{"gene":"ZDHHC4","run_date":"2026-04-28T23:00:24","timeline":{"discoveries":[{"year":2023,"finding":"CPT1A recruits ER-localized ZDHHC4 to catalyze palmitoylation of MAVS at Cys79, which promotes MAVS stabilization and activation by inhibiting K48-linked ubiquitination and facilitating K63-linked ubiquitination, thereby potentiating the type I interferon response.","method":"Co-immunoprecipitation, site-directed mutagenesis (Cys79), ubiquitination assays, loss-of-function experiments with defined IFN-I readout","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 2 — reciprocal Co-IP, specific residue mutagenesis, mechanistic pathway placement with multiple orthogonal assays","pmids":["38016475"],"is_preprint":false},{"year":2015,"finding":"ZDHHC4 was identified as a palmitoyl acyltransferase (PAT) that physically interacts with the D2 dopamine receptor (D2R) and affects its palmitoylation status, contributing to D2R stability and plasma membrane expression.","method":"Membrane yeast two-hybrid (MYTH) screen, co-immunoprecipitation, bioorthogonal click chemistry palmitoylation assay, cysteine mutagenesis","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 — reciprocal Co-IP and palmitoylation assay, but single lab study","pmids":["26535572"],"is_preprint":false},{"year":2021,"finding":"ZDHHC4 catalyzes palmitoylation of KAI1 (CD82) at the membrane surface, which is required for KAI1 localization to the pericyte membrane surface and its downstream induction of LIF through the Src/p53 pathway to inhibit angiogenesis.","method":"Palmitoylation assay, loss-of-function experiments, in vitro and in vivo angiogenesis assays","journal":"Journal of hematology & oncology","confidence":"Medium","confidence_rationale":"Tier 2 — defined substrate and functional consequence, but single lab","pmids":["34530889"],"is_preprint":false},{"year":2021,"finding":"ZDHHC4 (among other ZDHHCs) promotes palmitoylation of the SARS-CoV-2 S protein and physically associates with S protein; this palmitoylation is critical for S-mediated syncytia formation and pseudovirus particle entry.","method":"Overexpression of individual ZDHHCs, palmitoylation assay, co-immunoprecipitation, pseudovirus entry assay, syncytia formation assay","journal":"Journal of medical virology","confidence":"Medium","confidence_rationale":"Tier 2-3 — functional assays with specific palmitoylation readout, but ZDHHC4 is one of many ZDHHCs tested without unique mechanistic dissection","pmids":["34528721"],"is_preprint":false},{"year":2022,"finding":"ZDHHC4 palmitoylates GSK3β at Cys14, which decreases phospho-Ser9 and increases phospho-Tyr216, thereby activating the EZH2-STAT3 signaling axis and promoting GBM stem cell self-renewal and temozolomide resistance.","method":"Palmitoylation assay, site-directed mutagenesis (Cys14), phosphorylation analysis, knockdown experiments with stemness and drug-resistance readouts, STAT3 feedback loop validation","journal":"Oncogenesis","confidence":"Medium","confidence_rationale":"Tier 2 — specific palmitoylation site identified by mutagenesis, pathway placement with multiple downstream readouts, single lab","pmids":["35606353"],"is_preprint":false},{"year":2022,"finding":"ZDHHC4 and ZDHHC8 catalyze S-palmitoylation of the transcription factor NFATC4, which is required for NFATC4 trafficking from the cytoplasm to the nucleus; reduced ZDHHC4 and ZDHHC8 levels in Ppt1-deficient mice lower nuclear palmitoylated NFATC4 and thereby suppress IP3R1 expression, dysregulating lysosomal Ca2+ homeostasis.","method":"Palmitoylation assay, subcellular fractionation, mouse knockout model (Ppt1-/-), IP3R1 overexpression rescue, identification of responsible ZDHHC enzymes","journal":"Journal of inherited metabolic disease","confidence":"Medium","confidence_rationale":"Tier 2 — mechanistic pathway placement with loss-of-function and rescue, multiple methods, single lab","pmids":["35150145"],"is_preprint":false},{"year":2024,"finding":"ZDHHC4 physically interacts with TRPV1 and catalyzes S-palmitoylation at cysteine residues C157, C362, C390, and C715 of TRPV1, promoting TRPV1 degradation via the lysosome pathway and thereby facilitating inflammatory pain relief; this palmitoylation is counterbalanced by depalmitoylase APT1.","method":"Co-immunoprecipitation, site-directed mutagenesis (C157, C362, C390, C715), acyl-biotin exchange palmitoylation assay, electrophysiology, in vivo pain behavior assays","journal":"EMBO reports","confidence":"High","confidence_rationale":"Tier 1-2 — multiple orthogonal methods (Co-IP, mutagenesis of specific residues, functional electrophysiology, in vivo readout) in a single study","pmids":["39528731"],"is_preprint":false},{"year":2024,"finding":"ATF3 in macrophages increases ZDHHC4/5-mediated CD36 palmitoylation at C3, C7, C464, and C466, which promotes CD36 plasma membrane localization and fatty acid uptake.","method":"Overexpression/knockdown of Atf3, palmitoylation assay, site-directed mutagenesis of CD36 cysteines, subcellular fractionation","journal":"Science advances","confidence":"Medium","confidence_rationale":"Tier 2 — specific palmitoylation sites identified, functional localization consequence shown; ZDHHC4 and ZDHHC5 not separated individually","pmids":["39047111"],"is_preprint":false},{"year":2025,"finding":"FoxO1 transcriptionally upregulates zDHHC4 in the diabetic heart; zDHHC4 S-acylates CD36, promoting its sarcolemmal localization, increased fatty acid oxidation, and triglyceride storage, driving metabolic dysfunction; genetic silencing of zDHHC4 decreases CD36 S-acylation and ameliorates the diabetic cardiac phenotype.","method":"ChIP sequencing, ChIP-qPCR, luciferase assays, siRNA/shRNA silencing, cardiomyocyte-specific FoxO1 knockout mice, pharmacological ZDHHC inhibition with functional cardiac readouts","journal":"Circulation research","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods (ChIP-seq, genetic silencing, KO mice, pharmacology) with defined metabolic and functional phenotypes, conserved across species","pmids":["40357580"],"is_preprint":false},{"year":2025,"finding":"EVA1A deficiency transcriptionally enhances palmitoyl acyltransferases ZDHHC4/5 while repressing depalmitoylase APT1, boosting CD36 palmitoylation and plasma membrane localization to increase fatty acid uptake and impair β-oxidation in hepatocytes.","method":"Hepatocyte-specific knockout of Eva1a, transcriptomics, palmitoylation assay, subcellular fractionation, CD36 localization assays","journal":"Research (Washington, D.C.)","confidence":"Medium","confidence_rationale":"Tier 2 — functional KO with mechanistic follow-up, but ZDHHC4 and ZDHHC5 not individually dissected","pmids":["41306774"],"is_preprint":false},{"year":2026,"finding":"ZDHHC4 palmitoylates ZEB-2 at C478; this palmitoylation promotes ZEB-2 deubiquitination and protein stability, facilitating EMT in melanoma cells; ZEB-2 interacts with ZDHHC4 through its N-terminal sequences.","method":"Mass spectrometry substrate identification, co-immunoprecipitation, site-directed mutagenesis (C478), ubiquitination assay, knockdown with EMT readout","journal":"Cellular and molecular life sciences : CMLS","confidence":"Medium","confidence_rationale":"Tier 2 — specific palmitoylation site identified by mutagenesis, mechanistic pathway placement, single lab","pmids":["41603995"],"is_preprint":false},{"year":2026,"finding":"ZDHHC4 catalyzes palmitoylation of the cargo receptor CCDC50; the natural compound lactucopicrin directly binds His72 of ZDHHC4 to boost its enzymatic activity, enhancing CCDC50 palmitoylation and thereby promoting selective autophagic degradation of MAP2K4/MKK4, suppressing MAPK/JNK signaling and chondrocyte senescence in osteoarthritis.","method":"Acyl-biotin exchange palmitoylation assay, structural binding analysis (DARTS), mutagenesis (His72), autophagy assays, in vivo OA mouse model","journal":"Autophagy","confidence":"High","confidence_rationale":"Tier 1-2 — direct ligand-binding structural evidence, active-site residue mutagenesis, substrate identification, in vivo functional validation with multiple orthogonal methods","pmids":["41566717"],"is_preprint":false}],"current_model":"ZDHHC4 is a DHHC-family palmitoyl acyltransferase that S-palmitoylates multiple substrates—including MAVS (Cys79), GSK3β (Cys14), TRPV1 (C157/362/390/715), ZEB-2 (C478), CD36, KAI1, NFATC4, CCDC50, and SARS-CoV-2 S protein—at the ER membrane, and is recruited to substrates by adaptor proteins (e.g., CPT1A for MAVS) or regulated transcriptionally by FoxO1 and ATF3, with palmitoylation outcomes that control substrate stability (via ubiquitination), subcellular trafficking, channel activity, and downstream signaling in contexts ranging from innate immunity and pain to cardiac metabolism, neurodegeneration, and cancer."},"narrative":{"teleology":[{"year":2015,"claim":"Establishing ZDHHC4 as a functional palmitoyl acyltransferase: a MYTH screen identified ZDHHC4 as a PAT that interacts with and palmitoylates the D2 dopamine receptor, affecting D2R stability and surface expression—the first direct evidence linking ZDHHC4 to substrate palmitoylation.","evidence":"Membrane yeast two-hybrid screen, Co-IP, click chemistry palmitoylation assay, cysteine mutagenesis in heterologous cells","pmids":["26535572"],"confidence":"Medium","gaps":["Single lab study without independent replication","Specific palmitoylated cysteine(s) on D2R not mapped","No in vivo validation of D2R palmitoylation by ZDHHC4"]},{"year":2021,"claim":"Extending ZDHHC4 substrates to membrane proteins with distinct biological outputs: ZDHHC4 was shown to palmitoylate KAI1/CD82, controlling its membrane localization and enabling anti-angiogenic LIF induction via Src/p53, and separately to palmitoylate the SARS-CoV-2 spike protein to facilitate viral syncytia formation and entry.","evidence":"Palmitoylation assays, loss-of-function experiments, in vivo angiogenesis models (KAI1); ZDHHC overexpression panel, pseudovirus entry and syncytia assays (spike protein)","pmids":["34530889","34528721"],"confidence":"Medium","gaps":["For SARS-CoV-2 spike, ZDHHC4 was not individually dissected from other ZDHHC family members","Palmitoylation site(s) on KAI1 not mapped at residue level","No structural basis for substrate selectivity"]},{"year":2022,"claim":"Defining ZDHHC4 as a signaling-modulating enzyme through site-specific palmitoylation: ZDHHC4 palmitoylates GSK3β at Cys14 to shift its phosphorylation state and activate EZH2–STAT3 in GBM, and palmitoylates NFATC4 to enable its cytoplasm-to-nucleus trafficking, linking ZDHHC4 to both cancer stemness and lysosomal Ca²⁺ homeostasis in neurodegeneration.","evidence":"Cys14 mutagenesis, phosphorylation analysis, stemness/drug-resistance readouts (GSK3β); subcellular fractionation, Ppt1-KO mice, rescue experiments (NFATC4)","pmids":["35606353","35150145"],"confidence":"Medium","gaps":["ZDHHC4 versus ZDHHC8 individual contributions to NFATC4 palmitoylation not fully resolved","In vivo GBM model validation of ZDHHC4 dependence lacking","Structural basis for Cys14 selectivity on GSK3β unknown"]},{"year":2023,"claim":"Revealing an adaptor-dependent recruitment mechanism: CPT1A recruits ER-localized ZDHHC4 to palmitoylate MAVS at Cys79, switching ubiquitination from K48- to K63-linked chains and thereby stabilizing MAVS to amplify type I interferon signaling—the first demonstration that ZDHHC4 substrate access can be controlled by an adaptor protein.","evidence":"Reciprocal Co-IP, Cys79 mutagenesis, ubiquitination assays, IFN-I functional readout","pmids":["38016475"],"confidence":"High","gaps":["Whether CPT1A-mediated recruitment is a general mechanism for other ZDHHC4 substrates is untested","Structural details of CPT1A–ZDHHC4 interaction unresolved"]},{"year":2024,"claim":"Demonstrating that ZDHHC4 controls ion channel turnover and pain: ZDHHC4 palmitoylates TRPV1 at four cysteines to promote its lysosomal degradation, counterbalanced by depalmitoylase APT1, establishing a palmitoylation–depalmitoylation cycle that regulates inflammatory pain in vivo.","evidence":"Co-IP, quadruple-cysteine mutagenesis, acyl-biotin exchange, electrophysiology, in vivo pain behavior assays","pmids":["39528731"],"confidence":"High","gaps":["Whether palmitoylation at each individual cysteine contributes equally to TRPV1 degradation is unresolved","Mechanism by which palmitoylation targets TRPV1 to lysosomes not defined"]},{"year":2024,"claim":"Establishing transcriptional regulation of ZDHHC4 as a metabolic control point: FoxO1 transcriptionally upregulates ZDHHC4 in diabetic cardiomyocytes, and ATF3 does so in macrophages, with both converging on CD36 palmitoylation to drive its plasma membrane localization and fatty acid uptake, directly linking ZDHHC4 expression to metabolic disease.","evidence":"ChIP-seq/qPCR, luciferase assays, cardiomyocyte-specific FoxO1-KO mice, pharmacological ZDHHC inhibition (FoxO1); Atf3 overexpression/knockdown, CD36 cysteine mutagenesis, subcellular fractionation (ATF3)","pmids":["40357580","39047111"],"confidence":"High","gaps":["Whether FoxO1 and ATF3 regulate ZDHHC4 in a tissue-specific or generalizable manner is unknown","Relative contribution of ZDHHC4 versus ZDHHC5 to CD36 palmitoylation not individually quantified"]},{"year":2025,"claim":"Confirming ZDHHC4 as a hepatic regulator of CD36-dependent lipid uptake through a parallel pathway: EVA1A deficiency upregulates ZDHHC4/5 while repressing APT1, boosting CD36 palmitoylation and membrane localization to increase fatty acid uptake in hepatocytes.","evidence":"Hepatocyte-specific Eva1a-KO, transcriptomics, palmitoylation and subcellular fractionation assays","pmids":["41306774"],"confidence":"Medium","gaps":["ZDHHC4 and ZDHHC5 not individually separated in this context","Downstream metabolic phenotype not fully dissected for ZDHHC4 alone"]},{"year":2026,"claim":"Expanding ZDHHC4 to EMT regulation and selective autophagy: ZDHHC4 palmitoylates ZEB-2 at C478 to promote its deubiquitination and stabilization (driving melanoma EMT), and palmitoylates CCDC50 to target MAP2K4 for autophagic degradation; the natural compound lactucopicrin directly binds His72 of ZDHHC4 to enhance its enzymatic activity.","evidence":"Mass spectrometry, Co-IP, C478 mutagenesis, EMT readout (ZEB-2); DARTS binding assay, His72 mutagenesis, in vivo osteoarthritis model (CCDC50/lactucopicrin)","pmids":["41603995","41566717"],"confidence":"High","gaps":["Whether lactucopicrin-mediated activation of ZDHHC4 is specific or affects other ZDHHCs is not addressed","Structural basis of His72 in ZDHHC4 catalytic mechanism not fully elucidated"]},{"year":null,"claim":"Unresolved: the structural basis for ZDHHC4 substrate selectivity, how adaptor recruitment generalizes across substrates, and whether ZDHHC4 has substrates beyond those already identified that explain its broad biological footprint.","evidence":"","pmids":[],"confidence":"Low","gaps":["No crystal or cryo-EM structure of ZDHHC4","No systematic unbiased palmitoyl-proteomics to define the full ZDHHC4 substrate repertoire","Catalytic mechanism and role of His72 versus the DHHC motif not structurally resolved"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[0,1,2,4,5,6,7,8,10,11]}],"localization":[{"term_id":"GO:0005783","term_label":"endoplasmic reticulum","supporting_discovery_ids":[0]}],"pathway":[{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[0]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[4,5]},{"term_id":"R-HSA-1430728","term_label":"Metabolism","supporting_discovery_ids":[7,8,9]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[0,4,6,10,11]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[11]}],"complexes":[],"partners":["CPT1A","MAVS","TRPV1","GSK3B","CD36","CCDC50","ZEB2","NFATC4"],"other_free_text":[]},"mechanistic_narrative":"ZDHHC4 is an ER-resident DHHC-family palmitoyl acyltransferase that S-palmitoylates a diverse array of substrates to control their stability, trafficking, and downstream signaling across innate immunity, pain sensation, cardiac metabolism, neurodegeneration, and cancer. It palmitoylates MAVS at Cys79 (recruited by CPT1A), stabilizing MAVS by switching ubiquitination from K48- to K63-linked chains to potentiate type I interferon signaling [PMID:38016475]; palmitoylates TRPV1 at multiple cysteines to promote its lysosomal degradation and attenuate inflammatory pain [PMID:39528731]; and palmitoylates GSK3β at Cys14 to activate EZH2–STAT3 signaling in glioblastoma stem cells [PMID:35606353]. ZDHHC4 also palmitoylates CD36 to drive its sarcolemmal localization and fatty acid uptake—a process transcriptionally upregulated by FoxO1 in the diabetic heart and by ATF3 in macrophages [PMID:40357580, PMID:39047111]—and palmitoylates ZEB-2, NFATC4, KAI1, and CCDC50, linking its activity to EMT, nuclear transcription factor trafficking, anti-angiogenesis, and selective autophagy [PMID:41603995, PMID:35150145, PMID:34530889, PMID:41566717]."},"prefetch_data":{"uniprot":{"accession":"Q9NPG8","full_name":"Palmitoyltransferase ZDHHC4","aliases":["Zinc finger DHHC domain-containing protein 4","DHHC-4","Zinc finger protein 374"],"length_aa":344,"mass_kda":39.8,"function":"Palmitoyltransferase that catalyzes the addition of palmitate onto protein substrates including the D(2) dopamine receptor DRD2, GSK3B or MAVS. Mediates GSK3B palmitoylation to prevent its AKT1-mediated phosphorylation leading to activation of the STAT3 signaling pathway (PubMed:35606353). Also catalyzes MAVS palmitoylation which promotes its stabilization and activation by inhibiting 'Lys-48'- but facilitating 'Lys-63'-linked ubiquitination (PubMed:38016475)","subcellular_location":"Endoplasmic reticulum membrane; Golgi apparatus membrane; Cell membrane","url":"https://www.uniprot.org/uniprotkb/Q9NPG8/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/ZDHHC4","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/ZDHHC4","total_profiled":1310},"omim":[{"mim_id":"621547","title":"ZDHHC PALMITOYLTRANSFERASE 4; ZDHHC4","url":"https://www.omim.org/entry/621547"},{"mim_id":"617150","title":"ZDHHC PALMITOYLTRANSFERASE 3; ZDHHC3","url":"https://www.omim.org/entry/617150"},{"mim_id":"608784","title":"ZDHHC PALMITOYLTRANSFERASE 8; ZDHHC8","url":"https://www.omim.org/entry/608784"},{"mim_id":"605599","title":"LYSOPHOSPHOLIPASE I; LYPLA1","url":"https://www.omim.org/entry/605599"},{"mim_id":"605004","title":"GLYCOGEN SYNTHASE KINASE 3-BETA; GSK3B","url":"https://www.omim.org/entry/605004"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/ZDHHC4"},"hgnc":{"alias_symbol":["FLJ10479","ZNF374"],"prev_symbol":[]},"alphafold":{"accession":"Q9NPG8","domains":[{"cath_id":"-","chopping":"72-122_191-296_319-332","consensus_level":"medium","plddt":89.0337,"start":72,"end":332},{"cath_id":"-","chopping":"125-173","consensus_level":"medium","plddt":94.4065,"start":125,"end":173}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NPG8","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NPG8-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NPG8-F1-predicted_aligned_error_v6.png","plddt_mean":84.0},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=ZDHHC4","jax_strain_url":"https://www.jax.org/strain/search?query=ZDHHC4"},"sequence":{"accession":"Q9NPG8","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9NPG8.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9NPG8/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NPG8"}},"corpus_meta":[{"pmid":"38016475","id":"PMC_38016475","title":"CPT1A induction following epigenetic perturbation promotes MAVS palmitoylation and activation to potentiate antitumor immunity.","date":"2023","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/38016475","citation_count":51,"is_preprint":false},{"pmid":"34528721","id":"PMC_34528721","title":"Palmitoylation of SARS-CoV-2 S protein is critical for S-mediated syncytia formation and virus entry.","date":"2021","source":"Journal of medical virology","url":"https://pubmed.ncbi.nlm.nih.gov/34528721","citation_count":49,"is_preprint":false},{"pmid":"26535572","id":"PMC_26535572","title":"Effect of C-Terminal S-Palmitoylation on D2 Dopamine Receptor Trafficking and Stability.","date":"2015","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/26535572","citation_count":42,"is_preprint":false},{"pmid":"34530889","id":"PMC_34530889","title":"KAI1(CD82) is a key molecule to control angiogenesis and switch angiogenic milieu to quiescent state.","date":"2021","source":"Journal of hematology & oncology","url":"https://pubmed.ncbi.nlm.nih.gov/34530889","citation_count":40,"is_preprint":false},{"pmid":"39047111","id":"PMC_39047111","title":"Atf3-mediated metabolic reprogramming in hepatic macrophage orchestrates metabolic dysfunction-associated steatohepatitis.","date":"2024","source":"Science advances","url":"https://pubmed.ncbi.nlm.nih.gov/39047111","citation_count":28,"is_preprint":false},{"pmid":"35606353","id":"PMC_35606353","title":"GSK3β palmitoylation mediated by ZDHHC4 promotes tumorigenicity of glioblastoma stem cells in temozolomide-resistant glioblastoma through the EZH2-STAT3 axis.","date":"2022","source":"Oncogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/35606353","citation_count":25,"is_preprint":false},{"pmid":"27956064","id":"PMC_27956064","title":"Novel genes in brain tissues of EAE-induced normal and obese mice: Upregulation of metal ion-binding protein genes in obese-EAE mice.","date":"2016","source":"Neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/27956064","citation_count":20,"is_preprint":false},{"pmid":"35150145","id":"PMC_35150145","title":"Ppt1-deficiency dysregulates lysosomal Ca++ homeostasis contributing to pathogenesis in a mouse model of CLN1 disease.","date":"2022","source":"Journal of inherited metabolic disease","url":"https://pubmed.ncbi.nlm.nih.gov/35150145","citation_count":16,"is_preprint":false},{"pmid":"39563863","id":"PMC_39563863","title":"Research progress on S-palmitoylation modification mediated by the ZDHHC family in glioblastoma.","date":"2024","source":"Frontiers in cell and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/39563863","citation_count":8,"is_preprint":false},{"pmid":"40357580","id":"PMC_40357580","title":"FoxO1-zDHHC4-CD36 S-Acylation Axis Drives Metabolic Dysfunction in Diabetes.","date":"2025","source":"Circulation research","url":"https://pubmed.ncbi.nlm.nih.gov/40357580","citation_count":7,"is_preprint":false},{"pmid":"39528731","id":"PMC_39528731","title":"Palmitoylation by ZDHHC4 inhibits TRPV1-mediated nociception.","date":"2024","source":"EMBO reports","url":"https://pubmed.ncbi.nlm.nih.gov/39528731","citation_count":4,"is_preprint":false},{"pmid":"41306774","id":"PMC_41306774","title":"EVA1A Regulates Hepatic Lipid Homeostasis by Modulating CD36 Expression and Its Palmitoylation.","date":"2025","source":"Research (Washington, D.C.)","url":"https://pubmed.ncbi.nlm.nih.gov/41306774","citation_count":1,"is_preprint":false},{"pmid":"41603995","id":"PMC_41603995","title":"ZDHHC4-mediated ZEB-2 S-palmitoylation promotes melanoma epithelial-mesenchymal transition via deubiquitinating and stabilizing ZEB-2.","date":"2026","source":"Cellular and molecular life sciences : CMLS","url":"https://pubmed.ncbi.nlm.nih.gov/41603995","citation_count":0,"is_preprint":false},{"pmid":"40753321","id":"PMC_40753321","title":"ZDHHC4 Influences Obsessive-Compulsive Disorder Risk Through Imaging-Derived Phenotypes: A Mendelian Randomization Study.","date":"2025","source":"Journal of molecular neuroscience : MN","url":"https://pubmed.ncbi.nlm.nih.gov/40753321","citation_count":0,"is_preprint":false},{"pmid":"41049004","id":"PMC_41049004","title":"Inflammatory Bowel Disease Mediates the Causal Relationship Between Gut Microbiota and Colorectal Cancer: Identification of Therapeutic Targets and Predictive Modeling.","date":"2025","source":"Journal of Cancer","url":"https://pubmed.ncbi.nlm.nih.gov/41049004","citation_count":0,"is_preprint":false},{"pmid":"41566717","id":"PMC_41566717","title":"Lactucopicrin promotes the autophagic degradation of MAP2K4/MKK4 by mediating CCDC50 palmitoylation to alleviate osteoarthritis progression.","date":"2026","source":"Autophagy","url":"https://pubmed.ncbi.nlm.nih.gov/41566717","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":10177,"output_tokens":3068,"usd":0.038275},"stage2":{"model":"claude-opus-4-6","input_tokens":6432,"output_tokens":2992,"usd":0.16044},"total_usd":0.198715,"stage1_batch_id":"msgbatch_01BparvgRZ9S2LMidvaoo86K","stage2_batch_id":"msgbatch_01NkBfv9SVZWthhS7j6mU6G3","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2023,\n      \"finding\": \"CPT1A recruits ER-localized ZDHHC4 to catalyze palmitoylation of MAVS at Cys79, which promotes MAVS stabilization and activation by inhibiting K48-linked ubiquitination and facilitating K63-linked ubiquitination, thereby potentiating the type I interferon response.\",\n      \"method\": \"Co-immunoprecipitation, site-directed mutagenesis (Cys79), ubiquitination assays, loss-of-function experiments with defined IFN-I readout\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal Co-IP, specific residue mutagenesis, mechanistic pathway placement with multiple orthogonal assays\",\n      \"pmids\": [\"38016475\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"ZDHHC4 was identified as a palmitoyl acyltransferase (PAT) that physically interacts with the D2 dopamine receptor (D2R) and affects its palmitoylation status, contributing to D2R stability and plasma membrane expression.\",\n      \"method\": \"Membrane yeast two-hybrid (MYTH) screen, co-immunoprecipitation, bioorthogonal click chemistry palmitoylation assay, cysteine mutagenesis\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal Co-IP and palmitoylation assay, but single lab study\",\n      \"pmids\": [\"26535572\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"ZDHHC4 catalyzes palmitoylation of KAI1 (CD82) at the membrane surface, which is required for KAI1 localization to the pericyte membrane surface and its downstream induction of LIF through the Src/p53 pathway to inhibit angiogenesis.\",\n      \"method\": \"Palmitoylation assay, loss-of-function experiments, in vitro and in vivo angiogenesis assays\",\n      \"journal\": \"Journal of hematology & oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — defined substrate and functional consequence, but single lab\",\n      \"pmids\": [\"34530889\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"ZDHHC4 (among other ZDHHCs) promotes palmitoylation of the SARS-CoV-2 S protein and physically associates with S protein; this palmitoylation is critical for S-mediated syncytia formation and pseudovirus particle entry.\",\n      \"method\": \"Overexpression of individual ZDHHCs, palmitoylation assay, co-immunoprecipitation, pseudovirus entry assay, syncytia formation assay\",\n      \"journal\": \"Journal of medical virology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 — functional assays with specific palmitoylation readout, but ZDHHC4 is one of many ZDHHCs tested without unique mechanistic dissection\",\n      \"pmids\": [\"34528721\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"ZDHHC4 palmitoylates GSK3β at Cys14, which decreases phospho-Ser9 and increases phospho-Tyr216, thereby activating the EZH2-STAT3 signaling axis and promoting GBM stem cell self-renewal and temozolomide resistance.\",\n      \"method\": \"Palmitoylation assay, site-directed mutagenesis (Cys14), phosphorylation analysis, knockdown experiments with stemness and drug-resistance readouts, STAT3 feedback loop validation\",\n      \"journal\": \"Oncogenesis\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — specific palmitoylation site identified by mutagenesis, pathway placement with multiple downstream readouts, single lab\",\n      \"pmids\": [\"35606353\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"ZDHHC4 and ZDHHC8 catalyze S-palmitoylation of the transcription factor NFATC4, which is required for NFATC4 trafficking from the cytoplasm to the nucleus; reduced ZDHHC4 and ZDHHC8 levels in Ppt1-deficient mice lower nuclear palmitoylated NFATC4 and thereby suppress IP3R1 expression, dysregulating lysosomal Ca2+ homeostasis.\",\n      \"method\": \"Palmitoylation assay, subcellular fractionation, mouse knockout model (Ppt1-/-), IP3R1 overexpression rescue, identification of responsible ZDHHC enzymes\",\n      \"journal\": \"Journal of inherited metabolic disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — mechanistic pathway placement with loss-of-function and rescue, multiple methods, single lab\",\n      \"pmids\": [\"35150145\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"ZDHHC4 physically interacts with TRPV1 and catalyzes S-palmitoylation at cysteine residues C157, C362, C390, and C715 of TRPV1, promoting TRPV1 degradation via the lysosome pathway and thereby facilitating inflammatory pain relief; this palmitoylation is counterbalanced by depalmitoylase APT1.\",\n      \"method\": \"Co-immunoprecipitation, site-directed mutagenesis (C157, C362, C390, C715), acyl-biotin exchange palmitoylation assay, electrophysiology, in vivo pain behavior assays\",\n      \"journal\": \"EMBO reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — multiple orthogonal methods (Co-IP, mutagenesis of specific residues, functional electrophysiology, in vivo readout) in a single study\",\n      \"pmids\": [\"39528731\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"ATF3 in macrophages increases ZDHHC4/5-mediated CD36 palmitoylation at C3, C7, C464, and C466, which promotes CD36 plasma membrane localization and fatty acid uptake.\",\n      \"method\": \"Overexpression/knockdown of Atf3, palmitoylation assay, site-directed mutagenesis of CD36 cysteines, subcellular fractionation\",\n      \"journal\": \"Science advances\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — specific palmitoylation sites identified, functional localization consequence shown; ZDHHC4 and ZDHHC5 not separated individually\",\n      \"pmids\": [\"39047111\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"FoxO1 transcriptionally upregulates zDHHC4 in the diabetic heart; zDHHC4 S-acylates CD36, promoting its sarcolemmal localization, increased fatty acid oxidation, and triglyceride storage, driving metabolic dysfunction; genetic silencing of zDHHC4 decreases CD36 S-acylation and ameliorates the diabetic cardiac phenotype.\",\n      \"method\": \"ChIP sequencing, ChIP-qPCR, luciferase assays, siRNA/shRNA silencing, cardiomyocyte-specific FoxO1 knockout mice, pharmacological ZDHHC inhibition with functional cardiac readouts\",\n      \"journal\": \"Circulation research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods (ChIP-seq, genetic silencing, KO mice, pharmacology) with defined metabolic and functional phenotypes, conserved across species\",\n      \"pmids\": [\"40357580\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"EVA1A deficiency transcriptionally enhances palmitoyl acyltransferases ZDHHC4/5 while repressing depalmitoylase APT1, boosting CD36 palmitoylation and plasma membrane localization to increase fatty acid uptake and impair β-oxidation in hepatocytes.\",\n      \"method\": \"Hepatocyte-specific knockout of Eva1a, transcriptomics, palmitoylation assay, subcellular fractionation, CD36 localization assays\",\n      \"journal\": \"Research (Washington, D.C.)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — functional KO with mechanistic follow-up, but ZDHHC4 and ZDHHC5 not individually dissected\",\n      \"pmids\": [\"41306774\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"ZDHHC4 palmitoylates ZEB-2 at C478; this palmitoylation promotes ZEB-2 deubiquitination and protein stability, facilitating EMT in melanoma cells; ZEB-2 interacts with ZDHHC4 through its N-terminal sequences.\",\n      \"method\": \"Mass spectrometry substrate identification, co-immunoprecipitation, site-directed mutagenesis (C478), ubiquitination assay, knockdown with EMT readout\",\n      \"journal\": \"Cellular and molecular life sciences : CMLS\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — specific palmitoylation site identified by mutagenesis, mechanistic pathway placement, single lab\",\n      \"pmids\": [\"41603995\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"ZDHHC4 catalyzes palmitoylation of the cargo receptor CCDC50; the natural compound lactucopicrin directly binds His72 of ZDHHC4 to boost its enzymatic activity, enhancing CCDC50 palmitoylation and thereby promoting selective autophagic degradation of MAP2K4/MKK4, suppressing MAPK/JNK signaling and chondrocyte senescence in osteoarthritis.\",\n      \"method\": \"Acyl-biotin exchange palmitoylation assay, structural binding analysis (DARTS), mutagenesis (His72), autophagy assays, in vivo OA mouse model\",\n      \"journal\": \"Autophagy\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — direct ligand-binding structural evidence, active-site residue mutagenesis, substrate identification, in vivo functional validation with multiple orthogonal methods\",\n      \"pmids\": [\"41566717\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"ZDHHC4 is a DHHC-family palmitoyl acyltransferase that S-palmitoylates multiple substrates—including MAVS (Cys79), GSK3β (Cys14), TRPV1 (C157/362/390/715), ZEB-2 (C478), CD36, KAI1, NFATC4, CCDC50, and SARS-CoV-2 S protein—at the ER membrane, and is recruited to substrates by adaptor proteins (e.g., CPT1A for MAVS) or regulated transcriptionally by FoxO1 and ATF3, with palmitoylation outcomes that control substrate stability (via ubiquitination), subcellular trafficking, channel activity, and downstream signaling in contexts ranging from innate immunity and pain to cardiac metabolism, neurodegeneration, and cancer.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"ZDHHC4 is an ER-resident DHHC-family palmitoyl acyltransferase that S-palmitoylates a diverse array of substrates to control their stability, trafficking, and downstream signaling across innate immunity, pain sensation, cardiac metabolism, neurodegeneration, and cancer. It palmitoylates MAVS at Cys79 (recruited by CPT1A), stabilizing MAVS by switching ubiquitination from K48- to K63-linked chains to potentiate type I interferon signaling [PMID:38016475]; palmitoylates TRPV1 at multiple cysteines to promote its lysosomal degradation and attenuate inflammatory pain [PMID:39528731]; and palmitoylates GSK3β at Cys14 to activate EZH2–STAT3 signaling in glioblastoma stem cells [PMID:35606353]. ZDHHC4 also palmitoylates CD36 to drive its sarcolemmal localization and fatty acid uptake—a process transcriptionally upregulated by FoxO1 in the diabetic heart and by ATF3 in macrophages [PMID:40357580, PMID:39047111]—and palmitoylates ZEB-2, NFATC4, KAI1, and CCDC50, linking its activity to EMT, nuclear transcription factor trafficking, anti-angiogenesis, and selective autophagy [PMID:41603995, PMID:35150145, PMID:34530889, PMID:41566717].\",\n  \"teleology\": [\n    {\n      \"year\": 2015,\n      \"claim\": \"Establishing ZDHHC4 as a functional palmitoyl acyltransferase: a MYTH screen identified ZDHHC4 as a PAT that interacts with and palmitoylates the D2 dopamine receptor, affecting D2R stability and surface expression—the first direct evidence linking ZDHHC4 to substrate palmitoylation.\",\n      \"evidence\": \"Membrane yeast two-hybrid screen, Co-IP, click chemistry palmitoylation assay, cysteine mutagenesis in heterologous cells\",\n      \"pmids\": [\"26535572\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab study without independent replication\", \"Specific palmitoylated cysteine(s) on D2R not mapped\", \"No in vivo validation of D2R palmitoylation by ZDHHC4\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Extending ZDHHC4 substrates to membrane proteins with distinct biological outputs: ZDHHC4 was shown to palmitoylate KAI1/CD82, controlling its membrane localization and enabling anti-angiogenic LIF induction via Src/p53, and separately to palmitoylate the SARS-CoV-2 spike protein to facilitate viral syncytia formation and entry.\",\n      \"evidence\": \"Palmitoylation assays, loss-of-function experiments, in vivo angiogenesis models (KAI1); ZDHHC overexpression panel, pseudovirus entry and syncytia assays (spike protein)\",\n      \"pmids\": [\"34530889\", \"34528721\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"For SARS-CoV-2 spike, ZDHHC4 was not individually dissected from other ZDHHC family members\", \"Palmitoylation site(s) on KAI1 not mapped at residue level\", \"No structural basis for substrate selectivity\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Defining ZDHHC4 as a signaling-modulating enzyme through site-specific palmitoylation: ZDHHC4 palmitoylates GSK3β at Cys14 to shift its phosphorylation state and activate EZH2–STAT3 in GBM, and palmitoylates NFATC4 to enable its cytoplasm-to-nucleus trafficking, linking ZDHHC4 to both cancer stemness and lysosomal Ca²⁺ homeostasis in neurodegeneration.\",\n      \"evidence\": \"Cys14 mutagenesis, phosphorylation analysis, stemness/drug-resistance readouts (GSK3β); subcellular fractionation, Ppt1-KO mice, rescue experiments (NFATC4)\",\n      \"pmids\": [\"35606353\", \"35150145\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"ZDHHC4 versus ZDHHC8 individual contributions to NFATC4 palmitoylation not fully resolved\", \"In vivo GBM model validation of ZDHHC4 dependence lacking\", \"Structural basis for Cys14 selectivity on GSK3β unknown\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Revealing an adaptor-dependent recruitment mechanism: CPT1A recruits ER-localized ZDHHC4 to palmitoylate MAVS at Cys79, switching ubiquitination from K48- to K63-linked chains and thereby stabilizing MAVS to amplify type I interferon signaling—the first demonstration that ZDHHC4 substrate access can be controlled by an adaptor protein.\",\n      \"evidence\": \"Reciprocal Co-IP, Cys79 mutagenesis, ubiquitination assays, IFN-I functional readout\",\n      \"pmids\": [\"38016475\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether CPT1A-mediated recruitment is a general mechanism for other ZDHHC4 substrates is untested\", \"Structural details of CPT1A–ZDHHC4 interaction unresolved\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Demonstrating that ZDHHC4 controls ion channel turnover and pain: ZDHHC4 palmitoylates TRPV1 at four cysteines to promote its lysosomal degradation, counterbalanced by depalmitoylase APT1, establishing a palmitoylation–depalmitoylation cycle that regulates inflammatory pain in vivo.\",\n      \"evidence\": \"Co-IP, quadruple-cysteine mutagenesis, acyl-biotin exchange, electrophysiology, in vivo pain behavior assays\",\n      \"pmids\": [\"39528731\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether palmitoylation at each individual cysteine contributes equally to TRPV1 degradation is unresolved\", \"Mechanism by which palmitoylation targets TRPV1 to lysosomes not defined\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Establishing transcriptional regulation of ZDHHC4 as a metabolic control point: FoxO1 transcriptionally upregulates ZDHHC4 in diabetic cardiomyocytes, and ATF3 does so in macrophages, with both converging on CD36 palmitoylation to drive its plasma membrane localization and fatty acid uptake, directly linking ZDHHC4 expression to metabolic disease.\",\n      \"evidence\": \"ChIP-seq/qPCR, luciferase assays, cardiomyocyte-specific FoxO1-KO mice, pharmacological ZDHHC inhibition (FoxO1); Atf3 overexpression/knockdown, CD36 cysteine mutagenesis, subcellular fractionation (ATF3)\",\n      \"pmids\": [\"40357580\", \"39047111\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether FoxO1 and ATF3 regulate ZDHHC4 in a tissue-specific or generalizable manner is unknown\", \"Relative contribution of ZDHHC4 versus ZDHHC5 to CD36 palmitoylation not individually quantified\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Confirming ZDHHC4 as a hepatic regulator of CD36-dependent lipid uptake through a parallel pathway: EVA1A deficiency upregulates ZDHHC4/5 while repressing APT1, boosting CD36 palmitoylation and membrane localization to increase fatty acid uptake in hepatocytes.\",\n      \"evidence\": \"Hepatocyte-specific Eva1a-KO, transcriptomics, palmitoylation and subcellular fractionation assays\",\n      \"pmids\": [\"41306774\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"ZDHHC4 and ZDHHC5 not individually separated in this context\", \"Downstream metabolic phenotype not fully dissected for ZDHHC4 alone\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Expanding ZDHHC4 to EMT regulation and selective autophagy: ZDHHC4 palmitoylates ZEB-2 at C478 to promote its deubiquitination and stabilization (driving melanoma EMT), and palmitoylates CCDC50 to target MAP2K4 for autophagic degradation; the natural compound lactucopicrin directly binds His72 of ZDHHC4 to enhance its enzymatic activity.\",\n      \"evidence\": \"Mass spectrometry, Co-IP, C478 mutagenesis, EMT readout (ZEB-2); DARTS binding assay, His72 mutagenesis, in vivo osteoarthritis model (CCDC50/lactucopicrin)\",\n      \"pmids\": [\"41603995\", \"41566717\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether lactucopicrin-mediated activation of ZDHHC4 is specific or affects other ZDHHCs is not addressed\", \"Structural basis of His72 in ZDHHC4 catalytic mechanism not fully elucidated\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Unresolved: the structural basis for ZDHHC4 substrate selectivity, how adaptor recruitment generalizes across substrates, and whether ZDHHC4 has substrates beyond those already identified that explain its broad biological footprint.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No crystal or cryo-EM structure of ZDHHC4\", \"No systematic unbiased palmitoyl-proteomics to define the full ZDHHC4 substrate repertoire\", \"Catalytic mechanism and role of His72 versus the DHHC motif not structurally resolved\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [0, 1, 2, 4, 5, 6, 7, 8, 10, 11]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005783\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [4, 5]},\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [7, 8, 9]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [0, 4, 6, 10, 11]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [11]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"CPT1A\",\n      \"MAVS\",\n      \"TRPV1\",\n      \"GSK3B\",\n      \"CD36\",\n      \"CCDC50\",\n      \"ZEB2\",\n      \"NFATC4\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}