{"gene":"CDCA7","run_date":"2026-06-09T22:57:18","timeline":{"discoveries":[{"year":2018,"finding":"CDCA7 and HELLS form a stoichiometric bipartite nucleosome remodeling complex on chromatin; CDCA7 is essential for loading HELLS onto chromatin, and the HELLS-CDCA7 complex together possesses nucleosome remodeling activity that neither protein has alone. ICF-mutant CDCA7 fails to recruit the complex to chromatin.","method":"Xenopus egg extract chromatin proteomics, co-immunoprecipitation, in vitro nucleosome remodeling assay, Aurora B inhibitor treatment, ICF patient mutation analysis","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstitution of nucleosome remodeling activity, complemented by chromatin proteomic profiling and mutant analysis; multiple orthogonal methods in a single rigorous study","pmids":["29339483"],"is_preprint":false},{"year":2024,"finding":"The C-terminal zinc-finger domain (HMZF/zf-4CXXC_R1) of CDCA7 is an evolutionarily conserved hemimethylated CpG sensor. Cryo-EM structural analysis of the CDCA7-nucleosome complex reveals this domain recognizes hemimethylated CpG in the outward-facing DNA major groove within the nucleosome core particle. CDCA7 recruits HELLS to hemimethylated chromatin and facilitates UHRF1-mediated H3 ubiquitylation associated with replication-uncoupled maintenance DNA methylation. ICF disease mutations abolish hemimethylated DNA binding.","method":"Cryo-EM structure determination, in vitro DNA binding assays (hemimethylated vs. unmethylated vs. fully methylated CpG), ICF mutant functional analysis, UHRF1 ubiquitylation assay","journal":"Science advances","confidence":"High","confidence_rationale":"Tier 1 / Strong — cryo-EM structure with functional validation by mutagenesis and in vitro binding assays; replicated by independent preprint (PMID:38187757) with consistent results","pmids":["39178260","38187757"],"is_preprint":false},{"year":2024,"finding":"The CDCA7 C-terminal cysteine-rich domain (CRD) adopts a unique zinc-binding structure that recognizes a CpG dyad in a non-B DNA formed by two sequence motifs, with strand-specific preference for hemimethylated CpG. ICF mutant CDCA7 loses this binding. During S phase, CDCA7 concentrates in constitutive heterochromatin foci in a CRD-dependent manner, and this localization can be outcompeted by exogenous hemi-methylated non-B DNA.","method":"Structural determination of CRD zinc-binding fold, in vitro DNA binding assays with non-B DNA substrates, live-cell imaging of S-phase localization, ICF mutant analysis, competition assay with exogenous DNA","journal":"Science advances","confidence":"High","confidence_rationale":"Tier 1 / Strong — structural characterization combined with in vitro binding and cell-based localization assays with mutagenesis; replicated by preprint (PMID:38168392)","pmids":["39178265","38168392"],"is_preprint":false},{"year":2024,"finding":"The central region of CDCA7 is critical for binding HELLS, activating HELLS ATPase activity, and enabling nucleosome sliding by the HELLS-CDCA7 complex. The N-terminal region tends to inhibit ATPase activity. The C-terminal 4CXXC-type zinc finger domain confers preference for hemimethylated CpG DNA in HELLS-CDCA7 ATPase activity, and replication-dependent pericentromeric heterochromatin foci formation of CDCA7 with HELLS requires an intact zinc finger domain.","method":"In vitro ATPase assay, nucleosome sliding assay, domain-deletion/mutant CDCA7 proteins, live-cell imaging of replication foci in mouse embryonic stem cells","journal":"Nucleic acids research","confidence":"High","confidence_rationale":"Tier 1 / Strong — reconstituted in vitro enzymatic assays with multiple domain mutants plus corroborating cell imaging; multiple orthogonal methods in single study","pmids":["39142653"],"is_preprint":false},{"year":2018,"finding":"CDCA7 co-immunoprecipitates with C-NHEJ proteins Ku80 and Ku70 as well as HELLS; this interaction is sensitive to nuclease treatment and to an ICF3 mutation in CDCA7 that impairs chromatin binding. Loss of CDCA7 or HELLS compromises classical NHEJ activity and delays Ku80 accumulation at DNA damage sites.","method":"Co-immunoprecipitation (nuclease-sensitive), live-cell imaging of Ku80 recruitment to laser-induced DNA damage, NHEJ reporter assay in CDCA7/HELLS-deficient HEK293 cells","journal":"The Journal of clinical investigation","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal co-IP with nuclease sensitivity control, functional NHEJ assay, and live imaging; multiple orthogonal methods in single study","pmids":["30307408"],"is_preprint":false},{"year":2012,"finding":"AKT phosphorylates CDCA7 at threonine 163, promoting its binding to 14-3-3 proteins, dissociation from MYC, and cytoplasmic sequestration. Dephosphorylated CDCA7 associates with MYC in the nucleus and sensitizes cells to apoptosis upon serum withdrawal; CDCA7 knockdown reduces MYC-dependent apoptosis, and CDCA7 co-expression reduces MYC-mediated transformation of fibroblasts.","method":"In vitro kinase assay (AKT phosphorylation of Thr163), co-immunoprecipitation (CDCA7-MYC, CDCA7-14-3-3), phospho-mimetic/phospho-dead mutants, subcellular fractionation, apoptosis assays, Rat1a transformation assay","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — in vitro kinase assay plus multiple co-IP and functional cellular assays; multiple orthogonal methods in single study","pmids":["23166294"],"is_preprint":false},{"year":2001,"finding":"CDCA7 (JPO1) is a direct transcriptional target of c-Myc, encodes a 47-kDa nuclear protein, and complements a transformation-defective Myc Box II mutant in the Rat1a fibroblast transformation assay, establishing a functional genetic link between c-Myc and CDCA7.","method":"Representational difference analysis to identify Myc-responsive gene, inducible c-Myc systems, Rat1a transformation assay, clonogenicity assay in human lymphoblastoid cells","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis (complementation assay) and functional cellular assays; single lab but multiple assays","pmids":["11598121"],"is_preprint":false},{"year":2006,"finding":"CDCA7 is a direct transcriptional target of E2F1 (and E2F2/E2F4 but not E2F5/E2F6), as demonstrated by E2F-responsive element requirement in the CDCA7 promoter and ChIP showing E2F1/E2F2/E2F4 binding. The C-terminal cysteine-rich region of CDCA7 harbors intrinsic transcriptional activator activity in a mammalian one-hybrid assay.","method":"Adenoviral E2F1 overexpression, promoter-reporter assays with deletion constructs, chromatin immunoprecipitation (ChIP) of E2F binding, mammalian one-hybrid transcriptional activity assay","journal":"Biochimica et biophysica acta","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP plus promoter reporter with deletion analysis plus one-hybrid assay; single lab with multiple orthogonal methods","pmids":["16580749"],"is_preprint":false},{"year":2016,"finding":"ZBTB24 directly controls transcription of CDCA7: ZBTB24 is enriched at the CDCA7 promoter by ChIP, loss of functional Zbtb24 in mouse ESCs downregulates Cdca7 as the top affected gene, and ectopic ZBTB24 expression restores Cdca7 levels. CDCA7 protein levels are reduced in patients with ZBTB24 nonsense mutations.","method":"Zbtb24 BTB-domain deletion mouse model, transcriptome analysis (RNAseq), ChIP at CDCA7 promoter, ectopic ZBTB24 rescue experiment, patient lymphoblastoid cell analysis","journal":"Human molecular genetics","confidence":"High","confidence_rationale":"Tier 2 / Strong — ChIP showing direct promoter occupancy, genetic KO with transcriptome-wide readout, rescue experiment, and patient validation; multiple orthogonal methods replicated in vivo and in patients","pmids":["27466202"],"is_preprint":false},{"year":2014,"finding":"CDCA7 is a direct Notch transcriptional target in the aorta-gonad-mesonephros (AGM): RBPj and Notch1 are recruited to the Cdca7 promoter by ChIP. CDCA7 expression is restricted to hematopoietic clusters in the aorta and is required for hematopoietic stem cell emergence; its knockdown in zebrafish reduces HSC formation, and its downregulation in AGM cells promotes hematopoietic differentiation and loss of progenitors.","method":"ChIP-on-chip for RBPj in AGM, ChIP for Notch1, shRNA knockdown in AGM cells, zebrafish loss-of-function experiment (morpholinos), human ESC hematopoietic differentiation with Notch inhibition","journal":"The Journal of experimental medicine","confidence":"High","confidence_rationale":"Tier 2 / Strong — direct ChIP evidence of Notch/RBPj occupancy, orthogonal in vivo loss-of-function in zebrafish, and human ESC differentiation assay; multiple systems and methods","pmids":["25385755"],"is_preprint":false},{"year":2020,"finding":"The CDCA7/HELLS complex is required for the accumulation of proteins on nascent DNA, including the DNMT1/UHRF1 maintenance methylation complex and R-loop resolution factors. Loss of CDCA7/HELLS leads to increased transcription and aberrant DNA:RNA hybrid (R-loop) formation at pericentromeric repeats, and ectopic RNASEH1 expression reduces DNA damage accumulation at these loci in ICF mutant cells.","method":"iPOND (nascent DNA proteomics), R-loop detection (S9.6 antibody), RNASEH1 rescue experiment in CDCA7/HELLS-deficient cells, γH2AX quantification","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — nascent DNA proteomics plus R-loop detection and RNASEH1 genetic rescue; single lab with multiple orthogonal methods","pmids":["33082427"],"is_preprint":false},{"year":2019,"finding":"CDCA7 knockdown in lymphoma cells disrupts actomyosin and tubulin cytoskeleton polarization required for migration, increases filamentous actin formation, and induces myosin activation, impairing migration on fibronectin without affecting adhesion. Inhibitors of actin polymerization, myosin II, or ROCK restore migration capacity of CDCA7-silenced lymphoma cells.","method":"shRNA knockdown, matrigel transwell invasion assay, mouse xenograft invasion model, zebrafish invasion model, phalloidin staining of F-actin, myosin phosphorylation assays, pharmacological rescue with actin/myosin/ROCK inhibitors","journal":"Haematologica","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — KD with defined cytoskeletal phenotype and pharmacological rescue; multiple assays in single lab","pmids":["31221787"],"is_preprint":false},{"year":2021,"finding":"CDCA7 directly binds the CCNA2 (Cyclin A2) gene promoter and upregulates its expression; knockdown of CCNA2 reverses the proliferative phenotype induced by CDCA7 overexpression in esophageal squamous cell carcinoma cells.","method":"ChIP assay (CDCA7 binding to CCNA2 promoter), luciferase reporter assay, CCNA2 rescue/knockdown experiments, cell cycle analysis","journal":"Frontiers in oncology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP and luciferase reporter showing direct promoter binding, with genetic rescue; single lab, multiple orthogonal methods","pmids":["34737951"],"is_preprint":false},{"year":2022,"finding":"CDCA7 transcriptionally regulates Smad4 and Smad7 expression to modulate the TGF-β signaling pathway and promote EMT in esophageal squamous cell carcinoma; dual-luciferase reporter assays and rescue experiments established this regulatory link.","method":"Dual-luciferase reporter assay, ChIP, CDCA7 knockdown/overexpression, rescue assay with Smad4/Smad7, EMT marker Western blot","journal":"Cancer science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — luciferase reporter plus ChIP plus genetic rescue; single lab, multiple methods","pmids":["36056599"],"is_preprint":false},{"year":2019,"finding":"ZBTB24 regulates human T-cell apoptosis via CDCA7: ZBTB24 deficiency reduces CDCA7 expression, and CDCA7 knockdown phenocopies ZBTB24 loss (increased TRAIL/TRAIL-R expression, apoptosis). Overexpression of CDCA7 rescues the increased apoptosis in ZBTB24-depleted Jurkat T cells.","method":"shRNA knockdown of ZBTB24 and CDCA7 in Jurkat and primary T cells, CDCA7 overexpression rescue, TRAIL/TRAIL-R expression analysis, flow cytometry apoptosis assay","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis by rescue experiment, knockdown phenocopy; single lab with multiple cell types","pmids":["31030944"],"is_preprint":false},{"year":2021,"finding":"CDCA7 physically interacts with EZH2 in ovarian cancer cells, as verified by co-immunoprecipitation; CDCA7 knockdown suppresses EZH2 expression and reduces in vitro angiogenesis.","method":"Co-immunoprecipitation (CDCA7–EZH2), shRNA knockdown, HUVEC tube formation assay","journal":"Bioengineered","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single co-IP assay without reciprocal pull-down, single lab, limited mechanistic follow-up","pmids":["34551671"],"is_preprint":false},{"year":2025,"finding":"CDCA7 interacts with STAT3 and affects STAT3-dependent transcriptional regulation of hexokinase 2 (HK2), thereby promoting aerobic glycolysis in pancreatic cancer cells.","method":"Co-immunoprecipitation (CDCA7–STAT3), luciferase reporter for HK2 promoter, CDCA7 knockdown/overexpression with metabolic assays","journal":"Cell death & disease","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single co-IP without mutagenesis or reciprocal pull-down, single lab","pmids":["39905019"],"is_preprint":false},{"year":2024,"finding":"METTL3 enhances CDCA7 mRNA stability via N6-methyladenosine (m6A) modification, as demonstrated by methylated RNA immunoprecipitation (MeRIP) confirming METTL3-dependent m6A on CDCA7 mRNA in colon adenocarcinoma cells.","method":"Methylated RNA immunoprecipitation (MeRIP), METTL3 knockdown with CDCA7 mRNA stability measurement","journal":"Pathology, research and practice","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single MeRIP assay, single lab, limited mechanistic follow-up beyond mRNA stability","pmids":["38959625"],"is_preprint":false},{"year":2024,"finding":"CDCA7 loss in vivo causes large aberrantly hypomethylated domains overlapping the B genomic compartment without affecting H3K9me3 deposition. In brain tissue, CDCA7 acts as a transcriptional repressor of clustered protocadherin isoforms via DNA methylation; hypomethylation at protocadherin loci upon CDCA7 loss is accompanied by gain of H3K4me3 and increased CTCF binding.","method":"Pathogenic Cdca7 missense knock-in mouse, whole-genome bisulfite sequencing across multiple tissues, H3K9me3/H3K4me3 ChIP, CTCF ChIP, transcriptome analysis","journal":"Science advances","confidence":"High","confidence_rationale":"Tier 2 / Strong — in vivo genetic model with whole-genome methylation profiling, multiple histone mark ChIPs, and CTCF binding assays; orthogonal methods in a single rigorous study","pmids":["38335290"],"is_preprint":false},{"year":2025,"finding":"CDCA7 and HELLS constitute a ZBTB24-CDCA7-HELLS axis that suppresses totipotent 2C-like reprogramming in mESCs by maintaining DNA methylation of the Dux cluster. CDCA7 is enriched at the Dux cluster chromatin and recruits HELLS there; disruption leads to Dux hypomethylation, derepression, and upregulation of 2C-specific genes, reversible by site-specific re-methylation at the Dux promoter.","method":"Genetic KO of ZBTB24, CDCA7, and HELLS in mESCs, ChIP (CDCA7 enrichment at Dux cluster), bisulfite sequencing, dCas9-targeted methylation rescue, transcriptome analysis","journal":"Nucleic acids research","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic epistasis across three genes, direct ChIP localization, targeted methylation rescue experiment; multiple orthogonal methods","pmids":["40226918"],"is_preprint":false},{"year":2026,"finding":"CDCA7 exhibits two distinct DNA-binding modes and subcellular localization patterns: diffuse nuclear distribution in interphase (binding CG-rich promoter regions) and pericentromeric heterochromatin localization in late S phase (hemimethylated DNA-dependent). An ICF-causing G294V mutation abolishes both CG-rich DNA binding and heterochromatin localization. CDCA7 recruits LSH/HELLS to both CG-rich promoters in interphase and heterochromatin domains in late S phase, and both proteins can regulate transcription independently of DNA methylation.","method":"Genome-wide CDCA7 ChIP-seq, in vitro DNA binding assays (CG-rich vs. hemimethylated probes), live-cell imaging of cell-cycle-dependent localization, whole-genome DNA methylation analysis, transcriptome analysis, G294V ICF mutant comparison","journal":"Nucleic acids research","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — genome-wide ChIP-seq, in vitro binding assays, cell-cycle-resolved live imaging, methylome profiling, and transcriptome analysis with ICF mutant controls; multiple orthogonal methods in single rigorous study","pmids":["42234582"],"is_preprint":false},{"year":2026,"finding":"CDCA7 physically interacts with HELLS in gastric cancer cells (co-immunoprecipitation), promotes HELLS recruitment to chromatin (ChIP), and knockdown of CDCA7 reduces global 5hmC/5mC levels and histone methylation marks H3K9me3 and H4K20me3; HELLS overexpression partially reverses these effects and the antiproliferative/proapoptotic consequences of CDCA7 knockdown.","method":"Co-immunoprecipitation (CDCA7-HELLS), ChIP (HELLS chromatin recruitment), dot blot for 5mC/5hmC, Western blot for histone marks, HELLS overexpression rescue in CDCA7 KD cells","journal":"Oncology research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP plus ChIP plus epigenetic mark quantification plus genetic rescue; single lab, multiple methods","pmids":["42065057"],"is_preprint":false},{"year":2026,"finding":"CDCA7 transcriptionally activates autophagy-related genes ULK1, ATG2A, and ATG3 in breast cancer drug-tolerant persister (DTP) cells, as identified by ChIP-seq and validated by dual-luciferase assay with site-directed mutagenesis of binding sites. CDCA7 knockdown reduces autophagic flux and restores chemosensitivity.","method":"ChIP-seq (CDCA7 binding at ULK1, ATG2A, ATG3), dual-luciferase assay with mutagenesis, transmission electron microscopy of autolysosomes, mRFP-GFP-LC3 autophagic flux assay, drug resistance (CCK-8), in vivo xenograft","journal":"Frontiers in immunology","confidence":"Medium","confidence_rationale":"Tier 1–2 / Moderate — ChIP-seq with mutagenesis and luciferase validation plus autophagy flux assays; single lab with multiple orthogonal methods","pmids":["41890763"],"is_preprint":false}],"current_model":"CDCA7 is a hemimethylated CpG-sensing adaptor protein whose evolutionarily conserved C-terminal zinc-finger domain (HMZF/zf-4CXXC_R1) recognizes hemimethylated CpG within nucleosomal and linker DNA; it recruits the SNF2-family ATPase HELLS/LSH to form a bipartite nucleosome remodeling complex that repositions nucleosomes (especially H1-containing heterochromatic nucleosomes) to allow DNMT1/UHRF1 access for maintenance DNA methylation at pericentromeric and juxta-centromeric repeats, while also facilitating Ku70/Ku80-dependent non-homologous end joining, suppressing R-loop-associated DNA damage, and—in interphase—binding CG-rich promoters to regulate transcription (including protocadherin isoform choice and Dux silencing); CDCA7 expression is driven by c-Myc and E2F1, and its activity is modulated by AKT-mediated phosphorylation at Thr163 which sequesters it to the cytoplasm via 14-3-3 binding, while its expression is itself transcriptionally controlled by ZBTB24; mutations in CDCA7 that impair chromatin/hemimethylated DNA binding cause ICF syndrome types 3 by abolishing HELLS recruitment and DNA methylation maintenance."},"narrative":{"mechanistic_narrative":"CDCA7 is a hemimethylated CpG-sensing chromatin adaptor that, together with the SNF2-family ATPase HELLS/LSH, forms a bipartite nucleosome remodeling complex required for maintenance of DNA methylation at heterochromatin [PMID:29339483, PMID:39178260, PMID:38187757]. Its evolutionarily conserved C-terminal cysteine-rich/4CXXC-type zinc-finger domain folds into a unique zinc-binding structure that recognizes hemimethylated CpG within nucleosomal DNA, providing the specificity that directs the complex to replicating heterochromatin [PMID:39178260, PMID:38187757, PMID:39178265, PMID:38168392]. The central region of CDCA7 binds and activates the HELLS ATPase to drive nucleosome sliding, while the zinc-finger domain confers the complex's preference for hemimethylated CpG; both are needed for replication-dependent pericentromeric heterochromatin foci [PMID:39142653]. Through this activity CDCA7 promotes loading of the DNMT1/UHRF1 maintenance methylation machinery and facilitates UHRF1-mediated H3 ubiquitylation on nascent DNA, and its loss produces aberrant pericentromeric R-loops and DNA damage [PMID:39178260, PMID:38187757, PMID:33082427]. In vivo, CDCA7 loss causes large hypomethylated domains and acts as a DNA-methylation-dependent transcriptional repressor, silencing clustered protocadherin isoforms and, within a ZBTB24-CDCA7-HELLS axis, the Dux cluster to restrain 2C-like reprogramming [PMID:38335290, PMID:40226918]. CDCA7 also has methylation-independent roles, binding CG-rich promoters in interphase and recruiting HELLS there to regulate transcription, and supporting Ku70/Ku80-dependent classical non-homologous end joining [PMID:42234582, PMID:30307408]. CDCA7 expression is driven by c-Myc, E2F, and Notch and is directly controlled by ZBTB24, while AKT phosphorylation at Thr163 promotes 14-3-3 binding and cytoplasmic sequestration away from MYC [PMID:11598121, PMID:16580749, PMID:25385755, PMID:27466202, PMID:23166294]. ICF syndrome type 3 is caused by CDCA7 mutations that abolish hemimethylated DNA/chromatin binding and HELLS recruitment, disrupting maintenance methylation [PMID:39178260, PMID:38187757, PMID:39178265, PMID:38168392, PMID:42234582].","teleology":[{"year":2001,"claim":"Established the first functional context for CDCA7 by linking it genetically to c-Myc, framing it as a Myc-responsive nuclear effector relevant to transformation.","evidence":"Representational difference analysis, inducible c-Myc systems, and Rat1a transformation complementation assay","pmids":["11598121"],"confidence":"Medium","gaps":["Did not define a molecular activity for the protein","Mechanism of how CDCA7 complements Myc Box II function unresolved"]},{"year":2006,"claim":"Extended transcriptional control of CDCA7 to the cell-cycle machinery, identifying it as a direct E2F target and noting intrinsic transactivation potential in its C-terminal region.","evidence":"Adenoviral E2F1 overexpression, promoter-reporter deletion assays, ChIP, and mammalian one-hybrid assay","pmids":["16580749"],"confidence":"Medium","gaps":["One-hybrid transactivation not tied to endogenous target genes","No chromatin or DNA-binding mechanism defined"]},{"year":2012,"claim":"Showed how CDCA7 activity is post-translationally gated, revealing AKT-Thr163 phosphorylation as a switch controlling its partitioning between nuclear MYC association and cytoplasmic 14-3-3 sequestration.","evidence":"In vitro AKT kinase assay, phospho-mutants, co-IP, subcellular fractionation, and apoptosis/transformation assays","pmids":["23166294"],"confidence":"High","gaps":["Relationship of the MYC-apoptosis role to the later chromatin/methylation function unresolved","Upstream signals controlling Thr163 phosphorylation in vivo not defined"]},{"year":2014,"claim":"Added a developmental dimension, placing CDCA7 downstream of Notch as a direct RBPj/Notch1 target required for hematopoietic stem cell emergence.","evidence":"ChIP-on-chip and ChIP in AGM, shRNA knockdown, zebrafish morpholino loss-of-function, and human ESC differentiation","pmids":["25385755"],"confidence":"High","gaps":["Molecular function of CDCA7 in HSC emergence not mechanistically defined","Whether the chromatin remodeling activity underlies this role unknown"]},{"year":2016,"claim":"Connected CDCA7 to the ICF-syndrome regulatory network by establishing ZBTB24 as a direct transcriptional activator of CDCA7, linking two ICF disease genes in one pathway.","evidence":"Zbtb24 BTB-deletion mouse, RNA-seq, ChIP at the CDCA7 promoter, ectopic rescue, and patient cell analysis","pmids":["27466202"],"confidence":"High","gaps":["Did not establish the downstream biochemical role of CDCA7","How reduced CDCA7 produces ICF phenotypes not yet mechanistic"]},{"year":2018,"claim":"Defined the core biochemical function of CDCA7: it is required to load HELLS onto chromatin, and the CDCA7-HELLS complex possesses nucleosome remodeling activity absent in either protein alone, with ICF mutations abolishing recruitment.","evidence":"Xenopus egg extract chromatin proteomics, co-IP, in vitro nucleosome remodeling assay, and ICF mutant analysis","pmids":["29339483"],"confidence":"High","gaps":["DNA-sequence/modification specificity of recruitment not yet defined","Structural basis of CDCA7-nucleosome recognition unknown"]},{"year":2018,"claim":"Broadened CDCA7 function to genome stability, showing nuclease-sensitive association with Ku70/Ku80 and a requirement for CDCA7/HELLS in classical NHEJ.","evidence":"Nuclease-sensitive co-IP, laser-induced damage Ku80 recruitment imaging, and NHEJ reporter assays in knockout cells","pmids":["30307408"],"confidence":"High","gaps":["Whether NHEJ defect is direct or secondary to chromatin remodeling unresolved","Mechanism by which CDCA7/HELLS accelerates Ku recruitment unclear"]},{"year":2020,"claim":"Linked CDCA7/HELLS remodeling to maintenance methylation and R-loop suppression, showing the complex enables accumulation of DNMT1/UHRF1 and R-loop resolution factors on nascent DNA.","evidence":"iPOND nascent-DNA proteomics, S9.6 R-loop detection, and RNASEH1 rescue of DNA damage in ICF-mutant cells","pmids":["33082427"],"confidence":"Medium","gaps":["Causal order between methylation loss and R-loop accumulation not fully resolved","Single-lab nascent-DNA proteomics"]},{"year":2024,"claim":"Resolved the molecular specificity of CDCA7: cryo-EM and structural studies showed the C-terminal zinc-finger/CRD recognizes hemimethylated CpG in nucleosomal and non-B DNA, explaining how the complex is targeted to replicating heterochromatin and how ICF mutations disrupt it.","evidence":"Cryo-EM of the CDCA7-nucleosome complex, CRD zinc-fold determination, hemimethylated-CpG binding assays, S-phase localization imaging, and UHRF1 ubiquitylation assays","pmids":["39178260","39178265"],"confidence":"High","gaps":["How hemimethylated-DNA sensing is coupled to HELLS ATPase activation not fully integrated","In vivo dynamics of foci formation only partially defined"]},{"year":2024,"claim":"Dissected the domain architecture functionally, assigning HELLS binding and ATPase activation to the central region, autoinhibition to the N-terminus, and hemimethylated-CpG preference to the zinc finger.","evidence":"In vitro ATPase and nucleosome sliding assays with CDCA7 domain mutants plus replication-foci imaging in mESCs","pmids":["39142653"],"confidence":"High","gaps":["Conformational basis of N-terminal autoinhibition not structurally defined","Regulation of the autoinhibition in cells unknown"]},{"year":2024,"claim":"Demonstrated the in vivo consequences of CDCA7 loss as a methylation-dependent transcriptional repressor, with hypomethylation of B-compartment and protocadherin loci independent of H3K9me3.","evidence":"Pathogenic Cdca7 missense knock-in mouse, whole-genome bisulfite sequencing, histone-mark and CTCF ChIP, and transcriptomics","pmids":["38335290"],"confidence":"High","gaps":["Mechanism linking protocadherin hypomethylation to CTCF/H3K4me3 gain not fully causal","Tissue-specific selectivity of affected domains unexplained"]},{"year":2025,"claim":"Defined a ZBTB24-CDCA7-HELLS axis maintaining Dux-cluster methylation to suppress 2C-like totipotent reprogramming, integrating CDCA7's upstream regulation with its chromatin-targeting role.","evidence":"Genetic KO of ZBTB24/CDCA7/HELLS in mESCs, CDCA7 ChIP at Dux, bisulfite sequencing, and dCas9-targeted re-methylation rescue","pmids":["40226918"],"confidence":"High","gaps":["How CDCA7 selects the Dux locus among many repeats unclear","Relative contributions of CDCA7 vs HELLS to silencing not separated"]},{"year":2026,"claim":"Distinguished two cell-cycle-dependent DNA-binding modes, showing interphase CG-rich promoter binding versus late-S-phase hemimethylated heterochromatin localization, with both directing HELLS recruitment and methylation-independent transcriptional effects.","evidence":"Genome-wide CDCA7 ChIP-seq, in vitro CG-rich vs hemimethylated binding assays, cell-cycle-resolved live imaging, methylome and transcriptome analysis, and G294V ICF mutant","pmids":["42234582"],"confidence":"High","gaps":["Switch mechanism between the two binding modes not defined","Functional importance of interphase promoter binding in disease unclear"]},{"year":null,"claim":"It remains unresolved how CDCA7's conserved chromatin-remodeling/methylation function mechanistically gives rise to its many reported context-specific oncogenic transcriptional activities (e.g., CCNA2, TGF-β/Smad, STAT3/HK2, autophagy genes) and partner interactions (EZH2, STAT3).","evidence":"","pmids":[],"confidence":"Low","gaps":["Several cancer-context partnerships rest on single non-reciprocal co-IPs","Whether these transcriptional effects are direct or downstream of methylation changes is untested","Integration of cytoskeletal/migration phenotype with chromatin function unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0003677","term_label":"DNA binding","supporting_discovery_ids":[1,2,20]},{"term_id":"GO:0140657","term_label":"ATP-dependent activity","supporting_discovery_ids":[0,3]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,1,3]},{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[18,20]}],"localization":[{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[5,6,20]},{"term_id":"GO:0000228","term_label":"nuclear chromosome","supporting_discovery_ids":[2,3,20]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[5]}],"pathway":[{"term_id":"R-HSA-4839726","term_label":"Chromatin organization","supporting_discovery_ids":[0,1,3,18,20]},{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[18,19,20]},{"term_id":"R-HSA-73894","term_label":"DNA Repair","supporting_discovery_ids":[4,10]}],"complexes":["CDCA7-HELLS nucleosome remodeling complex"],"partners":["HELLS","UHRF1","DNMT1","XRCC6","XRCC5","MYC","YWHA","EZH2"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9BWT1","full_name":"Cell division cycle-associated protein 7","aliases":["Protein JPO1"],"length_aa":371,"mass_kda":42.6,"function":"Participates in MYC-mediated cell transformation and apoptosis; induces anchorage-independent growth and clonogenicity in lymphoblastoid cells. Insufficient to induce tumorigenicity when overexpressed but contributes to MYC-mediated tumorigenesis (PubMed:11598121, PubMed:15994934, PubMed:23166294). Also functions as a critical cofactor for the chromatin remodeler HELLS, facilitating its recruitment to specific genomic regions to maintain DNA methylation patterns and heterochromatin integrity. Recognizes hemimethylated CpG within nucleosomes where it recruits HELLS to remodel chromatin and facilitate access of de novo DNA methyltransferases to heterochromatic regions, enabling proper establishment of DNA methylation patterns (PubMed:30307408, PubMed:39178260). May play a role as transcriptional regulator (PubMed:16580749)","subcellular_location":"Nucleus; Cytoplasm; Chromosome","url":"https://www.uniprot.org/uniprotkb/Q9BWT1/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/CDCA7","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/CDCA7","total_profiled":1310},"omim":[{"mim_id":"616911","title":"IMMUNODEFICIENCY-CENTROMERIC INSTABILITY-FACIAL ANOMALIES SYNDROME 4; ICF4","url":"https://www.omim.org/entry/616911"},{"mim_id":"616910","title":"IMMUNODEFICIENCY-CENTROMERIC INSTABILITY-FACIAL ANOMALIES SYNDROME 3; ICF3","url":"https://www.omim.org/entry/616910"},{"mim_id":"609937","title":"CELL DIVISION CYCLE-ASSOCIATED PROTEIN 7; CDCA7","url":"https://www.omim.org/entry/609937"},{"mim_id":"609685","title":"CELL DIVISION CYCLE-ASSOCIATED PROTEIN 7-LIKE; CDCA7L","url":"https://www.omim.org/entry/609685"},{"mim_id":"603946","title":"HELICASE, LYMPHOID-SPECIFIC; HELLS","url":"https://www.omim.org/entry/603946"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Nucleoplasm","reliability":"Supported"},{"location":"Cytosol","reliability":"Supported"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"intestine","ntpm":26.5},{"tissue":"lymphoid tissue","ntpm":41.6}],"url":"https://www.proteinatlas.org/search/CDCA7"},"hgnc":{"alias_symbol":["FLJ14736","JPO1"],"prev_symbol":[]},"alphafold":{"accession":"Q9BWT1","domains":[{"cath_id":"-","chopping":"248-370","consensus_level":"medium","plddt":92.4315,"start":248,"end":370}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9BWT1","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9BWT1-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9BWT1-F1-predicted_aligned_error_v6.png","plddt_mean":68.75},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=CDCA7","jax_strain_url":"https://www.jax.org/strain/search?query=CDCA7"},"sequence":{"accession":"Q9BWT1","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9BWT1.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9BWT1/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9BWT1"}},"corpus_meta":[{"pmid":"26216346","id":"PMC_26216346","title":"Mutations in CDCA7 and HELLS cause immunodeficiency-centromeric instability-facial anomalies syndrome.","date":"2015","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/26216346","citation_count":151,"is_preprint":false},{"pmid":"29339483","id":"PMC_29339483","title":"HELLS and CDCA7 comprise a bipartite nucleosome remodeling complex defective in ICF syndrome.","date":"2018","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/29339483","citation_count":109,"is_preprint":false},{"pmid":"30307408","id":"PMC_30307408","title":"CDCA7 and HELLS mutations undermine nonhomologous end joining in centromeric instability syndrome.","date":"2018","source":"The Journal of clinical investigation","url":"https://pubmed.ncbi.nlm.nih.gov/30307408","citation_count":75,"is_preprint":false},{"pmid":"31332696","id":"PMC_31332696","title":"Long noncoding RNA FGD5-AS1 promotes colorectal cancer cell proliferation, migration, and invasion through upregulating CDCA7 via sponging miR-302e.","date":"2019","source":"In vitro cellular & developmental biology. 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ICF-mutant CDCA7 fails to recruit the complex to chromatin.\",\n      \"method\": \"Xenopus egg extract chromatin proteomics, co-immunoprecipitation, in vitro nucleosome remodeling assay, Aurora B inhibitor treatment, ICF patient mutation analysis\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstitution of nucleosome remodeling activity, complemented by chromatin proteomic profiling and mutant analysis; multiple orthogonal methods in a single rigorous study\",\n      \"pmids\": [\"29339483\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"The C-terminal zinc-finger domain (HMZF/zf-4CXXC_R1) of CDCA7 is an evolutionarily conserved hemimethylated CpG sensor. Cryo-EM structural analysis of the CDCA7-nucleosome complex reveals this domain recognizes hemimethylated CpG in the outward-facing DNA major groove within the nucleosome core particle. CDCA7 recruits HELLS to hemimethylated chromatin and facilitates UHRF1-mediated H3 ubiquitylation associated with replication-uncoupled maintenance DNA methylation. ICF disease mutations abolish hemimethylated DNA binding.\",\n      \"method\": \"Cryo-EM structure determination, in vitro DNA binding assays (hemimethylated vs. unmethylated vs. fully methylated CpG), ICF mutant functional analysis, UHRF1 ubiquitylation assay\",\n      \"journal\": \"Science advances\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — cryo-EM structure with functional validation by mutagenesis and in vitro binding assays; replicated by independent preprint (PMID:38187757) with consistent results\",\n      \"pmids\": [\"39178260\", \"38187757\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"The CDCA7 C-terminal cysteine-rich domain (CRD) adopts a unique zinc-binding structure that recognizes a CpG dyad in a non-B DNA formed by two sequence motifs, with strand-specific preference for hemimethylated CpG. ICF mutant CDCA7 loses this binding. During S phase, CDCA7 concentrates in constitutive heterochromatin foci in a CRD-dependent manner, and this localization can be outcompeted by exogenous hemi-methylated non-B DNA.\",\n      \"method\": \"Structural determination of CRD zinc-binding fold, in vitro DNA binding assays with non-B DNA substrates, live-cell imaging of S-phase localization, ICF mutant analysis, competition assay with exogenous DNA\",\n      \"journal\": \"Science advances\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — structural characterization combined with in vitro binding and cell-based localization assays with mutagenesis; replicated by preprint (PMID:38168392)\",\n      \"pmids\": [\"39178265\", \"38168392\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"The central region of CDCA7 is critical for binding HELLS, activating HELLS ATPase activity, and enabling nucleosome sliding by the HELLS-CDCA7 complex. The N-terminal region tends to inhibit ATPase activity. The C-terminal 4CXXC-type zinc finger domain confers preference for hemimethylated CpG DNA in HELLS-CDCA7 ATPase activity, and replication-dependent pericentromeric heterochromatin foci formation of CDCA7 with HELLS requires an intact zinc finger domain.\",\n      \"method\": \"In vitro ATPase assay, nucleosome sliding assay, domain-deletion/mutant CDCA7 proteins, live-cell imaging of replication foci in mouse embryonic stem cells\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — reconstituted in vitro enzymatic assays with multiple domain mutants plus corroborating cell imaging; multiple orthogonal methods in single study\",\n      \"pmids\": [\"39142653\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"CDCA7 co-immunoprecipitates with C-NHEJ proteins Ku80 and Ku70 as well as HELLS; this interaction is sensitive to nuclease treatment and to an ICF3 mutation in CDCA7 that impairs chromatin binding. Loss of CDCA7 or HELLS compromises classical NHEJ activity and delays Ku80 accumulation at DNA damage sites.\",\n      \"method\": \"Co-immunoprecipitation (nuclease-sensitive), live-cell imaging of Ku80 recruitment to laser-induced DNA damage, NHEJ reporter assay in CDCA7/HELLS-deficient HEK293 cells\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal co-IP with nuclease sensitivity control, functional NHEJ assay, and live imaging; multiple orthogonal methods in single study\",\n      \"pmids\": [\"30307408\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"AKT phosphorylates CDCA7 at threonine 163, promoting its binding to 14-3-3 proteins, dissociation from MYC, and cytoplasmic sequestration. Dephosphorylated CDCA7 associates with MYC in the nucleus and sensitizes cells to apoptosis upon serum withdrawal; CDCA7 knockdown reduces MYC-dependent apoptosis, and CDCA7 co-expression reduces MYC-mediated transformation of fibroblasts.\",\n      \"method\": \"In vitro kinase assay (AKT phosphorylation of Thr163), co-immunoprecipitation (CDCA7-MYC, CDCA7-14-3-3), phospho-mimetic/phospho-dead mutants, subcellular fractionation, apoptosis assays, Rat1a transformation assay\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — in vitro kinase assay plus multiple co-IP and functional cellular assays; multiple orthogonal methods in single study\",\n      \"pmids\": [\"23166294\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"CDCA7 (JPO1) is a direct transcriptional target of c-Myc, encodes a 47-kDa nuclear protein, and complements a transformation-defective Myc Box II mutant in the Rat1a fibroblast transformation assay, establishing a functional genetic link between c-Myc and CDCA7.\",\n      \"method\": \"Representational difference analysis to identify Myc-responsive gene, inducible c-Myc systems, Rat1a transformation assay, clonogenicity assay in human lymphoblastoid cells\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis (complementation assay) and functional cellular assays; single lab but multiple assays\",\n      \"pmids\": [\"11598121\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"CDCA7 is a direct transcriptional target of E2F1 (and E2F2/E2F4 but not E2F5/E2F6), as demonstrated by E2F-responsive element requirement in the CDCA7 promoter and ChIP showing E2F1/E2F2/E2F4 binding. The C-terminal cysteine-rich region of CDCA7 harbors intrinsic transcriptional activator activity in a mammalian one-hybrid assay.\",\n      \"method\": \"Adenoviral E2F1 overexpression, promoter-reporter assays with deletion constructs, chromatin immunoprecipitation (ChIP) of E2F binding, mammalian one-hybrid transcriptional activity assay\",\n      \"journal\": \"Biochimica et biophysica acta\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP plus promoter reporter with deletion analysis plus one-hybrid assay; single lab with multiple orthogonal methods\",\n      \"pmids\": [\"16580749\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"ZBTB24 directly controls transcription of CDCA7: ZBTB24 is enriched at the CDCA7 promoter by ChIP, loss of functional Zbtb24 in mouse ESCs downregulates Cdca7 as the top affected gene, and ectopic ZBTB24 expression restores Cdca7 levels. CDCA7 protein levels are reduced in patients with ZBTB24 nonsense mutations.\",\n      \"method\": \"Zbtb24 BTB-domain deletion mouse model, transcriptome analysis (RNAseq), ChIP at CDCA7 promoter, ectopic ZBTB24 rescue experiment, patient lymphoblastoid cell analysis\",\n      \"journal\": \"Human molecular genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — ChIP showing direct promoter occupancy, genetic KO with transcriptome-wide readout, rescue experiment, and patient validation; multiple orthogonal methods replicated in vivo and in patients\",\n      \"pmids\": [\"27466202\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"CDCA7 is a direct Notch transcriptional target in the aorta-gonad-mesonephros (AGM): RBPj and Notch1 are recruited to the Cdca7 promoter by ChIP. CDCA7 expression is restricted to hematopoietic clusters in the aorta and is required for hematopoietic stem cell emergence; its knockdown in zebrafish reduces HSC formation, and its downregulation in AGM cells promotes hematopoietic differentiation and loss of progenitors.\",\n      \"method\": \"ChIP-on-chip for RBPj in AGM, ChIP for Notch1, shRNA knockdown in AGM cells, zebrafish loss-of-function experiment (morpholinos), human ESC hematopoietic differentiation with Notch inhibition\",\n      \"journal\": \"The Journal of experimental medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — direct ChIP evidence of Notch/RBPj occupancy, orthogonal in vivo loss-of-function in zebrafish, and human ESC differentiation assay; multiple systems and methods\",\n      \"pmids\": [\"25385755\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"The CDCA7/HELLS complex is required for the accumulation of proteins on nascent DNA, including the DNMT1/UHRF1 maintenance methylation complex and R-loop resolution factors. Loss of CDCA7/HELLS leads to increased transcription and aberrant DNA:RNA hybrid (R-loop) formation at pericentromeric repeats, and ectopic RNASEH1 expression reduces DNA damage accumulation at these loci in ICF mutant cells.\",\n      \"method\": \"iPOND (nascent DNA proteomics), R-loop detection (S9.6 antibody), RNASEH1 rescue experiment in CDCA7/HELLS-deficient cells, γH2AX quantification\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — nascent DNA proteomics plus R-loop detection and RNASEH1 genetic rescue; single lab with multiple orthogonal methods\",\n      \"pmids\": [\"33082427\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"CDCA7 knockdown in lymphoma cells disrupts actomyosin and tubulin cytoskeleton polarization required for migration, increases filamentous actin formation, and induces myosin activation, impairing migration on fibronectin without affecting adhesion. Inhibitors of actin polymerization, myosin II, or ROCK restore migration capacity of CDCA7-silenced lymphoma cells.\",\n      \"method\": \"shRNA knockdown, matrigel transwell invasion assay, mouse xenograft invasion model, zebrafish invasion model, phalloidin staining of F-actin, myosin phosphorylation assays, pharmacological rescue with actin/myosin/ROCK inhibitors\",\n      \"journal\": \"Haematologica\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KD with defined cytoskeletal phenotype and pharmacological rescue; multiple assays in single lab\",\n      \"pmids\": [\"31221787\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"CDCA7 directly binds the CCNA2 (Cyclin A2) gene promoter and upregulates its expression; knockdown of CCNA2 reverses the proliferative phenotype induced by CDCA7 overexpression in esophageal squamous cell carcinoma cells.\",\n      \"method\": \"ChIP assay (CDCA7 binding to CCNA2 promoter), luciferase reporter assay, CCNA2 rescue/knockdown experiments, cell cycle analysis\",\n      \"journal\": \"Frontiers in oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP and luciferase reporter showing direct promoter binding, with genetic rescue; single lab, multiple orthogonal methods\",\n      \"pmids\": [\"34737951\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"CDCA7 transcriptionally regulates Smad4 and Smad7 expression to modulate the TGF-β signaling pathway and promote EMT in esophageal squamous cell carcinoma; dual-luciferase reporter assays and rescue experiments established this regulatory link.\",\n      \"method\": \"Dual-luciferase reporter assay, ChIP, CDCA7 knockdown/overexpression, rescue assay with Smad4/Smad7, EMT marker Western blot\",\n      \"journal\": \"Cancer science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — luciferase reporter plus ChIP plus genetic rescue; single lab, multiple methods\",\n      \"pmids\": [\"36056599\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"ZBTB24 regulates human T-cell apoptosis via CDCA7: ZBTB24 deficiency reduces CDCA7 expression, and CDCA7 knockdown phenocopies ZBTB24 loss (increased TRAIL/TRAIL-R expression, apoptosis). Overexpression of CDCA7 rescues the increased apoptosis in ZBTB24-depleted Jurkat T cells.\",\n      \"method\": \"shRNA knockdown of ZBTB24 and CDCA7 in Jurkat and primary T cells, CDCA7 overexpression rescue, TRAIL/TRAIL-R expression analysis, flow cytometry apoptosis assay\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis by rescue experiment, knockdown phenocopy; single lab with multiple cell types\",\n      \"pmids\": [\"31030944\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"CDCA7 physically interacts with EZH2 in ovarian cancer cells, as verified by co-immunoprecipitation; CDCA7 knockdown suppresses EZH2 expression and reduces in vitro angiogenesis.\",\n      \"method\": \"Co-immunoprecipitation (CDCA7–EZH2), shRNA knockdown, HUVEC tube formation assay\",\n      \"journal\": \"Bioengineered\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single co-IP assay without reciprocal pull-down, single lab, limited mechanistic follow-up\",\n      \"pmids\": [\"34551671\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"CDCA7 interacts with STAT3 and affects STAT3-dependent transcriptional regulation of hexokinase 2 (HK2), thereby promoting aerobic glycolysis in pancreatic cancer cells.\",\n      \"method\": \"Co-immunoprecipitation (CDCA7–STAT3), luciferase reporter for HK2 promoter, CDCA7 knockdown/overexpression with metabolic assays\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single co-IP without mutagenesis or reciprocal pull-down, single lab\",\n      \"pmids\": [\"39905019\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"METTL3 enhances CDCA7 mRNA stability via N6-methyladenosine (m6A) modification, as demonstrated by methylated RNA immunoprecipitation (MeRIP) confirming METTL3-dependent m6A on CDCA7 mRNA in colon adenocarcinoma cells.\",\n      \"method\": \"Methylated RNA immunoprecipitation (MeRIP), METTL3 knockdown with CDCA7 mRNA stability measurement\",\n      \"journal\": \"Pathology, research and practice\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single MeRIP assay, single lab, limited mechanistic follow-up beyond mRNA stability\",\n      \"pmids\": [\"38959625\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CDCA7 loss in vivo causes large aberrantly hypomethylated domains overlapping the B genomic compartment without affecting H3K9me3 deposition. In brain tissue, CDCA7 acts as a transcriptional repressor of clustered protocadherin isoforms via DNA methylation; hypomethylation at protocadherin loci upon CDCA7 loss is accompanied by gain of H3K4me3 and increased CTCF binding.\",\n      \"method\": \"Pathogenic Cdca7 missense knock-in mouse, whole-genome bisulfite sequencing across multiple tissues, H3K9me3/H3K4me3 ChIP, CTCF ChIP, transcriptome analysis\",\n      \"journal\": \"Science advances\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — in vivo genetic model with whole-genome methylation profiling, multiple histone mark ChIPs, and CTCF binding assays; orthogonal methods in a single rigorous study\",\n      \"pmids\": [\"38335290\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"CDCA7 and HELLS constitute a ZBTB24-CDCA7-HELLS axis that suppresses totipotent 2C-like reprogramming in mESCs by maintaining DNA methylation of the Dux cluster. CDCA7 is enriched at the Dux cluster chromatin and recruits HELLS there; disruption leads to Dux hypomethylation, derepression, and upregulation of 2C-specific genes, reversible by site-specific re-methylation at the Dux promoter.\",\n      \"method\": \"Genetic KO of ZBTB24, CDCA7, and HELLS in mESCs, ChIP (CDCA7 enrichment at Dux cluster), bisulfite sequencing, dCas9-targeted methylation rescue, transcriptome analysis\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic epistasis across three genes, direct ChIP localization, targeted methylation rescue experiment; multiple orthogonal methods\",\n      \"pmids\": [\"40226918\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"CDCA7 exhibits two distinct DNA-binding modes and subcellular localization patterns: diffuse nuclear distribution in interphase (binding CG-rich promoter regions) and pericentromeric heterochromatin localization in late S phase (hemimethylated DNA-dependent). An ICF-causing G294V mutation abolishes both CG-rich DNA binding and heterochromatin localization. CDCA7 recruits LSH/HELLS to both CG-rich promoters in interphase and heterochromatin domains in late S phase, and both proteins can regulate transcription independently of DNA methylation.\",\n      \"method\": \"Genome-wide CDCA7 ChIP-seq, in vitro DNA binding assays (CG-rich vs. hemimethylated probes), live-cell imaging of cell-cycle-dependent localization, whole-genome DNA methylation analysis, transcriptome analysis, G294V ICF mutant comparison\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — genome-wide ChIP-seq, in vitro binding assays, cell-cycle-resolved live imaging, methylome profiling, and transcriptome analysis with ICF mutant controls; multiple orthogonal methods in single rigorous study\",\n      \"pmids\": [\"42234582\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"CDCA7 physically interacts with HELLS in gastric cancer cells (co-immunoprecipitation), promotes HELLS recruitment to chromatin (ChIP), and knockdown of CDCA7 reduces global 5hmC/5mC levels and histone methylation marks H3K9me3 and H4K20me3; HELLS overexpression partially reverses these effects and the antiproliferative/proapoptotic consequences of CDCA7 knockdown.\",\n      \"method\": \"Co-immunoprecipitation (CDCA7-HELLS), ChIP (HELLS chromatin recruitment), dot blot for 5mC/5hmC, Western blot for histone marks, HELLS overexpression rescue in CDCA7 KD cells\",\n      \"journal\": \"Oncology research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP plus ChIP plus epigenetic mark quantification plus genetic rescue; single lab, multiple methods\",\n      \"pmids\": [\"42065057\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"CDCA7 transcriptionally activates autophagy-related genes ULK1, ATG2A, and ATG3 in breast cancer drug-tolerant persister (DTP) cells, as identified by ChIP-seq and validated by dual-luciferase assay with site-directed mutagenesis of binding sites. CDCA7 knockdown reduces autophagic flux and restores chemosensitivity.\",\n      \"method\": \"ChIP-seq (CDCA7 binding at ULK1, ATG2A, ATG3), dual-luciferase assay with mutagenesis, transmission electron microscopy of autolysosomes, mRFP-GFP-LC3 autophagic flux assay, drug resistance (CCK-8), in vivo xenograft\",\n      \"journal\": \"Frontiers in immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — ChIP-seq with mutagenesis and luciferase validation plus autophagy flux assays; single lab with multiple orthogonal methods\",\n      \"pmids\": [\"41890763\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"CDCA7 is a hemimethylated CpG-sensing adaptor protein whose evolutionarily conserved C-terminal zinc-finger domain (HMZF/zf-4CXXC_R1) recognizes hemimethylated CpG within nucleosomal and linker DNA; it recruits the SNF2-family ATPase HELLS/LSH to form a bipartite nucleosome remodeling complex that repositions nucleosomes (especially H1-containing heterochromatic nucleosomes) to allow DNMT1/UHRF1 access for maintenance DNA methylation at pericentromeric and juxta-centromeric repeats, while also facilitating Ku70/Ku80-dependent non-homologous end joining, suppressing R-loop-associated DNA damage, and—in interphase—binding CG-rich promoters to regulate transcription (including protocadherin isoform choice and Dux silencing); CDCA7 expression is driven by c-Myc and E2F1, and its activity is modulated by AKT-mediated phosphorylation at Thr163 which sequesters it to the cytoplasm via 14-3-3 binding, while its expression is itself transcriptionally controlled by ZBTB24; mutations in CDCA7 that impair chromatin/hemimethylated DNA binding cause ICF syndrome types 3 by abolishing HELLS recruitment and DNA methylation maintenance.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"CDCA7 is a hemimethylated CpG-sensing chromatin adaptor that, together with the SNF2-family ATPase HELLS/LSH, forms a bipartite nucleosome remodeling complex required for maintenance of DNA methylation at heterochromatin [#0, #1]. Its evolutionarily conserved C-terminal cysteine-rich/4CXXC-type zinc-finger domain folds into a unique zinc-binding structure that recognizes hemimethylated CpG within nucleosomal DNA, providing the specificity that directs the complex to replicating heterochromatin [#1, #2]. The central region of CDCA7 binds and activates the HELLS ATPase to drive nucleosome sliding, while the zinc-finger domain confers the complex's preference for hemimethylated CpG; both are needed for replication-dependent pericentromeric heterochromatin foci [#3]. Through this activity CDCA7 promotes loading of the DNMT1/UHRF1 maintenance methylation machinery and facilitates UHRF1-mediated H3 ubiquitylation on nascent DNA, and its loss produces aberrant pericentromeric R-loops and DNA damage [#1, #10]. In vivo, CDCA7 loss causes large hypomethylated domains and acts as a DNA-methylation-dependent transcriptional repressor, silencing clustered protocadherin isoforms and, within a ZBTB24-CDCA7-HELLS axis, the Dux cluster to restrain 2C-like reprogramming [#18, #19]. CDCA7 also has methylation-independent roles, binding CG-rich promoters in interphase and recruiting HELLS there to regulate transcription, and supporting Ku70/Ku80-dependent classical non-homologous end joining [#20, #4]. CDCA7 expression is driven by c-Myc, E2F, and Notch and is directly controlled by ZBTB24, while AKT phosphorylation at Thr163 promotes 14-3-3 binding and cytoplasmic sequestration away from MYC [#6, #7, #9, #8, #5]. ICF syndrome type 3 is caused by CDCA7 mutations that abolish hemimethylated DNA/chromatin binding and HELLS recruitment, disrupting maintenance methylation [#1, #2, #20].\",\n  \"teleology\": [\n    {\n      \"year\": 2001,\n      \"claim\": \"Established the first functional context for CDCA7 by linking it genetically to c-Myc, framing it as a Myc-responsive nuclear effector relevant to transformation.\",\n      \"evidence\": \"Representational difference analysis, inducible c-Myc systems, and Rat1a transformation complementation assay\",\n      \"pmids\": [\"11598121\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Did not define a molecular activity for the protein\", \"Mechanism of how CDCA7 complements Myc Box II function unresolved\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Extended transcriptional control of CDCA7 to the cell-cycle machinery, identifying it as a direct E2F target and noting intrinsic transactivation potential in its C-terminal region.\",\n      \"evidence\": \"Adenoviral E2F1 overexpression, promoter-reporter deletion assays, ChIP, and mammalian one-hybrid assay\",\n      \"pmids\": [\"16580749\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"One-hybrid transactivation not tied to endogenous target genes\", \"No chromatin or DNA-binding mechanism defined\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Showed how CDCA7 activity is post-translationally gated, revealing AKT-Thr163 phosphorylation as a switch controlling its partitioning between nuclear MYC association and cytoplasmic 14-3-3 sequestration.\",\n      \"evidence\": \"In vitro AKT kinase assay, phospho-mutants, co-IP, subcellular fractionation, and apoptosis/transformation assays\",\n      \"pmids\": [\"23166294\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Relationship of the MYC-apoptosis role to the later chromatin/methylation function unresolved\", \"Upstream signals controlling Thr163 phosphorylation in vivo not defined\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Added a developmental dimension, placing CDCA7 downstream of Notch as a direct RBPj/Notch1 target required for hematopoietic stem cell emergence.\",\n      \"evidence\": \"ChIP-on-chip and ChIP in AGM, shRNA knockdown, zebrafish morpholino loss-of-function, and human ESC differentiation\",\n      \"pmids\": [\"25385755\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular function of CDCA7 in HSC emergence not mechanistically defined\", \"Whether the chromatin remodeling activity underlies this role unknown\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Connected CDCA7 to the ICF-syndrome regulatory network by establishing ZBTB24 as a direct transcriptional activator of CDCA7, linking two ICF disease genes in one pathway.\",\n      \"evidence\": \"Zbtb24 BTB-deletion mouse, RNA-seq, ChIP at the CDCA7 promoter, ectopic rescue, and patient cell analysis\",\n      \"pmids\": [\"27466202\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not establish the downstream biochemical role of CDCA7\", \"How reduced CDCA7 produces ICF phenotypes not yet mechanistic\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Defined the core biochemical function of CDCA7: it is required to load HELLS onto chromatin, and the CDCA7-HELLS complex possesses nucleosome remodeling activity absent in either protein alone, with ICF mutations abolishing recruitment.\",\n      \"evidence\": \"Xenopus egg extract chromatin proteomics, co-IP, in vitro nucleosome remodeling assay, and ICF mutant analysis\",\n      \"pmids\": [\"29339483\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"DNA-sequence/modification specificity of recruitment not yet defined\", \"Structural basis of CDCA7-nucleosome recognition unknown\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Broadened CDCA7 function to genome stability, showing nuclease-sensitive association with Ku70/Ku80 and a requirement for CDCA7/HELLS in classical NHEJ.\",\n      \"evidence\": \"Nuclease-sensitive co-IP, laser-induced damage Ku80 recruitment imaging, and NHEJ reporter assays in knockout cells\",\n      \"pmids\": [\"30307408\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether NHEJ defect is direct or secondary to chromatin remodeling unresolved\", \"Mechanism by which CDCA7/HELLS accelerates Ku recruitment unclear\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Linked CDCA7/HELLS remodeling to maintenance methylation and R-loop suppression, showing the complex enables accumulation of DNMT1/UHRF1 and R-loop resolution factors on nascent DNA.\",\n      \"evidence\": \"iPOND nascent-DNA proteomics, S9.6 R-loop detection, and RNASEH1 rescue of DNA damage in ICF-mutant cells\",\n      \"pmids\": [\"33082427\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Causal order between methylation loss and R-loop accumulation not fully resolved\", \"Single-lab nascent-DNA proteomics\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Resolved the molecular specificity of CDCA7: cryo-EM and structural studies showed the C-terminal zinc-finger/CRD recognizes hemimethylated CpG in nucleosomal and non-B DNA, explaining how the complex is targeted to replicating heterochromatin and how ICF mutations disrupt it.\",\n      \"evidence\": \"Cryo-EM of the CDCA7-nucleosome complex, CRD zinc-fold determination, hemimethylated-CpG binding assays, S-phase localization imaging, and UHRF1 ubiquitylation assays\",\n      \"pmids\": [\"39178260\", \"39178265\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How hemimethylated-DNA sensing is coupled to HELLS ATPase activation not fully integrated\", \"In vivo dynamics of foci formation only partially defined\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Dissected the domain architecture functionally, assigning HELLS binding and ATPase activation to the central region, autoinhibition to the N-terminus, and hemimethylated-CpG preference to the zinc finger.\",\n      \"evidence\": \"In vitro ATPase and nucleosome sliding assays with CDCA7 domain mutants plus replication-foci imaging in mESCs\",\n      \"pmids\": [\"39142653\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Conformational basis of N-terminal autoinhibition not structurally defined\", \"Regulation of the autoinhibition in cells unknown\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Demonstrated the in vivo consequences of CDCA7 loss as a methylation-dependent transcriptional repressor, with hypomethylation of B-compartment and protocadherin loci independent of H3K9me3.\",\n      \"evidence\": \"Pathogenic Cdca7 missense knock-in mouse, whole-genome bisulfite sequencing, histone-mark and CTCF ChIP, and transcriptomics\",\n      \"pmids\": [\"38335290\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism linking protocadherin hypomethylation to CTCF/H3K4me3 gain not fully causal\", \"Tissue-specific selectivity of affected domains unexplained\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Defined a ZBTB24-CDCA7-HELLS axis maintaining Dux-cluster methylation to suppress 2C-like totipotent reprogramming, integrating CDCA7's upstream regulation with its chromatin-targeting role.\",\n      \"evidence\": \"Genetic KO of ZBTB24/CDCA7/HELLS in mESCs, CDCA7 ChIP at Dux, bisulfite sequencing, and dCas9-targeted re-methylation rescue\",\n      \"pmids\": [\"40226918\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How CDCA7 selects the Dux locus among many repeats unclear\", \"Relative contributions of CDCA7 vs HELLS to silencing not separated\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Distinguished two cell-cycle-dependent DNA-binding modes, showing interphase CG-rich promoter binding versus late-S-phase hemimethylated heterochromatin localization, with both directing HELLS recruitment and methylation-independent transcriptional effects.\",\n      \"evidence\": \"Genome-wide CDCA7 ChIP-seq, in vitro CG-rich vs hemimethylated binding assays, cell-cycle-resolved live imaging, methylome and transcriptome analysis, and G294V ICF mutant\",\n      \"pmids\": [\"42234582\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Switch mechanism between the two binding modes not defined\", \"Functional importance of interphase promoter binding in disease unclear\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unresolved how CDCA7's conserved chromatin-remodeling/methylation function mechanistically gives rise to its many reported context-specific oncogenic transcriptional activities (e.g., CCNA2, TGF-\\u03b2/Smad, STAT3/HK2, autophagy genes) and partner interactions (EZH2, STAT3).\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Several cancer-context partnerships rest on single non-reciprocal co-IPs\", \"Whether these transcriptional effects are direct or downstream of methylation changes is untested\", \"Integration of cytoskeletal/migration phenotype with chromatin function unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0003677\", \"supporting_discovery_ids\": [1, 2, 20]},\n      {\"term_id\": \"GO:0140657\", \"supporting_discovery_ids\": [0, 3]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 1, 3]},\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [18, 20]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [5, 6, 20]},\n      {\"term_id\": \"GO:0000228\", \"supporting_discovery_ids\": [2, 3, 20]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [5]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-4839726\", \"supporting_discovery_ids\": [0, 1, 3, 18, 20]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [18, 19, 20]},\n      {\"term_id\": \"R-HSA-73894\", \"supporting_discovery_ids\": [4, 10]}\n    ],\n    \"complexes\": [\"CDCA7-HELLS nucleosome remodeling complex\"],\n    \"partners\": [\"HELLS\", \"UHRF1\", \"DNMT1\", \"XRCC6\", \"XRCC5\", \"MYC\", \"YWHA\", \"EZH2\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":8,"faith_total":8,"faith_pct":100.0}}