{"gene":"CYTL1","run_date":"2026-06-09T22:57:19","timeline":{"discoveries":[{"year":2007,"finding":"CYTL1 promotes chondrogenic differentiation of mouse limb bud mesenchymal cells acting as an autocrine factor, operating via stimulation of Sox9 transcriptional activity and induction of IGF-1 expression; exogenous CYTL1 protein or lentivirus-mediated overexpression both induced chondrogenesis in micromass culture.","method":"Exogenous protein treatment, lentiviral overexpression, micromass culture of mesenchymal cells, reporter assays for Sox9 activity, gene expression analysis","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (exogenous protein, genetic overexpression, reporter assay), replicated in vivo and in vitro, two independent approaches converging on same mechanism","pmids":["17644814"],"is_preprint":false},{"year":2011,"finding":"Cytl1 knockout mice show normal chondrogenesis, cartilage/bone development, and endochondral ossification, but exhibit augmented osteoarthritic cartilage destruction upon destabilization of the medial meniscus, establishing a role in cartilage homeostasis maintenance rather than development.","method":"Cytl1-/- knockout mouse generation, destabilization of medial meniscus OA model, histological and ultrastructural analysis of cartilage","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic knockout with defined in vivo phenotypic readout, complemented by human OA tissue expression data","pmids":["21652695"],"is_preprint":false},{"year":2011,"finding":"Computational structural modeling predicts CYTL1 adopts an IL8-like chemokine fold similar to CCL2 (MCP-1) rather than a 4-helical cytokine fold, and identifies structural features in CYTL1 necessary for signaling through the chemokine receptor CCR2.","method":"Molecular modeling, structure-based computational analysis, comparison with CCL2 3D structure","journal":"Proteins","confidence":"Low","confidence_rationale":"Tier 4 / Weak — computational prediction only, no experimental structural or binding validation performed","pmids":["21322034"],"is_preprint":false},{"year":2017,"finding":"Recombinant human CYTL1 produced in bacteria shows predominantly beta-sheet secondary structure (chemokine-like) by circular dichroism, promotes calcium flux in chondrocytes, but unlike chemokines shows limited affinity to proteoglycans.","method":"Recombinant protein production in bacteria, circular dichroism spectroscopy, calcium flux assay in chondrocytes, proteoglycan binding assay","journal":"Cytokine","confidence":"Medium","confidence_rationale":"Tier 1-2 / Moderate — in vitro biochemical assays with recombinant protein, multiple biophysical methods, single lab","pmids":["28478073"],"is_preprint":false},{"year":2019,"finding":"Recombinant CYTL1 chemoattracts human monocytes via the CCR2/ERK pathway, and CYTL1 (recombinant or overexpressed) inhibits tumor cell migration and invasion accompanied by decreased STAT3 phosphorylation, with inhibition of tumor metastasis confirmed in experimental and spontaneous breast cancer mouse models.","method":"Recombinant CYTL1 treatment, CYTL1-overexpressing tumor cell lines, migration/invasion assays, STAT3 phosphorylation measurement, experimental and spontaneous metastasis mouse models","journal":"Oncoimmunology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (recombinant protein, genetic overexpression, in vivo models), single lab","pmids":["31069137"],"is_preprint":false},{"year":2016,"finding":"CYTL1 expression in mouse endometrium is regulated by ovarian hormones and peaks during embryo implantation; CYTL1 enhances proliferation of endometrial cells, stimulates endometrial secretion of LIF and HB-EGF, and enhances adhesion of endometrial cells to JAR spheroids in cell-cell adhesion assays.","method":"Mouse endometrial expression analysis (mRNA and protein), cell proliferation assays, ELISA for LIF and HB-EGF secretion, cell-cell adhesion assays with HEC-1-A/RL95-2 and JAR spheroids","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2-3 / Moderate — multiple functional assays in cell culture with defined readouts, single lab","pmids":["26800213"],"is_preprint":false},{"year":2020,"finding":"CYTL1 promotes chemotactic activity and phagocytosis of E. coli by LPS-stimulated neutrophils through CCR2-mediated action and phosphorylation of Akt (protein kinase B), and increases release of reactive oxygen species in activated neutrophils.","method":"In vivo and in vitro neutrophil phagocytosis assays, western blotting for Akt phosphorylation, chemotaxis assays, ROS measurement, LPS-stimulated neutrophil model","journal":"Inflammation","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo and in vitro assays, multiple functional readouts, single lab","pmids":["31823178"],"is_preprint":false},{"year":2020,"finding":"Recombinant CYTL1 (rCytl1) increases expression of Sox9 and Col2α1 while stabilizing Col1α1 in cultured chondrocytes, promoting redifferentiation; Cytl1 is expressed at all stages of cartilage development and shares expression patterns with other chondrogenic factors.","method":"Recombinant protein treatment of cultured chondrocytes, quantitative RT-PCR for chondrogenic markers, embryonic and adult cartilage expression analysis","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2-3 / Moderate — recombinant protein functional assay with multiple markers, single lab","pmids":["32736681"],"is_preprint":false},{"year":2022,"finding":"Intracellular CYTL1 (lacking signal peptide, ΔCYTL1) competitively binds the N-terminal sequence of NDUFV1 to block MDM2-mediated proteasomal degradation of NDUFV1, thereby stabilizing NDUFV1; stabilized NDUFV1 increases NAD+ levels and interacts with Src to attenuate LDHA phosphorylation at tyrosine 10, reducing lactate production and glycolytic reprogramming in breast cancer.","method":"ΔCYTL1 overexpression (signal-peptide-deleted construct), Co-IP/pulldown for CYTL1-NDUFV1 interaction, MDM2 ubiquitination/proteasome degradation assays, glucose uptake/lactate production assays, NDUFV1-Src interaction, LDHA phosphorylation assays, in vitro and in vivo tumor growth/metastasis assays","journal":"Signal transduction and targeted therapy","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — multiple orthogonal mechanistic methods (protein interaction, ubiquitination/degradation assay, phosphorylation assay, metabolic readouts, in vivo validation), single lab with comprehensive mechanistic follow-up","pmids":["35115484"],"is_preprint":false},{"year":2024,"finding":"CYTL1 activates Nrf2 by promoting autophagic degradation of Keap1; Nrf2 subsequently transactivates CBS (cystathionine β-synthase) expression, supporting de novo cysteine synthesis via the transsulfuration pathway; breast cancer cells with low CYTL1 have suppressed CBS activity and are hypersensitive to system xc- blockade-induced ferroptosis.","method":"CYTL1 knockdown/overexpression in breast cancer cells, autophagy flux assays for Keap1 degradation, Nrf2 reporter/target gene assays, CBS expression and activity measurement, system xc- inhibitor (ferroptosis) assays in vitro and in vivo, CBS silencing rescue experiment","journal":"Redox biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (genetic manipulation, pathway rescue, in vivo validation), mechanistic chain experimentally established across multiple steps","pmids":["38211443"],"is_preprint":false},{"year":2024,"finding":"CYTL1 regulates TGF-β/CCN2 (connective tissue growth factor) axis to promote M2 macrophage polarization; CYTL1 ablation reduces M2 macrophage polarization and alleviates bleomycin-induced lung injury and fibrosis in mice, and inhibition of the TGF-β pathway phenocopies CYTL1 ablation.","method":"CYTL1 knockdown/ablation in bleomycin-induced lung fibrosis mouse model, macrophage polarization assays (M2 markers), TGF-β pathway inhibition, CCN2 expression analysis","journal":"Clinical and experimental pharmacology & physiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo genetic ablation with defined cellular and molecular readouts, pathway inhibitor confirmation, single lab","pmids":["39103233"],"is_preprint":false},{"year":2024,"finding":"In neuro-endothelial cells (NECs) of resistance arterioles, Cytl1 mediates vasodilation and increased blood flow acutely, and promotes capillary density and clonal EC remodeling chronically; silencing NEC Cytl1 gene expression causes arteriolar vasoconstriction, reduced vascular density and organ blood flow, increased vascular permeability and immune cell homing.","method":"Intravital multiphoton imaging of optogenetic mouse models with NEC gain/loss-of-function, Cytl1 in vivo silencing, Cytl1 protein administration, single-cell transcriptome analysis","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct in vivo functional imaging with gain/loss-of-function, preprint not yet peer-reviewed","pmids":["bio_10.1101_2024.09.30.615824"],"is_preprint":true}],"current_model":"CYTL1 is a small secreted cytokine-like protein that acts as an autocrine chondrogenic factor (stimulating Sox9, IGF-1, Col2α1) and cartilage homeostasis factor (loss accelerates OA), signals through CCR2/ERK to chemoattract monocytes and activate neutrophils via Akt phosphorylation, and intracellularly (as a signal-peptide-lacking isoform) suppresses glycolytic reprogramming in breast cancer by competitively binding NDUFV1 to block MDM2-mediated degradation and by activating Nrf2 through autophagic Keap1 degradation to support CBS-dependent cysteine synthesis; it also modulates TGF-β/CCN2-driven macrophage polarization in fibrosis, and in vascular neuro-endothelial cells mediates Nos1-dependent vasodilation and angiogenic remodeling."},"narrative":{"mechanistic_narrative":"CYTL1 is a small secreted chemokine-like protein that functions in cartilage homeostasis, immune cell chemotaxis, and tumor metabolic regulation [PMID:17644814, PMID:21652695, PMID:35115484]. As an autocrine factor it promotes chondrogenic differentiation by stimulating Sox9 transcriptional activity and inducing IGF-1, and it drives chondrocyte redifferentiation through induction of Sox9 and Col2α1 [PMID:17644814, PMID:32736681]; while dispensable for cartilage and bone development, CYTL1 maintains adult cartilage homeostasis, as its loss accelerates osteoarthritic cartilage destruction after joint destabilization [PMID:21652695]. Biophysically the secreted protein adopts a predominantly beta-sheet, chemokine-like fold and elicits calcium flux in chondrocytes [PMID:28478073], and it signals through the chemokine receptor CCR2 to chemoattract monocytes via ERK and to activate neutrophil chemotaxis, phagocytosis, and ROS release via Akt phosphorylation [PMID:31069137, PMID:31823178]. A signal-peptide-lacking intracellular isoform (ΔCYTL1) suppresses glycolytic reprogramming in breast cancer by competitively binding the N-terminus of NDUFV1 to block MDM2-mediated proteasomal degradation, stabilizing NDUFV1 to raise NAD+ and attenuate Src-dependent LDHA phosphorylation [PMID:35115484]; CYTL1 additionally activates Nrf2 through autophagic Keap1 degradation, transactivating CBS to support transsulfuration-dependent cysteine synthesis and protect cells from ferroptosis [PMID:38211443]. CYTL1 further modulates the TGF-β/CCN2 axis to promote M2 macrophage polarization in lung fibrosis [PMID:39103233].","teleology":[{"year":2007,"claim":"Established CYTL1 as a functional autocrine chondrogenic factor rather than an orphan cytokine-like sequence, defining its first molecular activity.","evidence":"Exogenous protein and lentiviral overexpression in mesenchymal micromass culture with Sox9 reporter and IGF-1 expression readouts","pmids":["17644814"],"confidence":"High","gaps":["Receptor mediating the chondrogenic signal not identified","Mechanism linking CYTL1 to Sox9 activation unresolved"]},{"year":2011,"claim":"Distinguished CYTL1's developmental dispensability from its homeostatic role, showing it protects mature cartilage rather than building it.","evidence":"Cytl1-/- knockout mice analyzed in a destabilization of the medial meniscus osteoarthritis model with histology","pmids":["21652695"],"confidence":"High","gaps":["Molecular pathway protecting cartilage not defined","No connection to a receptor or downstream effector established"]},{"year":2011,"claim":"Proposed a structural basis for receptor engagement by predicting an IL8/CCL2-like chemokine fold and CCR2-relevant features.","evidence":"Computational molecular modeling compared with CCL2 structure","pmids":["21322034"],"confidence":"Low","gaps":["Computational prediction only, no experimental structure or binding validation","CCR2 binding not demonstrated experimentally here"]},{"year":2017,"claim":"Provided experimental biophysical support for a chemokine-like fold and demonstrated functional signaling output in chondrocytes.","evidence":"Bacterially produced recombinant CYTL1 analyzed by circular dichroism, calcium flux, and proteoglycan binding assays","pmids":["28478073"],"confidence":"Medium","gaps":["High-resolution structure absent","Limited proteoglycan affinity leaves signaling/localization mechanism unclear"]},{"year":2016,"claim":"Extended CYTL1 function to reproductive tissue, implicating it in endometrial receptivity and embryo implantation.","evidence":"Hormone-regulated endometrial expression analysis with proliferation, LIF/HB-EGF secretion, and spheroid adhesion assays","pmids":["26800213"],"confidence":"Medium","gaps":["Receptor and signaling pathway in endometrium not identified","Single-lab cell-based assays"]},{"year":2019,"claim":"Defined a CCR2/ERK signaling axis for monocyte chemoattraction and revealed an anti-metastatic role via STAT3 suppression.","evidence":"Recombinant and overexpressed CYTL1 in migration/invasion assays, STAT3 phosphorylation, and breast cancer metastasis mouse models","pmids":["31069137"],"confidence":"Medium","gaps":["Direct CYTL1-CCR2 binding not biochemically confirmed","Link between chemoattraction and tumor suppression incompletely connected"]},{"year":2020,"claim":"Generalized CCR2-mediated signaling to neutrophils and added Akt phosphorylation and ROS output to the effector repertoire.","evidence":"LPS-stimulated neutrophil phagocytosis, chemotaxis, Akt western blot, and ROS assays in vivo and in vitro","pmids":["31823178"],"confidence":"Medium","gaps":["Direct receptor binding not shown","Relationship between ERK and Akt branches downstream of CCR2 unclear"]},{"year":2020,"claim":"Reinforced the chondrogenic mechanism by showing recombinant CYTL1 drives chondrocyte redifferentiation across cartilage developmental stages.","evidence":"Recombinant protein treatment of chondrocytes with qRT-PCR for Sox9, Col2α1, Col1α1 and developmental expression analysis","pmids":["32736681"],"confidence":"Medium","gaps":["Receptor/signaling intermediates not identified","In vivo relevance to redifferentiation untested"]},{"year":2022,"claim":"Revealed an intracellular, receptor-independent function of a signal-peptide-lacking isoform that suppresses tumor glycolysis through protein stabilization.","evidence":"ΔCYTL1 overexpression with Co-IP, MDM2 ubiquitination/degradation assays, NDUFV1-Src interaction, LDHA phosphorylation, and tumor models","pmids":["35115484"],"confidence":"High","gaps":["Regulation switching between secreted and intracellular isoforms unknown","Structural basis of NDUFV1 N-terminal competition not resolved"]},{"year":2024,"claim":"Connected CYTL1 to redox/ferroptosis control via a Keap1-Nrf2-CBS axis supporting cysteine biosynthesis.","evidence":"CYTL1 knockdown/overexpression with autophagy flux, Nrf2 target assays, CBS activity, ferroptosis assays, and CBS silencing rescue in vitro and in vivo","pmids":["38211443"],"confidence":"High","gaps":["Mechanism by which CYTL1 promotes autophagic Keap1 degradation unclear","Whether this is intracellular or secreted CYTL1 not delineated"]},{"year":2024,"claim":"Implicated CYTL1 in fibrotic remodeling through TGF-β/CCN2-driven M2 macrophage polarization.","evidence":"CYTL1 ablation in bleomycin-induced lung fibrosis with M2 marker, CCN2 analysis, and TGF-β inhibitor phenocopy","pmids":["39103233"],"confidence":"Medium","gaps":["Direct molecular target of CYTL1 in the TGF-β/CCN2 axis unknown","Cell type producing CYTL1 in fibrosis not defined"]},{"year":2024,"claim":"Extended CYTL1 function to vascular control, mediating arteriolar vasodilation and angiogenic remodeling from neuro-endothelial cells.","evidence":"Intravital multiphoton imaging of optogenetic mice with NEC gain/loss-of-function and single-cell transcriptomics (preprint)","pmids":["bio_10.1101_2024.09.30.615824"],"confidence":"Medium","gaps":["Preprint not yet peer-reviewed","Molecular receptor/effector for vasodilation not defined in available evidence"]},{"year":null,"claim":"How CYTL1 partitions between a secreted CCR2-dependent ligand and an intracellular isoform, and whether a single receptor mediates its diverse chemotactic, chondrogenic, and vascular effects, remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No experimental demonstration of direct CYTL1-CCR2 binding in the corpus","Regulation of isoform choice (secreted vs. signal-peptide-lacking) uncharacterized","No high-resolution structure of CYTL1 or its complexes"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0048018","term_label":"receptor ligand activity","supporting_discovery_ids":[0,4,6]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[8,9]},{"term_id":"GO:0140313","term_label":"molecular sequestering activity","supporting_discovery_ids":[8]}],"localization":[{"term_id":"GO:0005576","term_label":"extracellular region","supporting_discovery_ids":[0,3,4]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[8]}],"pathway":[{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[0,1,7]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[4,6,10]},{"term_id":"R-HSA-1430728","term_label":"Metabolism","supporting_discovery_ids":[8,9]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[4,6]}],"complexes":[],"partners":["NDUFV1","CCR2","KEAP1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9NRR1","full_name":"Cytokine-like protein 1","aliases":["Protein C17"],"length_aa":136,"mass_kda":15.6,"function":"","subcellular_location":"Secreted","url":"https://www.uniprot.org/uniprotkb/Q9NRR1/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/CYTL1","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/CYTL1","total_profiled":1310},"omim":[{"mim_id":"607930","title":"CYTOKINE-LIKE PROTEIN 1; CYTL1","url":"https://www.omim.org/entry/607930"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Nuclear membrane","reliability":"Approved"},{"location":"Endoplasmic reticulum","reliability":"Approved"}],"tissue_specificity":"Tissue enriched","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"blood vessel","ntpm":198.0}],"url":"https://www.proteinatlas.org/search/CYTL1"},"hgnc":{"alias_symbol":["C17","C4orf4"],"prev_symbol":[]},"alphafold":{"accession":"Q9NRR1","domains":[{"cath_id":"-","chopping":"26-115","consensus_level":"high","plddt":92.9166,"start":26,"end":115}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NRR1","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NRR1-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NRR1-F1-predicted_aligned_error_v6.png","plddt_mean":85.56},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=CYTL1","jax_strain_url":"https://www.jax.org/strain/search?query=CYTL1"},"sequence":{"accession":"Q9NRR1","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9NRR1.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9NRR1/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NRR1"}},"corpus_meta":[{"pmid":"17644814","id":"PMC_17644814","title":"Cytokine-like 1 (Cytl1) regulates the chondrogenesis of mesenchymal cells.","date":"2007","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/17644814","citation_count":65,"is_preprint":false},{"pmid":"21652695","id":"PMC_21652695","title":"Cytokine-like 1 knock-out mice (Cytl1-/-) show normal cartilage and bone development but exhibit augmented osteoarthritic cartilage destruction.","date":"2011","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/21652695","citation_count":48,"is_preprint":false},{"pmid":"31089746","id":"PMC_31089746","title":"Protein Cytl1: its role in chondrogenesis, cartilage homeostasis, and disease.","date":"2019","source":"Cellular and molecular life sciences : CMLS","url":"https://pubmed.ncbi.nlm.nih.gov/31089746","citation_count":26,"is_preprint":false},{"pmid":"35115484","id":"PMC_35115484","title":"Intracellular CYTL1, a novel tumor suppressor, stabilizes NDUFV1 to inhibit metabolic reprogramming in breast cancer.","date":"2022","source":"Signal transduction and targeted therapy","url":"https://pubmed.ncbi.nlm.nih.gov/35115484","citation_count":25,"is_preprint":false},{"pmid":"31069137","id":"PMC_31069137","title":"CYTL1 inhibits tumor metastasis with decreasing STAT3 phosphorylation.","date":"2019","source":"Oncoimmunology","url":"https://pubmed.ncbi.nlm.nih.gov/31069137","citation_count":20,"is_preprint":false},{"pmid":"22613542","id":"PMC_22613542","title":"ANKRD7 and CYTL1 are novel risk genes for alcohol drinking behavior.","date":"2012","source":"Chinese medical journal","url":"https://pubmed.ncbi.nlm.nih.gov/22613542","citation_count":17,"is_preprint":false},{"pmid":"21322034","id":"PMC_21322034","title":"Identification of CCR2-binding features in Cytl1 by a CCL2-like chemokine model.","date":"2011","source":"Proteins","url":"https://pubmed.ncbi.nlm.nih.gov/21322034","citation_count":15,"is_preprint":false},{"pmid":"26800213","id":"PMC_26800213","title":"Cytokine-Like Protein 1(Cytl1): A Potential Molecular Mediator in Embryo Implantation.","date":"2016","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/26800213","citation_count":13,"is_preprint":false},{"pmid":"28478073","id":"PMC_28478073","title":"Biochemical and biophysical characterization of cytokine-like protein 1 (CYTL1).","date":"2017","source":"Cytokine","url":"https://pubmed.ncbi.nlm.nih.gov/28478073","citation_count":10,"is_preprint":false},{"pmid":"32736681","id":"PMC_32736681","title":"The involvement of cytokine-like 1 (Cytl1) in chondrogenesis and cartilage metabolism.","date":"2020","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/32736681","citation_count":10,"is_preprint":false},{"pmid":"38211443","id":"PMC_38211443","title":"Breast cancer cells have an increased ferroptosis risk induced by system xc- blockade after deliberately downregulating CYTL1 to mediate malignancy.","date":"2024","source":"Redox biology","url":"https://pubmed.ncbi.nlm.nih.gov/38211443","citation_count":8,"is_preprint":false},{"pmid":"31823178","id":"PMC_31823178","title":"CYTL1 Promotes the Activation of Neutrophils in a Sepsis Model.","date":"2020","source":"Inflammation","url":"https://pubmed.ncbi.nlm.nih.gov/31823178","citation_count":7,"is_preprint":false},{"pmid":"39103233","id":"PMC_39103233","title":"Down-regulation of CYTL1 attenuates bleomycin-induced pulmonary fibrosis in mice by inhibiting M2 macrophage polarization via the TGF-β/CCN2 axis.","date":"2024","source":"Clinical and experimental pharmacology & physiology","url":"https://pubmed.ncbi.nlm.nih.gov/39103233","citation_count":4,"is_preprint":false},{"pmid":"37745303","id":"PMC_37745303","title":"Bioinformatics-based analysis reveals elevated CYTL1 as a potential therapeutic target for BRAF-mutated melanoma.","date":"2023","source":"Frontiers in cell and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/37745303","citation_count":3,"is_preprint":false},{"pmid":"34511992","id":"PMC_34511992","title":"The Clinical Significance of Changes in Serum New Cytokine CYTL1 in Patients with Knee Osteoarthritis.","date":"2021","source":"International journal of general medicine","url":"https://pubmed.ncbi.nlm.nih.gov/34511992","citation_count":2,"is_preprint":false},{"pmid":"40853611","id":"PMC_40853611","title":"Multi-omics integration reveals CYTL1 and H6PD as key regulators of tumor metabolism in mesothelioma.","date":"2025","source":"Genes & genomics","url":"https://pubmed.ncbi.nlm.nih.gov/40853611","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2024.09.30.615824","title":"Neuronally differentiated endothelial cell subtype regulates organ blood flow and immune balance","date":"2024-10-02","source":"bioRxiv","url":"https://doi.org/10.1101/2024.09.30.615824","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":10545,"output_tokens":3196,"usd":0.039788,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":10717,"output_tokens":3852,"usd":0.074942,"stage2_stop_reason":"end_turn"},"total_usd":0.11473,"stage1_batch_id":"msgbatch_01ACAxJQabDsht39JXeWG22d","stage2_batch_id":"msgbatch_01QEU22A9U5sbPTgsMzTmvSf","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2007,\n      \"finding\": \"CYTL1 promotes chondrogenic differentiation of mouse limb bud mesenchymal cells acting as an autocrine factor, operating via stimulation of Sox9 transcriptional activity and induction of IGF-1 expression; exogenous CYTL1 protein or lentivirus-mediated overexpression both induced chondrogenesis in micromass culture.\",\n      \"method\": \"Exogenous protein treatment, lentiviral overexpression, micromass culture of mesenchymal cells, reporter assays for Sox9 activity, gene expression analysis\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (exogenous protein, genetic overexpression, reporter assay), replicated in vivo and in vitro, two independent approaches converging on same mechanism\",\n      \"pmids\": [\"17644814\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Cytl1 knockout mice show normal chondrogenesis, cartilage/bone development, and endochondral ossification, but exhibit augmented osteoarthritic cartilage destruction upon destabilization of the medial meniscus, establishing a role in cartilage homeostasis maintenance rather than development.\",\n      \"method\": \"Cytl1-/- knockout mouse generation, destabilization of medial meniscus OA model, histological and ultrastructural analysis of cartilage\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic knockout with defined in vivo phenotypic readout, complemented by human OA tissue expression data\",\n      \"pmids\": [\"21652695\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Computational structural modeling predicts CYTL1 adopts an IL8-like chemokine fold similar to CCL2 (MCP-1) rather than a 4-helical cytokine fold, and identifies structural features in CYTL1 necessary for signaling through the chemokine receptor CCR2.\",\n      \"method\": \"Molecular modeling, structure-based computational analysis, comparison with CCL2 3D structure\",\n      \"journal\": \"Proteins\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 4 / Weak — computational prediction only, no experimental structural or binding validation performed\",\n      \"pmids\": [\"21322034\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Recombinant human CYTL1 produced in bacteria shows predominantly beta-sheet secondary structure (chemokine-like) by circular dichroism, promotes calcium flux in chondrocytes, but unlike chemokines shows limited affinity to proteoglycans.\",\n      \"method\": \"Recombinant protein production in bacteria, circular dichroism spectroscopy, calcium flux assay in chondrocytes, proteoglycan binding assay\",\n      \"journal\": \"Cytokine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — in vitro biochemical assays with recombinant protein, multiple biophysical methods, single lab\",\n      \"pmids\": [\"28478073\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Recombinant CYTL1 chemoattracts human monocytes via the CCR2/ERK pathway, and CYTL1 (recombinant or overexpressed) inhibits tumor cell migration and invasion accompanied by decreased STAT3 phosphorylation, with inhibition of tumor metastasis confirmed in experimental and spontaneous breast cancer mouse models.\",\n      \"method\": \"Recombinant CYTL1 treatment, CYTL1-overexpressing tumor cell lines, migration/invasion assays, STAT3 phosphorylation measurement, experimental and spontaneous metastasis mouse models\",\n      \"journal\": \"Oncoimmunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (recombinant protein, genetic overexpression, in vivo models), single lab\",\n      \"pmids\": [\"31069137\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"CYTL1 expression in mouse endometrium is regulated by ovarian hormones and peaks during embryo implantation; CYTL1 enhances proliferation of endometrial cells, stimulates endometrial secretion of LIF and HB-EGF, and enhances adhesion of endometrial cells to JAR spheroids in cell-cell adhesion assays.\",\n      \"method\": \"Mouse endometrial expression analysis (mRNA and protein), cell proliferation assays, ELISA for LIF and HB-EGF secretion, cell-cell adhesion assays with HEC-1-A/RL95-2 and JAR spheroids\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 / Moderate — multiple functional assays in cell culture with defined readouts, single lab\",\n      \"pmids\": [\"26800213\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"CYTL1 promotes chemotactic activity and phagocytosis of E. coli by LPS-stimulated neutrophils through CCR2-mediated action and phosphorylation of Akt (protein kinase B), and increases release of reactive oxygen species in activated neutrophils.\",\n      \"method\": \"In vivo and in vitro neutrophil phagocytosis assays, western blotting for Akt phosphorylation, chemotaxis assays, ROS measurement, LPS-stimulated neutrophil model\",\n      \"journal\": \"Inflammation\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo and in vitro assays, multiple functional readouts, single lab\",\n      \"pmids\": [\"31823178\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Recombinant CYTL1 (rCytl1) increases expression of Sox9 and Col2α1 while stabilizing Col1α1 in cultured chondrocytes, promoting redifferentiation; Cytl1 is expressed at all stages of cartilage development and shares expression patterns with other chondrogenic factors.\",\n      \"method\": \"Recombinant protein treatment of cultured chondrocytes, quantitative RT-PCR for chondrogenic markers, embryonic and adult cartilage expression analysis\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 / Moderate — recombinant protein functional assay with multiple markers, single lab\",\n      \"pmids\": [\"32736681\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Intracellular CYTL1 (lacking signal peptide, ΔCYTL1) competitively binds the N-terminal sequence of NDUFV1 to block MDM2-mediated proteasomal degradation of NDUFV1, thereby stabilizing NDUFV1; stabilized NDUFV1 increases NAD+ levels and interacts with Src to attenuate LDHA phosphorylation at tyrosine 10, reducing lactate production and glycolytic reprogramming in breast cancer.\",\n      \"method\": \"ΔCYTL1 overexpression (signal-peptide-deleted construct), Co-IP/pulldown for CYTL1-NDUFV1 interaction, MDM2 ubiquitination/proteasome degradation assays, glucose uptake/lactate production assays, NDUFV1-Src interaction, LDHA phosphorylation assays, in vitro and in vivo tumor growth/metastasis assays\",\n      \"journal\": \"Signal transduction and targeted therapy\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — multiple orthogonal mechanistic methods (protein interaction, ubiquitination/degradation assay, phosphorylation assay, metabolic readouts, in vivo validation), single lab with comprehensive mechanistic follow-up\",\n      \"pmids\": [\"35115484\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CYTL1 activates Nrf2 by promoting autophagic degradation of Keap1; Nrf2 subsequently transactivates CBS (cystathionine β-synthase) expression, supporting de novo cysteine synthesis via the transsulfuration pathway; breast cancer cells with low CYTL1 have suppressed CBS activity and are hypersensitive to system xc- blockade-induced ferroptosis.\",\n      \"method\": \"CYTL1 knockdown/overexpression in breast cancer cells, autophagy flux assays for Keap1 degradation, Nrf2 reporter/target gene assays, CBS expression and activity measurement, system xc- inhibitor (ferroptosis) assays in vitro and in vivo, CBS silencing rescue experiment\",\n      \"journal\": \"Redox biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (genetic manipulation, pathway rescue, in vivo validation), mechanistic chain experimentally established across multiple steps\",\n      \"pmids\": [\"38211443\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CYTL1 regulates TGF-β/CCN2 (connective tissue growth factor) axis to promote M2 macrophage polarization; CYTL1 ablation reduces M2 macrophage polarization and alleviates bleomycin-induced lung injury and fibrosis in mice, and inhibition of the TGF-β pathway phenocopies CYTL1 ablation.\",\n      \"method\": \"CYTL1 knockdown/ablation in bleomycin-induced lung fibrosis mouse model, macrophage polarization assays (M2 markers), TGF-β pathway inhibition, CCN2 expression analysis\",\n      \"journal\": \"Clinical and experimental pharmacology & physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo genetic ablation with defined cellular and molecular readouts, pathway inhibitor confirmation, single lab\",\n      \"pmids\": [\"39103233\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"In neuro-endothelial cells (NECs) of resistance arterioles, Cytl1 mediates vasodilation and increased blood flow acutely, and promotes capillary density and clonal EC remodeling chronically; silencing NEC Cytl1 gene expression causes arteriolar vasoconstriction, reduced vascular density and organ blood flow, increased vascular permeability and immune cell homing.\",\n      \"method\": \"Intravital multiphoton imaging of optogenetic mouse models with NEC gain/loss-of-function, Cytl1 in vivo silencing, Cytl1 protein administration, single-cell transcriptome analysis\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct in vivo functional imaging with gain/loss-of-function, preprint not yet peer-reviewed\",\n      \"pmids\": [\"bio_10.1101_2024.09.30.615824\"],\n      \"is_preprint\": true\n    }\n  ],\n  \"current_model\": \"CYTL1 is a small secreted cytokine-like protein that acts as an autocrine chondrogenic factor (stimulating Sox9, IGF-1, Col2α1) and cartilage homeostasis factor (loss accelerates OA), signals through CCR2/ERK to chemoattract monocytes and activate neutrophils via Akt phosphorylation, and intracellularly (as a signal-peptide-lacking isoform) suppresses glycolytic reprogramming in breast cancer by competitively binding NDUFV1 to block MDM2-mediated degradation and by activating Nrf2 through autophagic Keap1 degradation to support CBS-dependent cysteine synthesis; it also modulates TGF-β/CCN2-driven macrophage polarization in fibrosis, and in vascular neuro-endothelial cells mediates Nos1-dependent vasodilation and angiogenic remodeling.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"CYTL1 is a small secreted chemokine-like protein that functions in cartilage homeostasis, immune cell chemotaxis, and tumor metabolic regulation [#0, #1, #8]. As an autocrine factor it promotes chondrogenic differentiation by stimulating Sox9 transcriptional activity and inducing IGF-1, and it drives chondrocyte redifferentiation through induction of Sox9 and Col2\\u03b11 [#0, #7]; while dispensable for cartilage and bone development, CYTL1 maintains adult cartilage homeostasis, as its loss accelerates osteoarthritic cartilage destruction after joint destabilization [#1]. Biophysically the secreted protein adopts a predominantly beta-sheet, chemokine-like fold and elicits calcium flux in chondrocytes [#3], and it signals through the chemokine receptor CCR2 to chemoattract monocytes via ERK and to activate neutrophil chemotaxis, phagocytosis, and ROS release via Akt phosphorylation [#4, #6]. A signal-peptide-lacking intracellular isoform (\\u0394CYTL1) suppresses glycolytic reprogramming in breast cancer by competitively binding the N-terminus of NDUFV1 to block MDM2-mediated proteasomal degradation, stabilizing NDUFV1 to raise NAD+ and attenuate Src-dependent LDHA phosphorylation [#8]; CYTL1 additionally activates Nrf2 through autophagic Keap1 degradation, transactivating CBS to support transsulfuration-dependent cysteine synthesis and protect cells from ferroptosis [#9]. CYTL1 further modulates the TGF-\\u03b2/CCN2 axis to promote M2 macrophage polarization in lung fibrosis [#10].\",\n  \"teleology\": [\n    {\n      \"year\": 2007,\n      \"claim\": \"Established CYTL1 as a functional autocrine chondrogenic factor rather than an orphan cytokine-like sequence, defining its first molecular activity.\",\n      \"evidence\": \"Exogenous protein and lentiviral overexpression in mesenchymal micromass culture with Sox9 reporter and IGF-1 expression readouts\",\n      \"pmids\": [\"17644814\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Receptor mediating the chondrogenic signal not identified\", \"Mechanism linking CYTL1 to Sox9 activation unresolved\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Distinguished CYTL1's developmental dispensability from its homeostatic role, showing it protects mature cartilage rather than building it.\",\n      \"evidence\": \"Cytl1-/- knockout mice analyzed in a destabilization of the medial meniscus osteoarthritis model with histology\",\n      \"pmids\": [\"21652695\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular pathway protecting cartilage not defined\", \"No connection to a receptor or downstream effector established\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Proposed a structural basis for receptor engagement by predicting an IL8/CCL2-like chemokine fold and CCR2-relevant features.\",\n      \"evidence\": \"Computational molecular modeling compared with CCL2 structure\",\n      \"pmids\": [\"21322034\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Computational prediction only, no experimental structure or binding validation\", \"CCR2 binding not demonstrated experimentally here\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Provided experimental biophysical support for a chemokine-like fold and demonstrated functional signaling output in chondrocytes.\",\n      \"evidence\": \"Bacterially produced recombinant CYTL1 analyzed by circular dichroism, calcium flux, and proteoglycan binding assays\",\n      \"pmids\": [\"28478073\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"High-resolution structure absent\", \"Limited proteoglycan affinity leaves signaling/localization mechanism unclear\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Extended CYTL1 function to reproductive tissue, implicating it in endometrial receptivity and embryo implantation.\",\n      \"evidence\": \"Hormone-regulated endometrial expression analysis with proliferation, LIF/HB-EGF secretion, and spheroid adhesion assays\",\n      \"pmids\": [\"26800213\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Receptor and signaling pathway in endometrium not identified\", \"Single-lab cell-based assays\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Defined a CCR2/ERK signaling axis for monocyte chemoattraction and revealed an anti-metastatic role via STAT3 suppression.\",\n      \"evidence\": \"Recombinant and overexpressed CYTL1 in migration/invasion assays, STAT3 phosphorylation, and breast cancer metastasis mouse models\",\n      \"pmids\": [\"31069137\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct CYTL1-CCR2 binding not biochemically confirmed\", \"Link between chemoattraction and tumor suppression incompletely connected\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Generalized CCR2-mediated signaling to neutrophils and added Akt phosphorylation and ROS output to the effector repertoire.\",\n      \"evidence\": \"LPS-stimulated neutrophil phagocytosis, chemotaxis, Akt western blot, and ROS assays in vivo and in vitro\",\n      \"pmids\": [\"31823178\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct receptor binding not shown\", \"Relationship between ERK and Akt branches downstream of CCR2 unclear\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Reinforced the chondrogenic mechanism by showing recombinant CYTL1 drives chondrocyte redifferentiation across cartilage developmental stages.\",\n      \"evidence\": \"Recombinant protein treatment of chondrocytes with qRT-PCR for Sox9, Col2\\u03b11, Col1\\u03b11 and developmental expression analysis\",\n      \"pmids\": [\"32736681\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Receptor/signaling intermediates not identified\", \"In vivo relevance to redifferentiation untested\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Revealed an intracellular, receptor-independent function of a signal-peptide-lacking isoform that suppresses tumor glycolysis through protein stabilization.\",\n      \"evidence\": \"\\u0394CYTL1 overexpression with Co-IP, MDM2 ubiquitination/degradation assays, NDUFV1-Src interaction, LDHA phosphorylation, and tumor models\",\n      \"pmids\": [\"35115484\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Regulation switching between secreted and intracellular isoforms unknown\", \"Structural basis of NDUFV1 N-terminal competition not resolved\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Connected CYTL1 to redox/ferroptosis control via a Keap1-Nrf2-CBS axis supporting cysteine biosynthesis.\",\n      \"evidence\": \"CYTL1 knockdown/overexpression with autophagy flux, Nrf2 target assays, CBS activity, ferroptosis assays, and CBS silencing rescue in vitro and in vivo\",\n      \"pmids\": [\"38211443\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism by which CYTL1 promotes autophagic Keap1 degradation unclear\", \"Whether this is intracellular or secreted CYTL1 not delineated\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Implicated CYTL1 in fibrotic remodeling through TGF-\\u03b2/CCN2-driven M2 macrophage polarization.\",\n      \"evidence\": \"CYTL1 ablation in bleomycin-induced lung fibrosis with M2 marker, CCN2 analysis, and TGF-\\u03b2 inhibitor phenocopy\",\n      \"pmids\": [\"39103233\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct molecular target of CYTL1 in the TGF-\\u03b2/CCN2 axis unknown\", \"Cell type producing CYTL1 in fibrosis not defined\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Extended CYTL1 function to vascular control, mediating arteriolar vasodilation and angiogenic remodeling from neuro-endothelial cells.\",\n      \"evidence\": \"Intravital multiphoton imaging of optogenetic mice with NEC gain/loss-of-function and single-cell transcriptomics (preprint)\",\n      \"pmids\": [\"bio_10.1101_2024.09.30.615824\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Preprint not yet peer-reviewed\", \"Molecular receptor/effector for vasodilation not defined in available evidence\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How CYTL1 partitions between a secreted CCR2-dependent ligand and an intracellular isoform, and whether a single receptor mediates its diverse chemotactic, chondrogenic, and vascular effects, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No experimental demonstration of direct CYTL1-CCR2 binding in the corpus\", \"Regulation of isoform choice (secreted vs. signal-peptide-lacking) uncharacterized\", \"No high-resolution structure of CYTL1 or its complexes\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0048018\", \"supporting_discovery_ids\": [0, 4, 6]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [8, 9]},\n      {\"term_id\": \"GO:0140313\", \"supporting_discovery_ids\": [8]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005576\", \"supporting_discovery_ids\": [0, 3, 4]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [8]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [0, 1, 7]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [4, 6, 10]},\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [8, 9]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [4, 6]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"NDUFV1\", \"CCR2\", \"KEAP1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"faith_supported":4,"faith_total":5,"faith_pct":80.0}}