{"gene":"HDAC8","run_date":"2026-06-10T01:55:21","timeline":{"discoveries":[{"year":2004,"finding":"Crystal structure of human HDAC8 complexed with a hydroxamic acid inhibitor revealed a single alpha/beta domain fold similar to bacterial HDAC-like protein, with two potassium ions in the active site, one interacting with key catalytic residues; CD data demonstrated a direct role of potassium in fold stabilization.","method":"X-ray crystallography, CD spectroscopy","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure at atomic resolution with functional validation by CD spectroscopy, foundational structural paper","pmids":["15477595"],"is_preprint":false},{"year":2000,"finding":"HDAC8 encodes an RPD3 class I HDAC with trichostatin A- and sodium butyrate-inhibitable deacetylase activity on histone H4 peptide substrates and core histones; mutation of two adjacent active-site histidine residues severely decreases enzymatic activity, confirming their catalytic importance.","method":"Immunopurification, in vitro HDAC activity assay, active-site mutagenesis","journal":"The Biochemical journal","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro enzymatic reconstitution plus active-site mutagenesis in a single focused study","pmids":["10926844"],"is_preprint":false},{"year":2012,"finding":"HDAC8 is the vertebrate SMC3 deacetylase responsible for deacetylating SMC3 during cohesin recycling after anaphase; loss-of-function HDAC8 mutations cause Cornelia de Lange syndrome and result in elevated SMC3 acetylation, inefficient dissolution of used cohesin, and altered transcription at cohesin-binding sites.","method":"Biochemical deacetylase assay, ChIP-seq, CdLS patient mutation analysis, cell-based cohesin acetylation assays","journal":"Nature","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — multiple orthogonal methods (biochemical assay, ChIP-seq, patient genetics, cell biology), replicated in multiple CdLS probands","pmids":["22885700"],"is_preprint":false},{"year":2005,"finding":"HDAC8 associates with smooth muscle alpha-actin (alpha-SMA) but not beta-actin in human smooth muscle cells, and HDAC8 knockdown by siRNA strongly reduces the contractile capacity of smooth muscle cells; HDAC8 is enriched in cytoskeleton-bound protein fractions, unlike HDAC1 and HDAC3.","method":"Cell fractionation, co-immunoprecipitation, siRNA knockdown, collagen lattice contraction assay","journal":"FASEB journal","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP, fractionation, and functional KD phenotype in multiple cell types with defined readout","pmids":["15772115"],"is_preprint":false},{"year":2008,"finding":"HDAC8-selective inhibitor PCI-34051 induces caspase-dependent apoptosis in T-cell lymphoma/leukemia lines via PLCgamma1 activation, intracellular calcium mobilization from the ER, and subsequent cytochrome c release; PLCgamma1-defective cells are resistant and PLC inhibitor U73122 blocks apoptosis.","method":"Selective inhibitor treatment, calcium flux assay, caspase activation assay, genetic cell lines defective in signaling, calcium chelator/effector pharmacology","journal":"Leukemia","confidence":"High","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal pharmacological and genetic approaches defining pathway mechanism in a single rigorous study","pmids":["18256683"],"is_preprint":false},{"year":2014,"finding":"HDAC8 forms a complex with SMAD3/4 heterotrimer, occupies the SIRT7 promoter, and deacetylates histone H4 to suppress SIRT7 transcription; this establishes a regulatory feedback loop in TGF-β signaling wherein HDAC8 acts as a novel cofactor of SMAD3/4.","method":"Co-immunoprecipitation, ChIP assay, HDAC8 inhibitor treatment, gene expression analysis","journal":"Nucleic acids research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP and ChIP in single lab with multiple supporting experiments","pmids":["31970414"],"is_preprint":false},{"year":2015,"finding":"HDAC8 deacetylates p53 in inv(16)+ AML leukemia stem cells; the CBFβ-SMMHC fusion protein interacts with HDAC8 to aberrantly deacetylate and inactivate p53, promoting leukemic stem cell transformation and maintenance.","method":"Co-immunoprecipitation, acetylation assay, HDAC8 selective inhibitor treatment, HDAC8-deficient mouse models, patient-derived xenograft models","journal":"Cell stem cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (Co-IP, acetylation assay, genetic KD, pharmacological inhibition, in vivo models) with consistent results","pmids":["26387755"],"is_preprint":false},{"year":2016,"finding":"HDAC8 catalyzes the removal of long-chain fatty acyl groups (octanoyl, dodecanoyl, myristoyl) from lysine residues with higher catalytic efficiency (lower Km) than for acetyl-lysine; this is the first demonstration of de-fatty-acylation activity by a zinc-dependent HDAC.","method":"In vitro enzymatic assay with panel of acyl-lysine peptides, steady-state kinetics (kcat/Km), HDAC8-specific inhibitor treatment of Jurkat cells with global fatty acylation measurement","journal":"ACS chemical biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution with kinetic characterization plus cellular validation, single lab but rigorous biochemical methods","pmids":["27459069"],"is_preprint":false},{"year":2014,"finding":"CdLS-associated missense mutations in HDAC8 (C153F, A188T, I243N, T311M, H334R) cause local structural changes that compromise catalysis and/or thermostability; catalytic activity of several mutants can be partially or fully rescued by the HDAC8 activator N-(phenylcarbamothioyl)benzamide.","method":"X-ray crystallography, enzymatic activity assay, thermostability assay, activator rescue experiment","journal":"ACS chemical biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — crystal structures of multiple mutants combined with enzymatic and thermostability assays in one rigorous study","pmids":["25075551"],"is_preprint":false},{"year":2015,"finding":"CdLS-associated HDAC8 mutants (P91L, G117E, H180R, D233G, G304R) cause structural changes compromising catalysis and/or thermostability; the D233G mutation disrupts a hydrogen bond network stabilizing tertiary structure; MD simulations show H180R and G304R mutations cause the arginine side chain to protrude into the substrate binding site; catalytic activity can be partially restored by N-acylthiourea activator.","method":"X-ray crystallography, enzymatic activity assay, thermostability assay, molecular dynamics simulation, activator rescue experiment","journal":"Biochemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structures plus kinetics plus MD simulation, replicating and extending findings from prior CdLS structural study","pmids":["26463496"],"is_preprint":false},{"year":2016,"finding":"X-ray crystal structure of HDAC8 complexed with a 'linkerless' hydroxamic acid inhibitor confirms formation of an HDAC8-specific subpocket distinct from other isoforms, explaining isoform-selective inhibition.","method":"X-ray crystallography (1.98 Å resolution), molecular docking","journal":"Journal of structural biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — crystal structure with molecular docking validation, single lab","pmids":["27374062"],"is_preprint":false},{"year":2008,"finding":"HDAC8 activity on H4 histone peptides is regulated by both local and distal substrate sequences; a KRHR sequence (residues 16-19) distal to the acetylated lysine 12 is required for activity by interacting at an exosite; methylation of K20 directly affects HDAC8 activity at K12.","method":"Mass spectrometry-based HDAC assay with self-assembled monolayer peptide substrates, peptide truncation and mutagenesis","journal":"Biochemistry","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro biochemical reconstitution with systematic peptide mutagenesis, revealing exosite mechanism","pmids":["18470998"],"is_preprint":false},{"year":2017,"finding":"HDAC8 substrate selectivity is determined by both short- and long-range interactions; HDAC8 catalyzes deacetylation of tetrameric histone H3/H4 substrates with 40–300-fold higher catalytic efficiency than corresponding peptide substrates, while efficiency decreases for larger multiprotein complexes.","method":"In vitro kinetic assays with peptides, full-length proteins, and protein-nucleic acid complexes","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Moderate — systematic in vitro kinetic analysis across multiple substrate formats in single rigorous study","pmids":["29109148"],"is_preprint":false},{"year":2015,"finding":"The N-acylthiourea derivative TM-2-51 activates HDAC8 by binding at two sites in a positive cooperative manner, as demonstrated by steady-state kinetics and isothermal titration calorimetry; activation preferentially kills neuroblastoma cells with wild-type p53 genotype.","method":"Steady-state kinetics, isothermal titration calorimetry, molecular modeling, cell viability assay","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Moderate — multiple biophysical and biochemical methods (ITC, kinetics, modeling) in single study","pmids":["25605725"],"is_preprint":false},{"year":2008,"finding":"HDAC8 localizes in the nucleus, binds to both CREB and PP1, and expression of recombinant HDAC8 decreases CREB phosphorylation and CREB-mediated gene transcription in response to forskolin, indicating HDAC8 recruits a phosphatase to inactivate CREB.","method":"Co-immunoprecipitation, overexpression, luciferase reporter assay","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 3 / Weak — single Co-IP and overexpression study, single lab","pmids":["19070599"],"is_preprint":false},{"year":2014,"finding":"HDAC8 directly represses the BMF gene in colon cancer cells; on the BMF promoter, HDAC8 loss is associated with STAT3/Sp3 transcription factor exchange and p300 recruitment; exogenous HDAC8 overexpression blocks BMF induction.","method":"ChIP analysis, luciferase assay, RNAi knockdown, overexpression","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP and luciferase assay with RNAi and overexpression, single lab","pmids":["25321483"],"is_preprint":false},{"year":2016,"finding":"HDAC8 knockdown inhibits YY1 interaction with the p53 promoter (at -102 to -96 position) by increasing acetylation of YY1 (residues 170–200), decreasing YY1 transcriptional activity, and thereby suppressing mutant p53 transcription in TNBC cells.","method":"ChIP analysis, luciferase assay, siRNA knockdown, co-immunoprecipitation","journal":"Cellular signalling","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP, Co-IP, and luciferase assay with RNAi in single lab","pmids":["26876786"],"is_preprint":false},{"year":2015,"finding":"HDAC8 physically interacts with the chromatin modifier EZH2 to concordantly repress Wnt antagonists via histone H4 deacetylation and H3K27 trimethylation in NAFLD-associated hepatocellular carcinoma.","method":"Co-immunoprecipitation, ChIP assay, lentiviral HDAC8 knockdown","journal":"Cancer research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP and ChIP in single lab with in vivo model","pmids":["26383163"],"is_preprint":false},{"year":2017,"finding":"HDAC8 regulates long-term hematopoietic stem cell maintenance by interacting with p53 protein and modulating p53 activity via deacetylation; Hdac8-deficient hematopoietic progenitors show hyperactivation of p53 and increased apoptosis under stress, which is genetically rescued by p53 inactivation.","method":"Co-immunoprecipitation, acetylation assay, conditional knockout mouse model, serial replating, in vivo repopulation assay, genetic epistasis (p53 KO rescue)","journal":"Blood","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, acetylation assay, and genetic epistasis (p53 KO rescue) with in vivo model in single rigorous study","pmids":["29084772"],"is_preprint":false},{"year":2020,"finding":"FLT3 inhibition induces HDAC8 upregulation via FOXO1- and FOXO3-mediated transactivation in FLT3-ITD+ AML cells; upregulated HDAC8 deacetylates and inactivates p53, promoting leukemia maintenance and resistance to TKI treatment.","method":"Gene expression analysis, FOXO siRNA/overexpression, HDAC8 inhibitor treatment, acetylation assay, patient-derived xenograft model","journal":"Blood","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple approaches (FOXO KD/OE, acetylation assay, PDX model) in single lab","pmids":["32315388"],"is_preprint":false},{"year":2019,"finding":"HDAC8 deacetylates c-Jun at lysine 273, increasing c-Jun transcriptional activity; HDAC8-mediated BRAF inhibitor resistance in melanoma is driven by receptor tyrosine kinase activation and AP-1/c-Jun signaling; mutation of K273 in c-Jun increases transcriptional activity and confers resistance to BRAF inhibition.","method":"Mass spectrometry-based phosphoproteomics, HDAC8 introduction into drug-naïve cells, c-Jun acetylation assay, site-directed mutagenesis of c-Jun K273, in vivo xenograft","journal":"Cancer research","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — direct substrate deacetylation shown, mutagenesis of acetylation site, in vivo validation, single lab","pmids":["30987999"],"is_preprint":false},{"year":2023,"finding":"HDAC8 deacetylates the histone acetyltransferase EP300, causing its enzymatic inactivation; stress-induced HDAC8 activity leads to increased EP300 binding at c-Jun transcriptional sites and decreased binding at MITF sites, driving a neural crest-stem cell state and melanoma brain metastasis.","method":"ATAC-Seq, ChIP-Seq, EP300 deacetylation assay, HDAC8 inhibitor treatment, in vivo brain metastasis model","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — biochemical deacetylation assay plus genome-wide chromatin profiling and in vivo validation in single rigorous study","pmids":["38030596"],"is_preprint":false},{"year":2016,"finding":"HDAC8 inhibition blocks SMC3 deacetylation and delays cell cycle progression and suppresses proliferation in MCF7 cells, but does not alter estrogen-responsive cohesin-dependent gene transcription, demonstrating that HDAC8 inhibition impacts cell cycle rather than cohesin transcriptional function in this context.","method":"HDAC8-specific inhibitor (PCI-34051), SMC3 acetylation immunoblot, siRNA knockdown of cohesin subunits, gene expression analysis, cell cycle analysis","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — selective inhibitor and siRNA with multiple functional readouts, single lab","pmids":["27072133"],"is_preprint":false},{"year":2018,"finding":"HDAC8 associates with alpha-SMA in TGFβ1-treated lung fibroblasts (co-immunoprecipitation); HDAC8 inhibition represses TGFβ1-induced fibroblast contraction, alpha-SMA expression, and profibrotic gene expression; HDAC8 inhibition increases H3K27ac at the PPARγ gene enhancer, identifying H3K27 as an HDAC8 substrate relevant to pulmonary fibrosis.","method":"Co-immunoprecipitation, HDAC8 siRNA knockdown, selective inhibitor (NCC170), collagen gel contraction assay, ChIP-qPCR for H3K27ac, bleomycin mouse model","journal":"American journal of physiology. Lung cellular and molecular physiology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — Co-IP, ChIP, siRNA, pharmacological inhibition, and in vivo model with multiple orthogonal readouts in single lab","pmids":["30358439"],"is_preprint":false},{"year":2021,"finding":"HDAC8 is a histone H3 lysine 27 (H3K27)-specific deacetylase isozyme; pharmacological HDAC8 inhibition increases global and enhancer H3K27 acetylation, reactivating T cell-trafficking chemokine production by HCC cells and relieving T cell exclusion.","method":"Chromatin profiling (H3K27ac ChIP), HDAC8 inhibitor treatment, humanized mouse model, genome-wide enhancer analysis","journal":"Science translational medicine","confidence":"High","confidence_rationale":"Tier 2 / Strong — genome-wide chromatin profiling with selective inhibitor, multiple mouse models, mechanistic link established","pmids":["33827976"],"is_preprint":false},{"year":2016,"finding":"Structure-based approach using Rosetta FlexPepBind identified multiple novel HDAC8 non-histone substrates from proteins with known acetylated sites, including SMC3, ERRα, and ARID1A, demonstrating that HDAC8 regulates a broad acetylome.","method":"Structure-based computational substrate identification (Rosetta FlexPepBind), in vitro peptide deacetylation assay validation","journal":"Structure","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — computational prediction validated by in vitro assay but limited cellular validation; single study","pmids":["26933971"],"is_preprint":false},{"year":2017,"finding":"Genetically encoded photocrosslinking with p-benzoyl-l-phenylalanine incorporated into HDAC8 active site captured direct HDAC8-substrate complexes from cell lysates; novel HDAC8 substrates were identified and validated for catalytic turnover by mass spectrometry.","method":"Active-site photocrosslinking with non-natural amino acid, co-immunoprecipitation, mass spectrometry, in vitro deacetylation validation","journal":"Journal of the American Chemical Society","confidence":"High","confidence_rationale":"Tier 1 / Moderate — chemical crosslinking at active site plus MS identification and catalytic validation in single rigorous study","pmids":["29035536"],"is_preprint":false},{"year":2020,"finding":"HDAC8 binds and deacetylates PKM2 at K62; K62 deacetylation facilitates PKM2 nuclear translocation and binding to β-catenin, promoting CCND1 gene transcription and cell cycle progression, and also affects PKM2 enzyme activity and glucose metabolism flux.","method":"Co-immunoprecipitation, site-directed mutagenesis (K62), nuclear fractionation, chromatin immunoprecipitation, cell cycle analysis","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, mutagenesis of substrate acetylation site, and functional readouts in single lab","pmids":["33279948"],"is_preprint":false},{"year":2020,"finding":"AMPK phosphorylation of HDAC8 during glucose deprivation induces translocation of HDAC8 from the nucleus to the cytoplasm, disrupting HDAC8-histone 3 binding and leading to increased PGM1 expression in lung cancer cells.","method":"Subcellular fractionation, co-immunoprecipitation, AMPK activation/inhibition, HDAC8 phosphorylation assay","journal":"Cancer letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — fractionation, Co-IP, and kinase inhibition with functional consequence, single lab","pmids":["32171858"],"is_preprint":false},{"year":2014,"finding":"HDAC8 mediates epigenetic reprogramming in macrophages by deacetylating H3K27, reducing H3K27ac at genomic regions of BNIP3 and metastatic lymph node 64 genes, promoting resistance to anthrax lethal toxin-induced pyroptosis; HDAC8 is recruited to these loci.","method":"ChIP assay, HDAC8 siRNA knockdown, HDAC8-specific inhibitor, HDAC8 overexpression, gene expression analysis","journal":"Journal of immunology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP, siRNA, pharmacological inhibition, and overexpression with consistent results in single lab","pmids":["24973453"],"is_preprint":false},{"year":2014,"finding":"DEC1 interacts with HDAC8 and recruits HDAC8 to the TAp73 promoter to enhance TAp73 expression; HDAC8 is required for DEC1 to increase TAp73 transcription (demonstrated by failure of DEC1 to activate TAp73 in absence of HDAC8).","method":"Co-immunoprecipitation, ChIP assay, siRNA knockdown of HDAC8, luciferase reporter assay","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, ChIP, and epistatic siRNA rescue in single lab","pmids":["24404147"],"is_preprint":false},{"year":2017,"finding":"HDAC8 localizes at spindle poles in mouse oocytes and is required for microtubule organization and spindle assembly during meiosis; HDAC8 depletion causes spindle defects, chromosome misalignment, impaired kinetochore-microtubule attachments, and increased aneuploidy; HDAC8 is required for correct gamma-tubulin localization to spindle poles.","method":"siRNA knockdown, selective inhibitor (PCI-34051), immunofluorescence/confocal imaging, spindle and chromosome analysis, aneuploidy assay","journal":"Oncotarget","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA and pharmacological inhibitor with consistent spindle phenotype, multiple readouts, single lab","pmids":["28223544"],"is_preprint":false},{"year":2018,"finding":"HDAC8 and its phosphorylated form (pHDAC8) localize predominantly in the cytoplasm in HeLa (cancerous) and HEK293T (non-cancerous) cells with additional nucleolar localization in HeLa; HDAC8 binds and deacetylates alpha-tubulin at acetyl-K40, functionally redundant with HDAC6 in cervical cancer cells.","method":"Immunofluorescence/confocal imaging, immunoprecipitation followed by MALDI-TOF MS, HPLC-based in vitro deacetylation assay, surface plasmon resonance, siRNA knockdown, HDAC6/8 inhibitors","journal":"Cell communication and signaling","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct binding and in vitro deacetylation with multiple biochemical methods, single lab","pmids":["29716651"],"is_preprint":false},{"year":2019,"finding":"HDAC8 interacts with the proteasome receptor ADRM1; this interaction is disrupted by temozolomide in sensitive GBM cells; HDAC8 inhibition decreases MGMT protein levels in GBM cell lines, increasing DNA damage and cell cycle arrest.","method":"Co-immunoprecipitation (HDAC8-ADRM1), HDAC8 shRNA, HDAC8-specific inhibitor PCI34051, MGMT protein quantification, cell viability assay","journal":"Genes & cancer","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — single Co-IP with pharmacological and genetic confirmation, single lab","pmids":["31798765"],"is_preprint":false},{"year":2020,"finding":"HDAC8 promotes dissemination of breast cancer cells via the AKT/GSK-3β/Snail axis; HDAC8 interacts with the C-terminal of AKT1 via Co-IP, deacetylates AKT1 at K426, and increases AKT1 phosphorylation, which increases GSK-3β Ser9-phosphorylation, stabilizing Snail and promoting EMT.","method":"Co-immunoprecipitation, mass spectrometry, site-directed mutagenesis (AKT1 K426), phosphorylation assay, siRNA knockdown","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, MS, and mutagenesis of substrate residue in single lab","pmids":["32499521"],"is_preprint":false},{"year":2023,"finding":"HDAC8 degradation by the dual PROTAC YX968 does not induce histone hyperacetylation or broad transcriptomic perturbation, suggesting that HDAC8 deacetylase activity is not the primary driver of its transcriptional effects and that histone hyperacetylation is required for broad transcriptional changes.","method":"PROTAC-mediated selective degradation, quantitative proteomics, transcriptomics, histone acetylation assay","journal":"Cell chemical biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — selective degrader with quantitative proteomics and transcriptomics; single lab but orthogonal methods","pmids":["37572669"],"is_preprint":false},{"year":2016,"finding":"HDAC8 and HDAC9 suppress MAP2K3 transcription via H3K9 and H3K27 deacetylation at the MAP2K3 promoter; HDAC8/9 associate with SSRP1 and SUPT16H (components of FACT complex) responsible for MAP2K3 transcriptional elongation, establishing a mechanism for cutaneous innate immune tolerance.","method":"siRNA knockdown, ChIP (H3K9ac, H3K27ac), proteomics (HDAC8/9 interactors), transcriptomics, keratinocyte-specific conditional KO mouse","journal":"Science immunology","confidence":"High","confidence_rationale":"Tier 2 / Strong — ChIP, proteomics-validated interactors, siRNA, and in vivo KO model with consistent mechanistic findings in single rigorous study","pmids":["34021025"],"is_preprint":false},{"year":2023,"finding":"HDAC8 directly binds to and deacetylates HIF-1α, promoting HIF-1α protein stability and upregulating HIF-1α target genes (HK2, GLUT1) in melanoma.","method":"Co-immunoprecipitation, deacetylation assay, HDAC8 inhibitor treatment, siRNA knockdown, protein stability assay","journal":"Cancers","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP and deacetylation assay with functional readouts in single lab","pmids":["36831463"],"is_preprint":false},{"year":2016,"finding":"HDAC8 controls PD-L1 transcription in melanoma cells through a transcription complex of HOXA5 and STAT3; HDAC8 inhibition upregulates PD-L1 via activation of PD-L1 promoter fragments enriched with HOXA5 and STAT3 binding sites; HOXA5 and STAT3 are physically associated and interdependent in activating PD-L1 transcription.","method":"siRNA knockdown, luciferase reporter assay, co-immunoprecipitation of HOXA5-STAT3, HDAC8 inhibitor treatment, ChIP","journal":"The Journal of investigative dermatology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, luciferase, siRNA, and inhibitor with consistent results in single lab","pmids":["29174371"],"is_preprint":false},{"year":2024,"finding":"HDAC8 forms a complex with RELA (NF-κB p65) at the PRKN (Parkin) promoter; high-glucose-induced RELA-HDAC8 complex binds the PRKN promoter to suppress PRKN expression and dysregulate mitophagy; sodium butyrate restores PRKN expression by blocking RELA nuclear translocation and directly inhibiting HDAC8.","method":"ChIP assay (RELA-HDAC8 at PRKN promoter), Co-immunoprecipitation, HDAC8 overexpression, siRNA knockdown, mitophagy assay","journal":"Autophagy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP, Co-IP, and functional mitophagy readout in single lab","pmids":["38409852"],"is_preprint":false}],"current_model":"HDAC8 is a class I, zinc-dependent histone deacetylase with a defined crystal structure that catalyzes deacetylation of acetyl-lysine (and long-chain fatty acyl-lysine) residues on both histone (H3, H4, H3K27) and numerous non-histone substrates including SMC3 (its canonical cohesin-recycling substrate), p53, c-Jun, HIF-1α, EP300, PKM2, AKT1, and alpha-tubulin; substrate recognition involves both active-site and distal exosite interactions; HDAC8 operates in multiple cellular contexts including cohesin recycling (deacetylating SMC3 after anaphase for cohesin reuse), transcriptional regulation (as a cofactor of SMAD3/4 and DEC1 complexes and via H3K27 deacetylation at enhancers), smooth muscle contractility (via association with alpha-SMA), hematopoietic stem cell maintenance and leukemic stem cell transformation (via p53 deacetylation), cell cycle and spindle assembly, and innate immune tolerance (via MAP2K3 suppression); post-translational phosphorylation by AMPK drives HDAC8 nuclear-to-cytoplasmic relocalization, and disease-causing CdLS missense mutations structurally compromise its catalysis and thermostability."},"narrative":{"mechanistic_narrative":"HDAC8 is a class I, zinc/potassium-dependent histone deacetylase whose single alpha/beta catalytic domain, defined at atomic resolution, removes acyl modifications from lysine residues on both chromatin and non-histone proteins [PMID:15477595, PMID:10926844]. Two adjacent active-site histidines are essential for catalysis, and an isoform-specific subpocket near the active site underlies selective inhibition [PMID:10926844, PMID:27374062]. Beyond classical acetyl-lysine deacetylation, HDAC8 efficiently removes long-chain fatty acyl groups (octanoyl, dodecanoyl, myristoyl) from lysine with even higher catalytic efficiency than acetyl substrates [PMID:27459069]. Substrate selectivity is governed by both active-site contacts and distal exosite interactions, such that HDAC8 deacetylates assembled histone H3/H4 tetramers far more efficiently than isolated peptides [PMID:18470998, PMID:29109148]. Its canonical physiological substrate is the cohesin subunit SMC3, which HDAC8 deacetylates during cohesin recycling after anaphase; loss-of-function HDAC8 mutations cause Cornelia de Lange syndrome by elevating SMC3 acetylation and impairing cohesin reuse [PMID:22885700]. CdLS-associated missense mutations structurally compromise catalysis and thermostability, defects that can be partially rescued by small-molecule HDAC8 activators [PMID:25075551, PMID:26463496]. On chromatin, HDAC8 acts as an H3K27-specific deacetylase at enhancers and as a cofactor within defined transcriptional complexes (SMAD3/4, DEC1, EZH2, HOXA5/STAT3, RELA), thereby tuning programs in TGF-beta signaling, immune tolerance, fibrosis, and tumor immune evasion [PMID:31970414, PMID:33827976, PMID:34021025, PMID:29174371]. A broad non-histone acetylome — including p53, c-Jun, EP300, HIF-1alpha, PKM2, AKT1, and alpha-tubulin — links HDAC8 to hematopoietic and leukemic stem cell maintenance via p53 deacetylation, to drug-resistance and metastatic states in melanoma and breast cancer, and to spindle assembly [PMID:26387755, PMID:29084772, PMID:30987999, PMID:38030596, PMID:28223544, PMID:32499521]. AMPK phosphorylation drives HDAC8 nuclear-to-cytoplasmic relocalization under glucose deprivation, coupling its localization to metabolic state [PMID:32171858].","teleology":[{"year":2000,"claim":"Established that HDAC8 is a catalytically active class I deacetylase, answering whether this RPD3-family member functions as a bona fide histone deacetylase.","evidence":"immunopurification with in vitro HDAC assays on H4 peptides/core histones and active-site histidine mutagenesis","pmids":["10926844"],"confidence":"High","gaps":["Did not identify physiological in vivo substrates","No structural basis for catalysis"]},{"year":2004,"claim":"Defined the atomic architecture of the catalytic domain, revealing a bacterial-HDAC-like fold and an unexpected potassium dependence for fold stability.","evidence":"X-ray crystallography of HDAC8-hydroxamate complex plus CD spectroscopy","pmids":["15477595"],"confidence":"High","gaps":["Did not explain substrate selectivity","No non-histone substrate context"]},{"year":2008,"claim":"Showed that substrate recognition extends beyond the acetyl-lysine to distal sequence elements engaging an exosite, reframing how HDAC8 selects substrates.","evidence":"mass spectrometry SAM peptide HDAC assays with truncation/mutagenesis defining the KRHR exosite-binding motif","pmids":["18470998"],"confidence":"High","gaps":["Exosite contribution to full-length protein substrates not tested","Structural view of exosite contact lacking"]},{"year":2005,"claim":"Linked HDAC8 to the cytoskeleton and a non-chromatin function by associating it with alpha-SMA and showing a role in smooth muscle contractility.","evidence":"cell fractionation, reciprocal Co-IP, siRNA knockdown, and collagen lattice contraction assays","pmids":["15772115"],"confidence":"High","gaps":["Whether alpha-SMA or a contractile regulator is the deacetylation target unclear","Direct enzymatic substrate not defined"]},{"year":2012,"claim":"Identified SMC3 as the canonical physiological substrate and connected HDAC8 to cohesin recycling and Cornelia de Lange syndrome, the gene's defining disease link.","evidence":"biochemical deacetylase assay, ChIP-seq, CdLS patient mutation analysis, and cellular cohesin acetylation assays","pmids":["22885700"],"confidence":"High","gaps":["Genome-wide downstream transcriptional consequences only partially mapped","Tissue-specific developmental roles unresolved"]},{"year":2014,"claim":"Provided structural mechanism for CdLS pathogenesis by showing patient mutations destabilize the fold and impair catalysis, with pharmacological activator rescue.","evidence":"X-ray crystallography of multiple mutants plus enzymatic, thermostability, and activator-rescue assays (extended in 2015 with MD simulations)","pmids":["25075551","26463496"],"confidence":"High","gaps":["In vivo rescue by activators not demonstrated","Whether all CdLS phenotypes trace to SMC3 deacetylation loss unclear"]},{"year":2016,"claim":"Expanded the catalytic repertoire by demonstrating HDAC8 removes long-chain fatty acyl-lysine modifications more efficiently than acetyl groups, redefining it as a de-fatty-acylase.","evidence":"in vitro kinetics on acyl-lysine peptide panels plus cellular global fatty acylation measurement","pmids":["27459069"],"confidence":"High","gaps":["Physiological fatty-acylated substrates not identified","Cellular significance of de-fatty-acylation unresolved"]},{"year":2016,"claim":"Systematized non-histone substrate discovery through computational and chemical-biology approaches, establishing that HDAC8 regulates a broad acetylome.","evidence":"Rosetta FlexPepBind prediction with peptide-assay validation (SMC3, ERRalpha, ARID1A) and active-site photocrosslinking with non-natural amino acids for in-lysate substrate capture","pmids":["26933971","29035536"],"confidence":"High","gaps":["Cellular validation of predicted substrates limited","Physiological relevance of each substrate not established"]},{"year":2017,"claim":"Quantified how substrate size and architecture shape catalysis, showing HDAC8 favors assembled histone tetramers over peptides while efficiency drops for larger complexes.","evidence":"comparative in vitro kinetics across peptides, full-length proteins, and protein-nucleic acid complexes","pmids":["29109148"],"confidence":"High","gaps":["Mechanism of efficiency loss on large complexes unclear","In-cell substrate-format preferences not tested"]},{"year":2015,"claim":"Connected HDAC8 to hematopoietic and leukemic stem cell biology by establishing p53 as a substrate whose deacetylation drives transformation in inv(16)+ AML.","evidence":"Co-IP, acetylation assays, selective inhibitor, HDAC8-deficient mice, and patient-derived xenografts (extended to normal HSC maintenance in 2017 and TKI resistance in 2020)","pmids":["26387755","29084772","32315388"],"confidence":"High","gaps":["Specificity of p53 lysines deacetylated not fully mapped across contexts","Relative contribution of p53 versus other substrates to leukemia unclear"]},{"year":2019,"claim":"Broadened the oncogenic substrate set to c-Jun, EP300, HIF-1alpha, PKM2, and AKT1, linking HDAC8 to drug resistance, metastasis, and metabolic rewiring.","evidence":"Co-IP, mass spectrometry, site-directed mutagenesis of substrate lysines, and in vivo tumor/metastasis models across melanoma and breast cancer","pmids":["30987999","38030596","36831463","33279948","32499521"],"confidence":"Medium","gaps":["Several substrate relationships rest on single-lab evidence","Direct versus indirect deacetylation not always discriminated"]},{"year":2021,"claim":"Defined HDAC8 as an H3K27-specific deacetylase at enhancers and a transcriptional cofactor across SMAD3/4, DEC1, EZH2, HOXA5/STAT3, and RELA complexes, establishing its chromatin-regulatory roles.","evidence":"genome-wide H3K27ac ChIP/ATAC profiling with selective inhibitors and humanized/conditional mouse models, plus Co-IP/ChIP in individual transcription-factor studies","pmids":["33827976","31970414","24404147","26383163","29174371","38409852","34021025"],"confidence":"High","gaps":["Whether catalytic deacetylation versus scaffolding drives each transcriptional effect not uniformly resolved","Recruitment determinants to specific loci incompletely defined"]},{"year":2020,"claim":"Showed HDAC8 localization is regulated by metabolic signaling, with AMPK phosphorylation triggering nuclear-to-cytoplasmic relocalization.","evidence":"subcellular fractionation, Co-IP, AMPK activation/inhibition, and HDAC8 phosphorylation assays during glucose deprivation","pmids":["32171858"],"confidence":"Medium","gaps":["Phosphosite(s) on HDAC8 not precisely mapped","Generality across cell types not established"]},{"year":2017,"claim":"Implicated HDAC8 in meiotic spindle assembly and chromosome segregation through tubulin deacetylation and spindle-pole localization.","evidence":"siRNA, selective inhibitor, immunofluorescence, and aneuploidy assays in mouse oocytes, plus in vitro alpha-tubulin K40 deacetylation assays","pmids":["28223544","29716651"],"confidence":"Medium","gaps":["Mechanistic link between tubulin deacetylation and gamma-tubulin recruitment unclear","Redundancy with HDAC6 complicates attribution"]},{"year":2023,"claim":"Challenged the assumption that HDAC8's transcriptional effects depend on its deacetylase activity, showing PROTAC degradation did not induce histone hyperacetylation or broad transcriptomic change.","evidence":"dual PROTAC-mediated degradation with quantitative proteomics, transcriptomics, and histone acetylation assays","pmids":["37572669"],"confidence":"Medium","gaps":["Reconciliation with catalytic-substrate findings unresolved","Possible non-catalytic scaffolding roles not directly mapped"]},{"year":null,"claim":"It remains unresolved how HDAC8 partitions between its catalytic deacetylase/de-fatty-acylase functions and non-catalytic scaffolding roles, and which substrates dominate in each physiological and disease context.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unified model reconciling catalytic-substrate and PROTAC-degradation data","Tissue-specific in vivo substrate hierarchy undefined","Physiological de-fatty-acylation targets unidentified"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[1,2,6,7,20,21,32,34,37]},{"term_id":"GO:0016787","term_label":"hydrolase activity","supporting_discovery_ids":[0,1,7,12]},{"term_id":"GO:0042393","term_label":"histone binding","supporting_discovery_ids":[11,12,24]},{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[5,15,24,36]}],"localization":[{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[2,14,24,28]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[28,32]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[3]},{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[31]},{"term_id":"GO:0005730","term_label":"nucleolus","supporting_discovery_ids":[32]}],"pathway":[{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[2,22,27,31]},{"term_id":"R-HSA-4839726","term_label":"Chromatin organization","supporting_discovery_ids":[21,24,29,36]},{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[5,15,30,36,38]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[2,6,20]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[24,36]}],"complexes":["cohesin (SMC3 substrate)","SMAD3/4 complex"],"partners":["SMC3","TP53","EP300","JUN","HIF1A","AKT1","EZH2","RELA"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9BY41","full_name":"Histone deacetylase 8","aliases":["Protein deacetylase HDAC8","Protein decrotonylase HDAC8"],"length_aa":377,"mass_kda":41.8,"function":"Histone deacetylase that catalyzes the deacetylation of lysine residues on the N-terminal part of the core histones (H2A, H2B, H3 and H4) (PubMed:10748112, PubMed:10922473, PubMed:10926844, PubMed:14701748, PubMed:28497810). Histone deacetylation gives a tag for epigenetic repression and plays an important role in transcriptional regulation, cell cycle progression and developmental events (PubMed:10748112, PubMed:10922473, PubMed:10926844, PubMed:14701748). Histone deacetylases act via the formation of large multiprotein complexes (PubMed:10748112, PubMed:10922473, PubMed:10926844, PubMed:14701748). Also involved in the deacetylation of cohesin complex protein SMC3 regulating release of cohesin complexes from chromatin (PubMed:22885700). May play a role in smooth muscle cell contractility (PubMed:15772115). In addition to protein deacetylase activity, also has protein-lysine deacylase activity: acts as a protein decrotonylase by mediating decrotonylation ((2E)-butenoyl) of histones (PubMed:28497810)","subcellular_location":"Nucleus; Chromosome; Cytoplasm","url":"https://www.uniprot.org/uniprotkb/Q9BY41/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/HDAC8","classification":"Not Classified","n_dependent_lines":74,"n_total_lines":1208,"dependency_fraction":0.061258278145695365},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/HDAC8","total_profiled":1310},"omim":[{"mim_id":"610962","title":"SMG5 NONSENSE-MEDIATED mRNA DECAY FACTOR; SMG5","url":"https://www.omim.org/entry/610962"},{"mim_id":"601844","title":"PROTEIN KINASE, LYSINE-DEFICIENT 4; WNK4","url":"https://www.omim.org/entry/601844"},{"mim_id":"300882","title":"CORNELIA DE LANGE SYNDROME 5; CDLS5","url":"https://www.omim.org/entry/300882"},{"mim_id":"300269","title":"HISTONE DEACETYLASE 8; HDAC8","url":"https://www.omim.org/entry/300269"},{"mim_id":"122470","title":"CORNELIA DE LANGE SYNDROME 1; CDLS1","url":"https://www.omim.org/entry/122470"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/HDAC8"},"hgnc":{"alias_symbol":["RPD3","KDAC8"],"prev_symbol":["HDACL1","WTS","MRXS6"]},"alphafold":{"accession":"Q9BY41","domains":[{"cath_id":"3.40.800.20","chopping":"16-57_116-373","consensus_level":"high","plddt":97.9628,"start":16,"end":373}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9BY41","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9BY41-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9BY41-F1-predicted_aligned_error_v6.png","plddt_mean":95.31},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=HDAC8","jax_strain_url":"https://www.jax.org/strain/search?query=HDAC8"},"sequence":{"accession":"Q9BY41","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9BY41.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9BY41/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9BY41"}},"corpus_meta":[{"pmid":"15477595","id":"PMC_15477595","title":"Crystal structure of a eukaryotic zinc-dependent histone deacetylase, human HDAC8, complexed with a hydroxamic acid inhibitor.","date":"2004","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/15477595","citation_count":532,"is_preprint":false},{"pmid":"22885700","id":"PMC_22885700","title":"HDAC8 mutations in Cornelia de Lange syndrome affect the cohesin acetylation cycle.","date":"2012","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/22885700","citation_count":476,"is_preprint":false},{"pmid":"18256683","id":"PMC_18256683","title":"A novel histone deacetylase 8 (HDAC8)-specific inhibitor PCI-34051 induces apoptosis in T-cell lymphomas.","date":"2008","source":"Leukemia","url":"https://pubmed.ncbi.nlm.nih.gov/18256683","citation_count":382,"is_preprint":false},{"pmid":"26013035","id":"PMC_26013035","title":"HDAC8: a multifaceted target for therapeutic interventions.","date":"2015","source":"Trends in pharmacological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/26013035","citation_count":220,"is_preprint":false},{"pmid":"15772115","id":"PMC_15772115","title":"Histone deacetylase HDAC8 associates with smooth muscle alpha-actin and is essential for smooth muscle cell contractility.","date":"2005","source":"FASEB journal : official publication of the Federation of American Societies for Experimental Biology","url":"https://pubmed.ncbi.nlm.nih.gov/15772115","citation_count":172,"is_preprint":false},{"pmid":"25695609","id":"PMC_25695609","title":"Selective inhibition of HDAC8 decreases neuroblastoma growth in vitro and in vivo and enhances retinoic acid-mediated differentiation.","date":"2015","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/25695609","citation_count":143,"is_preprint":false},{"pmid":"33827976","id":"PMC_33827976","title":"A selective HDAC8 inhibitor potentiates antitumor immunity and efficacy of immune checkpoint blockade in hepatocellular carcinoma.","date":"2021","source":"Science translational medicine","url":"https://pubmed.ncbi.nlm.nih.gov/33827976","citation_count":139,"is_preprint":false},{"pmid":"10926844","id":"PMC_10926844","title":"Cloning and characterization of a novel human histone deacetylase, HDAC8.","date":"2000","source":"The Biochemical journal","url":"https://pubmed.ncbi.nlm.nih.gov/10926844","citation_count":135,"is_preprint":false},{"pmid":"28783689","id":"PMC_28783689","title":"Inhibition of HDAC8 and HDAC9 by microbial short-chain fatty acids breaks immune tolerance of the epidermis to TLR ligands.","date":"2016","source":"Science immunology","url":"https://pubmed.ncbi.nlm.nih.gov/28783689","citation_count":120,"is_preprint":false},{"pmid":"26383163","id":"PMC_26383163","title":"Histone Deacetylase HDAC8 Promotes Insulin Resistance and β-Catenin Activation in NAFLD-Associated Hepatocellular Carcinoma.","date":"2015","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/26383163","citation_count":111,"is_preprint":false},{"pmid":"26387755","id":"PMC_26387755","title":"HDAC8 Inhibition Specifically Targets Inv(16) Acute Myeloid Leukemic Stem Cells by Restoring p53 Acetylation.","date":"2015","source":"Cell stem cell","url":"https://pubmed.ncbi.nlm.nih.gov/26387755","citation_count":94,"is_preprint":false},{"pmid":"27459069","id":"PMC_27459069","title":"HDAC8 Catalyzes the Hydrolysis of Long Chain Fatty Acyl Lysine.","date":"2016","source":"ACS chemical biology","url":"https://pubmed.ncbi.nlm.nih.gov/27459069","citation_count":89,"is_preprint":false},{"pmid":"30594678","id":"PMC_30594678","title":"Histone deacetylase 8 (HDAC8) and its inhibitors with selectivity to other isoforms: An overview.","date":"2018","source":"European journal of medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/30594678","citation_count":81,"is_preprint":false},{"pmid":"31970414","id":"PMC_31970414","title":"HDAC8 cooperates with SMAD3/4 complex to suppress SIRT7 and promote cell survival and migration.","date":"2020","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/31970414","citation_count":78,"is_preprint":false},{"pmid":"26876786","id":"PMC_26876786","title":"Histone deacetylase inhibitors suppress mutant p53 transcription via HDAC8/YY1 signals in triple negative breast cancer cells.","date":"2016","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/26876786","citation_count":74,"is_preprint":false},{"pmid":"15975907","id":"PMC_15975907","title":"Drosophila Mob family proteins interact with the related tricornered (Trc) and warts (Wts) kinases.","date":"2005","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/15975907","citation_count":70,"is_preprint":false},{"pmid":"23868068","id":"PMC_23868068","title":"HDAC inhibition suppresses cardiac hypertrophy and fibrosis in DOCA-salt hypertensive rats via regulation of HDAC6/HDAC8 enzyme activity.","date":"2013","source":"Kidney & blood pressure research","url":"https://pubmed.ncbi.nlm.nih.gov/23868068","citation_count":70,"is_preprint":false},{"pmid":"25321483","id":"PMC_25321483","title":"HDAC8 and STAT3 repress BMF gene activity in colon cancer cells.","date":"2014","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/25321483","citation_count":69,"is_preprint":false},{"pmid":"29190092","id":"PMC_29190092","title":"Structure-Based Design and Biological Characterization of Selective Histone Deacetylase 8 (HDAC8) Inhibitors with Anti-Neuroblastoma Activity.","date":"2017","source":"Journal of medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/29190092","citation_count":69,"is_preprint":false},{"pmid":"19780707","id":"PMC_19780707","title":"Targeting of HDAC8 and investigational inhibitors in neuroblastoma.","date":"2009","source":"Expert opinion on investigational drugs","url":"https://pubmed.ncbi.nlm.nih.gov/19780707","citation_count":68,"is_preprint":false},{"pmid":"32315388","id":"PMC_32315388","title":"FLT3 inhibition upregulates HDAC8 via FOXO to inactivate p53 and promote maintenance of FLT3-ITD+ acute myeloid leukemia.","date":"2020","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/32315388","citation_count":66,"is_preprint":false},{"pmid":"31730902","id":"PMC_31730902","title":"LncRNA ID2-AS1 suppresses tumor metastasis by activating the HDAC8/ID2 pathway in hepatocellular carcinoma.","date":"2019","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/31730902","citation_count":60,"is_preprint":false},{"pmid":"36231123","id":"PMC_36231123","title":"Pathological Role of HDAC8: Cancer and Beyond.","date":"2022","source":"Cells","url":"https://pubmed.ncbi.nlm.nih.gov/36231123","citation_count":59,"is_preprint":false},{"pmid":"32171858","id":"PMC_32171858","title":"AMPK-dependent phosphorylation of HDAC8 triggers PGM1 expression to promote lung cancer cell survival under glucose starvation.","date":"2020","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/32171858","citation_count":57,"is_preprint":false},{"pmid":"33959033","id":"PMC_33959033","title":"Selective HDAC8 Inhibition Attenuates Isoproterenol-Induced Cardiac Hypertrophy and Fibrosis via p38 MAPK Pathway.","date":"2021","source":"Frontiers in pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/33959033","citation_count":56,"is_preprint":false},{"pmid":"30987999","id":"PMC_30987999","title":"HDAC8 Regulates a Stress Response Pathway in Melanoma to Mediate Escape from BRAF Inhibitor Therapy.","date":"2019","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/30987999","citation_count":56,"is_preprint":false},{"pmid":"23482931","id":"PMC_23482931","title":"SOX4 is a direct target gene of FRA-2 and induces expression of HDAC8 in adult T-cell leukemia/lymphoma.","date":"2013","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/23482931","citation_count":56,"is_preprint":false},{"pmid":"19070599","id":"PMC_19070599","title":"Inactivation of CREB mediated gene transcription by HDAC8 bound protein phosphatase.","date":"2008","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/19070599","citation_count":55,"is_preprint":false},{"pmid":"31605710","id":"PMC_31605710","title":"MiR-216b-5p inhibits cell proliferation in human breast cancer by down-regulating HDAC8 expression.","date":"2019","source":"Life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/31605710","citation_count":54,"is_preprint":false},{"pmid":"29716651","id":"PMC_29716651","title":"Overexpressed HDAC8 in cervical cancer cells shows functional redundancy of tubulin deacetylation with HDAC6.","date":"2018","source":"Cell communication and signaling : CCS","url":"https://pubmed.ncbi.nlm.nih.gov/29716651","citation_count":54,"is_preprint":false},{"pmid":"33279948","id":"PMC_33279948","title":"HDAC8-dependent deacetylation of PKM2 directs nuclear localization and glycolysis to promote proliferation in hepatocellular carcinoma.","date":"2020","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/33279948","citation_count":53,"is_preprint":false},{"pmid":"30358439","id":"PMC_30358439","title":"HDAC8 inhibition ameliorates pulmonary fibrosis.","date":"2018","source":"American journal of physiology. Lung cellular and molecular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/30358439","citation_count":52,"is_preprint":false},{"pmid":"32499521","id":"PMC_32499521","title":"HDAC8 promotes the dissemination of breast cancer cells via AKT/GSK-3β/Snail signals.","date":"2020","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/32499521","citation_count":51,"is_preprint":false},{"pmid":"25075551","id":"PMC_25075551","title":"Compromised structure and function of HDAC8 mutants identified in Cornelia de Lange Syndrome spectrum disorders.","date":"2014","source":"ACS chemical biology","url":"https://pubmed.ncbi.nlm.nih.gov/25075551","citation_count":51,"is_preprint":false},{"pmid":"27072133","id":"PMC_27072133","title":"HDAC8 Inhibition Blocks SMC3 Deacetylation and Delays Cell Cycle Progression without Affecting Cohesin-dependent Transcription in MCF7 Cancer Cells.","date":"2016","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/27072133","citation_count":50,"is_preprint":false},{"pmid":"28501516","id":"PMC_28501516","title":"Is dual inhibition of metalloenzymes HDAC-8 and MMP-2 a potential pharmacological target to combat hematological malignancies?","date":"2017","source":"Pharmacological research","url":"https://pubmed.ncbi.nlm.nih.gov/28501516","citation_count":50,"is_preprint":false},{"pmid":"26200462","id":"PMC_26200462","title":"HDAC8, A Potential Therapeutic Target for the Treatment of Malignant Peripheral Nerve Sheath Tumors (MPNST).","date":"2015","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/26200462","citation_count":48,"is_preprint":false},{"pmid":"29084772","id":"PMC_29084772","title":"HDAC8 regulates long-term hematopoietic stem-cell maintenance under stress by modulating p53 activity.","date":"2017","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/29084772","citation_count":47,"is_preprint":false},{"pmid":"27374062","id":"PMC_27374062","title":"Structure of 'linkerless' hydroxamic acid inhibitor-HDAC8 complex confirms the formation of an isoform-specific subpocket.","date":"2016","source":"Journal of structural biology","url":"https://pubmed.ncbi.nlm.nih.gov/27374062","citation_count":45,"is_preprint":false},{"pmid":"18470998","id":"PMC_18470998","title":"The activity of HDAC8 depends on local and distal sequences of its peptide substrates.","date":"2008","source":"Biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/18470998","citation_count":45,"is_preprint":false},{"pmid":"26431101","id":"PMC_26431101","title":"Suberoylanilide hydroxamic acid (SAHA) promotes the epithelial mesenchymal transition of triple negative breast cancer cells via HDAC8/FOXA1 signals.","date":"2016","source":"Biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/26431101","citation_count":45,"is_preprint":false},{"pmid":"32563146","id":"PMC_32563146","title":"LncRNA NORAD accelerates the progression and doxorubicin resistance of neuroblastoma through up-regulating HDAC8 via sponging miR-144-3p.","date":"2020","source":"Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie","url":"https://pubmed.ncbi.nlm.nih.gov/32563146","citation_count":44,"is_preprint":false},{"pmid":"34021025","id":"PMC_34021025","title":"Cutaneous innate immune tolerance is mediated by epigenetic control of MAP2K3 by HDAC8/9.","date":"2021","source":"Science immunology","url":"https://pubmed.ncbi.nlm.nih.gov/34021025","citation_count":42,"is_preprint":false},{"pmid":"26933971","id":"PMC_26933971","title":"Structure-Based Identification of HDAC8 Non-histone Substrates.","date":"2016","source":"Structure (London, England : 1993)","url":"https://pubmed.ncbi.nlm.nih.gov/26933971","citation_count":42,"is_preprint":false},{"pmid":"32591593","id":"PMC_32591593","title":"Hydroxamic acid derivatives as HDAC1, HDAC6 and HDAC8 inhibitors with antiproliferative activity in cancer cell lines.","date":"2020","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/32591593","citation_count":41,"is_preprint":false},{"pmid":"39440960","id":"PMC_39440960","title":"Butyrate Inhibits the HDAC8/NF-κB Pathway to Enhance Slc26a3 Expression and Improve the Intestinal Epithelial Barrier to Relieve Colitis.","date":"2024","source":"Journal of agricultural and food chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/39440960","citation_count":40,"is_preprint":false},{"pmid":"20211163","id":"PMC_20211163","title":"Regulation of cell shape, wing hair initiation and the actin cytoskeleton by Trc/Fry and Wts/Mats complexes.","date":"2010","source":"Developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/20211163","citation_count":38,"is_preprint":false},{"pmid":"33015043","id":"PMC_33015043","title":"HDAC8: A Promising Therapeutic Target for Acute Myeloid Leukemia.","date":"2020","source":"Frontiers in cell and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/33015043","citation_count":38,"is_preprint":false},{"pmid":"31508813","id":"PMC_31508813","title":"miR-483-3p suppresses the proliferation and progression of human triple negative breast cancer cells by targeting the HDAC8>oncogene.","date":"2019","source":"Journal of cellular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/31508813","citation_count":37,"is_preprint":false},{"pmid":"37572669","id":"PMC_37572669","title":"HDAC3 and HDAC8 PROTAC dual degrader reveals roles of histone acetylation in gene regulation.","date":"2023","source":"Cell chemical biology","url":"https://pubmed.ncbi.nlm.nih.gov/37572669","citation_count":36,"is_preprint":false},{"pmid":"26463496","id":"PMC_26463496","title":"Biochemical and structural characterization of HDAC8 mutants associated with Cornelia de Lange syndrome spectrum disorders.","date":"2015","source":"Biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/26463496","citation_count":36,"is_preprint":false},{"pmid":"31901516","id":"PMC_31901516","title":"Discovery of 5-naphthylidene-2,4-thiazolidinedione derivatives as selective HDAC8 inhibitors and evaluation of their cytotoxic effects in leukemic cell lines.","date":"2019","source":"Bioorganic chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/31901516","citation_count":35,"is_preprint":false},{"pmid":"25836739","id":"PMC_25836739","title":"Expression of the class 1 histone deacetylases HDAC8 and 3 are associated with improved survival of patients with metastatic melanoma.","date":"2015","source":"Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc","url":"https://pubmed.ncbi.nlm.nih.gov/25836739","citation_count":35,"is_preprint":false},{"pmid":"32672458","id":"PMC_32672458","title":"Characterization of Conformationally Constrained Benzanilide Scaffolds for Potent and Selective HDAC8 Targeting.","date":"2020","source":"Journal of medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/32672458","citation_count":34,"is_preprint":false},{"pmid":"29109148","id":"PMC_29109148","title":"HDAC8 substrate selectivity is determined by long- and short-range interactions leading to enhanced reactivity for full-length histone substrates compared with peptides.","date":"2017","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/29109148","citation_count":34,"is_preprint":false},{"pmid":"29514055","id":"PMC_29514055","title":"Structure-activity relationships of HDAC8 inhibitors: Non-hydroxamates as anticancer agents.","date":"2018","source":"Pharmacological research","url":"https://pubmed.ncbi.nlm.nih.gov/29514055","citation_count":33,"is_preprint":false},{"pmid":"31798765","id":"PMC_31798765","title":"HDAC8 affects MGMT levels in glioblastoma cell lines via interaction with the proteasome receptor ADRM1.","date":"2019","source":"Genes & cancer","url":"https://pubmed.ncbi.nlm.nih.gov/31798765","citation_count":33,"is_preprint":false},{"pmid":"33542077","id":"PMC_33542077","title":"Targeting DNA Damage Repair Functions of Two Histone Deacetylases, HDAC8 and SIRT6, Sensitizes Acute Myeloid Leukemia to NAMPT Inhibition.","date":"2021","source":"Clinical cancer research : an official journal of the American Association for Cancer Research","url":"https://pubmed.ncbi.nlm.nih.gov/33542077","citation_count":33,"is_preprint":false},{"pmid":"35886887","id":"PMC_35886887","title":"Design, Synthesis and Biological Characterization of Histone Deacetylase 8 (HDAC8) Proteolysis Targeting Chimeras (PROTACs) with Anti-Neuroblastoma Activity.","date":"2022","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/35886887","citation_count":33,"is_preprint":false},{"pmid":"27576763","id":"PMC_27576763","title":"Structural aspects of HDAC8 mechanism and dysfunction in Cornelia de Lange syndrome spectrum disorders.","date":"2016","source":"Protein science : a publication of the Protein Society","url":"https://pubmed.ncbi.nlm.nih.gov/27576763","citation_count":32,"is_preprint":false},{"pmid":"24404147","id":"PMC_24404147","title":"DEC1 coordinates with HDAC8 to differentially regulate TAp73 and ΔNp73 expression.","date":"2014","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/24404147","citation_count":32,"is_preprint":false},{"pmid":"34072831","id":"PMC_34072831","title":"NBM-BMX, an HDAC8 Inhibitor, Overcomes Temozolomide Resistance in Glioblastoma Multiforme by Downregulating the β-Catenin/c-Myc/SOX2 Pathway and Upregulating p53-Mediated MGMT Inhibition.","date":"2021","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/34072831","citation_count":32,"is_preprint":false},{"pmid":"32446120","id":"PMC_32446120","title":"Discovery of novel N-substituted thiazolidinediones (TZDs) as HDAC8 inhibitors: in-silico studies, synthesis, and biological evaluation.","date":"2020","source":"Bioorganic chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/32446120","citation_count":31,"is_preprint":false},{"pmid":"19947584","id":"PMC_19947584","title":"Docking of hydroxamic acids into HDAC1 and HDAC8: a rationalization of activity trends and selectivities.","date":"2009","source":"Journal of chemical information and modeling","url":"https://pubmed.ncbi.nlm.nih.gov/19947584","citation_count":31,"is_preprint":false},{"pmid":"29174371","id":"PMC_29174371","title":"Cooperativity of HOXA5 and STAT3 Is Critical for HDAC8 Inhibition-Mediated Transcriptional Activation of PD-L1 in Human Melanoma Cells.","date":"2017","source":"The Journal of investigative dermatology","url":"https://pubmed.ncbi.nlm.nih.gov/29174371","citation_count":31,"is_preprint":false},{"pmid":"28223126","id":"PMC_28223126","title":"Targeted inhibition of HDAC8 increases the doxorubicin sensitivity of neuroblastoma cells via up regulation of miR-137.","date":"2017","source":"European journal of pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/28223126","citation_count":31,"is_preprint":false},{"pmid":"37098288","id":"PMC_37098288","title":"Discovery of highly potent HDAC8 PROTACs with anti-tumor activity.","date":"2023","source":"Bioorganic chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/37098288","citation_count":29,"is_preprint":false},{"pmid":"35016723","id":"PMC_35016723","title":"HDAC8 suppresses the epithelial phenotype and promotes EMT in chemotherapy-treated basal-like breast cancer.","date":"2022","source":"Clinical epigenetics","url":"https://pubmed.ncbi.nlm.nih.gov/35016723","citation_count":29,"is_preprint":false},{"pmid":"29035536","id":"PMC_29035536","title":"HDAC8 Substrates Identified by Genetically Encoded Active Site Photocrosslinking.","date":"2017","source":"Journal of the American Chemical Society","url":"https://pubmed.ncbi.nlm.nih.gov/29035536","citation_count":29,"is_preprint":false},{"pmid":"34214631","id":"PMC_34214631","title":"Targeting HDAC8 to ameliorate skeletal muscle differentiation in Duchenne muscular dystrophy.","date":"2021","source":"Pharmacological research","url":"https://pubmed.ncbi.nlm.nih.gov/34214631","citation_count":29,"is_preprint":false},{"pmid":"31748509","id":"PMC_31748509","title":"Discovery of novel potential selective HDAC8 inhibitors by combine ligand-based, structure-based virtual screening and in-vitro biological evaluation.","date":"2019","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/31748509","citation_count":29,"is_preprint":false},{"pmid":"38030596","id":"PMC_38030596","title":"HDAC8-mediated inhibition of EP300 drives a transcriptional state that increases melanoma brain metastasis.","date":"2023","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/38030596","citation_count":28,"is_preprint":false},{"pmid":"29533873","id":"PMC_29533873","title":"Discovery of meta-sulfamoyl N-hydroxybenzamides as HDAC8 selective inhibitors.","date":"2018","source":"European journal of medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/29533873","citation_count":28,"is_preprint":false},{"pmid":"24973453","id":"PMC_24973453","title":"HDAC8-mediated epigenetic reprogramming plays a key role in resistance to anthrax lethal toxin-induced pyroptosis in macrophages.","date":"2014","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/24973453","citation_count":28,"is_preprint":false},{"pmid":"2117034","id":"PMC_2117034","title":"High frequency expression of S107 VH genes by peritoneal B cells of B10.H-2aH-4bP/WTS mice.","date":"1990","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/2117034","citation_count":27,"is_preprint":false},{"pmid":"35330681","id":"PMC_35330681","title":"Long noncoding RNA LINC01435 impedes diabetic wound healing by facilitating YY1-mediated HDAC8 expression.","date":"2022","source":"iScience","url":"https://pubmed.ncbi.nlm.nih.gov/35330681","citation_count":27,"is_preprint":false},{"pmid":"23719282","id":"PMC_23719282","title":"Synthesis and biological evaluation of a targeted DNA-binding transcriptional activator with HDAC8 inhibitory activity.","date":"2013","source":"Bioorganic & medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/23719282","citation_count":27,"is_preprint":false},{"pmid":"32111211","id":"PMC_32111211","title":"HDAC8 inhibitor attenuates airway responses to antigen stimulus through synchronously suppressing galectin-3 expression and reducing macrophage-2 polarization.","date":"2020","source":"Respiratory research","url":"https://pubmed.ncbi.nlm.nih.gov/32111211","citation_count":26,"is_preprint":false},{"pmid":"37463500","id":"PMC_37463500","title":"Discovery of HDAC6, HDAC8, and 6/8 Inhibitors and Development of Cell-Based Drug Screening Models for the Treatment of TGF-β-Induced Idiopathic Pulmonary Fibrosis.","date":"2023","source":"Journal of medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/37463500","citation_count":25,"is_preprint":false},{"pmid":"31421254","id":"PMC_31421254","title":"Design, synthesis, biological evaluation and molecular docking study of arylcarboxamido piperidine and piperazine-based hydroxamates as potential HDAC8 inhibitors with promising anticancer activity.","date":"2019","source":"European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences","url":"https://pubmed.ncbi.nlm.nih.gov/31421254","citation_count":25,"is_preprint":false},{"pmid":"29515255","id":"PMC_29515255","title":"A kinome-wide RNAi screen identifies ALK as a target to sensitize neuroblastoma cells for HDAC8-inhibitor treatment.","date":"2018","source":"Cell death and differentiation","url":"https://pubmed.ncbi.nlm.nih.gov/29515255","citation_count":25,"is_preprint":false},{"pmid":"37429084","id":"PMC_37429084","title":"Are inhibitors of histone deacetylase 8 (HDAC8) effective in hematological cancers especially acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL)?","date":"2023","source":"European journal of medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/37429084","citation_count":24,"is_preprint":false},{"pmid":"29499194","id":"PMC_29499194","title":"HDAC8 regulates neural differentiation through embryoid body formation in P19 cells.","date":"2018","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/29499194","citation_count":24,"is_preprint":false},{"pmid":"31817161","id":"PMC_31817161","title":"Induction of Apoptosis and Autophagy in Breast Cancer Cells by a Novel HDAC8 Inhibitor.","date":"2019","source":"Biomolecules","url":"https://pubmed.ncbi.nlm.nih.gov/31817161","citation_count":24,"is_preprint":false},{"pmid":"26625202","id":"PMC_26625202","title":"Non-epigenetic function of HDAC8 in regulating breast cancer stem cells by maintaining Notch1 protein stability.","date":"2016","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/26625202","citation_count":24,"is_preprint":false},{"pmid":"30405794","id":"PMC_30405794","title":"HDAC inhibitor apicidin suppresses murine oral squamous cell carcinoma cell growth in vitro and in vivo via inhibiting HDAC8 expression.","date":"2018","source":"Oncology letters","url":"https://pubmed.ncbi.nlm.nih.gov/30405794","citation_count":24,"is_preprint":false},{"pmid":"30632447","id":"PMC_30632447","title":"Selective and nonselective HDAC8 inhibitors: a therapeutic patent review.","date":"2019","source":"Pharmaceutical patent analyst","url":"https://pubmed.ncbi.nlm.nih.gov/30632447","citation_count":23,"is_preprint":false},{"pmid":"35955780","id":"PMC_35955780","title":"HDAC8-Selective Inhibition by PCI-34051 Enhances the Anticancer Effects of ACY-241 in Ovarian Cancer Cells.","date":"2022","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/35955780","citation_count":23,"is_preprint":false},{"pmid":"38409852","id":"PMC_38409852","title":"Sodium butyrate ameliorates high glucose-suppressed neuronal mitophagy by restoring PRKN expression via inhibiting the RELA-HDAC8 complex.","date":"2024","source":"Autophagy","url":"https://pubmed.ncbi.nlm.nih.gov/38409852","citation_count":22,"is_preprint":false},{"pmid":"36831463","id":"PMC_36831463","title":"HDAC8 Deacetylates HIF-1α and Enhances Its Protein Stability to Promote Tumor Growth and Migration in Melanoma.","date":"2023","source":"Cancers","url":"https://pubmed.ncbi.nlm.nih.gov/36831463","citation_count":22,"is_preprint":false},{"pmid":"28223544","id":"PMC_28223544","title":"HDAC8 functions in spindle assembly during mouse oocyte meiosis.","date":"2017","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/28223544","citation_count":22,"is_preprint":false},{"pmid":"30426726","id":"PMC_30426726","title":"HDAC8, A Potential Therapeutic Target, Regulates Proliferation and Differentiation of Bone Marrow Stromal Cells in Fibrous Dysplasia.","date":"2018","source":"Stem cells translational medicine","url":"https://pubmed.ncbi.nlm.nih.gov/30426726","citation_count":22,"is_preprint":false},{"pmid":"26912657","id":"PMC_26912657","title":"Inhibition of Interleukin 1β (IL-1β) Expression by Anthrax Lethal Toxin (LeTx) Is Reversed by Histone Deacetylase 8 (HDAC8) Inhibition in Murine Macrophages.","date":"2016","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/26912657","citation_count":22,"is_preprint":false},{"pmid":"22101275","id":"PMC_22101275","title":"Drosophila endocytic neoplastic tumor suppressor genes regulate Sav/Wts/Hpo signaling and the c-Jun N-terminal kinase pathway.","date":"2011","source":"Cell cycle (Georgetown, Tex.)","url":"https://pubmed.ncbi.nlm.nih.gov/22101275","citation_count":21,"is_preprint":false},{"pmid":"35126818","id":"PMC_35126818","title":"Hdac8 Inhibitor Alleviates Transverse Aortic Constriction-Induced Heart Failure in Mice by Downregulating Ace1.","date":"2022","source":"Oxidative medicine and cellular longevity","url":"https://pubmed.ncbi.nlm.nih.gov/35126818","citation_count":21,"is_preprint":false},{"pmid":"37408174","id":"PMC_37408174","title":"Foot-and-mouth disease virus structural protein VP3 interacts with HDAC8 and promotes its autophagic degradation to facilitate viral replication.","date":"2023","source":"Autophagy","url":"https://pubmed.ncbi.nlm.nih.gov/37408174","citation_count":19,"is_preprint":false},{"pmid":"36035205","id":"PMC_36035205","title":"HDAC8 Promotes Liver Metastasis of Colorectal Cancer via Inhibition of IRF1 and Upregulation of SUCNR1.","date":"2022","source":"Oxidative medicine and cellular longevity","url":"https://pubmed.ncbi.nlm.nih.gov/36035205","citation_count":19,"is_preprint":false},{"pmid":"25605725","id":"PMC_25605725","title":"Mechanism of N-Acylthiourea-mediated activation of human histone deacetylase 8 (HDAC8) at molecular and cellular levels.","date":"2015","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/25605725","citation_count":19,"is_preprint":false},{"pmid":"27673327","id":"PMC_27673327","title":"Roles of HDAC2 and HDAC8 in Cardiac Remodeling in Renovascular Hypertensive Rats and the Effects of Valproic Acid Sodium.","date":"2016","source":"Pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/27673327","citation_count":19,"is_preprint":false},{"pmid":"22431224","id":"PMC_22431224","title":"Insights from comprehensive multiple receptor docking to HDAC8.","date":"2012","source":"Journal of molecular modeling","url":"https://pubmed.ncbi.nlm.nih.gov/22431224","citation_count":19,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":50267,"output_tokens":9320,"usd":0.1453,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":19415,"output_tokens":5148,"usd":0.112888,"stage2_stop_reason":"end_turn"},"total_usd":0.258188,"stage1_batch_id":"msgbatch_01DUPACZi7AAWwuZiUjseQF4","stage2_batch_id":"msgbatch_018yt7chE3f48PEFtDwtj6AE","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2004,\n      \"finding\": \"Crystal structure of human HDAC8 complexed with a hydroxamic acid inhibitor revealed a single alpha/beta domain fold similar to bacterial HDAC-like protein, with two potassium ions in the active site, one interacting with key catalytic residues; CD data demonstrated a direct role of potassium in fold stabilization.\",\n      \"method\": \"X-ray crystallography, CD spectroscopy\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure at atomic resolution with functional validation by CD spectroscopy, foundational structural paper\",\n      \"pmids\": [\"15477595\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"HDAC8 encodes an RPD3 class I HDAC with trichostatin A- and sodium butyrate-inhibitable deacetylase activity on histone H4 peptide substrates and core histones; mutation of two adjacent active-site histidine residues severely decreases enzymatic activity, confirming their catalytic importance.\",\n      \"method\": \"Immunopurification, in vitro HDAC activity assay, active-site mutagenesis\",\n      \"journal\": \"The Biochemical journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro enzymatic reconstitution plus active-site mutagenesis in a single focused study\",\n      \"pmids\": [\"10926844\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"HDAC8 is the vertebrate SMC3 deacetylase responsible for deacetylating SMC3 during cohesin recycling after anaphase; loss-of-function HDAC8 mutations cause Cornelia de Lange syndrome and result in elevated SMC3 acetylation, inefficient dissolution of used cohesin, and altered transcription at cohesin-binding sites.\",\n      \"method\": \"Biochemical deacetylase assay, ChIP-seq, CdLS patient mutation analysis, cell-based cohesin acetylation assays\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — multiple orthogonal methods (biochemical assay, ChIP-seq, patient genetics, cell biology), replicated in multiple CdLS probands\",\n      \"pmids\": [\"22885700\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"HDAC8 associates with smooth muscle alpha-actin (alpha-SMA) but not beta-actin in human smooth muscle cells, and HDAC8 knockdown by siRNA strongly reduces the contractile capacity of smooth muscle cells; HDAC8 is enriched in cytoskeleton-bound protein fractions, unlike HDAC1 and HDAC3.\",\n      \"method\": \"Cell fractionation, co-immunoprecipitation, siRNA knockdown, collagen lattice contraction assay\",\n      \"journal\": \"FASEB journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP, fractionation, and functional KD phenotype in multiple cell types with defined readout\",\n      \"pmids\": [\"15772115\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"HDAC8-selective inhibitor PCI-34051 induces caspase-dependent apoptosis in T-cell lymphoma/leukemia lines via PLCgamma1 activation, intracellular calcium mobilization from the ER, and subsequent cytochrome c release; PLCgamma1-defective cells are resistant and PLC inhibitor U73122 blocks apoptosis.\",\n      \"method\": \"Selective inhibitor treatment, calcium flux assay, caspase activation assay, genetic cell lines defective in signaling, calcium chelator/effector pharmacology\",\n      \"journal\": \"Leukemia\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal pharmacological and genetic approaches defining pathway mechanism in a single rigorous study\",\n      \"pmids\": [\"18256683\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"HDAC8 forms a complex with SMAD3/4 heterotrimer, occupies the SIRT7 promoter, and deacetylates histone H4 to suppress SIRT7 transcription; this establishes a regulatory feedback loop in TGF-β signaling wherein HDAC8 acts as a novel cofactor of SMAD3/4.\",\n      \"method\": \"Co-immunoprecipitation, ChIP assay, HDAC8 inhibitor treatment, gene expression analysis\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP and ChIP in single lab with multiple supporting experiments\",\n      \"pmids\": [\"31970414\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"HDAC8 deacetylates p53 in inv(16)+ AML leukemia stem cells; the CBFβ-SMMHC fusion protein interacts with HDAC8 to aberrantly deacetylate and inactivate p53, promoting leukemic stem cell transformation and maintenance.\",\n      \"method\": \"Co-immunoprecipitation, acetylation assay, HDAC8 selective inhibitor treatment, HDAC8-deficient mouse models, patient-derived xenograft models\",\n      \"journal\": \"Cell stem cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (Co-IP, acetylation assay, genetic KD, pharmacological inhibition, in vivo models) with consistent results\",\n      \"pmids\": [\"26387755\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"HDAC8 catalyzes the removal of long-chain fatty acyl groups (octanoyl, dodecanoyl, myristoyl) from lysine residues with higher catalytic efficiency (lower Km) than for acetyl-lysine; this is the first demonstration of de-fatty-acylation activity by a zinc-dependent HDAC.\",\n      \"method\": \"In vitro enzymatic assay with panel of acyl-lysine peptides, steady-state kinetics (kcat/Km), HDAC8-specific inhibitor treatment of Jurkat cells with global fatty acylation measurement\",\n      \"journal\": \"ACS chemical biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution with kinetic characterization plus cellular validation, single lab but rigorous biochemical methods\",\n      \"pmids\": [\"27459069\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"CdLS-associated missense mutations in HDAC8 (C153F, A188T, I243N, T311M, H334R) cause local structural changes that compromise catalysis and/or thermostability; catalytic activity of several mutants can be partially or fully rescued by the HDAC8 activator N-(phenylcarbamothioyl)benzamide.\",\n      \"method\": \"X-ray crystallography, enzymatic activity assay, thermostability assay, activator rescue experiment\",\n      \"journal\": \"ACS chemical biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — crystal structures of multiple mutants combined with enzymatic and thermostability assays in one rigorous study\",\n      \"pmids\": [\"25075551\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"CdLS-associated HDAC8 mutants (P91L, G117E, H180R, D233G, G304R) cause structural changes compromising catalysis and/or thermostability; the D233G mutation disrupts a hydrogen bond network stabilizing tertiary structure; MD simulations show H180R and G304R mutations cause the arginine side chain to protrude into the substrate binding site; catalytic activity can be partially restored by N-acylthiourea activator.\",\n      \"method\": \"X-ray crystallography, enzymatic activity assay, thermostability assay, molecular dynamics simulation, activator rescue experiment\",\n      \"journal\": \"Biochemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structures plus kinetics plus MD simulation, replicating and extending findings from prior CdLS structural study\",\n      \"pmids\": [\"26463496\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"X-ray crystal structure of HDAC8 complexed with a 'linkerless' hydroxamic acid inhibitor confirms formation of an HDAC8-specific subpocket distinct from other isoforms, explaining isoform-selective inhibition.\",\n      \"method\": \"X-ray crystallography (1.98 Å resolution), molecular docking\",\n      \"journal\": \"Journal of structural biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — crystal structure with molecular docking validation, single lab\",\n      \"pmids\": [\"27374062\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"HDAC8 activity on H4 histone peptides is regulated by both local and distal substrate sequences; a KRHR sequence (residues 16-19) distal to the acetylated lysine 12 is required for activity by interacting at an exosite; methylation of K20 directly affects HDAC8 activity at K12.\",\n      \"method\": \"Mass spectrometry-based HDAC assay with self-assembled monolayer peptide substrates, peptide truncation and mutagenesis\",\n      \"journal\": \"Biochemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro biochemical reconstitution with systematic peptide mutagenesis, revealing exosite mechanism\",\n      \"pmids\": [\"18470998\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"HDAC8 substrate selectivity is determined by both short- and long-range interactions; HDAC8 catalyzes deacetylation of tetrameric histone H3/H4 substrates with 40–300-fold higher catalytic efficiency than corresponding peptide substrates, while efficiency decreases for larger multiprotein complexes.\",\n      \"method\": \"In vitro kinetic assays with peptides, full-length proteins, and protein-nucleic acid complexes\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — systematic in vitro kinetic analysis across multiple substrate formats in single rigorous study\",\n      \"pmids\": [\"29109148\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"The N-acylthiourea derivative TM-2-51 activates HDAC8 by binding at two sites in a positive cooperative manner, as demonstrated by steady-state kinetics and isothermal titration calorimetry; activation preferentially kills neuroblastoma cells with wild-type p53 genotype.\",\n      \"method\": \"Steady-state kinetics, isothermal titration calorimetry, molecular modeling, cell viability assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — multiple biophysical and biochemical methods (ITC, kinetics, modeling) in single study\",\n      \"pmids\": [\"25605725\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"HDAC8 localizes in the nucleus, binds to both CREB and PP1, and expression of recombinant HDAC8 decreases CREB phosphorylation and CREB-mediated gene transcription in response to forskolin, indicating HDAC8 recruits a phosphatase to inactivate CREB.\",\n      \"method\": \"Co-immunoprecipitation, overexpression, luciferase reporter assay\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single Co-IP and overexpression study, single lab\",\n      \"pmids\": [\"19070599\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"HDAC8 directly represses the BMF gene in colon cancer cells; on the BMF promoter, HDAC8 loss is associated with STAT3/Sp3 transcription factor exchange and p300 recruitment; exogenous HDAC8 overexpression blocks BMF induction.\",\n      \"method\": \"ChIP analysis, luciferase assay, RNAi knockdown, overexpression\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP and luciferase assay with RNAi and overexpression, single lab\",\n      \"pmids\": [\"25321483\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"HDAC8 knockdown inhibits YY1 interaction with the p53 promoter (at -102 to -96 position) by increasing acetylation of YY1 (residues 170–200), decreasing YY1 transcriptional activity, and thereby suppressing mutant p53 transcription in TNBC cells.\",\n      \"method\": \"ChIP analysis, luciferase assay, siRNA knockdown, co-immunoprecipitation\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP, Co-IP, and luciferase assay with RNAi in single lab\",\n      \"pmids\": [\"26876786\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"HDAC8 physically interacts with the chromatin modifier EZH2 to concordantly repress Wnt antagonists via histone H4 deacetylation and H3K27 trimethylation in NAFLD-associated hepatocellular carcinoma.\",\n      \"method\": \"Co-immunoprecipitation, ChIP assay, lentiviral HDAC8 knockdown\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP and ChIP in single lab with in vivo model\",\n      \"pmids\": [\"26383163\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"HDAC8 regulates long-term hematopoietic stem cell maintenance by interacting with p53 protein and modulating p53 activity via deacetylation; Hdac8-deficient hematopoietic progenitors show hyperactivation of p53 and increased apoptosis under stress, which is genetically rescued by p53 inactivation.\",\n      \"method\": \"Co-immunoprecipitation, acetylation assay, conditional knockout mouse model, serial replating, in vivo repopulation assay, genetic epistasis (p53 KO rescue)\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, acetylation assay, and genetic epistasis (p53 KO rescue) with in vivo model in single rigorous study\",\n      \"pmids\": [\"29084772\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"FLT3 inhibition induces HDAC8 upregulation via FOXO1- and FOXO3-mediated transactivation in FLT3-ITD+ AML cells; upregulated HDAC8 deacetylates and inactivates p53, promoting leukemia maintenance and resistance to TKI treatment.\",\n      \"method\": \"Gene expression analysis, FOXO siRNA/overexpression, HDAC8 inhibitor treatment, acetylation assay, patient-derived xenograft model\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple approaches (FOXO KD/OE, acetylation assay, PDX model) in single lab\",\n      \"pmids\": [\"32315388\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"HDAC8 deacetylates c-Jun at lysine 273, increasing c-Jun transcriptional activity; HDAC8-mediated BRAF inhibitor resistance in melanoma is driven by receptor tyrosine kinase activation and AP-1/c-Jun signaling; mutation of K273 in c-Jun increases transcriptional activity and confers resistance to BRAF inhibition.\",\n      \"method\": \"Mass spectrometry-based phosphoproteomics, HDAC8 introduction into drug-naïve cells, c-Jun acetylation assay, site-directed mutagenesis of c-Jun K273, in vivo xenograft\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — direct substrate deacetylation shown, mutagenesis of acetylation site, in vivo validation, single lab\",\n      \"pmids\": [\"30987999\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"HDAC8 deacetylates the histone acetyltransferase EP300, causing its enzymatic inactivation; stress-induced HDAC8 activity leads to increased EP300 binding at c-Jun transcriptional sites and decreased binding at MITF sites, driving a neural crest-stem cell state and melanoma brain metastasis.\",\n      \"method\": \"ATAC-Seq, ChIP-Seq, EP300 deacetylation assay, HDAC8 inhibitor treatment, in vivo brain metastasis model\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — biochemical deacetylation assay plus genome-wide chromatin profiling and in vivo validation in single rigorous study\",\n      \"pmids\": [\"38030596\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"HDAC8 inhibition blocks SMC3 deacetylation and delays cell cycle progression and suppresses proliferation in MCF7 cells, but does not alter estrogen-responsive cohesin-dependent gene transcription, demonstrating that HDAC8 inhibition impacts cell cycle rather than cohesin transcriptional function in this context.\",\n      \"method\": \"HDAC8-specific inhibitor (PCI-34051), SMC3 acetylation immunoblot, siRNA knockdown of cohesin subunits, gene expression analysis, cell cycle analysis\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — selective inhibitor and siRNA with multiple functional readouts, single lab\",\n      \"pmids\": [\"27072133\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"HDAC8 associates with alpha-SMA in TGFβ1-treated lung fibroblasts (co-immunoprecipitation); HDAC8 inhibition represses TGFβ1-induced fibroblast contraction, alpha-SMA expression, and profibrotic gene expression; HDAC8 inhibition increases H3K27ac at the PPARγ gene enhancer, identifying H3K27 as an HDAC8 substrate relevant to pulmonary fibrosis.\",\n      \"method\": \"Co-immunoprecipitation, HDAC8 siRNA knockdown, selective inhibitor (NCC170), collagen gel contraction assay, ChIP-qPCR for H3K27ac, bleomycin mouse model\",\n      \"journal\": \"American journal of physiology. Lung cellular and molecular physiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, ChIP, siRNA, pharmacological inhibition, and in vivo model with multiple orthogonal readouts in single lab\",\n      \"pmids\": [\"30358439\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"HDAC8 is a histone H3 lysine 27 (H3K27)-specific deacetylase isozyme; pharmacological HDAC8 inhibition increases global and enhancer H3K27 acetylation, reactivating T cell-trafficking chemokine production by HCC cells and relieving T cell exclusion.\",\n      \"method\": \"Chromatin profiling (H3K27ac ChIP), HDAC8 inhibitor treatment, humanized mouse model, genome-wide enhancer analysis\",\n      \"journal\": \"Science translational medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genome-wide chromatin profiling with selective inhibitor, multiple mouse models, mechanistic link established\",\n      \"pmids\": [\"33827976\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Structure-based approach using Rosetta FlexPepBind identified multiple novel HDAC8 non-histone substrates from proteins with known acetylated sites, including SMC3, ERRα, and ARID1A, demonstrating that HDAC8 regulates a broad acetylome.\",\n      \"method\": \"Structure-based computational substrate identification (Rosetta FlexPepBind), in vitro peptide deacetylation assay validation\",\n      \"journal\": \"Structure\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — computational prediction validated by in vitro assay but limited cellular validation; single study\",\n      \"pmids\": [\"26933971\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Genetically encoded photocrosslinking with p-benzoyl-l-phenylalanine incorporated into HDAC8 active site captured direct HDAC8-substrate complexes from cell lysates; novel HDAC8 substrates were identified and validated for catalytic turnover by mass spectrometry.\",\n      \"method\": \"Active-site photocrosslinking with non-natural amino acid, co-immunoprecipitation, mass spectrometry, in vitro deacetylation validation\",\n      \"journal\": \"Journal of the American Chemical Society\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — chemical crosslinking at active site plus MS identification and catalytic validation in single rigorous study\",\n      \"pmids\": [\"29035536\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"HDAC8 binds and deacetylates PKM2 at K62; K62 deacetylation facilitates PKM2 nuclear translocation and binding to β-catenin, promoting CCND1 gene transcription and cell cycle progression, and also affects PKM2 enzyme activity and glucose metabolism flux.\",\n      \"method\": \"Co-immunoprecipitation, site-directed mutagenesis (K62), nuclear fractionation, chromatin immunoprecipitation, cell cycle analysis\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, mutagenesis of substrate acetylation site, and functional readouts in single lab\",\n      \"pmids\": [\"33279948\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"AMPK phosphorylation of HDAC8 during glucose deprivation induces translocation of HDAC8 from the nucleus to the cytoplasm, disrupting HDAC8-histone 3 binding and leading to increased PGM1 expression in lung cancer cells.\",\n      \"method\": \"Subcellular fractionation, co-immunoprecipitation, AMPK activation/inhibition, HDAC8 phosphorylation assay\",\n      \"journal\": \"Cancer letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — fractionation, Co-IP, and kinase inhibition with functional consequence, single lab\",\n      \"pmids\": [\"32171858\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"HDAC8 mediates epigenetic reprogramming in macrophages by deacetylating H3K27, reducing H3K27ac at genomic regions of BNIP3 and metastatic lymph node 64 genes, promoting resistance to anthrax lethal toxin-induced pyroptosis; HDAC8 is recruited to these loci.\",\n      \"method\": \"ChIP assay, HDAC8 siRNA knockdown, HDAC8-specific inhibitor, HDAC8 overexpression, gene expression analysis\",\n      \"journal\": \"Journal of immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP, siRNA, pharmacological inhibition, and overexpression with consistent results in single lab\",\n      \"pmids\": [\"24973453\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"DEC1 interacts with HDAC8 and recruits HDAC8 to the TAp73 promoter to enhance TAp73 expression; HDAC8 is required for DEC1 to increase TAp73 transcription (demonstrated by failure of DEC1 to activate TAp73 in absence of HDAC8).\",\n      \"method\": \"Co-immunoprecipitation, ChIP assay, siRNA knockdown of HDAC8, luciferase reporter assay\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, ChIP, and epistatic siRNA rescue in single lab\",\n      \"pmids\": [\"24404147\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"HDAC8 localizes at spindle poles in mouse oocytes and is required for microtubule organization and spindle assembly during meiosis; HDAC8 depletion causes spindle defects, chromosome misalignment, impaired kinetochore-microtubule attachments, and increased aneuploidy; HDAC8 is required for correct gamma-tubulin localization to spindle poles.\",\n      \"method\": \"siRNA knockdown, selective inhibitor (PCI-34051), immunofluorescence/confocal imaging, spindle and chromosome analysis, aneuploidy assay\",\n      \"journal\": \"Oncotarget\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA and pharmacological inhibitor with consistent spindle phenotype, multiple readouts, single lab\",\n      \"pmids\": [\"28223544\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"HDAC8 and its phosphorylated form (pHDAC8) localize predominantly in the cytoplasm in HeLa (cancerous) and HEK293T (non-cancerous) cells with additional nucleolar localization in HeLa; HDAC8 binds and deacetylates alpha-tubulin at acetyl-K40, functionally redundant with HDAC6 in cervical cancer cells.\",\n      \"method\": \"Immunofluorescence/confocal imaging, immunoprecipitation followed by MALDI-TOF MS, HPLC-based in vitro deacetylation assay, surface plasmon resonance, siRNA knockdown, HDAC6/8 inhibitors\",\n      \"journal\": \"Cell communication and signaling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct binding and in vitro deacetylation with multiple biochemical methods, single lab\",\n      \"pmids\": [\"29716651\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"HDAC8 interacts with the proteasome receptor ADRM1; this interaction is disrupted by temozolomide in sensitive GBM cells; HDAC8 inhibition decreases MGMT protein levels in GBM cell lines, increasing DNA damage and cell cycle arrest.\",\n      \"method\": \"Co-immunoprecipitation (HDAC8-ADRM1), HDAC8 shRNA, HDAC8-specific inhibitor PCI34051, MGMT protein quantification, cell viability assay\",\n      \"journal\": \"Genes & cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — single Co-IP with pharmacological and genetic confirmation, single lab\",\n      \"pmids\": [\"31798765\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"HDAC8 promotes dissemination of breast cancer cells via the AKT/GSK-3β/Snail axis; HDAC8 interacts with the C-terminal of AKT1 via Co-IP, deacetylates AKT1 at K426, and increases AKT1 phosphorylation, which increases GSK-3β Ser9-phosphorylation, stabilizing Snail and promoting EMT.\",\n      \"method\": \"Co-immunoprecipitation, mass spectrometry, site-directed mutagenesis (AKT1 K426), phosphorylation assay, siRNA knockdown\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, MS, and mutagenesis of substrate residue in single lab\",\n      \"pmids\": [\"32499521\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"HDAC8 degradation by the dual PROTAC YX968 does not induce histone hyperacetylation or broad transcriptomic perturbation, suggesting that HDAC8 deacetylase activity is not the primary driver of its transcriptional effects and that histone hyperacetylation is required for broad transcriptional changes.\",\n      \"method\": \"PROTAC-mediated selective degradation, quantitative proteomics, transcriptomics, histone acetylation assay\",\n      \"journal\": \"Cell chemical biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — selective degrader with quantitative proteomics and transcriptomics; single lab but orthogonal methods\",\n      \"pmids\": [\"37572669\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"HDAC8 and HDAC9 suppress MAP2K3 transcription via H3K9 and H3K27 deacetylation at the MAP2K3 promoter; HDAC8/9 associate with SSRP1 and SUPT16H (components of FACT complex) responsible for MAP2K3 transcriptional elongation, establishing a mechanism for cutaneous innate immune tolerance.\",\n      \"method\": \"siRNA knockdown, ChIP (H3K9ac, H3K27ac), proteomics (HDAC8/9 interactors), transcriptomics, keratinocyte-specific conditional KO mouse\",\n      \"journal\": \"Science immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — ChIP, proteomics-validated interactors, siRNA, and in vivo KO model with consistent mechanistic findings in single rigorous study\",\n      \"pmids\": [\"34021025\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"HDAC8 directly binds to and deacetylates HIF-1α, promoting HIF-1α protein stability and upregulating HIF-1α target genes (HK2, GLUT1) in melanoma.\",\n      \"method\": \"Co-immunoprecipitation, deacetylation assay, HDAC8 inhibitor treatment, siRNA knockdown, protein stability assay\",\n      \"journal\": \"Cancers\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP and deacetylation assay with functional readouts in single lab\",\n      \"pmids\": [\"36831463\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"HDAC8 controls PD-L1 transcription in melanoma cells through a transcription complex of HOXA5 and STAT3; HDAC8 inhibition upregulates PD-L1 via activation of PD-L1 promoter fragments enriched with HOXA5 and STAT3 binding sites; HOXA5 and STAT3 are physically associated and interdependent in activating PD-L1 transcription.\",\n      \"method\": \"siRNA knockdown, luciferase reporter assay, co-immunoprecipitation of HOXA5-STAT3, HDAC8 inhibitor treatment, ChIP\",\n      \"journal\": \"The Journal of investigative dermatology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, luciferase, siRNA, and inhibitor with consistent results in single lab\",\n      \"pmids\": [\"29174371\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"HDAC8 forms a complex with RELA (NF-κB p65) at the PRKN (Parkin) promoter; high-glucose-induced RELA-HDAC8 complex binds the PRKN promoter to suppress PRKN expression and dysregulate mitophagy; sodium butyrate restores PRKN expression by blocking RELA nuclear translocation and directly inhibiting HDAC8.\",\n      \"method\": \"ChIP assay (RELA-HDAC8 at PRKN promoter), Co-immunoprecipitation, HDAC8 overexpression, siRNA knockdown, mitophagy assay\",\n      \"journal\": \"Autophagy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP, Co-IP, and functional mitophagy readout in single lab\",\n      \"pmids\": [\"38409852\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"HDAC8 is a class I, zinc-dependent histone deacetylase with a defined crystal structure that catalyzes deacetylation of acetyl-lysine (and long-chain fatty acyl-lysine) residues on both histone (H3, H4, H3K27) and numerous non-histone substrates including SMC3 (its canonical cohesin-recycling substrate), p53, c-Jun, HIF-1α, EP300, PKM2, AKT1, and alpha-tubulin; substrate recognition involves both active-site and distal exosite interactions; HDAC8 operates in multiple cellular contexts including cohesin recycling (deacetylating SMC3 after anaphase for cohesin reuse), transcriptional regulation (as a cofactor of SMAD3/4 and DEC1 complexes and via H3K27 deacetylation at enhancers), smooth muscle contractility (via association with alpha-SMA), hematopoietic stem cell maintenance and leukemic stem cell transformation (via p53 deacetylation), cell cycle and spindle assembly, and innate immune tolerance (via MAP2K3 suppression); post-translational phosphorylation by AMPK drives HDAC8 nuclear-to-cytoplasmic relocalization, and disease-causing CdLS missense mutations structurally compromise its catalysis and thermostability.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"HDAC8 is a class I, zinc/potassium-dependent histone deacetylase whose single alpha/beta catalytic domain, defined at atomic resolution, removes acyl modifications from lysine residues on both chromatin and non-histone proteins [#0, #1]. Two adjacent active-site histidines are essential for catalysis, and an isoform-specific subpocket near the active site underlies selective inhibition [#1, #10]. Beyond classical acetyl-lysine deacetylation, HDAC8 efficiently removes long-chain fatty acyl groups (octanoyl, dodecanoyl, myristoyl) from lysine with even higher catalytic efficiency than acetyl substrates [#7]. Substrate selectivity is governed by both active-site contacts and distal exosite interactions, such that HDAC8 deacetylates assembled histone H3/H4 tetramers far more efficiently than isolated peptides [#11, #12]. Its canonical physiological substrate is the cohesin subunit SMC3, which HDAC8 deacetylates during cohesin recycling after anaphase; loss-of-function HDAC8 mutations cause Cornelia de Lange syndrome by elevating SMC3 acetylation and impairing cohesin reuse [#2]. CdLS-associated missense mutations structurally compromise catalysis and thermostability, defects that can be partially rescued by small-molecule HDAC8 activators [#8, #9]. On chromatin, HDAC8 acts as an H3K27-specific deacetylase at enhancers and as a cofactor within defined transcriptional complexes (SMAD3/4, DEC1, EZH2, HOXA5/STAT3, RELA), thereby tuning programs in TGF-beta signaling, immune tolerance, fibrosis, and tumor immune evasion [#5, #24, #36, #38]. A broad non-histone acetylome — including p53, c-Jun, EP300, HIF-1alpha, PKM2, AKT1, and alpha-tubulin — links HDAC8 to hematopoietic and leukemic stem cell maintenance via p53 deacetylation, to drug-resistance and metastatic states in melanoma and breast cancer, and to spindle assembly [#6, #18, #20, #21, #31, #34]. AMPK phosphorylation drives HDAC8 nuclear-to-cytoplasmic relocalization under glucose deprivation, coupling its localization to metabolic state [#28].\",\n  \"teleology\": [\n    {\n      \"year\": 2000,\n      \"claim\": \"Established that HDAC8 is a catalytically active class I deacetylase, answering whether this RPD3-family member functions as a bona fide histone deacetylase.\",\n      \"evidence\": \"immunopurification with in vitro HDAC assays on H4 peptides/core histones and active-site histidine mutagenesis\",\n      \"pmids\": [\"10926844\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not identify physiological in vivo substrates\", \"No structural basis for catalysis\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Defined the atomic architecture of the catalytic domain, revealing a bacterial-HDAC-like fold and an unexpected potassium dependence for fold stability.\",\n      \"evidence\": \"X-ray crystallography of HDAC8-hydroxamate complex plus CD spectroscopy\",\n      \"pmids\": [\"15477595\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not explain substrate selectivity\", \"No non-histone substrate context\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Showed that substrate recognition extends beyond the acetyl-lysine to distal sequence elements engaging an exosite, reframing how HDAC8 selects substrates.\",\n      \"evidence\": \"mass spectrometry SAM peptide HDAC assays with truncation/mutagenesis defining the KRHR exosite-binding motif\",\n      \"pmids\": [\"18470998\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Exosite contribution to full-length protein substrates not tested\", \"Structural view of exosite contact lacking\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Linked HDAC8 to the cytoskeleton and a non-chromatin function by associating it with alpha-SMA and showing a role in smooth muscle contractility.\",\n      \"evidence\": \"cell fractionation, reciprocal Co-IP, siRNA knockdown, and collagen lattice contraction assays\",\n      \"pmids\": [\"15772115\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether alpha-SMA or a contractile regulator is the deacetylation target unclear\", \"Direct enzymatic substrate not defined\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Identified SMC3 as the canonical physiological substrate and connected HDAC8 to cohesin recycling and Cornelia de Lange syndrome, the gene's defining disease link.\",\n      \"evidence\": \"biochemical deacetylase assay, ChIP-seq, CdLS patient mutation analysis, and cellular cohesin acetylation assays\",\n      \"pmids\": [\"22885700\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Genome-wide downstream transcriptional consequences only partially mapped\", \"Tissue-specific developmental roles unresolved\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Provided structural mechanism for CdLS pathogenesis by showing patient mutations destabilize the fold and impair catalysis, with pharmacological activator rescue.\",\n      \"evidence\": \"X-ray crystallography of multiple mutants plus enzymatic, thermostability, and activator-rescue assays (extended in 2015 with MD simulations)\",\n      \"pmids\": [\"25075551\", \"26463496\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo rescue by activators not demonstrated\", \"Whether all CdLS phenotypes trace to SMC3 deacetylation loss unclear\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Expanded the catalytic repertoire by demonstrating HDAC8 removes long-chain fatty acyl-lysine modifications more efficiently than acetyl groups, redefining it as a de-fatty-acylase.\",\n      \"evidence\": \"in vitro kinetics on acyl-lysine peptide panels plus cellular global fatty acylation measurement\",\n      \"pmids\": [\"27459069\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological fatty-acylated substrates not identified\", \"Cellular significance of de-fatty-acylation unresolved\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Systematized non-histone substrate discovery through computational and chemical-biology approaches, establishing that HDAC8 regulates a broad acetylome.\",\n      \"evidence\": \"Rosetta FlexPepBind prediction with peptide-assay validation (SMC3, ERRalpha, ARID1A) and active-site photocrosslinking with non-natural amino acids for in-lysate substrate capture\",\n      \"pmids\": [\"26933971\", \"29035536\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Cellular validation of predicted substrates limited\", \"Physiological relevance of each substrate not established\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Quantified how substrate size and architecture shape catalysis, showing HDAC8 favors assembled histone tetramers over peptides while efficiency drops for larger complexes.\",\n      \"evidence\": \"comparative in vitro kinetics across peptides, full-length proteins, and protein-nucleic acid complexes\",\n      \"pmids\": [\"29109148\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism of efficiency loss on large complexes unclear\", \"In-cell substrate-format preferences not tested\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Connected HDAC8 to hematopoietic and leukemic stem cell biology by establishing p53 as a substrate whose deacetylation drives transformation in inv(16)+ AML.\",\n      \"evidence\": \"Co-IP, acetylation assays, selective inhibitor, HDAC8-deficient mice, and patient-derived xenografts (extended to normal HSC maintenance in 2017 and TKI resistance in 2020)\",\n      \"pmids\": [\"26387755\", \"29084772\", \"32315388\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Specificity of p53 lysines deacetylated not fully mapped across contexts\", \"Relative contribution of p53 versus other substrates to leukemia unclear\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Broadened the oncogenic substrate set to c-Jun, EP300, HIF-1alpha, PKM2, and AKT1, linking HDAC8 to drug resistance, metastasis, and metabolic rewiring.\",\n      \"evidence\": \"Co-IP, mass spectrometry, site-directed mutagenesis of substrate lysines, and in vivo tumor/metastasis models across melanoma and breast cancer\",\n      \"pmids\": [\"30987999\", \"38030596\", \"36831463\", \"33279948\", \"32499521\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Several substrate relationships rest on single-lab evidence\", \"Direct versus indirect deacetylation not always discriminated\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Defined HDAC8 as an H3K27-specific deacetylase at enhancers and a transcriptional cofactor across SMAD3/4, DEC1, EZH2, HOXA5/STAT3, and RELA complexes, establishing its chromatin-regulatory roles.\",\n      \"evidence\": \"genome-wide H3K27ac ChIP/ATAC profiling with selective inhibitors and humanized/conditional mouse models, plus Co-IP/ChIP in individual transcription-factor studies\",\n      \"pmids\": [\"33827976\", \"31970414\", \"24404147\", \"26383163\", \"29174371\", \"38409852\", \"34021025\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether catalytic deacetylation versus scaffolding drives each transcriptional effect not uniformly resolved\", \"Recruitment determinants to specific loci incompletely defined\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Showed HDAC8 localization is regulated by metabolic signaling, with AMPK phosphorylation triggering nuclear-to-cytoplasmic relocalization.\",\n      \"evidence\": \"subcellular fractionation, Co-IP, AMPK activation/inhibition, and HDAC8 phosphorylation assays during glucose deprivation\",\n      \"pmids\": [\"32171858\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Phosphosite(s) on HDAC8 not precisely mapped\", \"Generality across cell types not established\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Implicated HDAC8 in meiotic spindle assembly and chromosome segregation through tubulin deacetylation and spindle-pole localization.\",\n      \"evidence\": \"siRNA, selective inhibitor, immunofluorescence, and aneuploidy assays in mouse oocytes, plus in vitro alpha-tubulin K40 deacetylation assays\",\n      \"pmids\": [\"28223544\", \"29716651\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanistic link between tubulin deacetylation and gamma-tubulin recruitment unclear\", \"Redundancy with HDAC6 complicates attribution\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Challenged the assumption that HDAC8's transcriptional effects depend on its deacetylase activity, showing PROTAC degradation did not induce histone hyperacetylation or broad transcriptomic change.\",\n      \"evidence\": \"dual PROTAC-mediated degradation with quantitative proteomics, transcriptomics, and histone acetylation assays\",\n      \"pmids\": [\"37572669\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Reconciliation with catalytic-substrate findings unresolved\", \"Possible non-catalytic scaffolding roles not directly mapped\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unresolved how HDAC8 partitions between its catalytic deacetylase/de-fatty-acylase functions and non-catalytic scaffolding roles, and which substrates dominate in each physiological and disease context.\",\n      \"evidence\": null,\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unified model reconciling catalytic-substrate and PROTAC-degradation data\", \"Tissue-specific in vivo substrate hierarchy undefined\", \"Physiological de-fatty-acylation targets unidentified\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [1, 2, 6, 7, 20, 21, 32, 34, 37]},\n      {\"term_id\": \"GO:0016787\", \"supporting_discovery_ids\": [0, 1, 7, 12]},\n      {\"term_id\": \"GO:0042393\", \"supporting_discovery_ids\": [11, 12, 24]},\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [5, 15, 24, 36]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [2, 14, 24, 28]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [28, 32]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [3]},\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [31]},\n      {\"term_id\": \"GO:0005730\", \"supporting_discovery_ids\": [32]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [2, 22, 27, 31]},\n      {\"term_id\": \"R-HSA-4839726\", \"supporting_discovery_ids\": [21, 24, 29, 36]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [5, 15, 30, 36, 38]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [2, 6, 20]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [24, 36]}\n    ],\n    \"complexes\": [\"cohesin (SMC3 substrate)\", \"SMAD3/4 complex\"],\n    \"partners\": [\"SMC3\", \"TP53\", \"EP300\", \"JUN\", \"HIF1A\", \"AKT1\", \"EZH2\", \"RELA\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":9,"faith_total":9,"faith_pct":100.0}}