{"gene":"EVC2","run_date":"2026-06-09T23:54:43","timeline":{"discoveries":[{"year":2012,"finding":"Hedgehog agonists promote physical association between Smoothened (Smo) and EVC2; this Smo-EVC2 complex forms specifically within the EvC zone, a distinct sub-compartment at the base of primary cilia. EVC2 mutants that localize to cilia but are displaced from the EvC zone act as dominant inhibitors of Hh signaling, and loss of EVC2 function blocks Hh signaling between Smo and the downstream regulators PKA and Suppressor of Fused, preventing Gli transcription factor activation.","method":"Co-immunoprecipitation, dominant-negative localization mutant analysis, epistasis with PKA/Sufu/Gli in cell-based assays, live imaging of ciliary localization","journal":"Developmental cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, dominant-negative ciliary displacement mutants, epistasis with multiple pathway components, replicated across three independent 2012 studies","pmids":["22981989"],"is_preprint":false},{"year":2012,"finding":"Hh stimulation induces binding of EVC/EVC2 to Smo in a manner that depends on phosphorylation of the Smo C-terminal intracellular tail; this binding is abolished in Kif3a-/- cilium-deficient cells, establishing that the interaction requires primary cilia. EVC/EVC2 acts upstream of Sufu to promote Gli activation, as EVC/EVC2 is dispensable for constitutive Gli activity in Sufu-/- cells.","method":"Co-immunoprecipitation, phosphorylation-dependent binding assay using Smo mutants, epistasis in Sufu-/- and Kif3a-/- cells","journal":"Cell research","confidence":"High","confidence_rationale":"Tier 2 / Strong — phosphorylation-dependent Co-IP with domain mutants, genetic epistasis in multiple KO cell lines, independent of and consistent with co-published studies","pmids":["22986504"],"is_preprint":false},{"year":2012,"finding":"EVC and EVC2 are mutually required for their localization to primary cilia and for maintaining normal protein levels of each other; in Evc2-deficient chondrocytes, Smo translocation to cilia is normal after Hh activation, but Gli3 recruitment to cilia tips is reduced and Sufu/Gli3 dissociation is impaired. Smo co-precipitates with the EVC/EVC2 complex. Expression of the Weyers-associated Evc2Δ43 mutant causes mislocalization of EVC/EVC2 within cilia and reproduces the Gli3 molecular defects. EVC silencing in Sufu-/- cells attenuates Hh pathway output, indicating EVC/EVC2 also promotes signaling downstream of Sufu.","method":"Mouse genetic models (Evc2-/-, Evc2Δ43 knock-in), Co-immunoprecipitation, immunofluorescence of ciliary Gli3/Sufu localization, Evc/Evc2 double mutant analysis","journal":"Human molecular genetics","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple mouse KO models, Co-IP, ciliary localization imaging, epistasis in Sufu-/- cells, replicated findings across multiple 2012 studies","pmids":["23026747"],"is_preprint":false},{"year":2011,"finding":"EVC2 and EVC directly interact (identified by yeast two-hybrid with EVC as bait, confirmed by co-immunoprecipitation). EVC2 and EVC co-localize at the basal body and on primary cilia, and their localization is co-dependent: ciliary/basal body localization requires co-transfection of both constructs. EVC2 is a transmembrane protein with an extracellular N-terminus and intracellular C-terminus, while EVC is fully intracellular. Full-length EVC2 (but not EVC) is also detected in the nucleus by Western blotting of nuclear fractions. EVC2 is a positive regulator of Hh pathway activation in response to Smo agonist purmorphamine.","method":"Yeast two-hybrid, co-immunoprecipitation, immunofluorescence/confocal microscopy, subcellular fractionation + Western blot, Hh reporter assay in Evc2-null cells","journal":"BMC biology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — yeast two-hybrid plus confirmatory Co-IP, direct localization imaging with functional consequence, nuclear fractionation; multiple orthogonal methods in one study","pmids":["21356043"],"is_preprint":false},{"year":2014,"finding":"EFCAB7 and IQCE form a complex that anchors the EVC-EVC2 complex to the EvC zone at the base of primary cilia. EFCAB7 directly binds to a C-terminal disordered region in EVC2 that is deleted in Weyers syndrome patients. EFCAB7 depletion causes mislocalization of EVC-EVC2 within cilia (phenocopying the Weyers cellular phenotype) and impairs GLI2 activation.","method":"Co-immunoprecipitation, immunofluorescence localization, siRNA knockdown, Hh pathway reporter assays, evolutionary/bioinformatic analysis of EVC2 C-terminal region","journal":"Developmental cell","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP, knockdown with defined localization and signaling phenotype, domain mapping of binding site; multiple orthogonal methods","pmids":["24582806"],"is_preprint":false},{"year":2009,"finding":"Expression of Weyers acrodental dysostosis EVC2 exon 22 missense mutations (but not a truncation mimicking Ellis-van Creveld) impairs Hedgehog signal transduction in NIH 3T3 cells, demonstrating a dominant-negative mechanism for Weyers mutations at the EVC2 C-terminus.","method":"Ectopic expression of murine Weyers EVC2 variants and truncation mutants in NIH 3T3 fibroblasts, Hh pathway reporter assays","journal":"Human mutation","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — single lab, cell-based functional assay with domain-specific mutants; mechanistically informative but not replicated independently","pmids":["19810119"],"is_preprint":false},{"year":2015,"finding":"Global and conditional knockout of Evc2/Limbin in mice eliminates ciliary localization of both EVC2 and EVC proteins, reduces Hedgehog signaling activity, and causes skeletal and oral defects. Cartilage-specific deletion recapitulates skeletal defects; neural crest-specific deletion causes incisor growth defects. Osteoblast-specific deletion does not cause overt skeletal changes.","method":"Conditional KO mouse generation (Cre-lox), immunofluorescence for ciliary localization, Hh signaling readouts (target gene expression), skeletal/histological phenotyping","journal":"Genesis","confidence":"High","confidence_rationale":"Tier 2 / Moderate — multiple conditional KO lines, direct ciliary localization experiment, signaling readouts in vivo; rigorous genetic dissection of tissue-specific roles","pmids":["26219237"],"is_preprint":false},{"year":2016,"finding":"In Evc2 mutant mouse growth plates, FGF signaling is elevated in addition to compromised Hh-PTHrP feedback. Elevated Fgf18 expression from the perichondrium upon Evc2 inactivation is a critical pathogenic contributor to limb dwarfism, as heterozygous deletion of Fgf18 in Evc2 mutants partially rescues the limb dwarfism phenotype.","method":"Evc2 mutant mouse analysis, in vivo and in vitro FGF signaling assays, genetic rescue (Fgf18+/- × Evc2-/- double mutant mice), gene expression analysis","journal":"PLoS genetics","confidence":"High","confidence_rationale":"Tier 2 / Moderate — genetic epistasis via double mutant rescue, in vivo and in vitro signaling assays, multiple tissue culture validations","pmids":["28027321"],"is_preprint":false},{"year":2017,"finding":"Dental mesenchymal-specific deletion of Evc2 phenocopies the tooth abnormalities (enamel hypoplasia, delayed ameloblast differentiation) seen in global Evc2 mutants, establishing that Evc2 function in dental mesenchyme is required for normal odontoblast differentiation and dental mesenchymal stem cell homeostasis. Delayed odontoblast differentiation secondarily impairs ameloblast differentiation.","method":"Conditional KO mice (dental mesenchyme-specific Cre), histology, ameloblast/odontoblast marker immunohistochemistry, stem cell marker analysis","journal":"Journal of dental research","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — single lab, clean conditional KO with defined cellular phenotype and marker analysis; functional pathway placement in tooth development","pmids":["28081373"],"is_preprint":false},{"year":2023,"finding":"The EVC-EVC2 complex is subject to ubiquitination and SUMOylation of the cytosolic tails. Monoubiquitination of EVC-EVC2 cytosolic tails reduces protein levels. SUMOylation with SUMO3 enhances EVC-EVC2 accumulation at the EvC zone, possibly via increased binding to the EFCAB7-IQCE complex. EvC zone targeting of EVC-EVC2 depends on two separate EFCAB7-binding motifs within EVC2's Weyers-deleted peptide. Proteomic screen of EVC interactome confirmed EVC2, IQCE, EFCAB7 as main interactors and identified USP7 as a novel interactor; however, USP7 (and USP48) are not responsible for EVC-EVC2 deubiquitination.","method":"Endogenous protein interactome (proteomics/MS), ubiquitination/SUMOylation assays, ciliary localization imaging, domain mapping of EFCAB7-binding motifs in EVC2","journal":"Frontiers in cell and developmental biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — mass spectrometry interactome, post-translational modification assays, localization imaging with functional consequence; single lab but multiple orthogonal methods","pmids":["37576597"],"is_preprint":false},{"year":2025,"finding":"EVC and EVC2 are aberrantly overexpressed in a subset of AML cells (particularly those with ASXL1 mutations or t(8;21)). Loss of EVC/EVC2 impairs leukemia cell proliferation, promotes differentiation, and blocks AML progression in vivo. The leukemogenic role is mediated through MYC pathway activation, independent of Hedgehog signaling. Elevated EVC/EVC2 expression is associated with gained AML1-ETO occupancy or enhanced chromatin interactions at EVC/EVC2 promoter regions.","method":"Functional knockdown/KO studies in AML cell lines, in vivo AML progression assays, MYC pathway readouts, chromatin interaction/occupancy analysis (ChIP/Hi-C type)","journal":"Leukemia","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — single study, loss-of-function with defined proliferation/differentiation phenotype in vivo, pathway placement via MYC readouts; novel non-Hh mechanism, not yet replicated","pmids":["41249566"],"is_preprint":false}],"current_model":"EVC2 is a ciliary transmembrane protein that forms a constitutive heterodimeric complex with EVC at the EvC zone (a signaling microdomain at the base of primary cilia), where—upon Hedgehog ligand stimulation—it physically associates with phosphorylated Smoothened to transduce the signal to Gli transcription factors by promoting Sufu/Gli3 dissociation and Gli nuclear activity; the complex is anchored at the EvC zone by EFCAB7-IQCE, regulated by monoubiquitination (which reduces protein levels) and SUMO3 modification (which enhances EvC zone accumulation), and is required in chondrocytes and dental mesenchyme for normal skeletal and tooth development partly by restraining FGF18 signaling; additionally, EVC/EVC2 overexpression in a subset of AML activates MYC signaling independently of its canonical Hedgehog role."},"narrative":{"mechanistic_narrative":"EVC2 is a ciliary transmembrane protein that transduces Hedgehog (Hh) signaling at the EvC zone, a specialized microdomain at the base of primary cilia [PMID:22981989]. EVC2 forms a constitutive, mutually dependent heterodimer with EVC: the two proteins require each other for ciliary and basal-body localization and for maintaining each other's protein levels [PMID:23026747, PMID:21356043]. Upon Hh stimulation, the EVC-EVC2 complex binds Smoothened in a manner that depends on phosphorylation of the Smo C-terminal tail and on intact primary cilia, and this binding occurs specifically within the EvC zone [PMID:22981989, PMID:22986504]. EVC2 acts downstream of Smo to promote Gli transcription factor activation, driving Gli3 recruitment to cilia tips and Sufu/Gli3 dissociation, with additional positive activity downstream of Sufu [PMID:22986504, PMID:23026747]. Anchoring at the EvC zone requires the EFCAB7-IQCE complex, which binds a C-terminal disordered region of EVC2 deleted in Weyers acrodental dysostosis; loss of this anchoring mislocalizes the complex within cilia and impairs Gli activation [PMID:24582806, PMID:37576597]. Weyers-associated EVC2 C-terminal mutations act as dominant-negative alleles that retain ciliary localization but are displaced from the EvC zone [PMID:22981989, PMID:19810119]. In vivo, Evc2 is required in chondrocytes and dental mesenchyme for skeletal and tooth development, in part by restraining perichondrial FGF18 signaling in the growth plate [PMID:26219237, PMID:28027321, PMID:28081373]. Independently of its Hedgehog role, EVC2 (with EVC) is aberrantly overexpressed in a subset of AML, where it promotes leukemia progression through MYC pathway activation [PMID:41249566].","teleology":[{"year":2009,"claim":"Established that the EVC2 C-terminus is functionally critical, showing Weyers missense mutations impair Hh signaling through a dominant-negative rather than simple loss-of-function mechanism.","evidence":"Ectopic expression of murine Weyers and Ellis-van Creveld-mimicking EVC2 variants in NIH 3T3 cells with Hh reporter assays","pmids":["19810119"],"confidence":"Medium","gaps":["Single lab, not independently replicated","Did not define the molecular partner or microdomain whose binding is disrupted","No structural basis for the dominant-negative effect"]},{"year":2011,"claim":"Defined the core physical interaction and topology, showing EVC2 and EVC directly bind and co-dependently localize to the basal body and primary cilium.","evidence":"Yeast two-hybrid, Co-IP, confocal immunofluorescence, subcellular fractionation, and Hh reporter assays in Evc2-null cells","pmids":["21356043"],"confidence":"High","gaps":["Functional significance of nuclear full-length EVC2 not established","Did not place EVC-EVC2 relative to Smo or Sufu in the pathway","Mechanism of co-dependent stabilization unresolved"]},{"year":2012,"claim":"Resolved where in the Hh pathway EVC-EVC2 acts, showing it binds phospho-Smo within the EvC zone in a cilium-dependent manner and promotes Gli activation by driving Sufu/Gli3 dissociation upstream and downstream of Sufu.","evidence":"Reciprocal Co-IP, phosphorylation-dependent Smo-binding assays, dominant-negative localization mutants, epistasis in Sufu-/-/Kif3a-/- cells, and Evc2-/- / Evc2Δ43 knock-in mice across three independent studies","pmids":["22981989","22986504","23026747"],"confidence":"High","gaps":["Molecular mechanism by which EVC-EVC2 triggers Sufu/Gli3 dissociation not defined","Structure of the Smo-EVC2 interface unknown","How the EvC zone is physically distinguished from the rest of the ciliary base unresolved"]},{"year":2014,"claim":"Identified the anchoring mechanism, showing EFCAB7-IQCE tethers EVC-EVC2 to the EvC zone via the same C-terminal region deleted in Weyers syndrome.","evidence":"Reciprocal Co-IP, domain mapping, siRNA depletion with localization and Hh reporter phenotypes","pmids":["24582806"],"confidence":"High","gaps":["How EFCAB7-IQCE itself is positioned at the EvC zone not established","Stoichiometry of the EVC-EVC2-EFCAB7-IQCE assembly unknown"]},{"year":2015,"claim":"Demonstrated the in vivo tissue requirements, showing Evc2 loss eliminates ciliary EVC/EVC2, reduces Hh signaling, and causes skeletal and oral defects with cartilage and neural-crest specificity.","evidence":"Global and tissue-specific conditional KO mice with ciliary localization imaging, Hh readouts, and skeletal phenotyping","pmids":["26219237"],"confidence":"High","gaps":["Osteoblast-specific deletion phenotype absent, leaving osteoblast role undefined","Cell-autonomous vs non-autonomous contributions not fully separated"]},{"year":2016,"claim":"Connected EVC2 loss to a downstream pathogenic mechanism, showing elevated perichondrial FGF18 contributes to limb dwarfism and is partially rescued genetically.","evidence":"Evc2 mutant mice, FGF signaling assays, and Fgf18+/- x Evc2-/- double-mutant genetic rescue","pmids":["28027321"],"confidence":"High","gaps":["Mechanism linking reduced Hh signaling to elevated Fgf18 not defined","Partial rescue implies additional pathogenic pathways"]},{"year":2017,"claim":"Localized the dental requirement, showing Evc2 acts in dental mesenchyme for odontoblast differentiation, with ameloblast defects arising secondarily.","evidence":"Dental mesenchyme-specific conditional KO mice with marker immunohistochemistry and stem-cell analysis","pmids":["28081373"],"confidence":"Medium","gaps":["Single lab","Signaling pathway mediating the mesenchymal effect not resolved"]},{"year":2023,"claim":"Revealed post-translational regulation of complex abundance and localization, showing monoubiquitination lowers EVC-EVC2 levels while SUMO3 modification enhances EvC zone accumulation.","evidence":"Endogenous interactome MS, ubiquitination/SUMOylation assays, localization imaging, and EFCAB7-binding motif mapping","pmids":["37576597"],"confidence":"Medium","gaps":["The deubiquitinase responsible was not identified (USP7/USP48 excluded)","The ubiquitin ligase and SUMO machinery acting on EVC-EVC2 unknown","Functional role of USP7 as an interactor undefined"]},{"year":2025,"claim":"Uncovered a Hedgehog-independent oncogenic role, showing EVC/EVC2 overexpression drives a subset of AML through MYC pathway activation.","evidence":"Knockdown/KO in AML cell lines, in vivo progression assays, MYC readouts, and chromatin occupancy/interaction analysis","pmids":["41249566"],"confidence":"Medium","gaps":["Single study, not replicated","Mechanism by which EVC/EVC2 activates MYC is undefined","Whether ciliary localization is involved in the AML role unknown"]},{"year":null,"claim":"How the EVC-EVC2 complex mechanistically couples phospho-Smo binding at the EvC zone to Sufu/Gli3 dissociation, and how this ciliary protein contributes to Hh-independent MYC activation in AML, remain unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model of the EVC-EVC2-Smo interface","Biochemical mechanism of Sufu/Gli3 release not established","MYC-activating mechanism in AML undefined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[0,1,2]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[3,4]}],"localization":[{"term_id":"GO:0005929","term_label":"cilium","supporting_discovery_ids":[0,2,3,6]},{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[3]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[3]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,1,2]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[6,7,8]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[10]}],"complexes":["EVC-EVC2 complex","EvC zone (EVC-EVC2-EFCAB7-IQCE)"],"partners":["EVC","SMO","EFCAB7","IQCE","USP7"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q86UK5","full_name":"Limbin","aliases":["Ellis-van Creveld syndrome protein 2","EVC2"],"length_aa":1308,"mass_kda":147.9,"function":"Component of the EvC complex that positively regulates ciliary Hedgehog (Hh) signaling. Plays a critical role in bone formation and skeletal development. May be involved in early embryonic morphogenesis","subcellular_location":"Cell membrane; Cytoplasm, cytoskeleton, cilium basal body; Cell projection, cilium; Cell projection, cilium membrane; Nucleus","url":"https://www.uniprot.org/uniprotkb/Q86UK5/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/EVC2","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/EVC2","total_profiled":1310},"omim":[{"mim_id":"619142","title":"CARDIOACROFACIAL DYSPLASIA 1; CAFD1","url":"https://www.omim.org/entry/619142"},{"mim_id":"618123","title":"POLYDACTYLY, POSTAXIAL, TYPE A8; PAPA8","url":"https://www.omim.org/entry/618123"},{"mim_id":"617632","title":"EF-HAND CALCIUM-BINDING DOMAIN-CONTAINING PROTEIN 7; EFCAB7","url":"https://www.omim.org/entry/617632"},{"mim_id":"617631","title":"IQ DOMAIN-CONTAINING PROTEIN E; IQCE","url":"https://www.omim.org/entry/617631"},{"mim_id":"617088","title":"SHORT-RIB THORACIC DYSPLASIA 15 WITH POLYDACTYLY; SRTD15","url":"https://www.omim.org/entry/617088"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Plasma membrane","reliability":"Approved"},{"location":"Microtubules","reliability":"Approved"},{"location":"Cytokinetic bridge","reliability":"Approved"},{"location":"Primary cilium","reliability":"Approved"},{"location":"Nucleoplasm","reliability":"Additional"},{"location":"Primary cilium tip","reliability":"Additional"},{"location":"Basal body","reliability":"Additional"},{"location":"Cytosol","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in many","driving_tissues":[],"url":"https://www.proteinatlas.org/search/EVC2"},"hgnc":{"alias_symbol":["LBN"],"prev_symbol":[]},"alphafold":{"accession":"Q86UK5","domains":[{"cath_id":"2.60.40.1510","chopping":"159-286","consensus_level":"high","plddt":77.97,"start":159,"end":286}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q86UK5","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q86UK5-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q86UK5-F1-predicted_aligned_error_v6.png","plddt_mean":72.69},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=EVC2","jax_strain_url":"https://www.jax.org/strain/search?query=EVC2"},"sequence":{"accession":"Q86UK5","fasta_url":"https://rest.uniprot.org/uniprotkb/Q86UK5.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q86UK5/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q86UK5"}},"corpus_meta":[{"pmid":"22981989","id":"PMC_22981989","title":"A Smoothened-Evc2 complex transduces the Hedgehog signal at primary cilia.","date":"2012","source":"Developmental cell","url":"https://pubmed.ncbi.nlm.nih.gov/22981989","citation_count":150,"is_preprint":false},{"pmid":"23026747","id":"PMC_23026747","title":"The ciliary Evc/Evc2 complex interacts with Smo and controls Hedgehog pathway activity in chondrocytes by regulating Sufu/Gli3 dissociation and Gli3 trafficking in primary cilia.","date":"2012","source":"Human molecular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/23026747","citation_count":94,"is_preprint":false},{"pmid":"22986504","id":"PMC_22986504","title":"Smoothened transduces Hedgehog signal by forming a complex with Evc/Evc2.","date":"2012","source":"Cell research","url":"https://pubmed.ncbi.nlm.nih.gov/22986504","citation_count":86,"is_preprint":false},{"pmid":"17024374","id":"PMC_17024374","title":"Sequencing EVC and EVC2 identifies mutations in two-thirds of Ellis-van Creveld syndrome patients.","date":"2006","source":"Human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/17024374","citation_count":80,"is_preprint":false},{"pmid":"12468274","id":"PMC_12468274","title":"A new gene, EVC2, is mutated in Ellis-van Creveld syndrome.","date":"2002","source":"Molecular genetics and metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/12468274","citation_count":80,"is_preprint":false},{"pmid":"24582806","id":"PMC_24582806","title":"EFCAB7 and IQCE regulate hedgehog signaling by tethering the EVC-EVC2 complex to the base of primary cilia.","date":"2014","source":"Developmental cell","url":"https://pubmed.ncbi.nlm.nih.gov/24582806","citation_count":77,"is_preprint":false},{"pmid":"21356043","id":"PMC_21356043","title":"Evc2 is a positive modulator of Hedgehog signalling that interacts with Evc at the cilia membrane and is also found in the nucleus.","date":"2011","source":"BMC biology","url":"https://pubmed.ncbi.nlm.nih.gov/21356043","citation_count":77,"is_preprint":false},{"pmid":"23220543","id":"PMC_23220543","title":"Novel and recurrent EVC and EVC2 mutations in Ellis-van Creveld syndrome and Weyers acrofacial dyostosis.","date":"2012","source":"European journal of medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/23220543","citation_count":59,"is_preprint":false},{"pmid":"19810119","id":"PMC_19810119","title":"Widening the mutation spectrum of EVC and EVC2: ectopic expression of Weyer variants in NIH 3T3 fibroblasts disrupts Hedgehog signaling.","date":"2009","source":"Human mutation","url":"https://pubmed.ncbi.nlm.nih.gov/19810119","citation_count":52,"is_preprint":false},{"pmid":"16404586","id":"PMC_16404586","title":"A novel heterozygous deletion in the EVC2 gene causes Weyers acrofacial dysostosis.","date":"2006","source":"Human 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Part A","url":"https://pubmed.ncbi.nlm.nih.gov/21815252","citation_count":20,"is_preprint":false},{"pmid":"29321360","id":"PMC_29321360","title":"Ellis-van Creveld syndrome and profound deafness resulted by sequence variants in the EVC/EVC2 and TMC1 genes.","date":"2017","source":"Journal of genetics","url":"https://pubmed.ncbi.nlm.nih.gov/29321360","citation_count":19,"is_preprint":false},{"pmid":"28027321","id":"PMC_28027321","title":"Elevated Fibroblast Growth Factor Signaling Is Critical for the Pathogenesis of the Dwarfism in Evc2/Limbin Mutant Mice.","date":"2016","source":"PLoS genetics","url":"https://pubmed.ncbi.nlm.nih.gov/28027321","citation_count":19,"is_preprint":false},{"pmid":"28081373","id":"PMC_28081373","title":"Loss of Function of Evc2 in Dental Mesenchyme Leads to Hypomorphic Enamel.","date":"2017","source":"Journal of dental 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physical association between Smoothened (Smo) and EVC2; this Smo-EVC2 complex forms specifically within the EvC zone, a distinct sub-compartment at the base of primary cilia. EVC2 mutants that localize to cilia but are displaced from the EvC zone act as dominant inhibitors of Hh signaling, and loss of EVC2 function blocks Hh signaling between Smo and the downstream regulators PKA and Suppressor of Fused, preventing Gli transcription factor activation.\",\n      \"method\": \"Co-immunoprecipitation, dominant-negative localization mutant analysis, epistasis with PKA/Sufu/Gli in cell-based assays, live imaging of ciliary localization\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, dominant-negative ciliary displacement mutants, epistasis with multiple pathway components, replicated across three independent 2012 studies\",\n      \"pmids\": [\"22981989\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Hh stimulation induces binding of EVC/EVC2 to Smo in a manner that depends on phosphorylation of the Smo C-terminal intracellular tail; this binding is abolished in Kif3a-/- cilium-deficient cells, establishing that the interaction requires primary cilia. EVC/EVC2 acts upstream of Sufu to promote Gli activation, as EVC/EVC2 is dispensable for constitutive Gli activity in Sufu-/- cells.\",\n      \"method\": \"Co-immunoprecipitation, phosphorylation-dependent binding assay using Smo mutants, epistasis in Sufu-/- and Kif3a-/- cells\",\n      \"journal\": \"Cell research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — phosphorylation-dependent Co-IP with domain mutants, genetic epistasis in multiple KO cell lines, independent of and consistent with co-published studies\",\n      \"pmids\": [\"22986504\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"EVC and EVC2 are mutually required for their localization to primary cilia and for maintaining normal protein levels of each other; in Evc2-deficient chondrocytes, Smo translocation to cilia is normal after Hh activation, but Gli3 recruitment to cilia tips is reduced and Sufu/Gli3 dissociation is impaired. Smo co-precipitates with the EVC/EVC2 complex. Expression of the Weyers-associated Evc2Δ43 mutant causes mislocalization of EVC/EVC2 within cilia and reproduces the Gli3 molecular defects. EVC silencing in Sufu-/- cells attenuates Hh pathway output, indicating EVC/EVC2 also promotes signaling downstream of Sufu.\",\n      \"method\": \"Mouse genetic models (Evc2-/-, Evc2Δ43 knock-in), Co-immunoprecipitation, immunofluorescence of ciliary Gli3/Sufu localization, Evc/Evc2 double mutant analysis\",\n      \"journal\": \"Human molecular genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple mouse KO models, Co-IP, ciliary localization imaging, epistasis in Sufu-/- cells, replicated findings across multiple 2012 studies\",\n      \"pmids\": [\"23026747\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"EVC2 and EVC directly interact (identified by yeast two-hybrid with EVC as bait, confirmed by co-immunoprecipitation). EVC2 and EVC co-localize at the basal body and on primary cilia, and their localization is co-dependent: ciliary/basal body localization requires co-transfection of both constructs. EVC2 is a transmembrane protein with an extracellular N-terminus and intracellular C-terminus, while EVC is fully intracellular. Full-length EVC2 (but not EVC) is also detected in the nucleus by Western blotting of nuclear fractions. EVC2 is a positive regulator of Hh pathway activation in response to Smo agonist purmorphamine.\",\n      \"method\": \"Yeast two-hybrid, co-immunoprecipitation, immunofluorescence/confocal microscopy, subcellular fractionation + Western blot, Hh reporter assay in Evc2-null cells\",\n      \"journal\": \"BMC biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — yeast two-hybrid plus confirmatory Co-IP, direct localization imaging with functional consequence, nuclear fractionation; multiple orthogonal methods in one study\",\n      \"pmids\": [\"21356043\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"EFCAB7 and IQCE form a complex that anchors the EVC-EVC2 complex to the EvC zone at the base of primary cilia. EFCAB7 directly binds to a C-terminal disordered region in EVC2 that is deleted in Weyers syndrome patients. EFCAB7 depletion causes mislocalization of EVC-EVC2 within cilia (phenocopying the Weyers cellular phenotype) and impairs GLI2 activation.\",\n      \"method\": \"Co-immunoprecipitation, immunofluorescence localization, siRNA knockdown, Hh pathway reporter assays, evolutionary/bioinformatic analysis of EVC2 C-terminal region\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP, knockdown with defined localization and signaling phenotype, domain mapping of binding site; multiple orthogonal methods\",\n      \"pmids\": [\"24582806\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Expression of Weyers acrodental dysostosis EVC2 exon 22 missense mutations (but not a truncation mimicking Ellis-van Creveld) impairs Hedgehog signal transduction in NIH 3T3 cells, demonstrating a dominant-negative mechanism for Weyers mutations at the EVC2 C-terminus.\",\n      \"method\": \"Ectopic expression of murine Weyers EVC2 variants and truncation mutants in NIH 3T3 fibroblasts, Hh pathway reporter assays\",\n      \"journal\": \"Human mutation\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — single lab, cell-based functional assay with domain-specific mutants; mechanistically informative but not replicated independently\",\n      \"pmids\": [\"19810119\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Global and conditional knockout of Evc2/Limbin in mice eliminates ciliary localization of both EVC2 and EVC proteins, reduces Hedgehog signaling activity, and causes skeletal and oral defects. Cartilage-specific deletion recapitulates skeletal defects; neural crest-specific deletion causes incisor growth defects. Osteoblast-specific deletion does not cause overt skeletal changes.\",\n      \"method\": \"Conditional KO mouse generation (Cre-lox), immunofluorescence for ciliary localization, Hh signaling readouts (target gene expression), skeletal/histological phenotyping\",\n      \"journal\": \"Genesis\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple conditional KO lines, direct ciliary localization experiment, signaling readouts in vivo; rigorous genetic dissection of tissue-specific roles\",\n      \"pmids\": [\"26219237\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"In Evc2 mutant mouse growth plates, FGF signaling is elevated in addition to compromised Hh-PTHrP feedback. Elevated Fgf18 expression from the perichondrium upon Evc2 inactivation is a critical pathogenic contributor to limb dwarfism, as heterozygous deletion of Fgf18 in Evc2 mutants partially rescues the limb dwarfism phenotype.\",\n      \"method\": \"Evc2 mutant mouse analysis, in vivo and in vitro FGF signaling assays, genetic rescue (Fgf18+/- × Evc2-/- double mutant mice), gene expression analysis\",\n      \"journal\": \"PLoS genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis via double mutant rescue, in vivo and in vitro signaling assays, multiple tissue culture validations\",\n      \"pmids\": [\"28027321\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Dental mesenchymal-specific deletion of Evc2 phenocopies the tooth abnormalities (enamel hypoplasia, delayed ameloblast differentiation) seen in global Evc2 mutants, establishing that Evc2 function in dental mesenchyme is required for normal odontoblast differentiation and dental mesenchymal stem cell homeostasis. Delayed odontoblast differentiation secondarily impairs ameloblast differentiation.\",\n      \"method\": \"Conditional KO mice (dental mesenchyme-specific Cre), histology, ameloblast/odontoblast marker immunohistochemistry, stem cell marker analysis\",\n      \"journal\": \"Journal of dental research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — single lab, clean conditional KO with defined cellular phenotype and marker analysis; functional pathway placement in tooth development\",\n      \"pmids\": [\"28081373\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"The EVC-EVC2 complex is subject to ubiquitination and SUMOylation of the cytosolic tails. Monoubiquitination of EVC-EVC2 cytosolic tails reduces protein levels. SUMOylation with SUMO3 enhances EVC-EVC2 accumulation at the EvC zone, possibly via increased binding to the EFCAB7-IQCE complex. EvC zone targeting of EVC-EVC2 depends on two separate EFCAB7-binding motifs within EVC2's Weyers-deleted peptide. Proteomic screen of EVC interactome confirmed EVC2, IQCE, EFCAB7 as main interactors and identified USP7 as a novel interactor; however, USP7 (and USP48) are not responsible for EVC-EVC2 deubiquitination.\",\n      \"method\": \"Endogenous protein interactome (proteomics/MS), ubiquitination/SUMOylation assays, ciliary localization imaging, domain mapping of EFCAB7-binding motifs in EVC2\",\n      \"journal\": \"Frontiers in cell and developmental biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — mass spectrometry interactome, post-translational modification assays, localization imaging with functional consequence; single lab but multiple orthogonal methods\",\n      \"pmids\": [\"37576597\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"EVC and EVC2 are aberrantly overexpressed in a subset of AML cells (particularly those with ASXL1 mutations or t(8;21)). Loss of EVC/EVC2 impairs leukemia cell proliferation, promotes differentiation, and blocks AML progression in vivo. The leukemogenic role is mediated through MYC pathway activation, independent of Hedgehog signaling. Elevated EVC/EVC2 expression is associated with gained AML1-ETO occupancy or enhanced chromatin interactions at EVC/EVC2 promoter regions.\",\n      \"method\": \"Functional knockdown/KO studies in AML cell lines, in vivo AML progression assays, MYC pathway readouts, chromatin interaction/occupancy analysis (ChIP/Hi-C type)\",\n      \"journal\": \"Leukemia\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — single study, loss-of-function with defined proliferation/differentiation phenotype in vivo, pathway placement via MYC readouts; novel non-Hh mechanism, not yet replicated\",\n      \"pmids\": [\"41249566\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"EVC2 is a ciliary transmembrane protein that forms a constitutive heterodimeric complex with EVC at the EvC zone (a signaling microdomain at the base of primary cilia), where—upon Hedgehog ligand stimulation—it physically associates with phosphorylated Smoothened to transduce the signal to Gli transcription factors by promoting Sufu/Gli3 dissociation and Gli nuclear activity; the complex is anchored at the EvC zone by EFCAB7-IQCE, regulated by monoubiquitination (which reduces protein levels) and SUMO3 modification (which enhances EvC zone accumulation), and is required in chondrocytes and dental mesenchyme for normal skeletal and tooth development partly by restraining FGF18 signaling; additionally, EVC/EVC2 overexpression in a subset of AML activates MYC signaling independently of its canonical Hedgehog role.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"EVC2 is a ciliary transmembrane protein that transduces Hedgehog (Hh) signaling at the EvC zone, a specialized microdomain at the base of primary cilia [#0]. EVC2 forms a constitutive, mutually dependent heterodimer with EVC: the two proteins require each other for ciliary and basal-body localization and for maintaining each other's protein levels [#2, #3]. Upon Hh stimulation, the EVC-EVC2 complex binds Smoothened in a manner that depends on phosphorylation of the Smo C-terminal tail and on intact primary cilia, and this binding occurs specifically within the EvC zone [#0, #1]. EVC2 acts downstream of Smo to promote Gli transcription factor activation, driving Gli3 recruitment to cilia tips and Sufu/Gli3 dissociation, with additional positive activity downstream of Sufu [#1, #2]. Anchoring at the EvC zone requires the EFCAB7-IQCE complex, which binds a C-terminal disordered region of EVC2 deleted in Weyers acrodental dysostosis; loss of this anchoring mislocalizes the complex within cilia and impairs Gli activation [#4, #9]. Weyers-associated EVC2 C-terminal mutations act as dominant-negative alleles that retain ciliary localization but are displaced from the EvC zone [#0, #5]. In vivo, Evc2 is required in chondrocytes and dental mesenchyme for skeletal and tooth development, in part by restraining perichondrial FGF18 signaling in the growth plate [#6, #7, #8]. Independently of its Hedgehog role, EVC2 (with EVC) is aberrantly overexpressed in a subset of AML, where it promotes leukemia progression through MYC pathway activation [#10].\",\n  \"teleology\": [\n    {\n      \"year\": 2009,\n      \"claim\": \"Established that the EVC2 C-terminus is functionally critical, showing Weyers missense mutations impair Hh signaling through a dominant-negative rather than simple loss-of-function mechanism.\",\n      \"evidence\": \"Ectopic expression of murine Weyers and Ellis-van Creveld-mimicking EVC2 variants in NIH 3T3 cells with Hh reporter assays\",\n      \"pmids\": [\"19810119\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Single lab, not independently replicated\", \"Did not define the molecular partner or microdomain whose binding is disrupted\", \"No structural basis for the dominant-negative effect\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Defined the core physical interaction and topology, showing EVC2 and EVC directly bind and co-dependently localize to the basal body and primary cilium.\",\n      \"evidence\": \"Yeast two-hybrid, Co-IP, confocal immunofluorescence, subcellular fractionation, and Hh reporter assays in Evc2-null cells\",\n      \"pmids\": [\"21356043\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Functional significance of nuclear full-length EVC2 not established\", \"Did not place EVC-EVC2 relative to Smo or Sufu in the pathway\", \"Mechanism of co-dependent stabilization unresolved\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Resolved where in the Hh pathway EVC-EVC2 acts, showing it binds phospho-Smo within the EvC zone in a cilium-dependent manner and promotes Gli activation by driving Sufu/Gli3 dissociation upstream and downstream of Sufu.\",\n      \"evidence\": \"Reciprocal Co-IP, phosphorylation-dependent Smo-binding assays, dominant-negative localization mutants, epistasis in Sufu-/-/Kif3a-/- cells, and Evc2-/- / Evc2\\u039443 knock-in mice across three independent studies\",\n      \"pmids\": [\"22981989\", \"22986504\", \"23026747\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Molecular mechanism by which EVC-EVC2 triggers Sufu/Gli3 dissociation not defined\", \"Structure of the Smo-EVC2 interface unknown\", \"How the EvC zone is physically distinguished from the rest of the ciliary base unresolved\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Identified the anchoring mechanism, showing EFCAB7-IQCE tethers EVC-EVC2 to the EvC zone via the same C-terminal region deleted in Weyers syndrome.\",\n      \"evidence\": \"Reciprocal Co-IP, domain mapping, siRNA depletion with localization and Hh reporter phenotypes\",\n      \"pmids\": [\"24582806\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"How EFCAB7-IQCE itself is positioned at the EvC zone not established\", \"Stoichiometry of the EVC-EVC2-EFCAB7-IQCE assembly unknown\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Demonstrated the in vivo tissue requirements, showing Evc2 loss eliminates ciliary EVC/EVC2, reduces Hh signaling, and causes skeletal and oral defects with cartilage and neural-crest specificity.\",\n      \"evidence\": \"Global and tissue-specific conditional KO mice with ciliary localization imaging, Hh readouts, and skeletal phenotyping\",\n      \"pmids\": [\"26219237\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Osteoblast-specific deletion phenotype absent, leaving osteoblast role undefined\", \"Cell-autonomous vs non-autonomous contributions not fully separated\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Connected EVC2 loss to a downstream pathogenic mechanism, showing elevated perichondrial FGF18 contributes to limb dwarfism and is partially rescued genetically.\",\n      \"evidence\": \"Evc2 mutant mice, FGF signaling assays, and Fgf18+/- x Evc2-/- double-mutant genetic rescue\",\n      \"pmids\": [\"28027321\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Mechanism linking reduced Hh signaling to elevated Fgf18 not defined\", \"Partial rescue implies additional pathogenic pathways\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Localized the dental requirement, showing Evc2 acts in dental mesenchyme for odontoblast differentiation, with ameloblast defects arising secondarily.\",\n      \"evidence\": \"Dental mesenchyme-specific conditional KO mice with marker immunohistochemistry and stem-cell analysis\",\n      \"pmids\": [\"28081373\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Single lab\", \"Signaling pathway mediating the mesenchymal effect not resolved\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Revealed post-translational regulation of complex abundance and localization, showing monoubiquitination lowers EVC-EVC2 levels while SUMO3 modification enhances EvC zone accumulation.\",\n      \"evidence\": \"Endogenous interactome MS, ubiquitination/SUMOylation assays, localization imaging, and EFCAB7-binding motif mapping\",\n      \"pmids\": [\"37576597\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"The deubiquitinase responsible was not identified (USP7/USP48 excluded)\", \"The ubiquitin ligase and SUMO machinery acting on EVC-EVC2 unknown\", \"Functional role of USP7 as an interactor undefined\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Uncovered a Hedgehog-independent oncogenic role, showing EVC/EVC2 overexpression drives a subset of AML through MYC pathway activation.\",\n      \"evidence\": \"Knockdown/KO in AML cell lines, in vivo progression assays, MYC readouts, and chromatin occupancy/interaction analysis\",\n      \"pmids\": [\"41249566\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Single study, not replicated\", \"Mechanism by which EVC/EVC2 activates MYC is undefined\", \"Whether ciliary localization is involved in the AML role unknown\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How the EVC-EVC2 complex mechanistically couples phospho-Smo binding at the EvC zone to Sufu/Gli3 dissociation, and how this ciliary protein contributes to Hh-independent MYC activation in AML, remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"No structural model of the EVC-EVC2-Smo interface\", \"Biochemical mechanism of Sufu/Gli3 release not established\", \"MYC-activating mechanism in AML undefined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [0, 1, 2]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [3, 4]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005929\", \"supporting_discovery_ids\": [0, 2, 3, 6]},\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [3]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [3]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 1, 2]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [6, 7, 8]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [10]}\n    ],\n    \"complexes\": [\"EVC-EVC2 complex\", \"EvC zone (EVC-EVC2-EFCAB7-IQCE)\"],\n    \"partners\": [\"EVC\", \"SMO\", \"EFCAB7\", \"IQCE\", \"USP7\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":8,"faith_total":8,"faith_pct":100.0}}