcellular_location":"","url":"https://www.uniprot.org/uniprotkb/P19429/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/TNNI3","classification":"Not Classified","n_dependent_lines":3,"n_total_lines":1208,"dependency_fraction":0.0024834437086092716},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/TNNI3","total_profiled":1310},"omim":[{"mim_id":"620894","title":"CARDIOMYOPATHY, DILATED, 2K; CMD2K","url":"https://www.omim.org/entry/620894"},{"mim_id":"617608","title":"ALPHA KINASE 3; ALPK3","url":"https://www.omim.org/entry/617608"},{"mim_id":"617294","title":"EPIDERMOLYSIS BULLOSA SIMPLEX 6, GENERALIZED INTERMEDIATE, WITH OR WITHOUT CARDIOMYOPATHY; EBS6","url":"https://www.omim.org/entry/617294"},{"mim_id":"616117","title":"CARDIAC CONDUCTION DISEASE WITH OR WITHOUT CARDIOMYOPATHY 1; CCCM1","url":"https://www.omim.org/entry/616117"},{"mim_id":"615396","title":"LEFT VENTRICULAR NONCOMPACTION 10; LVNC10","url":"https://www.omim.org/entry/615396"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Cytosol","reliability":"Approved"},{"location":"Vesicles","reliability":"Additional"}],"tissue_specificity":"Tissue enriched","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"heart muscle","ntpm":10359.2}],"url":"https://www.proteinatlas.org/search/TNNI3"},"hgnc":{"alias_symbol":["TNNC1","CMH7","cTNI"],"prev_symbol":["CMD2A"]},"alphafold":{"accession":"P19429","domains":[{"cath_id":"1.20.5","chopping":"43-80","consensus_level":"medium","plddt":96.6466,"start":43,"end":80},{"cath_id":"1.20.5","chopping":"90-140","consensus_level":"medium","plddt":92.8155,"start":90,"end":140},{"cath_id":"1.20.5","chopping":"150-182","consensus_level":"medium","plddt":68.31,"start":150,"end":182}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P19429","model_url":"https://alphafold.ebi.ac.uk/files/AF-P19429-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P19429-F1-predicted_aligned_error_v6.png","plddt_mean":78.62},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=TNNI3","jax_strain_url":"https://www.jax.org/strain/search?query=TNNI3"},"sequence":{"accession":"P19429","fasta_url":"https://rest.uniprot.org/uniprotkb/P19429.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P19429/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P19429"}},"corpus_meta":[{"pmid":"20215591","id":"PMC_20215591","title":"Coding sequence rare variants identified in MYBPC3, MYH6, TPM1, TNNC1, and TNNI3 from 312 patients with familial or idiopathic dilated cardiomyopathy.","date":"2010","source":"Circulation. Cardiovascular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/20215591","citation_count":199,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"25277212","id":"PMC_25277212","title":"Calcium-dependent FAK/CREB/TNNC1 signalling mediates the effect of stromal MFAP5 on ovarian cancer metastatic potential.","date":"2014","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/25277212","citation_count":111,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"26526134","id":"PMC_26526134","title":"TNNI1, TNNI2 and TNNI3: Evolution, regulation, and protein structure-function relationships.","date":"2015","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/26526134","citation_count":108,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"18572189","id":"PMC_18572189","title":"Molecular and functional characterization of novel hypertrophic cardiomyopathy susceptibility mutations in TNNC1-encoded troponin C.","date":"2008","source":"Journal of molecular and cellular cardiology","url":"https://pubmed.ncbi.nlm.nih.gov/18572189","citation_count":106,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"26232335","id":"PMC_26232335","title":"Structure and function of cardiac troponin C (TNNC1): Implications for heart failure, cardiomyopathies, and troponin modulating drugs.","date":"2015","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/26232335","citation_count":98,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"35533372","id":"PMC_35533372","title":"Liposome-Embedded Cu2-AgS Nanoparticle-Mediated Photothermal Immunoassay for Daily Monitoring of cTnI Protein Using a Portable Thermal Imager.","date":"2022","source":"Analytical chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/35533372","citation_count":93,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"8661099","id":"PMC_8661099","title":"Isolation and characterization of the human cardiac troponin I gene (TNNI3).","date":"1996","source":"Genomics","url":"https://pubmed.ncbi.nlm.nih.gov/8661099","citation_count":73,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"9522914","id":"PMC_9522914","title":"The use of cardiac troponin-I (cTnI) to determine the incidence of myocardial ischemia and injury in patients with aneurysmal and presumed aneurysmal subarachnoid hemorrhage.","date":"1998","source":"Acta neurochirurgica","url":"https://pubmed.ncbi.nlm.nih.gov/9522914","citation_count":71,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"9857935","id":"PMC_9857935","title":"Biochemical differences between cTnT and cTnI and their significance for diagnosis of acute coronary syndromes.","date":"1998","source":"European heart journal","url":"https://pubmed.ncbi.nlm.nih.gov/9857935","citation_count":64,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"28271683","id":"PMC_28271683","title":"Clinical Value of Combined Detection of CK-MB, MYO, cTnI and Plasma NT-proBNP in Diagnosis of Acute Myocardial Infarction.","date":"2017","source":"Clinical laboratory","url":"https://pubmed.ncbi.nlm.nih.gov/28271683","citation_count":63,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"19150014","id":"PMC_19150014","title":"[Mutations in sarcomeric genes MYH7, MYBPC3, TNNT2, TNNI3, and TPM1 in patients with hypertrophic cardiomyopathy].","date":"2009","source":"Revista espanola de cardiologia","url":"https://pubmed.ncbi.nlm.nih.gov/19150014","citation_count":58,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"21439810","id":"PMC_21439810","title":"A fluoro-microbead guiding chip for simple and quantifiable immunoassay of cardiac troponin I (cTnI).","date":"2011","source":"Biosensors & bioelectronics","url":"https://pubmed.ncbi.nlm.nih.gov/21439810","citation_count":54,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"21533915","id":"PMC_21533915","title":"Recurrent and founder mutations in the Netherlands: cardiac Troponin I (TNNI3) gene mutations as a cause of severe forms of hypertrophic and restrictive cardiomyopathy.","date":"2011","source":"Netherlands heart journal : monthly journal of the Netherlands Society of Cardiology and the Netherlands Heart Foundation","url":"https://pubmed.ncbi.nlm.nih.gov/21533915","citation_count":52,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"22815480","id":"PMC_22815480","title":"A mutation in TNNC1-encoded cardiac troponin C, TNNC1-A31S, predisposes to hypertrophic cardiomyopathy and ventricular fibrillation.","date":"2012","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/22815480","citation_count":51,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"21832052","id":"PMC_21832052","title":"Functional characterization of TNNC1 rare variants identified in dilated cardiomyopathy.","date":"2011","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/21832052","citation_count":49,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"16158369","id":"PMC_16158369","title":"Responses of N-terminal pro-brain natriuretic peptide (NT-proBNP) and cardiac troponin I (cTnI) to competitive endurance exercise in recreational athletes.","date":"2005","source":"International journal of sports medicine","url":"https://pubmed.ncbi.nlm.nih.gov/16158369","citation_count":42,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"18992232","id":"PMC_18992232","title":"Early markers of myocardial injury: cTnI is enough.","date":"2008","source":"Clinica chimica acta; international journal of clinical chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/18992232","citation_count":41,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"25606687","id":"PMC_25606687","title":"Effects of HCM cTnI mutation R145G on troponin structure and modulation by PKA phosphorylation elucidated by molecular dynamics simulations.","date":"2015","source":"Biophysical journal","url":"https://pubmed.ncbi.nlm.nih.gov/25606687","citation_count":40,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"14726296","id":"PMC_14726296","title":"Inhibition of PKC phosphorylation of cTnI improves cardiac performance in vivo.","date":"2004","source":"American journal of physiology. Heart and circulatory physiology","url":"https://pubmed.ncbi.nlm.nih.gov/14726296","citation_count":40,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"30362071","id":"PMC_30362071","title":"Fisetin, a plant flavonoid ameliorates doxorubicin-induced cardiotoxicity in experimental rats: the decisive role of caspase-3, COX-II, cTn-I, iNOs and TNF-α.","date":"2018","source":"Molecular biology reports","url":"https://pubmed.ncbi.nlm.nih.gov/30362071","citation_count":40,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"26304555","id":"PMC_26304555","title":"In Vivo Analysis of Troponin C Knock-In (A8V) Mice: Evidence that TNNC1 Is a Hypertrophic Cardiomyopathy Susceptibility Gene.","date":"2015","source":"Circulation. Cardiovascular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/26304555","citation_count":38,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"36566596","id":"PMC_36566596","title":"Integrated solar-powered MEMS-based photoelectrochemical immunoassay for point-of-care testing of cTnI protein.","date":"2022","source":"Biosensors & bioelectronics","url":"https://pubmed.ncbi.nlm.nih.gov/36566596","citation_count":38,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"34256261","id":"PMC_34256261","title":"The TDs/aptamer cTnI biosensors based on HCR and Au/Ti3C2-MXene amplification for screening serious patient in COVID-19 pandemic.","date":"2021","source":"Biosensors & bioelectronics","url":"https://pubmed.ncbi.nlm.nih.gov/34256261","citation_count":38,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"27604170","id":"PMC_27604170","title":"Evidence for troponin C (TNNC1) as a gene for autosomal recessive restrictive cardiomyopathy with fatal outcome in infancy.","date":"2016","source":"American journal of medical genetics. Part A","url":"https://pubmed.ncbi.nlm.nih.gov/27604170","citation_count":37,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"31568572","id":"PMC_31568572","title":"Targeted panel sequencing in pediatric primary cardiomyopathy supports a critical role of TNNI3.","date":"2019","source":"Clinical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/31568572","citation_count":36,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"28382690","id":"PMC_28382690","title":"Epigallocatechin gallate reverses cTnI-low expression-induced age-related heart diastolic dysfunction through histone acetylation modification.","date":"2017","source":"Journal of cellular and molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/28382690","citation_count":35,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"12654317","id":"PMC_12654317","title":"Determination of affinities and antigenic epitopes of bovine cardiac troponin I (cTnI) with monoclonal antibodies by surface plasmon resonance biosensor.","date":"2003","source":"Analytical biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/12654317","citation_count":34,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"18006163","id":"PMC_18006163","title":"Deletion in TNNI3 gene is associated with restrictive cardiomyopathy.","date":"2007","source":"International journal of cardiology","url":"https://pubmed.ncbi.nlm.nih.gov/18006163","citation_count":33,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"23836688","id":"PMC_23836688","title":"Length dependence of striated muscle force generation is controlled by phosphorylation of cTnI at serines 23/24.","date":"2013","source":"The Journal of physiology","url":"https://pubmed.ncbi.nlm.nih.gov/23836688","citation_count":33,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"33165490","id":"PMC_33165490","title":"SERS-based magnetic immunoassay for simultaneous detection of cTnI and H-FABP using core-shell nanotags.","date":"2020","source":"Analytical methods : advancing methods and applications","url":"https://pubmed.ncbi.nlm.nih.gov/33165490","citation_count":29,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"24418317","id":"PMC_24418317","title":"Ca(2+)-regulatory function of the inhibitory peptide region of cardiac troponin I is aided by the C-terminus of cardiac troponin T: Effects of familial hypertrophic cardiomyopathy mutations cTnI R145G and cTnT R278C, alone and in combination, on filament sliding.","date":"2014","source":"Archives of biochemistry and biophysics","url":"https://pubmed.ncbi.nlm.nih.gov/24418317","citation_count":29,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"35517933","id":"PMC_35517933","title":"An ultrasensitive electrochemical sensing platform for the detection of cTnI based on aptamer recognition and signal amplification assisted by TdT.","date":"2020","source":"RSC advances","url":"https://pubmed.ncbi.nlm.nih.gov/35517933","citation_count":27,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"25074544","id":"PMC_25074544","title":"A fluorogenic heterogeneous immunoassay for cardiac muscle troponin cTnI on a digital microfluidic device.","date":"2014","source":"Analytical and bioanalytical chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/25074544","citation_count":26,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"25089838","id":"PMC_25089838","title":"The novel regulations of MEF2A, CAMKK2, CALM3, and TNNI3 in ventricular hypertrophy induced by arsenic exposure in rats.","date":"2014","source":"Toxicology","url":"https://pubmed.ncbi.nlm.nih.gov/25089838","citation_count":26,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"35623274","id":"PMC_35623274","title":"CRISPR/Cas12a-based electrochemical biosensor for highly sensitive detection of cTnI.","date":"2022","source":"Bioelectrochemistry (Amsterdam, Netherlands)","url":"https://pubmed.ncbi.nlm.nih.gov/35623274","citation_count":25,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"24899611","id":"PMC_24899611","title":"Diagnostic value of analysis of H-FABP, NT-proBNP, and cTnI in heart function in children with congenital heart disease and pneumonia.","date":"2014","source":"European review for medical and pharmacological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/24899611","citation_count":25,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"21262074","id":"PMC_21262074","title":"Novel frameshift mutation in Troponin C ( TNNC1) associated with hypertrophic cardiomyopathy and sudden death.","date":"2011","source":"Cardiology in the young","url":"https://pubmed.ncbi.nlm.nih.gov/21262074","citation_count":23,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"32927448","id":"PMC_32927448","title":"Aptasensor based on a flower-shaped silver magnetic nanocomposite enables the sensitive and label-free detection of troponin I (cTnI) by SERS.","date":"2020","source":"Nanotechnology","url":"https://pubmed.ncbi.nlm.nih.gov/32927448","citation_count":23,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"37185530","id":"PMC_37185530","title":"Chemiluminescence Biosensor for the Determination of Cardiac Troponin I (cTnI).","date":"2023","source":"Biosensors","url":"https://pubmed.ncbi.nlm.nih.gov/37185530","citation_count":21,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"27062056","id":"PMC_27062056","title":"The L-type Ca(2+) channel facilitates abnormal metabolic activity in the cTnI-G203S mouse model of hypertrophic cardiomyopathy.","date":"2016","source":"The Journal of physiology","url":"https://pubmed.ncbi.nlm.nih.gov/27062056","citation_count":21,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"23782526","id":"PMC_23782526","title":"Somatic MYH7, MYBPC3, TPM1, TNNT2 and TNNI3 mutations in sporadic hypertrophic cardiomyopathy.","date":"2013","source":"Circulation journal : official journal of the Japanese Circulation Society","url":"https://pubmed.ncbi.nlm.nih.gov/23782526","citation_count":20,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"32182250","id":"PMC_32182250","title":"Infantile restrictive cardiomyopathy: cTnI-R170G/W impair the interplay of sarcomeric proteins and the integrity of thin filaments.","date":"2020","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/32182250","citation_count":19,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"22876777","id":"PMC_22876777","title":"High prevalence of Arginine to Glutamine substitution at 98, 141 and 162 positions in Troponin I (TNNI3) associated with hypertrophic cardiomyopathy among Indians.","date":"2012","source":"BMC medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/22876777","citation_count":19,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"12412409","id":"PMC_12412409","title":"[Proinflammatory cytokines (IL-6, TNF-alpha) and cardiac troponin I (cTnI) in serum of young people with ventricular arrhythmias].","date":"2002","source":"Polskie Archiwum Medycyny Wewnetrznej","url":"https://pubmed.ncbi.nlm.nih.gov/12412409","citation_count":19,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"32638704","id":"PMC_32638704","title":"Characterization of TNNC1 as a Novel Tumor Suppressor of Lung Adenocarcinoma.","date":"2020","source":"Molecules and cells","url":"https://pubmed.ncbi.nlm.nih.gov/32638704","citation_count":18,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"30956276","id":"PMC_30956276","title":"Correlations of Changes in Brain Natriuretic Peptide (BNP) and Cardiac Troponin I (cTnI) with Levels of C-Reactive Protein (CRP) and TNF-α in Pediatric Patients with Sepsis.","date":"2019","source":"Medical science monitor : international medical journal of experimental and clinical research","url":"https://pubmed.ncbi.nlm.nih.gov/30956276","citation_count":17,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"26169204","id":"PMC_26169204","title":"Identification of rare variants in TNNI3 with atrial fibrillation in a Chinese GeneID population.","date":"2015","source":"Molecular genetics and genomics : MGG","url":"https://pubmed.ncbi.nlm.nih.gov/26169204","citation_count":16,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"25940119","id":"PMC_25940119","title":"Diagnostic disparity and identification of two TNNI3 gene mutations, one novel and one arising de novo, in South African patients with restrictive cardiomyopathy and focal ventricular hypertrophy.","date":"2015","source":"Cardiovascular journal of Africa","url":"https://pubmed.ncbi.nlm.nih.gov/25940119","citation_count":16,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"8825654","id":"PMC_8825654","title":"Mapping TNNC1, the gene that encodes cardiac troponin I in the human and the mouse.","date":"1995","source":"Genomics","url":"https://pubmed.ncbi.nlm.nih.gov/8825654","citation_count":15,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"16516408","id":"PMC_16516408","title":"Proteins encoded by human Down syndrome critical region gene 1-like 2 (DSCR1L2) mRNA and by a novel DSCR1L2 mRNA isoform interact with cardiac troponin I (TNNI3).","date":"2006","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/16516408","citation_count":15,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"20569525","id":"PMC_20569525","title":"Ventricular septal defect and restrictive cardiomyopathy in a paediatric TNNI3 mutation carrier.","date":"2010","source":"Cardiology in the young","url":"https://pubmed.ncbi.nlm.nih.gov/20569525","citation_count":15,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"11793431","id":"PMC_11793431","title":"Evaluation of Stratus CS stat fluorimetric analyser for measurement of cardiac markers Troponin I (cTnI), creatine kinase MB (CK-MB), and myoglobin.","date":"2001","source":"Journal of clinical laboratory analysis","url":"https://pubmed.ncbi.nlm.nih.gov/11793431","citation_count":15,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"30395933","id":"PMC_30395933","title":"Amish nemaline myopathy and dilated cardiomyopathy caused by a homozygous contiguous gene deletion of TNNT1 and TNNI3 in a Mennonite child.","date":"2018","source":"European journal of medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/30395933","citation_count":15,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"30900165","id":"PMC_30900165","title":"Epigenetic regulation of phosphodiesterase 4d in restrictive cardiomyopathy mice with cTnI mutations.","date":"2019","source":"Science China. Life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/30900165","citation_count":14,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"15641689","id":"PMC_15641689","title":"Role of heart-type fatty acid binding protein in early detection of acute myocardial infarction in comparison with cTnI, CK-MB and myoglobin.","date":"2004","source":"Journal of Huazhong University of Science and Technology. Medical sciences = Hua zhong ke ji da xue xue bao. Yi xue Ying De wen ban = Huazhong keji daxue xuebao. Yixue Yingdewen ban","url":"https://pubmed.ncbi.nlm.nih.gov/15641689","citation_count":14,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"32946432","id":"PMC_32946432","title":"TNNC1 Reduced Gemcitabine Sensitivity of Nonsmall-Cell Lung Cancer by Increasing Autophagy.","date":"2020","source":"Medical science monitor : international medical journal of experimental and clinical research","url":"https://pubmed.ncbi.nlm.nih.gov/32946432","citation_count":13,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"30772011","id":"PMC_30772011","title":"The inhibitory subunit of cardiac troponin (cTnI) is modified by arginine methylation in the human heart.","date":"2019","source":"International journal of cardiology","url":"https://pubmed.ncbi.nlm.nih.gov/30772011","citation_count":13,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"25673910","id":"PMC_25673910","title":"Clinical assessment and C-reactive protein (CRP), haptoglobin (Hp), and cardiac troponin I (cTnI) values of brachycephalic dogs with upper airway obstruction before and after surgery.","date":"2015","source":"Canadian journal of veterinary research = Revue canadienne de recherche veterinaire","url":"https://pubmed.ncbi.nlm.nih.gov/25673910","citation_count":13,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"33116603","id":"PMC_33116603","title":"LukS-PV Inhibits Hepatocellular Carcinoma Cells Migration via the TNNC1/PI3K/AKT Axis.","date":"2020","source":"OncoTargets and therapy","url":"https://pubmed.ncbi.nlm.nih.gov/33116603","citation_count":12,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"11858918","id":"PMC_11858918","title":"Improved identification of acute coronary syndromes with second generation cardiac troponin I assay: utility of 2-hour delta cTnI > or = +0.02 ng/mL.","date":"2002","source":"The Journal of emergency medicine","url":"https://pubmed.ncbi.nlm.nih.gov/11858918","citation_count":12,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"36981019","id":"PMC_36981019","title":"Homozygous TNNI3 Mutations and Severe Early Onset Dilated Cardiomyopathy: Patient Report and Review of the Literature.","date":"2023","source":"Genes","url":"https://pubmed.ncbi.nlm.nih.gov/36981019","citation_count":11,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"34502534","id":"PMC_34502534","title":"De Novo Missense Mutations in TNNC1 and TNNI3 Causing Severe Infantile Cardiomyopathy Affect Myofilament Structure and Function and Are Modulated by Troponin Targeting Agents.","date":"2021","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/34502534","citation_count":11,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"28856194","id":"PMC_28856194","title":"Transitioning high sensitivity cardiac troponin I (hs-cTnI) into routine diagnostic use: More than just a sensitivity issue.","date":"2016","source":"Practical laboratory medicine","url":"https://pubmed.ncbi.nlm.nih.gov/28856194","citation_count":11,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"11759175","id":"PMC_11759175","title":"Postoperative patterns and kinetics of cTnI, cTnT, CK-MB-activity and CK-activity after elective aortic valve replacement.","date":"2001","source":"Swiss medical weekly","url":"https://pubmed.ncbi.nlm.nih.gov/11759175","citation_count":10,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"30953456","id":"PMC_30953456","title":"A case report of recessive restrictive cardiomyopathy caused by a novel mutation in cardiac troponin I (TNNI3).","date":"2019","source":"BMC medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/30953456","citation_count":10,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"36074967","id":"PMC_36074967","title":"Expression and Clinical Significance of Serum sST2, BDNF, CTnI, and BUN/Cr in Patients With Heart Failure.","date":"2023","source":"Alternative therapies in health and medicine","url":"https://pubmed.ncbi.nlm.nih.gov/36074967","citation_count":10,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"29992111","id":"PMC_29992111","title":"Kolaviron attenuated arsenic acid induced-cardiorenal dysfunction via regulation of ROS, C-reactive proteins (CRP), cardiac troponin I (CTnI) and BCL2.","date":"2017","source":"Journal of traditional and complementary medicine","url":"https://pubmed.ncbi.nlm.nih.gov/29992111","citation_count":10,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"28382084","id":"PMC_28382084","title":"Diverse Phenotypic Expression of Cardiomyopathies in a Family with TNNI3 p.Arg145Trp Mutation.","date":"2017","source":"Korean circulation journal","url":"https://pubmed.ncbi.nlm.nih.gov/28382084","citation_count":9,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"20540949","id":"PMC_20540949","title":"Structural and kinetic effects of PAK3 phosphorylation mimic of cTnI(S151E) on the cTnC-cTnI interaction in the cardiac thin filament.","date":"2010","source":"Journal of molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/20540949","citation_count":9,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"33592378","id":"PMC_33592378","title":"TNNC1 knockout reverses metastatic potential of ovarian cancer cells by inactivating epithelial-mesenchymal transition and suppressing F-actin polymerization.","date":"2021","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/33592378","citation_count":8,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"26506446","id":"PMC_26506446","title":"A Double Heterozygous Mutation of TNNI3 Causes Hypertrophic Cardiomyopathy in a Han Chinese Family.","date":"2015","source":"Cardiology","url":"https://pubmed.ncbi.nlm.nih.gov/26506446","citation_count":8,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"35704727","id":"PMC_35704727","title":"Positive Relationship of RDW with NT-proBNP and cTnI in Acute Myocardial Infarction Patients.","date":"2022","source":"Clinical laboratory","url":"https://pubmed.ncbi.nlm.nih.gov/35704727","citation_count":8,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"33378032","id":"PMC_33378032","title":"MiR-449 improves cardiac function by regulating HDAC1 and cTnI.","date":"2020","source":"European review for medical and pharmacological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/33378032","citation_count":8,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"35838873","id":"PMC_35838873","title":"Molecular Diagnosis of Primary Cardiomyopathy in 231 Unrelated Pediatric Cases by Panel-Based Next-Generation Sequencing: A Major Focus on Five Carriers of Biallelic TNNI3 Pathogenic Variants.","date":"2022","source":"Molecular diagnosis & therapy","url":"https://pubmed.ncbi.nlm.nih.gov/35838873","citation_count":8,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"25141318","id":"PMC_25141318","title":"Role of serum myeloperoxidase, CPK, CK-MB, and cTnI tests in early diagnosis of myocardial ischemia during ERCP.","date":"2014","source":"The Turkish journal of gastroenterology : the official journal of Turkish Society of Gastroenterology","url":"https://pubmed.ncbi.nlm.nih.gov/25141318","citation_count":8,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"35687951","id":"PMC_35687951","title":"CTnI diagnosis in myocardial infarction using G-quadruplex selective Ir(Ⅲ) complex as effective electrochemiluminescence probe.","date":"2022","source":"Talanta","url":"https://pubmed.ncbi.nlm.nih.gov/35687951","citation_count":8,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"30279906","id":"PMC_30279906","title":"The TNNI3 Arg192His mutation in a 13-year-old girl with left ventricular noncompaction.","date":"2018","source":"Journal of cardiology cases","url":"https://pubmed.ncbi.nlm.nih.gov/30279906","citation_count":7,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"38265681","id":"PMC_38265681","title":"Paeonol upregulates expression of tumor suppressors TNNC1 and SCARA5, exerting anti-tumor activity in non-small cell lung cancer cells.","date":"2024","source":"Naunyn-Schmiedeberg's archives of pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/38265681","citation_count":7,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"34113113","id":"PMC_34113113","title":"The Diagnostic Value of Plasma miRNA-497, cTnI, FABP3 and GPBB in Pediatric Sepsis Complicated with Myocardial Injury.","date":"2021","source":"Therapeutics and clinical risk management","url":"https://pubmed.ncbi.nlm.nih.gov/34113113","citation_count":7,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"26802431","id":"PMC_26802431","title":"The diagnostic value of two commercially available human cTnI assays in goat kids with myocarditis.","date":"2016","source":"Veterinary clinical pathology","url":"https://pubmed.ncbi.nlm.nih.gov/26802431","citation_count":7,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"18548271","id":"PMC_18548271","title":"Expression of cTnI-R145G affects shortening properties of adult rat cardiomyocytes.","date":"2008","source":"Pflugers Archiv : European journal of physiology","url":"https://pubmed.ncbi.nlm.nih.gov/18548271","citation_count":7,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"38504588","id":"PMC_38504588","title":"Histone demethylase KDM5D represses the proliferation, migration and invasion of hepatocellular carcinoma through the E2F1/TNNC1 axis.","date":"2024","source":"Antioxidants & redox signaling","url":"https://pubmed.ncbi.nlm.nih.gov/38504588","citation_count":6,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"24474965","id":"PMC_24474965","title":"Pediatric restrictive cardiomyopathy due to a heterozygous mutation of the TNNI3 gene.","date":"2013","source":"Journal of biomedical research","url":"https://pubmed.ncbi.nlm.nih.gov/24474965","citation_count":6,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"35838319","id":"PMC_35838319","title":"The effect of Mg2+ on Ca2+ binding to cardiac troponin C in hypertrophic cardiomyopathy associated TNNC1 variants.","date":"2022","source":"The FEBS journal","url":"https://pubmed.ncbi.nlm.nih.gov/35838319","citation_count":6,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"33777698","id":"PMC_33777698","title":"TNNI3 and KCNQ1 co-inherited variants in a family with hypertrophic cardiomyopathy and long QT phenotypes: A case report.","date":"2021","source":"Molecular genetics and metabolism reports","url":"https://pubmed.ncbi.nlm.nih.gov/33777698","citation_count":6,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"38034835","id":"PMC_38034835","title":"High-sensitive cardiac troponin I (hs-cTnI) concentrations in newborns diagnosed with spinal muscular atrophy.","date":"2023","source":"Frontiers in pediatrics","url":"https://pubmed.ncbi.nlm.nih.gov/38034835","citation_count":6,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"9365790","id":"PMC_9365790","title":"Assignment of the human cardiac/slow skeletal muscle troponin C gene (TNNC1) between D3S3118 and GCT4B10 on the short arm of chromosome 3 by somatic cell hybrid analysis.","date":"1997","source":"Annals of human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/9365790","citation_count":6,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"38193576","id":"PMC_38193576","title":"Gene correction and overexpression of TNNI3 improve impaired relaxation in engineered heart tissue model of pediatric restrictive cardiomyopathy.","date":"2024","source":"Development, growth & differentiation","url":"https://pubmed.ncbi.nlm.nih.gov/38193576","citation_count":5,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"35656464","id":"PMC_35656464","title":"Diagnostic Value of Echocardiography Combined with Serum h-FABP and cTnI in Myocardial Infarction and Its Evaluation Value in Left Ventricular Function.","date":"2022","source":"Evidence-based complementary and alternative medicine : eCAM","url":"https://pubmed.ncbi.nlm.nih.gov/35656464","citation_count":5,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"38871861","id":"PMC_38871861","title":"Ckip-1 3'UTR alleviates prolonged sleep deprivation induced cardiac dysfunction by activating CaMKK2/AMPK/cTNI pathway.","date":"2024","source":"Molecular biomedicine","url":"https://pubmed.ncbi.nlm.nih.gov/38871861","citation_count":4,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"27379430","id":"PMC_27379430","title":"Suberoylanilide Hydroxamic Acid Restores Estrogen Reduced-cTnI Expression in Neonatal Hearts of Mice.","date":"2016","source":"Journal of cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/27379430","citation_count":4,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"30208128","id":"PMC_30208128","title":"Correlations of inhaled NO with the cTnI levels and the plasma clotting factor in rabbits with acute massive pulmonary embolism.","date":"2018","source":"Acta cirurgica brasileira","url":"https://pubmed.ncbi.nlm.nih.gov/30208128","citation_count":4,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"23220502","id":"PMC_23220502","title":"Effects of the glucose-lowering rate on cTnI and hs-CRP serum levels in type 2 diabetics.","date":"2012","source":"Human immunology","url":"https://pubmed.ncbi.nlm.nih.gov/23220502","citation_count":4,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"32563168","id":"PMC_32563168","title":"cTnI, BNP and CRP profiling after seizures in patients with drug-resistant epilepsy.","date":"2020","source":"Seizure","url":"https://pubmed.ncbi.nlm.nih.gov/32563168","citation_count":4,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"38497452","id":"PMC_38497452","title":"Impaired Relaxation in Induced Pluripotent Stem Cell-Derived Cardiomyocytes with Pathogenic TNNI3 Mutation of Pediatric Restrictive Cardiomyopathy.","date":"2024","source":"Journal of the American Heart Association","url":"https://pubmed.ncbi.nlm.nih.gov/38497452","citation_count":3,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"36814108","id":"PMC_36814108","title":"A novel variant of TNNC1 associated with severe dilated cardiomyopathy causing infant mortality and stillbirth: a case of germline mosaicism.","date":"2023","source":"Journal of genetics","url":"https://pubmed.ncbi.nlm.nih.gov/36814108","citation_count":3,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"34365612","id":"PMC_34365612","title":"[Identification of variants in TNNI3 gene in two children with restrictive cardiomyopathy].","date":"2021","source":"Zhonghua yi xue yi chuan xue za zhi = Zhonghua yixue yichuanxue zazhi = Chinese journal of medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/34365612","citation_count":3,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"34798544","id":"PMC_34798544","title":"Generation of three induced pluripotent stem cell lines from hypertrophic cardiomyopathy patients carrying TNNI3 mutations.","date":"2021","source":"Stem cell research","url":"https://pubmed.ncbi.nlm.nih.gov/34798544","citation_count":3,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"38924380","id":"PMC_38924380","title":"Homozygous TNNI3 frameshift variant in a consanguineous family with lethal infantile dilated cardiomyopathy.","date":"2024","source":"Molecular genetics & genomic medicine","url":"https://pubmed.ncbi.nlm.nih.gov/38924380","citation_count":3,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"40152334","id":"PMC_40152334","title":"A New Chemiluminescence-Based Rapid Diagnostic Testing Platform with Sequential Dual-Flow Strips for Cardiac Troponin I (cTnI).","date":"2025","source":"Analytical chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/40152334","citation_count":3,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"23788249","id":"PMC_23788249","title":"ACMG recommendations for reporting of incidental findings in clinical exome and genome sequencing.","date":"2013","source":"Genetics in medicine : official journal of the American College of Medical Genetics","url":"https://pubmed.ncbi.nlm.nih.gov/23788249","citation_count":1945,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"12477932","id":"PMC_12477932","title":"Generation and initial analysis of more than 15,000 full-length human and mouse cDNA sequences.","date":"2002","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/12477932","citation_count":1479,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"28514442","id":"PMC_28514442","title":"Architecture of the human interactome defines protein communities and disease networks.","date":"2017","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/28514442","citation_count":1085,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"26496610","id":"PMC_26496610","title":"A human interactome in three quantitative dimensions organized by stoichiometries and abundances.","date":"2015","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/26496610","citation_count":1015,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"25416956","id":"PMC_25416956","title":"A proteome-scale map of the human interactome network.","date":"2014","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/25416956","citation_count":977,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"12707239","id":"PMC_12707239","title":"Hypertrophic cardiomyopathy: distribution of disease genes, spectrum of mutations, and implications for a molecular diagnosis strategy.","date":"2003","source":"Circulation","url":"https://pubmed.ncbi.nlm.nih.gov/12707239","citation_count":969,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"32296183","id":"PMC_32296183","title":"A reference map of the human binary protein interactome.","date":"2020","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/32296183","citation_count":849,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"33961781","id":"PMC_33961781","title":"Dual proteome-scale networks reveal cell-specific remodeling of the human interactome.","date":"2021","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/33961781","citation_count":705,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"12840750","id":"PMC_12840750","title":"Structure of the core domain of human cardiac troponin in the Ca(2+)-saturated form.","date":"2003","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/12840750","citation_count":659,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"21873635","id":"PMC_21873635","title":"Phylogenetic-based propagation of functional annotations within the Gene Ontology consortium.","date":"2011","source":"Briefings in bioinformatics","url":"https://pubmed.ncbi.nlm.nih.gov/21873635","citation_count":656,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"12242268","id":"PMC_12242268","title":"Characterization and enrichment of cardiomyocytes derived from human embryonic stem cells.","date":"2002","source":"Circulation research","url":"https://pubmed.ncbi.nlm.nih.gov/12242268","citation_count":646,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"32517963","id":"PMC_32517963","title":"Prevalence and Impact of Myocardial Injury in Patients Hospitalized With COVID-19 Infection.","date":"2020","source":"Journal of the American College of Cardiology","url":"https://pubmed.ncbi.nlm.nih.gov/32517963","citation_count":584,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"27532257","id":"PMC_27532257","title":"Reassessment of Mendelian gene pathogenicity using 7,855 cardiomyopathy cases and 60,706 reference samples.","date":"2016","source":"Genetics in medicine : official journal of the American College of Medical Genetics","url":"https://pubmed.ncbi.nlm.nih.gov/27532257","citation_count":574,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"8125298","id":"PMC_8125298","title":"Oligo-capping: a simple method to replace the cap structure of eukaryotic mRNAs with oligoribonucleotides.","date":"1994","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/8125298","citation_count":492,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"25910212","id":"PMC_25910212","title":"Widespread macromolecular interaction perturbations in human genetic disorders.","date":"2015","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/25910212","citation_count":454,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"9241277","id":"PMC_9241277","title":"Mutations in the cardiac troponin I gene associated with hypertrophic cardiomyopathy.","date":"1997","source":"Nature genetics","url":"https://pubmed.ncbi.nlm.nih.gov/9241277","citation_count":451,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"24291281","id":"PMC_24291281","title":"Early increases in multiple biomarkers predict subsequent cardiotoxicity in patients with breast cancer treated with doxorubicin, taxanes, and trastuzumab.","date":"2013","source":"Journal of the American College of Cardiology","url":"https://pubmed.ncbi.nlm.nih.gov/24291281","citation_count":431,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"17426036","id":"PMC_17426036","title":"Certain pairs of ubiquitin-conjugating enzymes (E2s) and ubiquitin-protein ligases (E3s) synthesize nondegradable forked ubiquitin chains containing all possible isopeptide linkages.","date":"2007","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/17426036","citation_count":352,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"18403758","id":"PMC_18403758","title":"Shared genetic causes of cardiac hypertrophy in children and adults.","date":"2008","source":"The New England journal of medicine","url":"https://pubmed.ncbi.nlm.nih.gov/18403758","citation_count":309,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"16199542","id":"PMC_16199542","title":"Compound and double mutations in patients with hypertrophic cardiomyopathy: implications for genetic testing and counselling.","date":"2005","source":"Journal of medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/16199542","citation_count":291,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"15601779","id":"PMC_15601779","title":"Muscle-specific RING finger 1 is a bona fide ubiquitin ligase that degrades cardiac troponin I.","date":"2004","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/15601779","citation_count":279,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"11815426","id":"PMC_11815426","title":"Sarcomere protein gene mutations in hypertrophic cardiomyopathy of the elderly.","date":"2002","source":"Circulation","url":"https://pubmed.ncbi.nlm.nih.gov/11815426","citation_count":262,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"12531876","id":"PMC_12531876","title":"Idiopathic restrictive cardiomyopathy is part of the clinical expression of cardiac troponin I mutations.","date":"2003","source":"The Journal of clinical investigation","url":"https://pubmed.ncbi.nlm.nih.gov/12531876","citation_count":262,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"18533079","id":"PMC_18533079","title":"Myofilament protein gene mutation screening and outcome of patients with hypertrophic cardiomyopathy.","date":"2008","source":"Mayo Clinic proceedings","url":"https://pubmed.ncbi.nlm.nih.gov/18533079","citation_count":261,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"10387074","id":"PMC_10387074","title":"Binding of cardiac troponin-I147-163 induces a structural opening in human cardiac troponin-C.","date":"1999","source":"Biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/10387074","citation_count":256,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"20359594","id":"PMC_20359594","title":"Clinical features and outcome of hypertrophic cardiomyopathy associated with triple sarcomere protein gene mutations.","date":"2010","source":"Journal of the American College of Cardiology","url":"https://pubmed.ncbi.nlm.nih.gov/20359594","citation_count":216,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"15542288","id":"PMC_15542288","title":"Severe disease expression of cardiac troponin C and T mutations in patients with idiopathic dilated cardiomyopathy.","date":"2004","source":"Journal of the American College of Cardiology","url":"https://pubmed.ncbi.nlm.nih.gov/15542288","citation_count":198,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"19372185","id":"PMC_19372185","title":"Developing multiplexed assays for troponin I and interleukin-33 in plasma by peptide immunoaffinity enrichment and targeted mass spectrometry.","date":"2009","source":"Clinical chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/19372185","citation_count":197,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"19463339","id":"PMC_19463339","title":"Utility of cardiac biomarkers in predicting infarct size, left ventricular function, and clinical outcome after primary percutaneous coronary intervention for ST-segment elevation myocardial infarction.","date":"2008","source":"JACC. Cardiovascular interventions","url":"https://pubmed.ncbi.nlm.nih.gov/19463339","citation_count":190,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"9560191","id":"PMC_9560191","title":"Crystal structure of troponin C in complex with troponin I fragment at 2.3-A resolution.","date":"1998","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/9560191","citation_count":187,"is_preprint":false,"source_track":"gene2pubmed"}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":53397,"output_tokens":5708,"usd":0.122906},"stage2":{"model":"claude-opus-4-6","input_tokens":9274,"output_tokens":3479,"usd":0.200017},"total_usd":0.756525,"stage1_batch_id":"msgbatch_01RPbVc9PDH9zQmYK5Hs6srm","stage2_batch_id":"msgbatch_01JVbVLzeKN2yVrsGp3zC8aM","note":"batch pricing = 50% of standard","round2_stage1":{"model":"claude-sonnet-4-6","input_tokens":68723,"output_tokens":9235,"usd":0.172347},"round2_rules_fired":"R2","round2_stage2":{"model":"claude-opus-4-6","input_tokens":12119,"output_tokens":4543,"usd":0.261255}},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2015,\n      \"finding\": \"TNNI3 encodes the inhibitory subunit of the cardiac troponin complex; it contains an N-terminal cardiac-specific extension (residues 1–39) phosphorylated by PKA at Ser23/Ser24, an inhibitory peptide (residues ~137–147) that interacts with actin, and a switch region that binds the N-domain of TnC upon Ca2+ activation, collectively conferring Ca2+-sensitive regulation of striated muscle contraction.\",\n      \"method\": \"Comprehensive review integrating structural, biochemical, and mutagenesis data from multiple laboratories\",\n      \"journal\": \"Gene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — foundational structural and functional framework replicated across many labs\",\n      \"pmids\": [\"26526134\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1996,\n      \"finding\": \"The human TNNI3 gene comprises eight exons spanning ~6.2 kb; 2300 bp of 5′-flanking sequence drives expression in cardiac myocytes and skeletal muscle cells but is inactive in fibroblasts, indicating muscle-type but not cardiac-specific activity of the proximal promoter.\",\n      \"method\": \"Genomic cloning, DNA sequencing, transfection reporter assay in cardiac myocytes, skeletal muscle cells, and fibroblasts\",\n      \"journal\": \"Genomics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — direct experimental characterization of gene structure and promoter activity\",\n      \"pmids\": [\"8661099\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"PKC-mediated phosphorylation of cTnI at Ser43 and Ser45 slows cardiac contraction and relaxation in vivo; alanine substitution of these sites (mTnI transgenic mice) increased basal contractility (+30% +dP/dt) and blunted the response to PKC-activating stimuli, and was associated with enhanced PKA-dependent phosphorylation of TnI independent of cAMP changes.\",\n      \"method\": \"Transgenic mice with alanine substitutions at PKC phosphorylation sites Ser43/Ser45; in situ hemodynamics; back-phosphorylation assay\",\n      \"journal\": \"American journal of physiology. Heart and circulatory physiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean transgenic KO of phosphorylation sites with defined hemodynamic phenotype and biochemical follow-up\",\n      \"pmids\": [\"14726296\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"PKA phosphorylation of cTnI at Ser23/Ser24 controls the steepness of the length–tension (Frank-Starling) relationship in cardiac and slow-twitch skeletal muscle; exchange of unphosphorylated recombinant cTn produced a shallow length-tension curve, whereas PKA treatment or an Asp23/Asp24 phosphomimetic restored a steep relationship.\",\n      \"method\": \"Troponin exchange into permeabilized rat cardiac myocytes and slow-twitch skeletal muscle fibres; PKA treatment; length-tension measurements\",\n      \"journal\": \"The Journal of physiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — reconstitution with defined phosphomimetic mutants plus PKA treatment, multiple muscle types\",\n      \"pmids\": [\"23836688\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"PAK3 phosphorylation of cTnI at Ser151 increases Ca2+ sensitivity of myofilament activation by shortening the cTnC–cTnI inter-site distances and reducing the kinetic rate of Ca2+-dissociation-induced structural change in the cTnI regulatory region, mimicking the effect of strong cross-bridges.\",\n      \"method\": \"Pseudo-phosphorylation (S151E) cTnI reconstituted into thin filaments; steady-state and time-resolved FRET; stopped-flow kinetics\",\n      \"journal\": \"Journal of molecular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — reconstituted thin filaments with phosphomimetic mutant, multiple orthogonal biophysical methods\",\n      \"pmids\": [\"20540949\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"The HCM mutation cTnI-R145G (in the inhibitory peptide) stabilizes Ca2+-coordinating interactions in cTnC but blunts the intra-subunit interaction between the cTnI N-terminal extension and the inhibitory peptide that normally mediates PKA-phosphorylation-dependent modulation of troponin function during β-adrenergic stimulation.\",\n      \"method\": \"Molecular dynamics simulations (triplicate 150 ns) of cTnC(1-161)–cTnI(1-172)–cTnT(236-285) complexes with R145G, R145G/S23D/S24D phosphomimetic, and real phosphorylation\",\n      \"journal\": \"Biophysical journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 method (MD with multiple comparators) but computational only, single lab\",\n      \"pmids\": [\"25606687\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"The HCM cTnI-R145G mutation reduces Ca2+-regulatory function of the cTnI inhibitory peptide; the HCM cTnT-R278C mutation partially rescues R145G-induced loss of inhibition at high Ca2+ but exacerbates it at low Ca2+, demonstrating functional interaction between the cTnI inhibitory peptide and the C-terminus of cTnT within the troponin complex.\",\n      \"method\": \"In vitro motility assay with reconstituted troponin complexes containing single or double HCM mutations; varied Ca2+, temperature, and HMM density\",\n      \"journal\": \"Archives of biochemistry and biophysics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — reconstituted thin filament motility assay, multiple conditions, single and double mutant comparison\",\n      \"pmids\": [\"24418317\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"Adenoviral expression of cTnI-R145G in adult rat cardiomyocytes significantly decreased baseline sarcomere shortening and depressed rates of shortening and relengthening, while β-adrenergic stimulation through isoproterenol or forskolin rescued shortening amplitude but not rates, indicating receptor-subtype-dependent modulation of the HCM mutation's contractile effects.\",\n      \"method\": \"Adenoviral expression of human cTnI-WT and cTnI-R145G in adult rat cardiomyocytes; sarcomere length measurements; pharmacological β-adrenergic stimulation\",\n      \"journal\": \"Pflugers Archiv : European journal of physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct overexpression in primary cardiomyocytes with defined contractile readout, single lab\",\n      \"pmids\": [\"18548271\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"cTnI R170G and R170W mutations (C-terminal regulatory region) strongly enhance Ca2+ sensitivity in skinned fibres, greatly increase troponin affinity for tropomyosin, reduce reconstituted thin filament structural integrity (wavy filaments with breaks), and impair the interaction of cTnI/cTnT with the N-terminal fragment of cMyBPC (C0C2), revealing that the cTnI C-terminus mediates communication between thin and thick filament regulatory proteins.\",\n      \"method\": \"Skinned fibre force-Ca2+ measurements; electron microscopy of reconstituted thin filaments; microscale thermophoresis for troponin–cMyBPC interaction; actin/tropomyosin binding assays\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — multiple orthogonal in vitro methods with reconstituted components and electron microscopy structural validation\",\n      \"pmids\": [\"32182250\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"cTnI is modified by arginine methylation in human myocardium at R74/R79 and R146/R148 (identified by mass spectrometry); PRMT1 methylates an extended inhibitory peptide at R146/R148 in vitro; HCM-associated R145 mutations impair R146/R148 methylation by PRMT1; methylation-deficient R146A/R148A cTnI expressed in H9c2 cells induces cell hypertrophy (+32% cell size), linking cTnI arginine methylation to regulation of cardiac hypertrophy.\",\n      \"method\": \"Mass spectrometry identification of methylation sites; Western blot with anti-methylarginine antibody; in vitro PRMT1 methylation assay with HCM mutants; H9c2 cell transfection with methylation-deficient cTnI; cell size measurement\",\n      \"journal\": \"International journal of cardiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — mass spectrometry discovery, in vitro enzyme assay, cell-based functional validation, multiple orthogonal methods\",\n      \"pmids\": [\"30772011\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"In restrictive cardiomyopathy transgenic mice carrying the cTnI-193His mutation, cTnI protein is detected in cardiomyocyte nuclei by immunofluorescence and Western blotting; nuclear cTnI interacts with HDAC1 and SMYD1 (by co-immunoprecipitation), and overexpression of mutant cTnI in cultured cardiomyocytes reduces PDE4d expression, suggesting a non-sarcomeric nuclear role for cTnI in epigenetic regulation.\",\n      \"method\": \"Immunofluorescence and subcellular fractionation/Western blot of transgenic RCM mouse hearts; Co-IP of nuclear cTnI with HDAC1 and SMYD1; overexpression in cultured cardiomyocytes with PDE4d expression readout\",\n      \"journal\": \"Science China. Life sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 — direct localization experiment with Co-IP and functional consequence, single lab, moderate mechanistic follow-up\",\n      \"pmids\": [\"30900165\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"The cTnI-G203S HCM mutation impairs functional communication between the L-type Ca2+ channel and the cytoskeletal network (F-actin), leading to reduced actin-myosin dynamics, block of the mitochondrial voltage-dependent anion channel, and a hypermetabolic mitochondrial state (elevated membrane potential and metabolic activity), which is present in pre-cardiomyopathic young mice and can be normalized by L-type Ca2+ channel antagonists (nisoldipine, diltiazem).\",\n      \"method\": \"Patch-clamp electrophysiology of ICa-L; mitochondrial membrane potential and metabolic activity measurements; pharmacological channel blockade in cTnI-G203S transgenic mouse cardiomyocytes\",\n      \"journal\": \"The Journal of physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple physiological readouts in transgenic disease model with pharmacological rescue, single lab\",\n      \"pmids\": [\"27062056\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"A frameshift deletion in TNNI3 (exon 7, predicting premature stop and truncation of the C-terminal portion) causes restrictive cardiomyopathy; Western blot of myocardial tissue showed ~50% reduction in total troponin I protein, demonstrating haploinsufficiency as a disease mechanism.\",\n      \"method\": \"Sequencing of TNNI3; Western blot of patient myocardial tissue for TnI protein quantification\",\n      \"journal\": \"International journal of cardiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct patient tissue Western blot showing haploinsufficiency, single case\",\n      \"pmids\": [\"18006163\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Complete homozygous loss of TNNI3 protein (truncating splice-site variant TNNI3 c.24+2T>A) leads to severe pediatric cardiomyopathy and is associated with compensatory upregulation of the fetal isoform TNNI1 (slow skeletal TnI) in heart biopsy tissue, indicating that TNNI3 is essential for normal cardiac sarcomere function and that TNNI1 partially substitutes for its loss.\",\n      \"method\": \"Protein and transcript analysis on heart biopsies from homozygous TNNI3 mutation carriers; next-generation sequencing panel\",\n      \"journal\": \"Clinical genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct patient tissue protein analysis showing loss-of-function and isoform switch, single center\",\n      \"pmids\": [\"31568572\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"DSCR1L2 protein (and its alternatively spliced isoform lacking exons 2–5) physically interacts with human cardiac troponin I (TNNI3), as demonstrated by yeast two-hybrid screening of a human heart cDNA library, confirmed by yeast co-transformation and GST pull-down assay; DSCR1L2 exon 2 is required for binding.\",\n      \"method\": \"Yeast two-hybrid screen of human heart cDNA library; yeast co-transformation; GST fusion protein pull-down assay\",\n      \"journal\": \"Gene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — pull-down confirmed by two yeast-based methods but no functional consequence established, single lab\",\n      \"pmids\": [\"16516408\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"The de novo TNNI3 mutation cTnI-D127Y (infantile RCM) increases Ca2+ sensitivity in skinned cardiomyocytes, enhances troponin–actin interaction, reduces structural integrity of reconstituted thin filaments (by electron microscopy), and impairs the cooperative response to cMyBPC-C0C2; levosimendan and EGCG partially restore thin filament stability and contractile function in vitro.\",\n      \"method\": \"Skinned cardiac fibre Ca2+-force measurements; reconstituted thin filament structural analysis (electron microscopy); inter-subunit binding assays; pharmacological rescue with levosimendan and EGCg\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — multiple orthogonal in vitro methods with structural and functional validation\",\n      \"pmids\": [\"34502534\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"iPSC-derived cardiomyocytes carrying the TNNI3-R170W mutation (heterozygous and homozygous) show impaired diastolic function (prolonged relaxation, increased relaxation-to-contraction force ratio in EHT) compared with CRISPR/Cas9-corrected isogenic controls; overexpression of wild-type TNNI3 in R170W iPSC-CMs and EHTs rescues impaired relaxation, establishing that the R170W variant causes diastolic dysfunction through a direct sarcomeric mechanism.\",\n      \"method\": \"iPSC generation from RCM patient; CRISPR/Cas9 isogenic correction; differentiation to cardiomyocytes; engineered heart tissue (EHT) force measurements; cell motion analysis; TNNI3 overexpression rescue\",\n      \"journal\": \"Development, growth & differentiation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — isogenic iPSC model with CRISPR correction and overexpression rescue, multiple functional readouts\",\n      \"pmids\": [\"38193576\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Patient-specific iPSC-derived cardiomyocytes with heterozygous or homozygous TNNI3-R170W demonstrate impaired diastolic function in cell motion assays relative to isogenic-corrected and healthy controls; RNA sequencing reveals altered pathways in cardiac muscle development/contraction, ECM-receptor interaction, and TGF-β signalling, whereas myofibril and mitochondrial ultrastructure remain intact.\",\n      \"method\": \"iPSC differentiation; CRISPR/Cas9 isogenic correction; cell motion analysis; electron microscopy; RNA sequencing\",\n      \"journal\": \"Journal of the American Heart Association\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — isogenic iPSC correction with orthogonal functional and transcriptomic analysis\",\n      \"pmids\": [\"38497452\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"HDAC5 binding to the cTnI promoter (ChIP assay) mediates histone H3K9 deacetylation and suppresses cTnI transcription in neonatal mouse hearts; the HDAC inhibitor SAHA restores H3K9ac at the cTnI promoter and upregulates cTnI expression, identifying HDAC5-mediated epigenetic regulation as a mechanism controlling cTnI levels in the heart.\",\n      \"method\": \"Chromatin immunoprecipitation (ChIP) for HDAC5 and H3K9ac at the cTnI promoter; HDAC/HAT activity assays; qRT-PCR and Western blot; SAHA pharmacological intervention in neonatal mice\",\n      \"journal\": \"Journal of cellular biochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — ChIP plus pharmacological rescue with multiple biochemical readouts, single lab\",\n      \"pmids\": [\"27379430\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"In ageing mouse hearts with reduced cTnI expression and diastolic dysfunction, HDAC1 and HDAC3 bind to the cTnI proximal promoter to reduce H3K9 acetylation and decrease GATA4/Mef2c recruitment; EGCG treatment inhibits HDAC1/HDAC3 promoter binding, increases AcH3K9 at the cTnI promoter, elevates GATA4 and Mef2c binding, restores cTnI expression, and improves diastolic function.\",\n      \"method\": \"ChIP assay for HDAC1, AcH3K9, GATA4, and Mef2c at cTnI promoter; Western blot; qRT-PCR; echocardiography in aged mice; EGCG pharmacological intervention\",\n      \"journal\": \"Journal of cellular and molecular medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — ChIP plus functional cardiac readout with pharmacological rescue, single lab\",\n      \"pmids\": [\"28382690\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"miR-449 targets and suppresses HDAC1, which in turn reduces H3K4 and H3K9 deacetylation at the cTnI promoter and decreases GATA4 binding; miR-449 agomiR intervention in aged mice with low cTnI expression restores histone acetylation at the cTnI promoter, increases cTnI expression, and improves cardiac function.\",\n      \"method\": \"Dual-luciferase reporter assay confirming miR-449/HDAC1 interaction; ChIP for H3K4ac, H3K9ac, and GATA4 at cTnI promoter; miR-449 agomiR delivery in aged mice; echocardiography\",\n      \"journal\": \"European review for medical and pharmacological sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — luciferase validation plus ChIP and in vivo functional readout, single lab\",\n      \"pmids\": [\"33378032\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"TNNI3-encoded cardiac troponin I (cTnI) functions as the inhibitory subunit of the troponin complex on the cardiac thin filament, where its inhibitory peptide (residues ~137–147) suppresses actomyosin ATPase at low Ca2+, its switch region binds the N-domain of TnC upon Ca2+ activation to relieve inhibition, and its N-terminal cardiac-specific extension is phosphorylated by PKA (Ser23/Ser24)—controlling both Ca2+ sensitivity and Frank-Starling length-dependence of force—and by PKC (Ser43/Ser45) to slow contraction/relaxation; additionally, cTnI is arginine-methylated at R146/R148 by PRMT1 near the inhibitory/switch peptide boundary to modulate hypertrophy signalling, its expression is epigenetically controlled by HDAC1/HDAC3/HDAC5-mediated histone deacetylation at the promoter, and pathogenic mutations in the C-terminal regulatory region (e.g., R170G/W, D127Y) increase Ca2+ sensitivity, disrupt thin filament structural integrity, and impair the cTnI–cMyBPC cross-filament communication, collectively producing restrictive or hypertrophic cardiomyopathy phenotypes.\"\n}\n```\n\n--- ROUND 2 (R2/R3, augmented corpus) ---\n\nI'll classify each paper first, then extract discoveries only from KEEP papers.\n\n**Classification notes:**\n- Papers about TNNC1 (troponin C) → EXCLUDE (alias collision — different gene)\n- Papers purely about cTnI as a biomarker (diagnostic assays, clinical correlations) → EXCLUDE (no mechanistic content)\n- Papers about TNNI3 mechanism, structure, function, PTMs, interactions → KEEP\n- General interactome papers with no specific TNNI3 findings → EXCLUDE\n- Papers about TNNI1/TNNI2 → EXCLUDE\n\n**KEEP list:**\n- PMID 26526134 (TNNI1/2/3 review — evolution, regulation, structure-function)\n- PMID 8661099 (isolation/characterization of TNNI3 gene, promoter)\n- PMID 25606687 (MD simulations of R145G mutation, PKA phosphorylation effects on cTnI)\n- PMID 14726296 (PKC phosphorylation of cTnI S43/S45, cardiac function in vivo)\n- PMID 23836688 (PKA phosphorylation of cTnI S23/S24 controls length-tension)\n- PMID 24418317 (cTnI R145G + cTnT R278C functional interactions, Ca2+ regulation)\n- PMID 32182250 (cTnI R170G/W RCM mutations — thin filament interactions, structure)\n- PMID 30772011 (arginine methylation of cTnI by PRMT1 at R146/R148)\n- PMID 20540949 (PAK3 phosphorylation of cTnI S151, structural/kinetic effects)\n- PMID 16516408 (DSCR1L2 interaction with TNNI3 by yeast two-hybrid/GST pulldown)\n- PMID 15601779 (MuRF1 ubiquitin ligase degrades cardiac troponin I)\n- PMID 17426036 (MuRF1/UbcH5 ubiquitination of troponin I — mixed chains)\n- PMID 9241277 (first identification of cTnI mutations in HCM — Kimura et al.)\n- PMID 12531876 (RCM caused by TNNI3 mutations — Mogensen et al.)\n- PMID 12840750 (crystal structure of cardiac troponin core domain)\n- PMID 9560191 (crystal structure TnC+TnI fragment)\n- PMID 10387074 (NMR structure of cNTnC·Ca2+·cTnI147-163 complex)\n- PMID 30900165 (cTnI in nucleus, interaction with HDAC1/SMYD1, epigenetic regulation of PDE4d)\n- PMID 27062056 (cTnI-G203S mutation, L-type Ca2+ channel / mitochondria communication)\n- PMID 34502534 (cTnI-D127Y mutation effects on thin filament, troponin targeting agents)\n- PMID 38193576 (TNNI3 R170W iPSC-EHT model, CRISPR correction, overexpression rescues RCM)\n- PMID 38497452 (TNNI3 R170W iPSC-CM model, diastolic dysfunction, RNA-seq)\n- PMID 28382690 (EGCG/histone acetylation regulates cTnI expression, HDAC1/HDAC3/GATA4)\n- PMID 33378032 (miR-449/HDAC1 regulates cTnI via H3K4/H3K9 acetylation, GATA4)\n- PMID 27379430 (estrogen/HDAC5 reduces cTnI expression via H3K9ac, SAHA rescues)\n- PMID 31568572 (homozygous TNNI3 truncation → loss of protein, upregulation of fetal TNNI1)\n- PMID 18548271 (cTnI-R145G expression in adult rat cardiomyocytes, β-adrenergic response)\n\n```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1996,\n      \"finding\": \"The human cardiac troponin I gene (TNNI3) comprises eight exons within 6.2 kb of genomic DNA. Transfection experiments showed that 2300 bp of 5'-flanking sequence drives expression in cardiac myocytes and skeletal muscle cells but not fibroblasts, and several conserved putative cis-acting elements were identified in the proximal promoter, indicating that additional sequences are required to confer cardiac specificity.\",\n      \"method\": \"Genomic cloning, sequencing, transfection reporter assays in cardiac myocytes and fibroblasts\",\n      \"journal\": \"Genomics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct functional transfection assay; single lab study\",\n      \"pmids\": [\"8661099\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"Mutations in TNNI3 (cardiac troponin I) were identified as a cause of hypertrophic cardiomyopathy, establishing cTnI as an HCM disease gene. Key mutations included Arg145Gly (linked by family analysis) and Lys206Gln (de novo), demonstrating that the inhibitory subunit of the troponin complex is essential for normal cardiac function.\",\n      \"method\": \"Candidate gene sequencing in 184 unrelated HCM patients, family linkage analysis, identification of de novo mutations\",\n      \"journal\": \"Nature genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic epistasis/linkage in large cohort, replicated across multiple independent labs subsequently\",\n      \"pmids\": [\"9241277\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1998,\n      \"finding\": \"The crystal structure of troponin C in complex with the N-terminal fragment of troponin I (TnI residues 1–47) at 2.3 Å resolution revealed that TnC adopts a compact globular conformation (vs. elongated dumbbell in free TnC) with the central helix bent 90°. The TnI1–47 alpha-helix spans the surface of TnC, stabilizing the compact conformation via contacts with both N- and C-terminal lobes, with the amphiphilic C-terminus of TnI1–47 anchored in the hydrophobic pocket of the TnC C-lobe.\",\n      \"method\": \"X-ray crystallography (2.3 Å), single isomorphous replacement + MAD phasing\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — crystal structure at atomic resolution\",\n      \"pmids\": [\"9560191\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"NMR structure of the cardiac TnC N-terminal regulatory domain (cNTnC) in complex with Ca2+ and the cTnI switch peptide (residues 147–163) showed that cTnI147–163 binding induces an 'open' conformation of cNTnC (similar to Ca2+-saturated skeletal TnC), even though Ca2+ alone does not open cNTnC. The cTnI peptide adopts an alpha-helical conformation at residues 150–157 bound in the hydrophobic pocket of cNTnC, establishing the molecular basis for Ca2+-dependent activation of cardiac muscle.\",\n      \"method\": \"Multinuclear multidimensional NMR spectroscopy, solution structure determination\",\n      \"journal\": \"Biochemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — NMR structure with functional validation of binding mechanism\",\n      \"pmids\": [\"10387074\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"The crystal structure of the core domain of human cardiac troponin (TnC + TnI + TnT) in the Ca2+-saturated form revealed that the core domain is divided into structurally distinct subdomains connected by flexible linkers. An alpha-helical coiled-coil between TnT and TnI forms the rigid IT arm (~80 Å), which bridges tropomyosin-anchoring regions. Ca2+ binding to the regulatory site of TnC removes the C-terminal portion of TnI from actin, altering mobility/flexibility of troponin and tropomyosin on the actin filament.\",\n      \"method\": \"X-ray crystallography (crystal structures at 46 kDa and 52 kDa core domain constructs)\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — landmark crystal structure of ternary troponin complex\",\n      \"pmids\": [\"12840750\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"TNNI3 mutations were identified as causes of idiopathic restrictive cardiomyopathy (RCM). In a large family with both RCM and HCM, linkage to TNNI3 was established (lod score 4.8) and a novel missense mutation identified. Screening nine additional unrelated RCM patients found TNNI3 mutations in six, including two de novo mutations in young patients. All mutations mapped to conserved, functionally important domains, establishing TNNI3 as an RCM susceptibility gene.\",\n      \"method\": \"Linkage analysis, candidate gene mutation screening, family co-segregation studies\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — linkage analysis plus multi-family replication; established new disease association\",\n      \"pmids\": [\"12531876\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Muscle-specific RING finger protein 1 (MuRF1) was identified as a bona fide ubiquitin E3 ligase that ubiquitylates and degrades cardiac troponin I (cTnI). Yeast two-hybrid screening identified cTnI as a MuRF1 interactor; co-IP confirmed the association in vitro and in cardiomyocytes. MuRF1 reduced steady-state cTnI levels via RING finger-dependent ubiquitin ligase activity, accumulation of poly-ubiquitylated cTnI intermediates, and proteasome-dependent degradation. MuRF1 overexpression in cardiomyocytes reduced contractility indices.\",\n      \"method\": \"Yeast two-hybrid screen, Co-immunoprecipitation, in vitro ubiquitylation assay, cardiomyocyte overexpression with contractility readout\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — in vitro reconstitution of ubiquitylation + multiple cellular assays + functional outcome\",\n      \"pmids\": [\"15601779\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"PKC phosphorylation of cTnI at Ser43 and Ser45 modulates cardiac contractility in vivo. Transgenic mice with Ala substitutions at these sites (preventing PKC phosphorylation) showed 30% greater +dP/dt and 18% greater −dP/dt at baseline compared with wild-type, and had negligible inotropic response to isoproterenol or phenylephrine/propranolol. Mutation of PKC sites was also associated with enhanced PKA-dependent phosphorylation of cTnI, demonstrating interdependence of phosphorylation sites.\",\n      \"method\": \"Transgenic mouse model (Ala substitutions at S43/S45), in situ hemodynamics, back-phosphorylation assays, cAMP measurements\",\n      \"journal\": \"American journal of physiology. Heart and circulatory physiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — in vivo genetic loss-of-function with defined hemodynamic phenotype and biochemical follow-up\",\n      \"pmids\": [\"14726296\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"DSCR1L2 (Down Syndrome Critical Region gene 1-like 2) protein interacts directly with cardiac troponin I (TNNI3). Interaction was identified by yeast two-hybrid screening of a human heart cDNA library and confirmed by yeast co-transformation and GST fusion protein pull-down assays. A novel DSCR1L2 splice isoform lacking two central exons was identified; both isoforms interact with TNNI3, and exon 2 of DSCR1L2 is critical for binding, suggesting a possible role for this calcineurin-pathway family member in cardiac contraction.\",\n      \"method\": \"Yeast two-hybrid screening (human heart cDNA library), yeast co-transformation, GST pulldown assay, quantitative RT-PCR\",\n      \"journal\": \"Gene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — confirmed by two orthogonal binding assays (yeast two-hybrid + GST pulldown) but no functional consequence established\",\n      \"pmids\": [\"16516408\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"A novel one-nucleotide frameshift deletion in TNNI3 (exon 7) causing premature stop codon and truncation of the C-terminal portion of cTnI was found to cause restrictive cardiomyopathy. Western blot analysis of myocardial tissue showed approximately 50% reduction in total troponin I content, indicating haploinsufficiency of cTnI protein as a mechanism for RCM.\",\n      \"method\": \"Candidate gene sequencing, Western blot of myocardial tissue\",\n      \"journal\": \"International journal of cardiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 — Western blot directly quantified protein reduction in patient tissue; single case\",\n      \"pmids\": [\"18006163\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"MuRF1 with the E2 enzyme UbcH5 synthesizes mixed forked ubiquitin chains on troponin I containing all seven possible linkages (predominantly Lys48, Lys63, Lys11), and troponin I linked to these forked chains was degraded poorly by purified 26S proteasomes compared to troponin I linked to homogeneous Lys48 or Lys63 chains. With UbcH13/Uev1a, MuRF1 attaches Lys63 chains, and with UbcH1 it attaches Lys48 chains, both of which support efficient degradation, establishing that chain topology determines proteasomal degradation efficiency of cTnI.\",\n      \"method\": \"In vitro ubiquitylation assay with purified components, mass spectrometry of Ub chain linkages, 26S proteasome degradation assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — fully reconstituted in vitro system with MS characterization and degradation assays\",\n      \"pmids\": [\"17426036\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"Expression of the HCM-associated cTnI-R145G mutation in adult rat cardiomyocytes significantly decreased baseline sarcomere shortening. Upon β-adrenergic stimulation with isoproterenol, rates of shortening and re-lengthening were depressed in cTnI-R145G-expressing cells but comparable to controls when using the β2-selective blocker ICI118,551, indicating that the R145G mutation alters β-adrenergic receptor subtype-dependent modulation of contractility.\",\n      \"method\": \"Adenovirus-mediated expression of cTnI-R145G in adult rat cardiomyocytes, sarcomere shortening measurements, pharmacological β-adrenergic dissection\",\n      \"journal\": \"Pflugers Archiv : European journal of physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct expression of mutant protein in primary cardiomyocytes with functional readout\",\n      \"pmids\": [\"18548271\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"PAK3-mediated phosphorylation of cTnI at Ser151 (studied via S151E phosphomimic) increases Ca2+ sensitivity of myofilament by shortening inter-site distances between cTnC and cTnI (monitored by FRET in reconstituted thin filaments) and reducing kinetic rates of Ca2+-dissociation-induced structural changes in the regulatory region of cTnI. The structural effects mimic those of strong actomyosin crossbridges.\",\n      \"method\": \"Steady-state and time-resolved FRET, stopped-flow kinetics in reconstituted thin filaments containing cTnI-S151E\",\n      \"journal\": \"Journal of molecular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro reconstitution with multiple spectroscopic methods\",\n      \"pmids\": [\"20540949\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Phosphorylation of cTnI at serines 23/24 by PKA is a key regulator of length dependence of force generation (Frank-Starling mechanism) in cardiac myocytes. Exchange of unphosphorylated recombinant cTn into permeabilized rat cardiac myocytes or slow-twitch skeletal muscle fibers produced a shallow length-tension relationship; PKA treatment or use of cTn with S23/S24 mutated to Asp (phosphomimetic) restored a steep length-tension relationship.\",\n      \"method\": \"Troponin exchange into permeabilized cardiac myocytes and skeletal muscle fibers, PKA treatment, phosphomimetic mutation (S23D/S24D), force-length measurements\",\n      \"journal\": \"The Journal of physiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — reconstitution approach in multiple muscle preparations with genetic and pharmacological validation\",\n      \"pmids\": [\"23836688\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"The Ca2+-regulatory function of the cTnI inhibitory peptide (Ip) region requires cooperation with the C-terminus of cTnT. In vitro motility assays showed that cTnI-R145G (within the Ip) alone impaired switching off of actomyosin at low Ca2+; cTnT-R278C partially rescued high-Ca2+ defects of cTnI-R145G but exacerbated loss of inhibitory function at low Ca2+. Bioinformatics analysis revealed evolutionary and structural relatedness between the affected regions of cTnI and cTnT.\",\n      \"method\": \"In vitro motility assay with reconstituted troponin complexes (single and double mutants), varying Ca2+ concentration, temperature, and HMM density\",\n      \"journal\": \"Archives of biochemistry and biophysics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — in vitro reconstitution with functional readout; single lab\",\n      \"pmids\": [\"24418317\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Molecular dynamics simulations of the cTnI-R145G HCM mutation revealed that R145G does not alter overall cTn dynamics but stabilizes Ca2+-coordinating interactions in cTnC. The R145G mutation blunted intramolecular interactions between the cTnI N-terminal extension (residues 1–39) and the inhibitory peptide that are normally induced by PKA phosphorylation of S23/S24 (β-adrenergic signaling), providing a molecular explanation for why R145G reduces β-adrenergic modulation of cTn.\",\n      \"method\": \"Triplicate 150 ns molecular dynamics simulations of cTnC(1–161)–cTnI(1–172)–cTnT(236–285) complexes with R145G, R145G/S23D/S24D, and R145G/pS23/pS24\",\n      \"journal\": \"Biophysical journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 4 — computational simulation, but replicated triplicates with explicit comparison to phosphomimetic states\",\n      \"pmids\": [\"25606687\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"TNNI3 encodes the inhibitory subunit of the troponin complex and has evolved from TNNI1 and TNNI2 (slow and fast skeletal isoforms) through vertebrate evolution. cTnI has a unique N-terminal cardiac-specific extension (residues 1–32) that is phosphorylated by PKA at S23/S24 and by PKC at S43/S45, and contains an inhibitory peptide region and a switch peptide that interact with cTnC. Developmental regulation switches expression from slow skeletal TNNI1 to TNNI3 postnatally in cardiac muscle.\",\n      \"method\": \"Comparative genomic and phylogenetic analysis, review of biochemical and PTM literature\",\n      \"journal\": \"Gene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3-4 — review synthesizing published experimental data; supported by extensive primary literature\",\n      \"pmids\": [\"26526134\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"In cardiomyocytes from cTnI-G203S HCM mice, L-type Ca2+ channel (ICaL) inactivation was significantly faster. Activation of ICaL caused a greater increase in mitochondrial membrane potential and metabolic activity in cTnI-G203S myocytes compared to wild-type. This hypermetabolic response resulted from impaired communication between ICaL and F-actin, involving reduced actin-myosin dynamics and block of the mitochondrial voltage-dependent anion channel. L-type Ca2+ channel antagonists normalized mitochondrial membrane potential.\",\n      \"method\": \"Patch-clamp electrophysiology, mitochondrial membrane potential measurements, metabolic activity assays, pharmacological manipulation (nisoldipine, diltiazem) in isolated cardiomyocytes from transgenic cTnI-G203S mice\",\n      \"journal\": \"The Journal of physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple electrophysiological and metabolic measurements in disease model with pharmacological rescue\",\n      \"pmids\": [\"27062056\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Estrogen treatment of neonatal mouse hearts increased HDAC activity and HDAC5 binding at the cTnI promoter, reduced histone H3K9 acetylation at the cTnI promoter, and decreased cTnI expression at both mRNA and protein levels. SAHA (an HDAC inhibitor) rescued cTnI expression by inhibiting HDAC5 binding and restoring H3K9 acetylation at the cTnI promoter, indicating that HDAC5-mediated histone deacetylation represses TNNI3 transcription.\",\n      \"method\": \"Chromatin immunoprecipitation (ChIP), colorimetric HDAC/HAT activity assays, RT-PCR, Western blot, pharmacological HDAC inhibition in neonatal mouse hearts\",\n      \"journal\": \"Journal of cellular biochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — ChIP with functional pharmacological rescue; single lab\",\n      \"pmids\": [\"27379430\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"EGCG (epigallocatechin-3-gallate) improved cardiac diastolic function in aged mice by reversing age-related low cTnI expression. Mechanistically, EGCG inhibited HDAC1 and HDAC3 expression and reduced HDAC1 binding to the cTnI proximal promoter, increased H3K9 acetylation at the promoter, and enhanced binding of transcription factors GATA4 and Mef2c to the cTnI promoter.\",\n      \"method\": \"ChIP assay, Western blot, echocardiography, HDAC1/HDAC3 expression analysis, transcription factor binding assay in aged mice treated with EGCG\",\n      \"journal\": \"Journal of cellular and molecular medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — ChIP and multiple molecular assays with in vivo functional readout; single lab\",\n      \"pmids\": [\"28382690\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"cTnI is modified by arginine methylation in human myocardium. Mass spectrometry identified methylation sites at R74/R79 and R146/R148 in human cardiac samples. PRMT1 methylated an extended cTnI inhibitory peptide at R146 and R148 in vitro. HCM-associated mutations at R145 hampered R146/R148 methylation by PRMT1 in vitro. Methylation-deficient R146A/R148A cTnI expressed in H9c2 cells induced a 32% increase in cell size (hypertrophy), and Western blot showed reduced arginine methylation levels in HCM vs. DCM biopsies and in a rat cardiac hypertrophy model.\",\n      \"method\": \"Western blot with anti-methylarginine antibody, mass spectrometry, in vitro PRMT1 methylation assay, cell size measurement in H9c2 cells transfected with methylation-deficient mutant\",\n      \"journal\": \"International journal of cardiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — mass spectrometry identification + in vitro enzyme assay + cell-based functional consequence; single lab\",\n      \"pmids\": [\"30772011\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"In restrictive cardiomyopathy mice carrying the cTnI-193His mutation, PDE4d expression was epigenetically downregulated. cTnI was detected in the nucleus of cardiomyocytes by immunofluorescence and Western blot. Co-immunoprecipitation demonstrated direct interaction between cTnI and SMYD1 (histone methyltransferase) and between cTnI and HDAC1. Overexpression of mutated cTnI in cultured cardiomyocytes reduced PDE4d expression, suggesting nuclear cTnI modulates histone modifications and gene expression via interaction with chromatin-modifying enzymes.\",\n      \"method\": \"Immunofluorescence, Western blot of nuclear fractions, Co-IP, ChIP, cardiomyocyte overexpression studies\",\n      \"journal\": \"Science China. Life sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 — multiple methods (nuclear fractionation, Co-IP, ChIP) but findings require independent replication\",\n      \"pmids\": [\"30900165\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Panel sequencing of 80 pediatric cardiomyopathy patients identified 5 pathogenic TNNI3 variants; one patient carried a homozygous truncating variant (c.24+2T>A). Protein and transcript analysis on heart biopsies from this individual showed complete absence of TNNI3 protein and compensatory upregulation of the fetal slow skeletal isoform TNNI1, demonstrating that total loss of cTnI triggers a developmental isoform switch.\",\n      \"method\": \"Next-generation sequencing panel, Western blot and transcript analysis of endomyocardial biopsy\",\n      \"journal\": \"Clinical genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct protein analysis of patient cardiac tissue demonstrating isoform compensation\",\n      \"pmids\": [\"31568572\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"RCM-associated cTnI mutations R170G and R170W (in the regulatory C-terminus) strongly enhanced Ca2+ sensitivity in skinned cardiac fibers. Both mutants increased affinity of the troponin complex for tropomyosin; R170G strengthened and R170W weakened actin binding. Electron microscopy of reconstituted thin filaments showed wavy, broken filaments with R170G/W. MyBP-C N-terminal fragment (cMyBPC-C0C2) that normally reduces troponin binding to actin and increases cooperativity of thin filament activation had no effect in the presence of R170G/W cTn, and microscale thermophoresis identified direct cMyBPC-C0C2 binding to cTnI and cTnT (but not cTnC).\",\n      \"method\": \"Skinned fiber Ca2+-sensitivity measurements, electron microscopy of reconstituted thin filaments, co-sedimentation assays, microscale thermophoresis\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — multiple orthogonal methods (reconstitution, EM, MST, fiber mechanics) in one study\",\n      \"pmids\": [\"32182250\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"miR-449 regulates cTnI expression in cardiomyocytes by targeting HDAC1. miR-449 sponges HDAC1, relieving HDAC1-mediated deacetylation of H3K4 and H3K9 at the cTnI promoter GATA element, which in turn recruits transcription factor GATA4 to increase TNNI3 transcription. In vivo, miR-449 agomiR intervention in aged mice with low cTnI expression restored cTnI levels and improved cardiac function.\",\n      \"method\": \"Dual-luciferase reporter assay (miR-449/HDAC1 binding site), biochemical analysis, in vivo miR-449 agomiR injection in aged mice, echocardiography\",\n      \"journal\": \"European review for medical and pharmacological sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — reporter assay + in vivo rescue with defined molecular mechanism; single lab\",\n      \"pmids\": [\"33378032\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"De novo cTnI-D127Y mutation (infantile restrictive cardiomyopathy) increased Ca2+ sensitivity and reduced maximum force in skinned cardiomyocytes. In reconstituted thin filaments, cTnI-D127Y showed increased actin/tropomyosin binding affinity and compromised structural integrity (EM). Levosimendan and EGCG (troponin-targeting agents) partially stabilized thin filament structure and improved contractile parameters in vitro for both cTnI-D127Y and the companion cTnC-G34S mutation.\",\n      \"method\": \"Skinned fiber mechanics, reconstituted thin filament assays, electron microscopy, protein interaction binding assays, pharmacological intervention (levosimendan, EGCg)\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1-2 — reconstitution plus EM plus pharmacological rescue; single lab\",\n      \"pmids\": [\"34502534\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"iPSC-derived cardiomyocytes and engineered heart tissues (EHT) from a patient carrying TNNI3 R170W showed prolonged tau (relaxation time constant) in Ca2+ kinetics and an increased ratio of relaxation force to contractile force. CRISPR/Cas9 isogenic correction of R170W reversed these defects. Overexpression of wild-type TNNI3 in R170W-iPSC-CMs and EHTs also rescued impaired relaxation, demonstrating that cTnI levels and function directly determine diastolic dysfunction in RCM.\",\n      \"method\": \"iPSC generation, CRISPR/Cas9 isogenic correction, EHT formation, Ca2+ kinetics imaging, force measurements, TNNI3 overexpression rescue\",\n      \"journal\": \"Development, growth & differentiation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — isogenic correction + overexpression rescue in patient-derived model with functional readout\",\n      \"pmids\": [\"38193576\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Patient-specific iPSC-derived cardiomyocytes heterozygous and homozygous for TNNI3 R170W showed impaired diastolic function (cell motion analysis) compared to CRISPR-corrected isogenic lines and three independent healthy controls. Intracellular Ca2+ oscillation and troponin I immunocytochemistry were not significantly altered, and myofibril/mitochondrial ultrastructure appeared intact by electron microscopy, but RNA-seq revealed altered pathways in cardiac muscle development/contraction, extracellular matrix-receptor interaction, and TGF-β signaling.\",\n      \"method\": \"iPSC-CM differentiation, CRISPR/Cas9 isogenic lines, cell motion analysis, Ca2+ imaging, immunocytochemistry, electron microscopy, RNA sequencing\",\n      \"journal\": \"Journal of the American Heart Association\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — isogenic comparison with multiple readouts; transcriptomic data are associative\",\n      \"pmids\": [\"38497452\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"TNNI3-encoded cardiac troponin I (cTnI) is the inhibitory subunit of the sarcomeric troponin complex, whose switch peptide (residues 147–163) binds the N-terminal regulatory domain of cTnC in a Ca2+-dependent manner (inducing the open cTnC conformation) while its inhibitory peptide (residues ~128–147) normally restrains actin-tropomyosin; the cardiac-specific N-terminal extension is phosphorylated by PKA at S23/S24 (increasing length-dependent Ca2+ sensitivity and Frank-Starling response) and by PKC at S43/S45 (reducing contractility), PAK3 at S151 (increasing Ca2+ sensitivity via altered cTnC–cTnI distances), and by PRMT1 at R146/R148 (arginine methylation that modulates inhibitory/switch peptide function); cTnI is degraded by the E3 ubiquitin ligase MuRF1 in a RING finger- and UbcH5-dependent manner through mixed-linkage polyubiquitin chains; a fraction of cTnI localizes to the cardiomyocyte nucleus where it interacts with HDAC1 and SMYD1 to epigenetically regulate target genes including PDE4d; and TNNI3 transcription is regulated by HDAC1/HDAC3/HDAC5-mediated histone deacetylation at its promoter, counteracted by GATA4 and MEF2C binding, with mutations throughout the gene causing HCM, RCM, or DCM depending on their functional impact on Ca2+ sensitivity and thin filament structure.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"TNNI3 encodes cardiac troponin I (cTnI), the inhibitory subunit of the cardiac troponin complex that confers Ca²⁺-dependent regulation of striated muscle contraction by suppressing actomyosin ATPase activity at low Ca²⁺ through its inhibitory peptide (residues ~137–147) and relieving this inhibition when its switch region engages the N-domain of troponin C upon Ca²⁺ binding [PMID:26526134, PMID:24418317]. PKA phosphorylation of the cardiac-specific N-terminal extension at Ser23/Ser24 governs the steepness of the Frank-Starling length–tension relationship, while PKC phosphorylation at Ser43/Ser45 slows contraction and relaxation, and PAK3 phosphorylation at Ser151 increases myofilament Ca²⁺ sensitivity, establishing cTnI as a central integrator of multiple kinase-dependent signaling inputs [PMID:23836688, PMID:14726296, PMID:20540949]. The cTnI C-terminal regulatory region mediates cross-filament communication with cardiac myosin-binding protein C, and arginine methylation of R146/R148 by PRMT1 near the inhibitory peptide modulates hypertrophic signaling [PMID:32182250, PMID:30772011]. Pathogenic TNNI3 mutations cause hypertrophic and restrictive cardiomyopathy through increased Ca²⁺ sensitivity, impaired thin filament integrity, and disrupted β-adrenergic modulation, while complete loss of TNNI3 leads to compensatory TNNI1 upregulation and severe pediatric cardiomyopathy [PMID:34502534, PMID:38193576, PMID:31568572].\",\n  \"teleology\": [\n    {\n      \"year\": 1996,\n      \"claim\": \"Establishing the genomic architecture of TNNI3 and the activity of its proximal promoter answered whether the gene's expression was transcriptionally restricted to cardiac tissue.\",\n      \"evidence\": \"Genomic cloning, sequencing, and reporter assays in cardiac myocytes, skeletal muscle cells, and fibroblasts showed eight exons over ~6.2 kb with a proximal promoter active in muscle but not fibroblasts\",\n      \"pmids\": [\"8661099\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Distal enhancers conferring cardiac specificity not identified\", \"Transcription factor occupancy at the promoter not mapped\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Demonstrating that PKC phosphorylation of cTnI at Ser43/Ser45 slows contraction and relaxation in vivo resolved the functional consequence of this kinase input on cardiac mechanics.\",\n      \"evidence\": \"Transgenic mice with Ser43/45Ala substitutions showed enhanced basal contractility and blunted PKC-stimulated responses, with increased PKA-site phosphorylation\",\n      \"pmids\": [\"14726296\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Cross-talk mechanism between PKC and PKA phosphorylation sites not structurally resolved\", \"Contribution under pathological load not tested\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Identification of a TNNI3 frameshift truncation causing restrictive cardiomyopathy with ~50% protein reduction established haploinsufficiency as a disease mechanism.\",\n      \"evidence\": \"Patient TNNI3 sequencing plus Western blot of explanted myocardium showing halved cTnI levels\",\n      \"pmids\": [\"18006163\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single case report; no rescue or allele-specific expression data\", \"Whether residual truncated protein exerts dominant-negative effects unclear\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Showing that PAK3 phosphorylation at Ser151 increases Ca²⁺ sensitivity by altering cTnC–cTnI inter-site distances added a third kinase pathway converging on cTnI regulatory dynamics.\",\n      \"evidence\": \"S151E phosphomimetic cTnI reconstituted into thin filaments assessed by steady-state FRET, time-resolved FRET, and stopped-flow kinetics\",\n      \"pmids\": [\"20540949\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo relevance of PAK3–cTnI axis in intact hearts not demonstrated\", \"Interplay with Ser23/24 and Ser43/45 phosphorylation not tested\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Establishing that PKA phosphorylation at Ser23/Ser24 controls the steepness of the length–tension relationship resolved how β-adrenergic signaling tunes the Frank-Starling mechanism.\",\n      \"evidence\": \"Troponin exchange into permeabilized cardiac and slow-twitch fibres with unphosphorylated, PKA-treated, and Asp23/24 phosphomimetic cTnI; length–tension curves measured\",\n      \"pmids\": [\"23836688\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis for length-dependent change in phosphorylated cTnI conformation unknown\", \"Whether slow-twitch fibre result extends to fast-twitch muscle not addressed\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Reconstitution of HCM double-mutant troponin (cTnI-R145G + cTnT-R278C) revealed functional cross-talk between the cTnI inhibitory peptide and the cTnT C-terminus, answering whether disease mutations interact within the complex.\",\n      \"evidence\": \"In vitro motility assay with single and double mutant reconstituted troponin at varied Ca²⁺, temperature, and HMM density\",\n      \"pmids\": [\"24418317\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of intermolecular rescue not resolved\", \"No in vivo double-mutant model\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Molecular dynamics simulations of cTnI-R145G with phosphomimetic Ser23/24 showed that the HCM mutation disrupts intra-subunit communication between the N-terminal extension and the inhibitory peptide, explaining blunted β-adrenergic responsiveness in mutation carriers.\",\n      \"evidence\": \"Triplicate 150-ns MD simulations of troponin core domain with R145G, R145G/S23D/S24D, and phosphorylated states\",\n      \"pmids\": [\"25606687\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Computational prediction awaits experimental validation by FRET or cross-linking\", \"Simulated domain shorter than full-length cTnI\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Identification of HDAC5 binding at the cTnI promoter and pharmacological rescue with SAHA established epigenetic control of cTnI expression through histone H3K9 deacetylation, later extended to HDAC1/HDAC3 in aging hearts.\",\n      \"evidence\": \"ChIP for HDAC5/HDAC1/HDAC3 and H3K9ac at cTnI promoter; SAHA and EGCG treatments in neonatal and aged mice with echocardiographic readout\",\n      \"pmids\": [\"27379430\", \"28382690\", \"33378032\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"All from a single research group; independent replication needed\", \"Direct demonstration that HDAC-mediated cTnI reduction is sufficient to cause diastolic dysfunction not shown\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Discovery that cTnI-G203S impairs L-type Ca²⁺ channel–cytoskeleton coupling and produces mitochondrial hypermetabolism in pre-cardiomyopathic hearts revealed a non-canonical sarcomeric mutation effect beyond myofilament Ca²⁺ sensitivity.\",\n      \"evidence\": \"Patch-clamp ICa-L recordings, mitochondrial membrane potential and metabolic assays, and Ca²⁺-channel antagonist rescue in transgenic G203S mouse cardiomyocytes\",\n      \"pmids\": [\"27062056\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism linking cytoskeletal disruption to VDAC blockade not identified\", \"Single transgenic model without isogenic control\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Identification of arginine methylation at R146/R148 by PRMT1 and demonstration that methylation-deficient cTnI induces cardiomyocyte hypertrophy introduced a new post-translational regulatory layer adjacent to the inhibitory peptide.\",\n      \"evidence\": \"Mass spectrometry of human myocardium; in vitro PRMT1 assay with HCM mutants; H9c2 cell size measurement with methylation-deficient cTnI\",\n      \"pmids\": [\"30772011\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Downstream signaling pathway from methylation to hypertrophy not mapped\", \"In vivo validation of methylation-deficient phenotype lacking\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Complete TNNI3 loss-of-function in humans with compensatory TNNI1 upregulation established that cTnI is essential for normal cardiac function and that fetal TnI partially but insufficiently substitutes.\",\n      \"evidence\": \"Protein and transcript analysis of heart biopsies from homozygous TNNI3 splice-site mutation carriers\",\n      \"pmids\": [\"31568572\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Degree of functional rescue by TNNI1 not quantified mechanically\", \"Single center; small number of patients\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Characterizing cTnI-R170G/W mutations showed the C-terminal regulatory region controls thin filament structural integrity and mediates cross-filament communication with cMyBPC, answering how these mutations cause restrictive cardiomyopathy.\",\n      \"evidence\": \"Skinned fibre Ca²⁺-force curves, electron microscopy of reconstituted thin filaments, and microscale thermophoresis for troponin–cMyBPC C0C2 binding\",\n      \"pmids\": [\"32182250\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Atomic-level structure of cTnI C-terminus–cMyBPC interface not determined\", \"Effect on thick filament regulation in situ not tested\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Isogenic iPSC-cardiomyocyte and engineered heart tissue models of TNNI3-R170W, with CRISPR correction and overexpression rescue, proved that this variant directly causes diastolic dysfunction through a sarcomeric mechanism and identified secondary transcriptomic changes in TGF-β and ECM pathways.\",\n      \"evidence\": \"Patient iPSC-CMs and CRISPR-corrected controls; EHT force measurements; cell motion analysis; RNA sequencing; TNNI3 overexpression rescue\",\n      \"pmids\": [\"38193576\", \"38497452\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether transcriptomic changes are cause or consequence of mechanical dysfunction not resolved\", \"Long-term remodeling in vivo not modeled\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Key open questions include the structural basis of cTnI C-terminus–cMyBPC interaction, how arginine methylation integrates with phosphorylation to regulate thin filament function in vivo, and whether epigenetic control of TNNI3 transcription is therapeutically targetable in heart failure.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No high-resolution structure of cTnI C-terminus bound to cMyBPC N-terminal domains\", \"In vivo role of PRMT1-mediated cTnI methylation uncharacterized\", \"HDAC-targeted transcriptional rescue of cTnI awaits clinical translation studies\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 2, 3, 4]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [0, 8, 15]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [0, 8, 15]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [10]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-397014\", \"supporting_discovery_ids\": [0, 3, 4, 8, 16]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [2, 3, 4, 9]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [8, 12, 13, 15, 16, 17]}\n    ],\n    \"complexes\": [\n      \"cardiac troponin complex (cTnI–cTnC–cTnT)\"\n    ],\n    \"partners\": [\n      \"TNNC1\",\n      \"TNNT2\",\n      \"MYBPC3\",\n      \"PRMT1\",\n      \"HDAC1\",\n      \"HDAC5\",\n      \"PAK3\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```\n\n--- ROUND 2 (R2/R3, augmented corpus) ---\n\n```json\n{\n  \"mechanistic_narrative\": \"TNNI3 encodes cardiac troponin I (cTnI), the inhibitory subunit of the sarcomeric troponin complex that governs Ca²⁺-dependent regulation of cardiac muscle contraction. The cTnI switch peptide (residues 147–163) binds the N-terminal regulatory domain of troponin C in a Ca²⁺-dependent manner to induce its open conformation, while the inhibitory peptide (~residues 128–147) restrains actin–tropomyosin at low Ca²⁺; these interactions, resolved by crystal and NMR structures of the ternary troponin complex, are modulated by PKA phosphorylation at S23/S24 (governing length-dependent activation and the Frank–Starling mechanism), PKC phosphorylation at S43/S45 (reducing contractility), PAK3 phosphorylation at S151 (increasing Ca²⁺ sensitivity), and PRMT1-mediated arginine methylation at R146/R148 [PMID:10387074, PMID:12840750, PMID:23836688, PMID:14726296, PMID:20540949, PMID:30772011]. cTnI is targeted for proteasomal degradation by the E3 ubiquitin ligase MuRF1, which assembles mixed-linkage polyubiquitin chains via UbcH5 whose topology determines degradation efficiency [PMID:15601779, PMID:17426036]. Mutations throughout TNNI3 cause hypertrophic, restrictive, or dilated cardiomyopathy by altering Ca²⁺ sensitivity, thin-filament integrity, and diastolic relaxation, as demonstrated by family linkage studies, reconstituted fiber mechanics, and CRISPR-corrected patient iPSC-cardiomyocyte models [PMID:9241277, PMID:12531876, PMID:32182250, PMID:38193576].\",\n  \"teleology\": [\n    {\n      \"year\": 1996,\n      \"claim\": \"Defining the genomic organization and cardiac-enriched promoter of TNNI3 established a framework for understanding its transcriptional regulation, revealing that 2.3 kb of 5′-flanking sequence drives expression in cardiac but not non-muscle cells, though full cardiac specificity required additional elements.\",\n      \"evidence\": \"Genomic cloning, sequencing, and transfection reporter assays in cardiac myocytes and fibroblasts\",\n      \"pmids\": [\"8661099\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Minimal regulatory elements sufficient for cardiac specificity not mapped\", \"No in vivo transgenic validation of promoter constructs\"]\n    },\n    {\n      \"year\": 1997,\n      \"claim\": \"Identification of TNNI3 mutations in HCM families established cTnI as a disease gene and demonstrated that the inhibitory subunit of the troponin complex is essential for normal cardiac function, opening the mechanistic question of how specific residue changes alter contractility.\",\n      \"evidence\": \"Candidate gene sequencing in 184 unrelated HCM patients with family linkage analysis\",\n      \"pmids\": [\"9241277\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular mechanism by which individual mutations (e.g., R145G) alter troponin function not yet determined\", \"No functional reconstitution at this stage\"]\n    },\n    {\n      \"year\": 1998,\n      \"claim\": \"The first crystal structure of a TnC–TnI fragment complex revealed that TnI binding converts TnC from an elongated to a compact globular conformation, providing the first atomic-level view of how the inhibitory subunit contacts both lobes of TnC.\",\n      \"evidence\": \"X-ray crystallography at 2.3 Å resolution with MAD phasing\",\n      \"pmids\": [\"9560191\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Only N-terminal TnI fragment (1–47) resolved; inhibitory and switch peptide regions not visualized\", \"No Ca²⁺-dependent conformational changes captured\"]\n    },\n    {\n      \"year\": 1999,\n      \"claim\": \"NMR resolution of the cTnI switch peptide (147–163) bound to the Ca²⁺-loaded cTnC N-terminal domain revealed that cTnI binding—not Ca²⁺ alone—induces the open conformation of cNTnC, establishing the molecular basis for Ca²⁺-dependent activation of cardiac muscle.\",\n      \"evidence\": \"Multinuclear multidimensional NMR spectroscopy and solution structure determination\",\n      \"pmids\": [\"10387074\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Isolated peptide–domain complex; behavior in the context of the full troponin–tropomyosin–actin filament not addressed\", \"Inhibitory peptide conformation not resolved simultaneously\"]\n    },\n    {\n      \"year\": 2003,\n      \"claim\": \"Two landmark advances: the crystal structure of the ternary cardiac troponin core domain revealed the IT arm coiled-coil architecture and Ca²⁺-dependent release of cTnI from actin, while genetic linkage and multi-family screening established TNNI3 as a cause of restrictive cardiomyopathy in addition to HCM.\",\n      \"evidence\": \"X-ray crystallography of ternary troponin core domain (Nature); linkage analysis (LOD 4.8) plus mutation screening in RCM families (JCI)\",\n      \"pmids\": [\"12840750\", \"12531876\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Full-length troponin on the thin filament not structurally resolved\", \"Mechanism by which the same gene causes HCM vs. RCM not clarified at structural level\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Parallel discoveries showed that cTnI is a substrate of the E3 ubiquitin ligase MuRF1 (linking proteasomal degradation to contractile protein turnover) and that PKC phosphorylation at S43/S45 attenuates contractility in vivo, with cross-talk between PKC and PKA phosphorylation sites.\",\n      \"evidence\": \"Yeast two-hybrid, co-IP, in vitro ubiquitylation, and cardiomyocyte contractility assays for MuRF1; transgenic S43A/S45A mice with in situ hemodynamics for PKC sites\",\n      \"pmids\": [\"15601779\", \"14726296\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological signals controlling MuRF1-mediated cTnI turnover rates in vivo unclear\", \"PKC isoform specificity for S43/S45 phosphorylation in intact heart not defined\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Reconstitution of MuRF1-mediated ubiquitylation with defined E2 enzymes revealed that ubiquitin chain topology (mixed forked chains via UbcH5 vs. homogeneous K48 or K63 chains) determines cTnI degradation efficiency by the 26S proteasome, adding a regulatory layer to sarcomeric protein quality control.\",\n      \"evidence\": \"In vitro ubiquitylation with purified components, mass spectrometry of chain linkages, 26S proteasome degradation assays\",\n      \"pmids\": [\"17426036\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo chain type preference on endogenous cTnI not confirmed\", \"Whether deubiquitinases selectively edit specific chain types on cTnI is unknown\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"PAK3-mediated phosphorylation of cTnI at S151 was shown to increase myofilament Ca²⁺ sensitivity by shortening inter-site cTnC–cTnI distances and slowing Ca²⁺-dissociation-induced structural transitions, mimicking the effect of strong crossbridges and identifying a third kinase-specific regulatory input on cTnI.\",\n      \"evidence\": \"Steady-state and time-resolved FRET, stopped-flow kinetics in reconstituted thin filaments with S151E phosphomimetic\",\n      \"pmids\": [\"20540949\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo contribution of PAK3 to cTnI phosphorylation at S151 under physiological stimuli not quantified\", \"Interplay between S151 phosphorylation and S23/S24 or S43/S45 not tested\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"PKA phosphorylation of cTnI at S23/S24 was established as the molecular switch for length-dependent Ca²⁺ sensitivity (Frank–Starling mechanism), as unphosphorylated troponin exchange abolished steep length–tension relationships that were restored by PKA treatment or phosphomimetic substitution.\",\n      \"evidence\": \"Troponin exchange in permeabilized cardiac myocytes and skeletal fibers, PKA treatment, S23D/S24D phosphomimetics, force-length measurements\",\n      \"pmids\": [\"23836688\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural mechanism by which N-terminal extension phosphorylation transmits length sensing to the regulatory domain not resolved\", \"Contribution of titin compliance to the observed effects not dissected\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"MD simulations of the R145G HCM mutation provided a molecular explanation for its blunted β-adrenergic response: R145G stabilizes Ca²⁺ coordination in cTnC and prevents PKA-phosphorylation-induced intramolecular contact between the N-terminal extension and the inhibitory peptide, consistent with experimental observation of impaired β-adrenergic modulation in R145G cardiomyocytes.\",\n      \"evidence\": \"Triplicate 150 ns molecular dynamics simulations of troponin subcomplexes with R145G and phosphomimetic variants; complemented by earlier cardiomyocyte sarcomere shortening data\",\n      \"pmids\": [\"25606687\", \"18548271\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Computational prediction; direct experimental validation of altered intramolecular contacts in full-length cTnI-R145G lacking\", \"Simulations limited to 150 ns timescale\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Epigenetic regulation of TNNI3 transcription was uncovered: HDAC5 binds the cTnI promoter and deacetylates H3K9, repressing transcription—an effect reversed by HDAC inhibitor SAHA—adding a transcriptional regulatory layer to cTnI biology.\",\n      \"evidence\": \"ChIP, HDAC activity assays, RT-PCR, Western blot, and HDAC inhibitor treatment in neonatal mouse hearts\",\n      \"pmids\": [\"27379430\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Physiological contexts in which HDAC5 is recruited to the TNNI3 promoter beyond estrogen exposure unclear\", \"Whether HDAC5 directly binds DNA or is recruited via a transcription factor complex not determined\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"HDAC1 and HDAC3 were further implicated in TNNI3 promoter repression, with GATA4 and MEF2C binding shown to activate transcription when HDAC-mediated deacetylation is relieved, defining a complete activator–repressor switch at the TNNI3 locus.\",\n      \"evidence\": \"ChIP, Western blot, echocardiography, and EGCG treatment in aged mice\",\n      \"pmids\": [\"28382690\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct HDAC1/3 recruitment mechanism not resolved\", \"Whether age-related cTnI decline is a cause or consequence of cardiac dysfunction not established\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Three concurrent advances expanded cTnI biology: (1) arginine methylation at R146/R148 by PRMT1 was discovered in human myocardium, with HCM mutations at R145 impairing this modification and methylation-deficient cTnI inducing hypertrophy; (2) nuclear cTnI was shown to interact with SMYD1 and HDAC1, epigenetically regulating PDE4d expression; (3) complete loss of cTnI protein in a patient with a homozygous truncating variant triggered compensatory re-expression of fetal TNNI1.\",\n      \"evidence\": \"Mass spectrometry, in vitro PRMT1 assays, H9c2 cell transfection (PMID:30772011); nuclear fractionation, co-IP, ChIP in RCM mice (PMID:30900165); Western blot/transcript analysis of endomyocardial biopsy (PMID:31568572)\",\n      \"pmids\": [\"30772011\", \"30900165\", \"31568572\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Nuclear cTnI findings require independent replication and quantification of nuclear vs. sarcomeric pools\", \"Functional consequence of R146/R148 methylation on thin filament Ca²⁺ regulation not tested in reconstituted systems\", \"Whether TNNI1 compensation is sufficient for long-term cardiac function unknown\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Reconstitution and EM studies of RCM-associated R170G/W mutations showed increased tropomyosin affinity, disrupted thin filament structural integrity, and loss of cMyBP-C regulatory modulation—demonstrating that a single cTnI residue change can simultaneously affect multiple protein–protein interfaces within the thin filament.\",\n      \"evidence\": \"Skinned fiber mechanics, reconstituted thin filament EM, co-sedimentation, microscale thermophoresis\",\n      \"pmids\": [\"32182250\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether loss of MyBP-C modulation is a general feature of RCM mutations or specific to R170 unclear\", \"In vivo consequence of altered thin filament integrity not assessed in animal model\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Patient-derived iPSC-cardiomyocyte and engineered heart tissue models of TNNI3 R170W RCM demonstrated that the mutation directly causes diastolic dysfunction (prolonged relaxation), which was reversed by CRISPR isogenic correction and wild-type TNNI3 overexpression, establishing a causal link from genotype to diastolic phenotype and validating gene augmentation as a potential therapeutic strategy.\",\n      \"evidence\": \"iPSC-CM generation, CRISPR/Cas9 isogenic correction, EHT formation, Ca²⁺ kinetics imaging, force measurements, TNNI3 overexpression rescue\",\n      \"pmids\": [\"38193576\", \"38497452\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Long-term in vivo efficacy of TNNI3 gene augmentation not tested\", \"Whether TNNI3 overexpression causes dominant-negative effects at supraphysiological levels not evaluated\", \"Transcriptomic changes (TGF-β, ECM pathways) observed in mutant cells await mechanistic dissection\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Key unresolved questions include: the structural basis for how individual TNNI3 mutations differentially cause HCM, RCM, or DCM; the physiological significance and regulation of nuclear cTnI; the in vivo dynamics and chain-type specificity of MuRF1-mediated cTnI turnover; and whether gene therapy or allele-specific silencing can correct TNNI3-associated cardiomyopathies.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No high-resolution cryo-EM structure of the full thin filament with disease-mutant troponin\", \"Nuclear cTnI interactome and gene regulatory targets not comprehensively mapped\", \"No clinical gene therapy trials for TNNI3 cardiomyopathies\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [3, 4, 12, 13, 14]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [4, 14, 23]},\n      {\"term_id\": \"GO:0005198\", \"supporting_discovery_ids\": [4, 23]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [4, 6, 23]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [21]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-397014\", \"supporting_discovery_ids\": [3, 4, 7, 12, 13, 14, 23, 26]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [7, 13, 17]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [6, 10, 20]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [1, 5, 9, 11, 25, 26]}\n    ],\n    \"complexes\": [\n      \"cardiac troponin complex (cTnC–cTnI–cTnT)\",\n      \"thin filament (troponin–tropomyosin–actin)\"\n    ],\n    \"partners\": [\n      \"TNNC1\",\n      \"TNNT2\",\n      \"TRIM63\",\n      \"HDAC1\",\n      \"SMYD1\",\n      \"PRMT1\",\n      \"MYBPC3\",\n      \"RCAN3\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}