{"gene":"CALD1","run_date":"2026-04-28T17:28:52","timeline":{"discoveries":[{"year":1992,"finding":"Smooth muscle caldesmon (CALD1) is phosphorylated in vitro by MAP kinase (p44mpk) at up to 2.0 mol phosphate/mol protein at Ser and Thr residues located predominantly in the C-terminal 10-kDa fragment that houses binding sites for calmodulin, tropomyosin, and F-actin. Phosphorylation attenuated caldesmon's interaction with actin but did not affect its binding to calmodulin or tropomyosin. Differentiated smooth muscle cells (chicken gizzard, rat aorta) contain endogenous MAP kinase that immunoprecipitates with caldesmon-directed kinase activity, establishing MAP kinase as a writer of caldesmon phosphorylation in smooth muscle.","method":"In vitro phosphorylation assay with purified sea-star p44mpk, tryptic peptide mapping of 32P-labeled caldesmon, actin/calmodulin/tropomyosin binding assays post-phosphorylation, immunoprecipitation of endogenous MAP kinase from smooth muscle extracts","journal":"The Journal of Biological Chemistry","confidence":"High","confidence_rationale":"Tier 1 — in vitro reconstitution with mutagenesis-equivalent peptide mapping and functional binding assays; replicated in two smooth muscle tissue types","pmids":["1331069"],"is_preprint":false},{"year":2000,"finding":"Kinase-dead PAK1 (K299R) stabilizes actin stress fibers in invasive breast cancer cells in part through persistent co-localization with caldesmon (CALD1) and tropomyosin on F-actin filaments. Loss of PAK1 activity prevents stress fiber disassembly and reduces invasiveness, placing caldesmon downstream of PAK1 signaling in cytoskeletal remodeling during cancer cell invasion.","method":"Overexpression of kinase-dead K299R PAK1 in MDA-MB435 and MDA-MB231 cells; immunofluorescence co-localization of caldesmon and tropomyosin with F-actin; focal adhesion and invasion assays","journal":"The Journal of Biological Chemistry","confidence":"Medium","confidence_rationale":"Tier 2 — clean loss-of-function (kinase-dead mutant) with defined cellular phenotype and co-localization; single lab","pmids":["10766836"],"is_preprint":false},{"year":2004,"finding":"Differential splicing of CALD1 exons occurs in glioma microvasculature versus normal brain microvasculature. Missplicing involving exons 1, 1+4, and 1'+4 is exclusive to tumor microvessels and results in upregulated caldesmon protein expression. This transcriptional/splicing change co-exists with downregulation of tight junction proteins occludin and ZO-1, implicating CALD1 missplicing as an epigenetic event contributing to endothelial barrier dysfunction in glioma.","method":"Laser-capture microdissection of tumor and normal microvessels, RT-PCR for splice variants, coupled in vitro transcription/translation assay (IVTT) for mutation scanning, immunohistochemistry for caldesmon, occludin, and ZO-1","journal":"The American Journal of Pathology","confidence":"Medium","confidence_rationale":"Tier 2 — direct tissue microdissection with RT-PCR splice-variant identification and protein-level confirmation; single lab with multiple orthogonal methods","pmids":["15161654"],"is_preprint":false},{"year":2007,"finding":"The low-molecular-weight caldesmon isoform (Hela l-CaD) localizes specifically to motility-related cell protrusions — filopodia, microspikes, lamellipodia, podosomes, membrane blebs, and membrane ruffles — in activated endothelial cells and endothelial progenitor cells within human tumor vasculature, implicating this isoform in EC/EPC migration during tumor angiogenesis and vasculogenesis.","method":"Immunohistochemistry on histologically preserved tumor microenvironment sections from multiple human tumor types; subcellular localization of Hela l-CaD to identified protrusion subtypes","journal":"Cell Adhesion & Migration","confidence":"Medium","confidence_rationale":"Tier 3 — direct localization experiment with functional context (protrusion subtypes tied to motility); single lab","pmids":["19329885"],"is_preprint":false},{"year":2008,"finding":"Tumor-specific alternative splicing of CALD1 is identified across colon, bladder, and prostate cancers, representing a general cancer-related splicing event. In silico protein predictions indicate that cancer-specific CALD1 splice variants encode proteins with potentially altered functions, suggesting involvement in cancer pathogenesis.","method":"GeneChip Human Exon 1.0 ST Array on 102 normal and cancer tissue samples; RT-PCR validation on independent set of 81 normal and tumor samples; in silico protein structure prediction","journal":"Molecular & Cellular Proteomics","confidence":"Medium","confidence_rationale":"Tier 2 — genome-wide discovery confirmed by RT-PCR in independent cohort; multi-organ replication","pmids":["18353764"],"is_preprint":false},{"year":2021,"finding":"CALD1 modulates glioma progression by facilitating tumor angiogenesis. Single-cell RNA-seq and bulk RNA-seq analyses demonstrate CALD1 upregulation in neoplastic cells, and histology/immunofluorescence shows CALD1 association with vessel architecture. CALD1 expression correlates with endothelial cell and pericyte infiltration in the tumor microenvironment.","method":"Bulk RNA-seq and single-cell RNA-seq (TCGA, CGGA databases), immunofluorescence, histological analysis of vessel architecture","journal":"Cancers","confidence":"Low","confidence_rationale":"Tier 3/4 — primarily bioinformatic and correlative; limited direct mechanistic experimentation","pmids":["34070840"],"is_preprint":false},{"year":2021,"finding":"CALD1 promotes PD-L1 (CD274) expression in bladder cancer via activation of the JAK/STAT signaling pathway. CALD1 overexpression upregulates PD-L1, and this effect is blocked by the specific JAK inhibitor Ruxolitinib, placing CALD1 upstream of JAK/STAT-mediated immune checkpoint regulation.","method":"WGCNA gene network analysis; CCK-8, flow cytometry, Transwell assays; nude mouse xenograft; JAK inhibitor (Ruxolitinib) rescue experiment; GSEA pathway analysis; correlation analysis with TIMER database","journal":"Annals of Translational Medicine","confidence":"Medium","confidence_rationale":"Tier 2 — pharmacological rescue (JAK inhibitor) places CALD1 in pathway; in vitro and in vivo evidence; single lab","pmids":["34733993"],"is_preprint":false},{"year":2021,"finding":"Linggui Zhugan decoction (LGZGD) improves insulin resistance in vivo (HFD rats) and in vitro (TNF-α-treated 3T3-L1 adipocytes) by downregulating Cald1 expression. Knockdown of Cald1 is associated with increased insulin-stimulated glucose uptake, identifying CALD1 as a novel regulator of insulin sensitivity in adipocytes.","method":"Gene microarray to identify DEGs in HFD vs. LGZGD-treated rats; qRT-PCR validation; 3H-2-DG glucose uptake assay in IR 3T3-L1 adipocytes; Oil Red O staining","journal":"Journal of Traditional Chinese Medicine","confidence":"Low","confidence_rationale":"Tier 3 — functional glucose uptake assay with CALD1 expression change, but mechanistic link is indirect; single lab, single method","pmids":["34708628"],"is_preprint":false},{"year":2022,"finding":"AHSA1 promotes proliferation and epithelial-mesenchymal transition (EMT) in hepatocellular carcinoma through an ERK/CALD1 axis. AHSA1 recruits ERK1/2 and promotes phosphorylation (inactivation) of CALD1; ERK1/2 phosphorylation inhibitor SCH772984 reverses the effect of AHSA1 on proliferation and EMT. CALD1 knockdown rescues the inhibition of proliferation and EMT caused by AHSA1 knockdown, placing CALD1 as a downstream effector of AHSA1-ERK signaling independent of HSP90 and MEK1/2.","method":"Gain- and loss-of-function studies in vitro and in vivo; ERK1/2 inhibitor (SCH772984) rescue; CALD1 knockdown epistasis; western blot for phospho-CALD1","journal":"Cancers","confidence":"Medium","confidence_rationale":"Tier 2 — epistasis (double knockdown rescue) and pharmacological inhibitor evidence establish pathway position; in vitro and in vivo; single lab","pmids":["36230524"],"is_preprint":false},{"year":2023,"finding":"miR-1278 directly targets CALD1 mRNA (confirmed by dual luciferase reporter assay) and negatively regulates its expression. CALD1 overexpression promotes gastric cancer cell viability and migration and activates the MAPK pathway (Ras, p-P38, p-ERK1/2). miR-1278 mimic partially rescues the pro-tumorigenic effects of CALD1 overexpression, placing CALD1 as a downstream target of miR-1278 in the miR-1278/CALD1/MAPK axis.","method":"Dual luciferase reporter assay (miR-1278 targeting CALD1 3'UTR); CCK-8 and Transwell migration assays; western blot for MAPK pathway proteins; tumor xenograft assay","journal":"Open Medicine","confidence":"Medium","confidence_rationale":"Tier 2 — direct 3'UTR luciferase validation of miRNA-target interaction plus epistasis rescue experiment; single lab","pmids":["38025524"],"is_preprint":false},{"year":2023,"finding":"METTL14-mediated m6A methylation regulates CALD1 mRNA levels in oral squamous cell carcinoma. METTL14 silencing depletes both mRNA and m6A levels of CALD1 (confirmed by MeRIP assay), and CALD1 overexpression rescues the growth and metastasis inhibition caused by METTL14 knockdown, identifying METTL14 as an m6A writer that stabilizes CALD1 mRNA to promote OSCC progression.","method":"MeRIP (m6A RNA immunoprecipitation) assay; qRT-PCR and western blot; colony formation and Transwell assays; tumorigenicity assay in vivo; CALD1 OE rescue of si-METTL14","journal":"Journal of Environmental Pathology, Toxicology and Oncology","confidence":"Medium","confidence_rationale":"Tier 2 — MeRIP directly measures m6A on CALD1 mRNA; epistasis rescue confirms functional relationship; single lab","pmids":["37017680"],"is_preprint":false},{"year":2024,"finding":"CALD1 promotes EMT and invasiveness in gastric cancer via the PI3K-Akt signaling pathway. CALD1 overexpression increases PI3K, p-AKT, and p-mTOR expression and decreases PTEN; PI3K-Akt inhibitor treatment attenuates CALD1-driven cell migration, invasion, and EMT marker changes. Animal xenograft experiments confirm that CALD1 inhibition slows tumor growth with corresponding changes in PI3K-Akt and EMT pathway proteins.","method":"CALD1 siRNA knockdown and overexpression in AGS and MKN45 GC cells; CCK-8, wound healing, Transwell assays; western blot for PI3K/p-AKT/p-mTOR/PTEN; PI3K-Akt inhibitor rescue; GC xenograft mouse model","journal":"World Journal of Gastrointestinal Oncology","confidence":"Medium","confidence_rationale":"Tier 2 — pharmacological pathway inhibitor rescue combined with in vitro and in vivo loss/gain-of-function; single lab","pmids":["38577446"],"is_preprint":false},{"year":2024,"finding":"Endothelin-1 (ET-1) increases high-molecular-weight CALD1 (a marker of the contractile phenotype of smooth muscle cells) in placental chorionic plate vessels and human umbilical vein smooth muscle cells (HUVSMC) via ETAR and ETBR receptors. This increase in CALD1 is associated with stronger vasoconstriction in preeclamptic placental veins, independent of CaV1.2 channel activity.","method":"Vasocontraction measurements (DMT) with ET-1, receptor antagonists, and CaV1.2 blockers; whole-cell patch clamp for CaV1.2 in single SMCs; PCR, western blot, ELISA for CALD1 and receptor expression; placental vessel explants and HUVSMC ET-1 treatment with/without antagonists","journal":"Placenta","confidence":"Medium","confidence_rationale":"Tier 2 — pharmacological receptor antagonism identifies ETAR/ETBR as upstream regulators of CALD1 in a functional vasoconstriction assay; multiple orthogonal methods; single lab","pmids":["39476475"],"is_preprint":false},{"year":2025,"finding":"CALD1 knockdown in SK-OV-3 ovarian cancer cells reduces F-actin stress fibers, loosens cytoskeletal structure, decreases Vinculin expression, and reduces focal adhesion number, resulting in enhanced cell migration and invasiveness. This establishes CALD1 as a regulator of cytoskeletal organization and focal adhesion formation that suppresses ovarian cancer cell invasion.","method":"Stable CALD1 shRNA knockdown in SK-OV-3 cells; Transwell invasion assay; immunofluorescence staining of F-actin and Vinculin; western blot","journal":"Translational Cancer Research","confidence":"Medium","confidence_rationale":"Tier 2 — clean KO with defined cytoskeletal phenotype (F-actin, Vinculin, focal adhesions) and functional invasiveness readout; single lab","pmids":["40104711"],"is_preprint":false},{"year":2025,"finding":"In Helicobacter pylori-stimulated cancer-associated fibroblasts (CAFs), TLR signaling suppresses miR-148a-5p, leading to upregulation of CALD1 in CAFs. CALD1 upregulation drives collagen VI secretion from CAFs, which interacts with tumoral SDC4 receptors to promote gastric cancer cell proliferation. miR-148a-5p agomir inhibits this pathway in vivo and enhances chemotherapy efficacy.","method":"microRNA and transcriptome analysis; single-cell sequencing; in vitro CAF-GC co-culture; cell-derived xenograft (CDX) and patient-derived xenograft (PDX) mouse models; miR-148a-5p agomir treatment","journal":"Cellular Oncology","confidence":"Medium","confidence_rationale":"Tier 2 — pathway dissected through miRNA manipulation, co-culture, and in vivo PDX/CDX models; single lab","pmids":["41171370"],"is_preprint":false}],"current_model":"CALD1 (caldesmon) is a cytoskeleton-associated actin-binding protein whose C-terminal domain (housing calmodulin, tropomyosin, and F-actin binding sites) is phosphorylated by MAP kinase and p34cdc2, which attenuates its actin interaction and thereby modulates smooth muscle contractility and stress fiber stability; in non-muscle cells, CALD1 localizes to motility protrusions and regulates focal adhesion number, F-actin stress fiber integrity, and cell invasion downstream of PAK1/ERK signaling, while its expression is controlled at the post-transcriptional level by m6A methylation (METTL14) and miRNAs (miR-1278, miR-148a-5p), and it acts as an upstream regulator of JAK/STAT, PI3K-Akt, and MAPK pathways to influence EMT and immune checkpoint expression in cancer contexts."},"narrative":{"teleology":[{"year":1992,"claim":"Identifying how caldesmon's actin-regulatory activity is switched off resolved a long-standing question about the kinase signal that controls smooth muscle thin-filament regulation: MAP kinase phosphorylates the C-terminal actin-binding domain and selectively weakens the caldesmon–actin interaction without disrupting calmodulin or tropomyosin binding.","evidence":"In vitro phosphorylation of purified chicken gizzard caldesmon by sea-star p44mpk; tryptic peptide mapping; actin/calmodulin/tropomyosin co-sedimentation assays; immunoprecipitation of endogenous MAP kinase from rat aorta smooth muscle","pmids":["1331069"],"confidence":"High","gaps":["In vivo phosphorylation sites not mapped at single-residue resolution in this study","Structural basis of how phosphorylation weakens actin binding not determined","Whether additional kinases (e.g. p34cdc2) phosphorylate the same or distinct sites in smooth muscle not addressed"]},{"year":2000,"claim":"Establishing that caldesmon participates in PAK1-dependent cytoskeletal remodeling in invasive breast cancer cells extended its role beyond smooth muscle contraction to cancer cell motility: kinase-dead PAK1 traps caldesmon and tropomyosin on stabilized stress fibers, blocking invasion.","evidence":"Overexpression of kinase-dead PAK1(K299R) in MDA-MB435 and MDA-MB231 cells; immunofluorescence co-localization of caldesmon/tropomyosin with F-actin; invasion assays","pmids":["10766836"],"confidence":"Medium","gaps":["Whether PAK1 directly phosphorylates caldesmon was not tested","Caldesmon was not knocked down to confirm it is required for PAK1-dependent invasion"]},{"year":2004,"claim":"Discovery of tumor-specific CALD1 splice variants in glioma and subsequently in colon, bladder, and prostate cancers revealed that alternative splicing diversifies caldesmon function in the tumor microenvironment and may alter its actin-regulatory properties.","evidence":"Laser-capture microdissection of glioma versus normal microvessels with RT-PCR splice-variant identification; GeneChip Exon Array across 102 samples with RT-PCR validation in 81 independent samples","pmids":["15161654","18353764"],"confidence":"Medium","gaps":["Functional consequences of cancer-specific splice variants on actin binding or contractility not experimentally validated","Whether splice variants are causative or merely correlative in tumor progression remains untested"]},{"year":2007,"claim":"Subcellular mapping of the low-molecular-weight caldesmon isoform to motility-associated protrusions (filopodia, lamellipodia, podosomes, membrane blebs) in tumor-associated endothelial cells specified the cellular compartment in which caldesmon operates during tumor angiogenesis.","evidence":"Immunohistochemistry on multiple human tumor types identifying l-CaD in endothelial and endothelial progenitor cell protrusion subtypes","pmids":["19329885"],"confidence":"Medium","gaps":["Loss-of-function in endothelial protrusions not performed","Whether l-CaD is required for protrusion formation or merely accumulates there is unknown"]},{"year":2021,"claim":"Placing CALD1 upstream of JAK/STAT-mediated PD-L1 expression in bladder cancer expanded its role from a cytoskeletal regulator to a modulator of immune checkpoint signaling, as pharmacological JAK inhibition (Ruxolitinib) blocked CALD1-driven PD-L1 upregulation.","evidence":"CALD1 overexpression/knockdown in bladder cancer cells; JAK inhibitor rescue; nude mouse xenograft; GSEA pathway analysis","pmids":["34733993"],"confidence":"Medium","gaps":["Direct physical interaction between CALD1 and JAK pathway components not demonstrated","Whether the JAK/STAT link operates in non-bladder cancer contexts is unknown"]},{"year":2022,"claim":"Epistasis experiments showed CALD1 is a functional effector downstream of the AHSA1–ERK axis in hepatocellular carcinoma EMT: CALD1 knockdown rescued the proliferation and EMT inhibition caused by AHSA1 depletion, and ERK inhibitor SCH772984 reversed AHSA1-driven CALD1 phosphorylation.","evidence":"Double knockdown epistasis (AHSA1 + CALD1); ERK1/2 inhibitor rescue; in vitro and in vivo HCC models; western blot for phospho-CALD1","pmids":["36230524"],"confidence":"Medium","gaps":["Specific ERK phosphorylation sites on CALD1 not mapped in this system","Whether CALD1's actin-binding function is required for the EMT phenotype not tested"]},{"year":2023,"claim":"Identification of post-transcriptional regulators of CALD1 — m6A methylation by METTL14 and direct miR-1278 targeting of the 3′UTR — established that CALD1 expression is tuned at the mRNA level in cancer, with functional consequences for MAPK pathway activation and tumor growth.","evidence":"MeRIP assay for m6A on CALD1 mRNA with METTL14 knockdown/rescue in OSCC; dual luciferase reporter assay validating miR-1278 binding to CALD1 3′UTR in gastric cancer; xenograft models","pmids":["37017680","38025524"],"confidence":"Medium","gaps":["Specific m6A sites on CALD1 mRNA not mapped at nucleotide resolution","Whether METTL14 and miR-1278 regulation operates simultaneously or in different cellular contexts is unknown"]},{"year":2024,"claim":"CALD1 was shown to drive EMT and invasion in gastric cancer specifically through PI3K-Akt-mTOR signaling, with pathway inhibitor rescue confirming directionality; separately, endothelin-1 was identified as an upstream inducer of high-molecular-weight CALD1 in placental smooth muscle, linking CALD1 to vasoconstriction in preeclampsia.","evidence":"PI3K-Akt inhibitor rescue of CALD1 overexpression phenotypes in GC cells and xenografts; ET-1 receptor antagonism in placental veins and HUVSMCs with vasocontraction measurements","pmids":["38577446","39476475"],"confidence":"Medium","gaps":["How CALD1, an actin-binding protein, activates PI3K-Akt remains mechanistically undefined","Whether ET-1-induced CALD1 upregulation is transcriptional or post-translational not determined"]},{"year":2025,"claim":"Two studies completed the picture of CALD1's context-dependent role: in ovarian cancer, CALD1 knockdown reduced F-actin stress fibers and focal adhesions, paradoxically increasing invasion; in gastric cancer CAFs, H. pylori-driven TLR signaling suppresses miR-148a-5p to upregulate CALD1, which drives collagen VI secretion and paracrine proliferative signaling through tumoral SDC4.","evidence":"Stable shRNA knockdown in SK-OV-3 with F-actin/Vinculin immunofluorescence and invasion assays; miR-148a-5p agomir in CAF-GC co-culture and PDX/CDX mouse models","pmids":["40104711","41171370"],"confidence":"Medium","gaps":["Opposing invasion phenotypes in ovarian cancer versus other tumor types not reconciled mechanistically","How CALD1 regulates collagen VI secretion in CAFs is unknown","Whether the miR-148a-5p/CALD1/collagen VI/SDC4 axis operates outside gastric cancer is untested"]},{"year":null,"claim":"A unifying structural and signaling model explaining how a single actin-binding protein exerts context-dependent pro- or anti-invasive effects across different cell types, and how its cytoskeletal function connects to downstream transcriptional pathway activation (JAK/STAT, PI3K-Akt, MAPK), remains to be established.","evidence":"","pmids":[],"confidence":"High","gaps":["No high-resolution structure of caldesmon bound to F-actin available","Direct mechanism linking CALD1 to activation of PI3K-Akt, JAK/STAT, and MAPK transcriptional programs is unknown","Relative contribution of individual phosphorylation sites (ERK, p34cdc2, PAK1) to distinct phenotypic outputs has not been dissected in a single system"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[0,1,3,13]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[6,8,9,11]}],"localization":[{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[0,1,3,13]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[3]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[1,13]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[6,8,9,11]},{"term_id":"R-HSA-397014","term_label":"Muscle contraction","supporting_discovery_ids":[0,12]},{"term_id":"R-HSA-1474244","term_label":"Extracellular matrix organization","supporting_discovery_ids":[14]}],"complexes":[],"partners":["ACTA2","TPM1","CALM1","PAK1","MAPK1","METTL14","AHSA1"],"other_free_text":[]},"mechanistic_narrative":"CALD1 (caldesmon) is an actin- and calmodulin-binding cytoskeletal regulatory protein that controls smooth muscle contractility and non-muscle cell motility by modulating F-actin stress fiber stability and focal adhesion dynamics. In smooth muscle, its C-terminal domain binds F-actin, tropomyosin, and calmodulin, and phosphorylation by MAP kinase or ERK1/2 attenuates its actin interaction, relieving actin-filament inhibition and permitting contraction or cytoskeletal remodeling [PMID:1331069, PMID:36230524]. In non-muscle and cancer contexts, CALD1 expression is regulated post-transcriptionally by METTL14-mediated m6A methylation and miRNAs (miR-1278, miR-148a-5p), and CALD1 signals through PI3K-Akt, MAPK, and JAK/STAT pathways to influence epithelial–mesenchymal transition, invasion, and immune checkpoint (PD-L1) expression [PMID:37017680, PMID:38025524, PMID:34733993, PMID:38577446]. Tumor-specific alternative splicing of CALD1 occurs across multiple cancer types and in glioma microvasculature, yielding isoforms with potentially altered function [PMID:18353764, PMID:15161654]."},"prefetch_data":{"uniprot":{"accession":"Q05682","full_name":"Caldesmon","aliases":[],"length_aa":793,"mass_kda":93.2,"function":"Actin- and myosin-binding protein implicated in the regulation of actomyosin interactions in smooth muscle and nonmuscle cells (could act as a bridge between myosin and actin filaments). Stimulates actin binding of tropomyosin which increases the stabilization of actin filament structure. In muscle tissues, inhibits the actomyosin ATPase by binding to F-actin. This inhibition is attenuated by calcium-calmodulin and is potentiated by tropomyosin. Interacts with actin, myosin, two molecules of tropomyosin and with calmodulin. Also plays an essential role during cellular mitosis and receptor capping. Involved in Schwann cell migration during peripheral nerve regeneration (By similarity)","subcellular_location":"Cytoplasm, cytoskeleton; Cytoplasm, myofibril; Cytoplasm, cytoskeleton, stress fiber","url":"https://www.uniprot.org/uniprotkb/Q05682/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/CALD1","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":true,"resolved_as":"","ensg_id":"ENSG00000122786","cell_line_id":"CID000958","localizations":[{"compartment":"cytoskeleton","grade":3},{"compartment":"cytoplasmic","grade":1}],"interactors":[{"gene":"CAPZB","stoichiometry":10.0},{"gene":"FLOT1","stoichiometry":0.2},{"gene":"AP2A2","stoichiometry":0.2},{"gene":"FLNB","stoichiometry":0.2},{"gene":"RPLP0;RPLP0P6","stoichiometry":0.2},{"gene":"RPLP1","stoichiometry":0.2},{"gene":"ATP1A1","stoichiometry":0.2},{"gene":"OAT","stoichiometry":0.2},{"gene":"VIM","stoichiometry":0.2},{"gene":"ANXA6","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/target/CID000958","total_profiled":1310},"omim":[{"mim_id":"608667","title":"NIPPED-B-LIKE; NIPBL","url":"https://www.omim.org/entry/608667"},{"mim_id":"114213","title":"CALDESMON 1; CALD1","url":"https://www.omim.org/entry/114213"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Enhanced","locations":[{"location":"Plasma membrane","reliability":"Enhanced"},{"location":"Actin filaments","reliability":"Enhanced"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/CALD1"},"hgnc":{"alias_symbol":["CDM","H-CAD","L-CAD","h-CD"],"prev_symbol":[]},"alphafold":{"accession":"Q05682","domains":[],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q05682","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q05682-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q05682-F1-predicted_aligned_error_v6.png","plddt_mean":64.44},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=CALD1","jax_strain_url":"https://www.jax.org/strain/search?query=CALD1"},"sequence":{"accession":"Q05682","fasta_url":"https://rest.uniprot.org/uniprotkb/Q05682.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q05682/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q05682"}},"corpus_meta":[{"pmid":"11518968","id":"PMC_11518968","title":"Haematopoietic cell-specific CDM family protein DOCK2 is essential for lymphocyte 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assay\",\n      \"journal\": \"Annals of Translational Medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — KD/OE with specific inhibitor rescue and defined pathway placement, single lab\",\n      \"pmids\": [\"34733993\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"AHSA1 recruits ERK1/2 and promotes phosphorylation and inactivation of CALD1; ERK1/2 phosphorylation inhibitor SCH772984 reverses AHSA1-driven proliferation and EMT, and CALD1 knockdown rescues the inhibition of proliferation/EMT caused by AHSA1 knockdown, placing CALD1 as a downstream effector of the AHSA1/ERK axis in HCC.\",\n      \"method\": \"Co-IP, siRNA knockdown, overexpression, ERK inhibitor treatment, Western blot, in vivo xenograft\",\n      \"journal\": \"Cancers\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — epistasis with pharmacological and genetic rescue, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"36230524\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"miR-1278 directly targets CALD1 mRNA (validated by dual luciferase reporter assay); CALD1 overexpression activates the MAPK pathway (Ras, p-P38, p-ERK1/2) to promote gastric cancer cell viability and migration, and miR-1278 mimic partially rescues CALD1 overexpression effects.\",\n      \"method\": \"Dual luciferase reporter assay, bioinformatics prediction, qRT-PCR, Western blot, CCK-8, Transwell, xenograft\",\n      \"journal\": \"Open Medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct target validation by luciferase assay with pathway rescue, single lab\",\n      \"pmids\": [\"38025524\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"METTL14 methyltransferase promotes CALD1 expression by increasing both mRNA levels and m6A methylation of CALD1 mRNA; METTL14 silencing depletes CALD1 m6A and mRNA, and CALD1 overexpression rescues the anti-tumor effects of METTL14 knockdown in oral squamous cell carcinoma.\",\n      \"method\": \"MeRIP assay (m6A-seq), siRNA knockdown, overexpression rescue, qRT-PCR, Western blot, colony formation, Transwell, xenograft\",\n      \"journal\": \"Journal of Environmental Pathology, Toxicology and Oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct m6A modification of CALD1 mRNA identified with MeRIP and functional rescue, single lab\",\n      \"pmids\": [\"37017680\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CALD1 (high-molecular-weight caldesmon), a marker of contractile smooth muscle phenotype, is increased in preeclampsia placental chorionic plate veins; ET-1 treatment increases CALD1 expression in placental explants and HUVSMCs via ETAR/ETBR receptors, and elevated CALD1 is associated with stronger vascular contraction in preeclampsia, independent of CaV1.2 channel activity.\",\n      \"method\": \"DMT vessel tone measurement, whole-cell patch clamp, Western blot, PCR, ELISA, placental explant culture, HUVSMC treatment with ET-1 and receptor antagonists\",\n      \"journal\": \"Placenta\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods linking ETAR/ETBR signaling to CALD1 induction and contractile phenotype, single lab\",\n      \"pmids\": [\"39476475\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CALD1 promotes epithelial-mesenchymal transition (EMT) and invasion in gastric cancer by activating the PI3K-Akt pathway; CALD1 overexpression increases PI3K, p-AKT, and p-mTOR while decreasing PTEN, and PI3K-Akt inhibitor treatment reverses CALD1-driven migration, invasion, and EMT marker changes.\",\n      \"method\": \"siRNA knockdown, CALD1 overexpression, PI3K-Akt inhibitor treatment, Western blot, qRT-PCR, wound healing, Transwell, CCK-8, xenograft\",\n      \"journal\": \"World Journal of Gastrointestinal Oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — KD/OE with pharmacological inhibitor rescue demonstrating pathway placement, single lab\",\n      \"pmids\": [\"38577446\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"CALD1 knockdown in SK-OV-3 ovarian cancer cells reduces F-actin stress fibers, loosens cytoskeletal structure, decreases Vinculin expression, reduces focal adhesions, and enhances cell invasion/migration, establishing CALD1 as a regulator of actin cytoskeletal organization and focal adhesion formation that restrains ovarian cancer cell motility.\",\n      \"method\": \"Stable CALD1 knockdown (shRNA), Transwell invasion assay, immunofluorescence of F-actin and Vinculin, Western blot, qRT-PCR\",\n      \"journal\": \"Translational Cancer Research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct loss-of-function with structural and functional readouts, single lab with multiple methods\",\n      \"pmids\": [\"40104711\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"In Hp-stimulated cancer-associated fibroblasts, miR-148a-5p targets CALD1 via a TLR/miR-148a-5p/CALD1/collagen VI signaling pathway; decreased miR-148a-5p leads to CALD1 upregulation in CAFs, which promotes collagen VI secretion that interacts with tumoral SDC4 receptors to drive gastric cancer cell proliferation.\",\n      \"method\": \"Single-cell RNA sequencing, miRNA/transcriptome analysis, molecular biology experiments, in vitro co-culture, CDX and PDX in vivo models, miR-148a-5p agomir treatment\",\n      \"journal\": \"Cellular Oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — pathway validated with multiple orthogonal methods and in vivo rescue, single lab\",\n      \"pmids\": [\"41171370\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Circ-0003746, a circRNA derived from two CALD1 exons, is overexpressed in bladder cancer and promotes EMT by sequestering miR-526b; luciferase reporter assays confirmed direct circ-0003746/miR-526b interaction, and circ-0003746 silencing suppressed proliferation and migration in vitro and in vivo.\",\n      \"method\": \"Luciferase reporter assay, siRNA knockdown, qRT-PCR, Western blot, in vivo tumor model\",\n      \"journal\": \"Experimental Cell Research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct molecular interaction validated by luciferase assay with in vivo functional confirmation, single lab\",\n      \"pmids\": [\"40818763\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"CALD1 (caldesmon) is an actin cytoskeleton-associated protein that regulates F-actin organization, focal adhesion formation, and cell motility; it is phosphorylated and inactivated by ERK1/2 (recruited via AHSA1), its mRNA is subject to m6A methylation by METTL14, it acts upstream of PI3K-Akt and JAK/STAT signaling pathways to promote EMT and immune checkpoint (PD-L1) expression, its expression in vascular smooth muscle is induced by ET-1 via ETAR/ETBR to enhance contractile phenotype, and a circular RNA derived from its locus (circ-0003746) acts as a miR-526b sponge to promote EMT in bladder cancer.\"\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:**\n- Papers about CALD1 (caldesmon 1): KEEP — papers [7], [12], [13], [16], [20], [25], [27] (PMID:38025524), [28] (PMID:39476475), [29] (PMID:37017680), [34], [35], [37], [38] from retrieved; gene2pubmed papers [29] (PMID:1331069), [27] (PMID:10766836), [24] (PMID:18353764)\n- Papers about CDM (various \"CDM\" acronyms unrelated to caldesmon): EXCLUDE — [1],[2],[3],[4],[5],[6],[8],[9],[10],[11],[14],[15],[17],[19],[21],[22],[24],[26],[31],[32],[33],[36],[39],[40],[41],[42],[43]\n- Paper [38] (circ-0003746 derived from CALD1 locus) = alt-locus product (circRNA) → EXCLUDE (case B)\n- Gene2pubmed large-scale proteomics/interactome papers: mostly not mechanistically about CALD1 specifically → evaluate carefully\n\n**KEEP list for extraction:**\n- PMID:15161654 (CALD1 splicing in glioma)\n- PMID:34070840 (CALD1 in glioma angiogenesis)\n- PMID:34733993 (CALD1/JAK-STAT/PD-L1 in bladder cancer)\n- PMID:19329885 (l-CaD in EC migration)\n- PMID:36230524 (AHSA1/ERK/CALD1 in HCC)\n- PMID:38577446 (CALD1/PI3K-Akt/EMT in gastric cancer)\n- PMID:38025524 (miR-1278/CALD1/MAPK in gastric cancer)\n- PMID:39476475 (ET-1/ETAR/ETBR/CALD1 in preeclampsia)\n- PMID:37017680 (METTL14/m6A/CALD1 in OSCC)\n- PMID:34708628 (Cald1 in insulin resistance)\n- PMID:40104711 (CALD1/F-actin/focal adhesion in ovarian cancer)\n- PMID:41171370 (miR-148a-5p/CALD1/collagen VI in GC-CAFs)\n- PMID:1331069 (MAP kinase phosphorylation of caldesmon)\n- PMID:10766836 (PAK1/caldesmon/stress fibers in breast cancer)\n- PMID:18353764 (alternative splicing of CALD1 in cancer)\n\n```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1992,\n      \"finding\": \"Smooth muscle caldesmon (CALD1) is phosphorylated in vitro by MAP kinase (p44mpk) at up to 2.0 mol phosphate/mol protein at Ser and Thr residues located predominantly in the C-terminal 10-kDa fragment that houses binding sites for calmodulin, tropomyosin, and F-actin. Phosphorylation attenuated caldesmon's interaction with actin but did not affect its binding to calmodulin or tropomyosin. Differentiated smooth muscle cells (chicken gizzard, rat aorta) contain endogenous MAP kinase that immunoprecipitates with caldesmon-directed kinase activity, establishing MAP kinase as a writer of caldesmon phosphorylation in smooth muscle.\",\n      \"method\": \"In vitro phosphorylation assay with purified sea-star p44mpk, tryptic peptide mapping of 32P-labeled caldesmon, actin/calmodulin/tropomyosin binding assays post-phosphorylation, immunoprecipitation of endogenous MAP kinase from smooth muscle extracts\",\n      \"journal\": \"The Journal of Biological Chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro reconstitution with mutagenesis-equivalent peptide mapping and functional binding assays; replicated in two smooth muscle tissue types\",\n      \"pmids\": [\"1331069\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"Kinase-dead PAK1 (K299R) stabilizes actin stress fibers in invasive breast cancer cells in part through persistent co-localization with caldesmon (CALD1) and tropomyosin on F-actin filaments. Loss of PAK1 activity prevents stress fiber disassembly and reduces invasiveness, placing caldesmon downstream of PAK1 signaling in cytoskeletal remodeling during cancer cell invasion.\",\n      \"method\": \"Overexpression of kinase-dead K299R PAK1 in MDA-MB435 and MDA-MB231 cells; immunofluorescence co-localization of caldesmon and tropomyosin with F-actin; focal adhesion and invasion assays\",\n      \"journal\": \"The Journal of Biological Chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — clean loss-of-function (kinase-dead mutant) with defined cellular phenotype and co-localization; single lab\",\n      \"pmids\": [\"10766836\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Differential splicing of CALD1 exons occurs in glioma microvasculature versus normal brain microvasculature. Missplicing involving exons 1, 1+4, and 1'+4 is exclusive to tumor microvessels and results in upregulated caldesmon protein expression. This transcriptional/splicing change co-exists with downregulation of tight junction proteins occludin and ZO-1, implicating CALD1 missplicing as an epigenetic event contributing to endothelial barrier dysfunction in glioma.\",\n      \"method\": \"Laser-capture microdissection of tumor and normal microvessels, RT-PCR for splice variants, coupled in vitro transcription/translation assay (IVTT) for mutation scanning, immunohistochemistry for caldesmon, occludin, and ZO-1\",\n      \"journal\": \"The American Journal of Pathology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct tissue microdissection with RT-PCR splice-variant identification and protein-level confirmation; single lab with multiple orthogonal methods\",\n      \"pmids\": [\"15161654\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"The low-molecular-weight caldesmon isoform (Hela l-CaD) localizes specifically to motility-related cell protrusions — filopodia, microspikes, lamellipodia, podosomes, membrane blebs, and membrane ruffles — in activated endothelial cells and endothelial progenitor cells within human tumor vasculature, implicating this isoform in EC/EPC migration during tumor angiogenesis and vasculogenesis.\",\n      \"method\": \"Immunohistochemistry on histologically preserved tumor microenvironment sections from multiple human tumor types; subcellular localization of Hela l-CaD to identified protrusion subtypes\",\n      \"journal\": \"Cell Adhesion & Migration\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — direct localization experiment with functional context (protrusion subtypes tied to motility); single lab\",\n      \"pmids\": [\"19329885\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"Tumor-specific alternative splicing of CALD1 is identified across colon, bladder, and prostate cancers, representing a general cancer-related splicing event. In silico protein predictions indicate that cancer-specific CALD1 splice variants encode proteins with potentially altered functions, suggesting involvement in cancer pathogenesis.\",\n      \"method\": \"GeneChip Human Exon 1.0 ST Array on 102 normal and cancer tissue samples; RT-PCR validation on independent set of 81 normal and tumor samples; in silico protein structure prediction\",\n      \"journal\": \"Molecular & Cellular Proteomics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genome-wide discovery confirmed by RT-PCR in independent cohort; multi-organ replication\",\n      \"pmids\": [\"18353764\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"CALD1 modulates glioma progression by facilitating tumor angiogenesis. Single-cell RNA-seq and bulk RNA-seq analyses demonstrate CALD1 upregulation in neoplastic cells, and histology/immunofluorescence shows CALD1 association with vessel architecture. CALD1 expression correlates with endothelial cell and pericyte infiltration in the tumor microenvironment.\",\n      \"method\": \"Bulk RNA-seq and single-cell RNA-seq (TCGA, CGGA databases), immunofluorescence, histological analysis of vessel architecture\",\n      \"journal\": \"Cancers\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3/4 — primarily bioinformatic and correlative; limited direct mechanistic experimentation\",\n      \"pmids\": [\"34070840\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"CALD1 promotes PD-L1 (CD274) expression in bladder cancer via activation of the JAK/STAT signaling pathway. CALD1 overexpression upregulates PD-L1, and this effect is blocked by the specific JAK inhibitor Ruxolitinib, placing CALD1 upstream of JAK/STAT-mediated immune checkpoint regulation.\",\n      \"method\": \"WGCNA gene network analysis; CCK-8, flow cytometry, Transwell assays; nude mouse xenograft; JAK inhibitor (Ruxolitinib) rescue experiment; GSEA pathway analysis; correlation analysis with TIMER database\",\n      \"journal\": \"Annals of Translational Medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — pharmacological rescue (JAK inhibitor) places CALD1 in pathway; in vitro and in vivo evidence; single lab\",\n      \"pmids\": [\"34733993\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Linggui Zhugan decoction (LGZGD) improves insulin resistance in vivo (HFD rats) and in vitro (TNF-α-treated 3T3-L1 adipocytes) by downregulating Cald1 expression. Knockdown of Cald1 is associated with increased insulin-stimulated glucose uptake, identifying CALD1 as a novel regulator of insulin sensitivity in adipocytes.\",\n      \"method\": \"Gene microarray to identify DEGs in HFD vs. LGZGD-treated rats; qRT-PCR validation; 3H-2-DG glucose uptake assay in IR 3T3-L1 adipocytes; Oil Red O staining\",\n      \"journal\": \"Journal of Traditional Chinese Medicine\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — functional glucose uptake assay with CALD1 expression change, but mechanistic link is indirect; single lab, single method\",\n      \"pmids\": [\"34708628\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"AHSA1 promotes proliferation and epithelial-mesenchymal transition (EMT) in hepatocellular carcinoma through an ERK/CALD1 axis. AHSA1 recruits ERK1/2 and promotes phosphorylation (inactivation) of CALD1; ERK1/2 phosphorylation inhibitor SCH772984 reverses the effect of AHSA1 on proliferation and EMT. CALD1 knockdown rescues the inhibition of proliferation and EMT caused by AHSA1 knockdown, placing CALD1 as a downstream effector of AHSA1-ERK signaling independent of HSP90 and MEK1/2.\",\n      \"method\": \"Gain- and loss-of-function studies in vitro and in vivo; ERK1/2 inhibitor (SCH772984) rescue; CALD1 knockdown epistasis; western blot for phospho-CALD1\",\n      \"journal\": \"Cancers\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — epistasis (double knockdown rescue) and pharmacological inhibitor evidence establish pathway position; in vitro and in vivo; single lab\",\n      \"pmids\": [\"36230524\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"miR-1278 directly targets CALD1 mRNA (confirmed by dual luciferase reporter assay) and negatively regulates its expression. CALD1 overexpression promotes gastric cancer cell viability and migration and activates the MAPK pathway (Ras, p-P38, p-ERK1/2). miR-1278 mimic partially rescues the pro-tumorigenic effects of CALD1 overexpression, placing CALD1 as a downstream target of miR-1278 in the miR-1278/CALD1/MAPK axis.\",\n      \"method\": \"Dual luciferase reporter assay (miR-1278 targeting CALD1 3'UTR); CCK-8 and Transwell migration assays; western blot for MAPK pathway proteins; tumor xenograft assay\",\n      \"journal\": \"Open Medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct 3'UTR luciferase validation of miRNA-target interaction plus epistasis rescue experiment; single lab\",\n      \"pmids\": [\"38025524\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"METTL14-mediated m6A methylation regulates CALD1 mRNA levels in oral squamous cell carcinoma. METTL14 silencing depletes both mRNA and m6A levels of CALD1 (confirmed by MeRIP assay), and CALD1 overexpression rescues the growth and metastasis inhibition caused by METTL14 knockdown, identifying METTL14 as an m6A writer that stabilizes CALD1 mRNA to promote OSCC progression.\",\n      \"method\": \"MeRIP (m6A RNA immunoprecipitation) assay; qRT-PCR and western blot; colony formation and Transwell assays; tumorigenicity assay in vivo; CALD1 OE rescue of si-METTL14\",\n      \"journal\": \"Journal of Environmental Pathology, Toxicology and Oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — MeRIP directly measures m6A on CALD1 mRNA; epistasis rescue confirms functional relationship; single lab\",\n      \"pmids\": [\"37017680\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CALD1 promotes EMT and invasiveness in gastric cancer via the PI3K-Akt signaling pathway. CALD1 overexpression increases PI3K, p-AKT, and p-mTOR expression and decreases PTEN; PI3K-Akt inhibitor treatment attenuates CALD1-driven cell migration, invasion, and EMT marker changes. Animal xenograft experiments confirm that CALD1 inhibition slows tumor growth with corresponding changes in PI3K-Akt and EMT pathway proteins.\",\n      \"method\": \"CALD1 siRNA knockdown and overexpression in AGS and MKN45 GC cells; CCK-8, wound healing, Transwell assays; western blot for PI3K/p-AKT/p-mTOR/PTEN; PI3K-Akt inhibitor rescue; GC xenograft mouse model\",\n      \"journal\": \"World Journal of Gastrointestinal Oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — pharmacological pathway inhibitor rescue combined with in vitro and in vivo loss/gain-of-function; single lab\",\n      \"pmids\": [\"38577446\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Endothelin-1 (ET-1) increases high-molecular-weight CALD1 (a marker of the contractile phenotype of smooth muscle cells) in placental chorionic plate vessels and human umbilical vein smooth muscle cells (HUVSMC) via ETAR and ETBR receptors. This increase in CALD1 is associated with stronger vasoconstriction in preeclamptic placental veins, independent of CaV1.2 channel activity.\",\n      \"method\": \"Vasocontraction measurements (DMT) with ET-1, receptor antagonists, and CaV1.2 blockers; whole-cell patch clamp for CaV1.2 in single SMCs; PCR, western blot, ELISA for CALD1 and receptor expression; placental vessel explants and HUVSMC ET-1 treatment with/without antagonists\",\n      \"journal\": \"Placenta\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — pharmacological receptor antagonism identifies ETAR/ETBR as upstream regulators of CALD1 in a functional vasoconstriction assay; multiple orthogonal methods; single lab\",\n      \"pmids\": [\"39476475\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"CALD1 knockdown in SK-OV-3 ovarian cancer cells reduces F-actin stress fibers, loosens cytoskeletal structure, decreases Vinculin expression, and reduces focal adhesion number, resulting in enhanced cell migration and invasiveness. This establishes CALD1 as a regulator of cytoskeletal organization and focal adhesion formation that suppresses ovarian cancer cell invasion.\",\n      \"method\": \"Stable CALD1 shRNA knockdown in SK-OV-3 cells; Transwell invasion assay; immunofluorescence staining of F-actin and Vinculin; western blot\",\n      \"journal\": \"Translational Cancer Research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — clean KO with defined cytoskeletal phenotype (F-actin, Vinculin, focal adhesions) and functional invasiveness readout; single lab\",\n      \"pmids\": [\"40104711\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"In Helicobacter pylori-stimulated cancer-associated fibroblasts (CAFs), TLR signaling suppresses miR-148a-5p, leading to upregulation of CALD1 in CAFs. CALD1 upregulation drives collagen VI secretion from CAFs, which interacts with tumoral SDC4 receptors to promote gastric cancer cell proliferation. miR-148a-5p agomir inhibits this pathway in vivo and enhances chemotherapy efficacy.\",\n      \"method\": \"microRNA and transcriptome analysis; single-cell sequencing; in vitro CAF-GC co-culture; cell-derived xenograft (CDX) and patient-derived xenograft (PDX) mouse models; miR-148a-5p agomir treatment\",\n      \"journal\": \"Cellular Oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — pathway dissected through miRNA manipulation, co-culture, and in vivo PDX/CDX models; single lab\",\n      \"pmids\": [\"41171370\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"CALD1 (caldesmon) is a cytoskeleton-associated actin-binding protein whose C-terminal domain (housing calmodulin, tropomyosin, and F-actin binding sites) is phosphorylated by MAP kinase and p34cdc2, which attenuates its actin interaction and thereby modulates smooth muscle contractility and stress fiber stability; in non-muscle cells, CALD1 localizes to motility protrusions and regulates focal adhesion number, F-actin stress fiber integrity, and cell invasion downstream of PAK1/ERK signaling, while its expression is controlled at the post-transcriptional level by m6A methylation (METTL14) and miRNAs (miR-1278, miR-148a-5p), and it acts as an upstream regulator of JAK/STAT, PI3K-Akt, and MAPK pathways to influence EMT and immune checkpoint expression in cancer contexts.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"CALD1 encodes caldesmon, an actin-binding protein that organizes F-actin stress fibers and focal adhesions, thereby regulating cell contractility and motility in both smooth muscle and non-muscle contexts [PMID:40104711, PMID:39476475]. CALD1 is phosphorylated and functionally inactivated by ERK1/2 downstream of the AHSA1 co-chaperone axis, and its overexpression activates the PI3K-Akt and MAPK pathways to promote epithelial-mesenchymal transition and cell invasion [PMID:36230524, PMID:38577446, PMID:38025524]. CALD1 also drives PD-L1 expression through JAK/STAT signaling and, in cancer-associated fibroblasts, promotes collagen VI secretion that signals through tumoral SDC4 receptors [PMID:34733993, PMID:41171370]. CALD1 mRNA is stabilized by METTL14-mediated m6A methylation and is subject to post-transcriptional repression by multiple microRNAs including miR-1278 and miR-148a-5p, while a circular RNA derived from the CALD1 locus (circ-0003746) independently promotes EMT by sponging miR-526b [PMID:37017680, PMID:38025524, PMID:40818763].\",\n  \"teleology\": [\n    {\n      \"year\": 2004,\n      \"claim\": \"The discovery that CALD1 undergoes differential exon splicing specifically in glioma microvessels — coinciding with loss of tight junction proteins — first linked caldesmon isoform switching to pathological vascular remodeling.\",\n      \"evidence\": \"Laser-capture microdissection with RT-PCR and in vitro transcription/translation in glioma versus normal brain microvasculature\",\n      \"pmids\": [\"15161654\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"No direct causal manipulation (knockdown/overexpression) was performed to test whether CALD1 splice variants drive tight junction loss\",\n        \"Mechanism by which caldesmon isoforms affect occludin/ZO-1 expression is unknown\"\n      ]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Spatial mapping of low-molecular-weight caldesmon to motility-associated membrane protrusions in tumor endothelial cells provided the first subcellular localization evidence implicating the l-CaD isoform in angiogenic cell migration.\",\n      \"evidence\": \"Immunohistochemistry with spatial localization in tumor vasculature\",\n      \"pmids\": [\"19329885\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"Localization-only study without functional perturbation; awaits knockdown or mutation-based validation\",\n        \"Isoform-specific contributions to migration versus adhesion were not dissected\"\n      ]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Establishing CALD1 as an upstream activator of JAK/STAT-dependent PD-L1 expression revealed an unexpected role for this cytoskeletal protein in immune checkpoint regulation in bladder cancer.\",\n      \"evidence\": \"siRNA knockdown, overexpression, Ruxolitinib (JAK inhibitor) rescue, flow cytometry, and xenograft in bladder cancer cells\",\n      \"pmids\": [\"34733993\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Molecular mechanism by which CALD1 activates JAK/STAT is unresolved — no direct binding to JAK kinases demonstrated\",\n        \"In vivo immune checkpoint functional consequence not tested in immunocompetent models\"\n      ]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Epistasis experiments placing CALD1 downstream of AHSA1-recruited ERK1/2 phosphorylation clarified how CALD1 is post-translationally inactivated, with CALD1 knockdown rescuing AHSA1-loss phenotypes and ERK inhibitors blocking the cascade.\",\n      \"evidence\": \"Co-IP, siRNA knockdown, overexpression, ERK inhibitor SCH772984, Western blot, and xenograft in HCC\",\n      \"pmids\": [\"36230524\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Specific ERK phosphorylation sites on CALD1 were not mapped in this study\",\n        \"Whether AHSA1–CALD1 interaction is direct or entirely ERK-mediated remains unclear\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Two studies converged to show CALD1 is both a direct microRNA target (miR-1278) and an activator of MAPK signaling (Ras, p-ERK1/2, p-P38), and that METTL14-dependent m6A methylation of CALD1 mRNA stabilizes its expression, revealing layered transcriptional and epitranscriptomic control of CALD1 abundance.\",\n      \"evidence\": \"Dual luciferase reporter assay for miR-1278 targeting; MeRIP for m6A on CALD1 mRNA; knockdown/overexpression rescue in gastric cancer and oral squamous cell carcinoma\",\n      \"pmids\": [\"38025524\", \"37017680\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Specific m6A sites on CALD1 mRNA and the reader protein mediating stabilization are not identified\",\n        \"Whether MAPK activation by CALD1 is direct or via cytoskeletal remodeling is unknown\"\n      ]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Demonstration that ET-1 induces high-molecular-weight CALD1 via ETAR/ETBR in vascular smooth muscle cells and placental veins, correlating with enhanced contraction in preeclampsia, extended CALD1's role to physiological smooth muscle contractility regulation.\",\n      \"evidence\": \"DMT vessel tone measurement, whole-cell patch clamp, Western blot, placental explant culture with ET-1 and receptor antagonists\",\n      \"pmids\": [\"39476475\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Whether CALD1 increase alone is sufficient to enhance contraction was not tested by CALD1-specific manipulation\",\n        \"Signaling intermediates between ETAR/ETBR and CALD1 transcriptional induction are not mapped\"\n      ]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Pharmacological inhibitor rescue confirmed CALD1 drives EMT and invasion in gastric cancer specifically through PI3K-Akt-mTOR pathway activation with concurrent PTEN suppression, defining a second major signaling axis downstream of CALD1.\",\n      \"evidence\": \"CALD1 overexpression/knockdown with PI3K-Akt inhibitor treatment, Western blot, Transwell, xenograft in gastric cancer\",\n      \"pmids\": [\"38577446\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Direct molecular link between CALD1 (an actin-binding protein) and PI3K activation is not established\",\n        \"Relative contribution of PI3K-Akt versus MAPK signaling to CALD1-driven EMT not compared\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Three studies expanded CALD1 biology: loss-of-function in ovarian cancer directly demonstrated CALD1 maintains F-actin stress fibers and focal adhesions (via Vinculin); in gastric CAFs, a TLR/miR-148a-5p/CALD1/collagen VI axis was defined as a paracrine EMT driver; and circ-0003746 from the CALD1 locus was shown to sponge miR-526b to promote EMT independently of CALD1 protein.\",\n      \"evidence\": \"shRNA-stable knockdown with F-actin/Vinculin immunofluorescence (ovarian cancer); scRNA-seq with miR-148a-5p agomir rescue in CDX/PDX models (gastric CAFs); luciferase reporter assay for circ-0003746/miR-526b with in vivo bladder cancer model\",\n      \"pmids\": [\"40104711\", \"41171370\", \"40818763\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Whether CALD1's cytoskeletal and signaling functions are separable has not been tested with domain mutants\",\n        \"The collagen VI–SDC4 axis downstream of CALD1 in CAFs requires validation in additional tumor types\",\n        \"Circ-0003746 functions are characterized only for miR-526b sponging; other targets not explored\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The mechanism by which an actin-binding protein activates multiple intracellular signaling cascades (PI3K-Akt, MAPK, JAK/STAT) remains the central unresolved question — whether CALD1 signals through cytoskeletal remodeling-dependent integrin/mechanotransduction or through direct protein interactions is unknown.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"No structural model of CALD1 signaling domains beyond actin/calmodulin binding\",\n        \"Isoform-specific (h-CaD vs l-CaD) contributions to signaling versus cytoskeletal functions have not been dissected with isoform-selective tools\",\n        \"No unbiased interactome or phosphoproteomics study connecting CALD1 to its downstream signaling effectors\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [1, 8]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [1, 8]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [2, 3, 4, 7]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [2]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"AHSA1\",\n      \"MAPK3\",\n      \"MAPK1\",\n      \"METTL14\",\n      \"VCL\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```\n\n--- ROUND 2 (R2/R3, augmented corpus) ---\n\n```json\n{\n  \"mechanistic_narrative\": \"CALD1 (caldesmon) is an actin- and calmodulin-binding cytoskeletal regulatory protein that controls smooth muscle contractility and non-muscle cell motility by modulating F-actin stress fiber stability and focal adhesion dynamics. In smooth muscle, its C-terminal domain binds F-actin, tropomyosin, and calmodulin, and phosphorylation by MAP kinase or ERK1/2 attenuates its actin interaction, relieving actin-filament inhibition and permitting contraction or cytoskeletal remodeling [PMID:1331069, PMID:36230524]. In non-muscle and cancer contexts, CALD1 expression is regulated post-transcriptionally by METTL14-mediated m6A methylation and miRNAs (miR-1278, miR-148a-5p), and CALD1 signals through PI3K-Akt, MAPK, and JAK/STAT pathways to influence epithelial–mesenchymal transition, invasion, and immune checkpoint (PD-L1) expression [PMID:37017680, PMID:38025524, PMID:34733993, PMID:38577446]. Tumor-specific alternative splicing of CALD1 occurs across multiple cancer types and in glioma microvasculature, yielding isoforms with potentially altered function [PMID:18353764, PMID:15161654].\",\n  \"teleology\": [\n    {\n      \"year\": 1992,\n      \"claim\": \"Identifying how caldesmon's actin-regulatory activity is switched off resolved a long-standing question about the kinase signal that controls smooth muscle thin-filament regulation: MAP kinase phosphorylates the C-terminal actin-binding domain and selectively weakens the caldesmon–actin interaction without disrupting calmodulin or tropomyosin binding.\",\n      \"evidence\": \"In vitro phosphorylation of purified chicken gizzard caldesmon by sea-star p44mpk; tryptic peptide mapping; actin/calmodulin/tropomyosin co-sedimentation assays; immunoprecipitation of endogenous MAP kinase from rat aorta smooth muscle\",\n      \"pmids\": [\"1331069\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"In vivo phosphorylation sites not mapped at single-residue resolution in this study\",\n        \"Structural basis of how phosphorylation weakens actin binding not determined\",\n        \"Whether additional kinases (e.g. p34cdc2) phosphorylate the same or distinct sites in smooth muscle not addressed\"\n      ]\n    },\n    {\n      \"year\": 2000,\n      \"claim\": \"Establishing that caldesmon participates in PAK1-dependent cytoskeletal remodeling in invasive breast cancer cells extended its role beyond smooth muscle contraction to cancer cell motility: kinase-dead PAK1 traps caldesmon and tropomyosin on stabilized stress fibers, blocking invasion.\",\n      \"evidence\": \"Overexpression of kinase-dead PAK1(K299R) in MDA-MB435 and MDA-MB231 cells; immunofluorescence co-localization of caldesmon/tropomyosin with F-actin; invasion assays\",\n      \"pmids\": [\"10766836\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Whether PAK1 directly phosphorylates caldesmon was not tested\",\n        \"Caldesmon was not knocked down to confirm it is required for PAK1-dependent invasion\"\n      ]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Discovery of tumor-specific CALD1 splice variants in glioma and subsequently in colon, bladder, and prostate cancers revealed that alternative splicing diversifies caldesmon function in the tumor microenvironment and may alter its actin-regulatory properties.\",\n      \"evidence\": \"Laser-capture microdissection of glioma versus normal microvessels with RT-PCR splice-variant identification; GeneChip Exon Array across 102 samples with RT-PCR validation in 81 independent samples\",\n      \"pmids\": [\"15161654\", \"18353764\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Functional consequences of cancer-specific splice variants on actin binding or contractility not experimentally validated\",\n        \"Whether splice variants are causative or merely correlative in tumor progression remains untested\"\n      ]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Subcellular mapping of the low-molecular-weight caldesmon isoform to motility-associated protrusions (filopodia, lamellipodia, podosomes, membrane blebs) in tumor-associated endothelial cells specified the cellular compartment in which caldesmon operates during tumor angiogenesis.\",\n      \"evidence\": \"Immunohistochemistry on multiple human tumor types identifying l-CaD in endothelial and endothelial progenitor cell protrusion subtypes\",\n      \"pmids\": [\"19329885\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Loss-of-function in endothelial protrusions not performed\",\n        \"Whether l-CaD is required for protrusion formation or merely accumulates there is unknown\"\n      ]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Placing CALD1 upstream of JAK/STAT-mediated PD-L1 expression in bladder cancer expanded its role from a cytoskeletal regulator to a modulator of immune checkpoint signaling, as pharmacological JAK inhibition (Ruxolitinib) blocked CALD1-driven PD-L1 upregulation.\",\n      \"evidence\": \"CALD1 overexpression/knockdown in bladder cancer cells; JAK inhibitor rescue; nude mouse xenograft; GSEA pathway analysis\",\n      \"pmids\": [\"34733993\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Direct physical interaction between CALD1 and JAK pathway components not demonstrated\",\n        \"Whether the JAK/STAT link operates in non-bladder cancer contexts is unknown\"\n      ]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Epistasis experiments showed CALD1 is a functional effector downstream of the AHSA1–ERK axis in hepatocellular carcinoma EMT: CALD1 knockdown rescued the proliferation and EMT inhibition caused by AHSA1 depletion, and ERK inhibitor SCH772984 reversed AHSA1-driven CALD1 phosphorylation.\",\n      \"evidence\": \"Double knockdown epistasis (AHSA1 + CALD1); ERK1/2 inhibitor rescue; in vitro and in vivo HCC models; western blot for phospho-CALD1\",\n      \"pmids\": [\"36230524\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Specific ERK phosphorylation sites on CALD1 not mapped in this system\",\n        \"Whether CALD1's actin-binding function is required for the EMT phenotype not tested\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Identification of post-transcriptional regulators of CALD1 — m6A methylation by METTL14 and direct miR-1278 targeting of the 3′UTR — established that CALD1 expression is tuned at the mRNA level in cancer, with functional consequences for MAPK pathway activation and tumor growth.\",\n      \"evidence\": \"MeRIP assay for m6A on CALD1 mRNA with METTL14 knockdown/rescue in OSCC; dual luciferase reporter assay validating miR-1278 binding to CALD1 3′UTR in gastric cancer; xenograft models\",\n      \"pmids\": [\"37017680\", \"38025524\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Specific m6A sites on CALD1 mRNA not mapped at nucleotide resolution\",\n        \"Whether METTL14 and miR-1278 regulation operates simultaneously or in different cellular contexts is unknown\"\n      ]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"CALD1 was shown to drive EMT and invasion in gastric cancer specifically through PI3K-Akt-mTOR signaling, with pathway inhibitor rescue confirming directionality; separately, endothelin-1 was identified as an upstream inducer of high-molecular-weight CALD1 in placental smooth muscle, linking CALD1 to vasoconstriction in preeclampsia.\",\n      \"evidence\": \"PI3K-Akt inhibitor rescue of CALD1 overexpression phenotypes in GC cells and xenografts; ET-1 receptor antagonism in placental veins and HUVSMCs with vasocontraction measurements\",\n      \"pmids\": [\"38577446\", \"39476475\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"How CALD1, an actin-binding protein, activates PI3K-Akt remains mechanistically undefined\",\n        \"Whether ET-1-induced CALD1 upregulation is transcriptional or post-translational not determined\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Two studies completed the picture of CALD1's context-dependent role: in ovarian cancer, CALD1 knockdown reduced F-actin stress fibers and focal adhesions, paradoxically increasing invasion; in gastric cancer CAFs, H. pylori-driven TLR signaling suppresses miR-148a-5p to upregulate CALD1, which drives collagen VI secretion and paracrine proliferative signaling through tumoral SDC4.\",\n      \"evidence\": \"Stable shRNA knockdown in SK-OV-3 with F-actin/Vinculin immunofluorescence and invasion assays; miR-148a-5p agomir in CAF-GC co-culture and PDX/CDX mouse models\",\n      \"pmids\": [\"40104711\", \"41171370\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Opposing invasion phenotypes in ovarian cancer versus other tumor types not reconciled mechanistically\",\n        \"How CALD1 regulates collagen VI secretion in CAFs is unknown\",\n        \"Whether the miR-148a-5p/CALD1/collagen VI/SDC4 axis operates outside gastric cancer is untested\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"A unifying structural and signaling model explaining how a single actin-binding protein exerts context-dependent pro- or anti-invasive effects across different cell types, and how its cytoskeletal function connects to downstream transcriptional pathway activation (JAK/STAT, PI3K-Akt, MAPK), remains to be established.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"No high-resolution structure of caldesmon bound to F-actin available\",\n        \"Direct mechanism linking CALD1 to activation of PI3K-Akt, JAK/STAT, and MAPK transcriptional programs is unknown\",\n        \"Relative contribution of individual phosphorylation sites (ERK, p34cdc2, PAK1) to distinct phenotypic outputs has not been dissected in a single system\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [0, 1, 3, 13]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [6, 8, 9, 11]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [0, 1, 3, 13]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [3]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [1, 13]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [6, 8, 9, 11]},\n      {\"term_id\": \"R-HSA-397014\", \"supporting_discovery_ids\": [0, 12]},\n      {\"term_id\": \"R-HSA-1474244\", \"supporting_discovery_ids\": [14]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"ACTA2\",\n      \"TPM1\",\n      \"CALM1\",\n      \"PAK1\",\n      \"MAPK1\",\n      \"METTL14\",\n      \"AHSA1\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}