{"gene":"CFL1","run_date":"2026-06-09T22:57:18","timeline":{"discoveries":[{"year":2019,"finding":"Mechanical forces from cardiac contractility regulate cardiomyocyte myofilament maturation through the VCL-SSH1-CFL axis: vinculin (VCL) recruits the phosphatase SSH1 and its effector cofilin (CFL) to regulate F-actin rearrangement and promote myofilament maturation in zebrafish cardiomyocytes.","method":"Interactome profiling (contracting vs. non-contracting cardiomyocytes), Co-IP, genetic loss-of-function, live imaging, epistasis analysis","journal":"Developmental cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal interactome, Co-IP, genetic epistasis, and functional phenotype across multiple orthogonal methods in a single rigorous study","pmids":["31495694"],"is_preprint":false},{"year":2017,"finding":"LIMK1 directly phosphorylates and inactivates CFL1; inhibition of LIMK1 reduces CFL1 expression and phosphorylation, thereby impairing CFL1-dependent F-actin/G-actin remodeling and suppressing breast cancer cell migration and invasion.","method":"Western blot for pCFL1/CFL1 following LIMK1 knockdown, migration/invasion assays, miRNA mimic transfection in TNBC cells","journal":"Oncotarget","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — multiple cell-based assays in single lab, consistent with known LIMK1-cofilin pathway but no in vitro kinase reconstitution","pmids":["29156719"],"is_preprint":false},{"year":2020,"finding":"SSH1, the canonical CFL (cofilin) phosphatase, dephosphorylates phospho-Ser403-SQSTM1/p62, impairing autophagic cargo clearance; this action is separable from SSH1-mediated CFL activation, demonstrating that SSH1 has a CFL-independent substrate.","method":"RNAi knockdown, overexpression with defined mutant constructs, genetically encoded fluorescent reporters, cell lines and primary neurons","journal":"Autophagy","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (RNAi, OE, mutant constructs, reporters) in cell lines and primary neurons, with genetic dissection of CFL-dependent vs. independent actions","pmids":["33044112"],"is_preprint":false},{"year":2021,"finding":"Hhex enhances the interaction of RHOGDIA with RHOA/CDC42, maintaining them in inactive forms, which blocks the RHOA/CDC42–LIMK1–pCFL1 signaling cascade, preserving CFL1 F-actin-severing activity and suppressing filopodium/lamellipodium formation and cell migration in lung cancer cells.","method":"Western blot, co-immunoprecipitation, wound-healing scratch assay, laser confocal imaging, overexpression/knockdown in NSCLC cells","journal":"Cell communication and signaling : CCS","confidence":"Medium","confidence_rationale":"Tier 2-3 / Moderate — Co-IP, functional migration assays, and pathway epistasis in single lab with multiple orthogonal methods","pmids":["34321041"],"is_preprint":false},{"year":2023,"finding":"HUNK kinase directly phosphorylates GEF-H1 at serine 645, which activates RhoA and triggers a cascade of LIMK-1/CFL-1 phosphorylation that stabilizes F-actin and inhibits EMT and metastasis in colorectal cancer cells.","method":"In vitro kinase assay (direct phosphorylation), Western blot for phospho-CFL-1, overexpression/knockdown, metastasis assays","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct in vitro kinase phosphorylation assay plus cell-based epistasis, single lab","pmids":["37193711"],"is_preprint":false},{"year":2023,"finding":"CFL1 promotes phosphoglycerate dehydrogenase (PHGDH) transcription to enhance serine synthesis and metabolism, increasing antioxidant production that scavenges sorafenib-induced ROS, thereby impairing sorafenib sensitivity in hepatocellular carcinoma.","method":"Transcriptome sequencing of sorafenib-sensitive vs. insensitive HCC patients, siRNA knockdown, metabolic assays, nanoparticle co-delivery experiments in vivo","journal":"Advanced science (Weinheim, Baden-Wurttemberg, Germany)","confidence":"Medium","confidence_rationale":"Tier 2-3 / Moderate — RNA-seq, siRNA loss-of-function, metabolic readouts, and in vivo validation in single lab","pmids":["37203277"],"is_preprint":false},{"year":2022,"finding":"Estradiol (E2) dose-dependently increases total CFL1 and phosphorylated CFL1 (pCFL1) expression in PBMCs and endocervical mucosa, and this induction is abrogated by LIMK1/2 inhibitor LIMKi3; CFL1 knockdown partially restores HIV-1 infection suppressed by E2, linking E2-mediated CFL1 phosphorylation to inhibition of HIV-1BaL infection.","method":"LIMK inhibitor treatment, CFL1 siRNA knockdown, protein expression by Western blot, HIV-1 infectivity assay in PBMCs and endocervical explants","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2-3 / Moderate — pharmacological inhibition, siRNA knockdown, and functional infection assay in two tissue models, single lab","pmids":["35418661"],"is_preprint":false},{"year":2024,"finding":"CFL1 overexpression in pSS bone marrow MSCs restores their migratory capacity by upregulating CCR1 expression; pharmacological inhibition of CCR1 suppresses the CFL1-induced migration and proliferation, placing CFL1 upstream of the CCL5/CCR1 axis.","method":"Lentivirus-mediated CFL1 overexpression, RNA-seq, Transwell migration assay, wound healing assay, CCR1 inhibitor rescue experiment, NOD mouse model","journal":"International immunopharmacology","confidence":"Medium","confidence_rationale":"Tier 2-3 / Moderate — RNA-seq pathway identification, pharmacological rescue epistasis, and in vivo validation, single lab","pmids":["38183912"],"is_preprint":false},{"year":2024,"finding":"Expression of a non-functional form of Cfl1 (nf-Cfl1) in connective tissue mast cells (CTMCs) via Mcpt5-Cre results in complete absence of CTMCs, demonstrating that Cfl1-dependent actin depolymerization is essential for CTMC formation; CTMCs lacking Cfl1 activity show impaired systemic anaphylaxis.","method":"Conditional knock-in of non-functional Cfl1 in CTMCs (Mcpt5-Cre-nf-Cfl1fl/fl), flow cytometry, passive systemic anaphylaxis model","journal":"Frontiers in immunology","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — genetic knock-in of non-functional protein with cell-type-specific Cre, clear phenotypic readout (complete CTMC absence), functional anaphylaxis assay","pmids":["41684538"],"is_preprint":false},{"year":2025,"finding":"Overexpression of Cfl1 in the polymorphic layer of the hippocampal dentate gyrus (PoDG) inhibits ML-DG synapses, increases motivation to seek alcohol, impairs extinction of alcohol-seeking, and decreases reward consumption during relapse; reducing Cfl1 levels has opposite effects, establishing hippocampal Cfl1 as a regulator of synaptic transmission and AUD-relevant behavior.","method":"Local viral overexpression/knockdown of Cfl in hippocampal DG subregions, in vivo two-photon imaging of synaptic activity, automated IntelliCage behavioral testing, RNA sequencing","journal":"Molecular psychiatry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — bidirectional (OE and KD) viral manipulation with synaptic electrophysiology and behavioral phenotype, single lab","pmids":["40931167"],"is_preprint":false},{"year":2026,"finding":"c-Myc transcriptionally activates CFL1 by binding E-boxes in the CFL1 promoter (confirmed by ChIP-qPCR); cofilin-1 upregulation mediates c-Myc-induced oncogene-induced senescence (OIS) and F-actin/nuclear G-actin accumulation in lung cancer cells; knockdown of cofilin-1 suppresses cMIS, and a physical interaction between c-Myc and cofilin-1 was detected by co-immunoprecipitation.","method":"ChIP-qPCR, cofilin-1 knockdown, c-Myc overexpression/truncation mutants, Co-IP, F-actin/G-actin assays, conditioned medium bystander assays","journal":"Cell death discovery","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP-qPCR identifies direct promoter binding, Co-IP shows physical interaction, loss-of-function rescues phenotype; multiple orthogonal methods, single lab","pmids":["41888102"],"is_preprint":false},{"year":2024,"finding":"Conditional knockout of ADF and Cfl1 in microglia (ADF/Cfl1-KO) causes increased accumulation of stabilized F-actin and altered microtubule dynamics, reduces microglial fine process motility, and impairs migration toward laser-induced lesions in vivo; microglial ADF/Cfl1-deficiency also decreases learning and memory in mice.","method":"Conditional KO mouse model, in vivo two-photon imaging, F-actin staining, microtubule dynamics assays, behavioral memory tests","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic KO with in vivo imaging and behavioral readouts; preprint, not yet peer-reviewed","pmids":["bio_10.1101_2024.09.27.615114"],"is_preprint":true},{"year":1996,"finding":"Human non-muscle cofilin (CFL1) was mapped to chromosome 11q13 by PCR in somatic cell hybrid panels and confirmed by FISH, while muscle-type cofilin (CFL2) was mapped to chromosome 14; CFL1 was identified as an actin-binding protein involved in actin-cofilin complex translocation from cytoplasm to nucleus.","method":"PCR in rodent-human somatic cell hybrids, FISH of genomic cosmid clones, radiation hybrid mapping","journal":"Annals of human genetics","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — multiple independent mapping methods (somatic cell hybrids, FISH, radiation hybrids) converging on same chromosomal locus","pmids":["8800436"],"is_preprint":false},{"year":2024,"finding":"CFL1 expression in nerves (rather than in tumor cells) within the head and neck squamous cell carcinoma (HNSCC) tumor microenvironment is associated with perineural invasion; HNSCC cells induce neuronal CFL1 expression, and conditional knockout of neuronal CFL1 impedes tumor-nerve interactions.","method":"Multiplex fluorescent immunohistochemistry, conditional neuronal CFL1 knockout, tumor-nerve co-culture models","journal":"Molecular carcinogenesis","confidence":"Medium","confidence_rationale":"Tier 2-3 / Moderate — conditional KO with functional tumor-nerve interaction readout and localization by multiplex IHC, single lab","pmids":["38353363"],"is_preprint":false},{"year":2024,"finding":"Imatinib therapy in CML upregulates CFL1 expression and activity (cofilin, P-cofilin) and increases F-actin (including branched F-actin), indicating that imatinib affects F-actin remodeling in CML cells by regulating CFL1.","method":"Western blot for CFL1 and P-cofilin in CML patient samples and cell lines pre/post imatinib, F-actin staining","journal":"Journal of Cancer","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single-lab observational protein measurements with limited mechanistic dissection","pmids":["38495482"],"is_preprint":false}],"current_model":"CFL1 (non-muscle cofilin-1) is an actin-severing and depolymerizing protein whose activity is tightly regulated by phosphorylation: LIMK1/2 phosphorylates and inactivates CFL1, while SSH1 dephosphorylates and activates it; mechanical forces transmitted through vinculin (VCL) recruit SSH1 to activate CFL1 for F-actin rearrangement during cardiomyocyte myofilament maturation; CFL1 is also transcriptionally activated by c-Myc (via direct E-box binding) to drive oncogene-induced senescence, promotes serine synthesis metabolism to confer chemoresistance, regulates cell migration via the RHOA/CDC42–LIMK1 axis downstream of Hhex and upstream of CCR1, and is essential for connective tissue mast cell formation and microglial cytoskeletal dynamics and motility."},"narrative":{"mechanistic_narrative":"CFL1 (non-muscle cofilin-1) is an actin-binding protein that severs and depolymerizes F-actin to drive cytoskeletal remodeling underlying cell migration, tissue morphogenesis, and synaptic function [PMID:8800436, PMID:41684538]. Its activity is gated by reversible phosphorylation: LIMK1 directly phosphorylates and inactivates CFL1, blocking its F-actin/G-actin remodeling activity [PMID:29156719], while the phosphatase SSH1 dephosphorylates and reactivates it [PMID:33044112]. This kinase-phosphatase switch sits downstream of RHOA/CDC42 signaling, where upstream regulators such as Hhex (via RHOGDIA sequestration of RHOA/CDC42) and HUNK (via GEF-H1/RhoA activation) tune the RHOA/CDC42-LIMK1-pCFL1 cascade to control filopodium/lamellipodium formation, EMT, and metastasis [PMID:34321041, PMID:37193711]. Mechanical forces transmitted through vinculin recruit SSH1 and cofilin to remodel F-actin during cardiomyocyte myofilament maturation, embedding CFL1 in mechanotransduction [PMID:31495694]. CFL1-dependent actin depolymerization is genetically essential for connective tissue mast cell formation and systemic anaphylaxis [PMID:41684538] and, together with ADF, for microglial process motility, migration, and learning-related behavior [PMID:bio_10.1101_2024.09.27.615114]. Beyond its cytoskeletal role, CFL1 acts as a transcriptional driver: c-Myc binds E-boxes in the CFL1 promoter to upregulate cofilin-1 and induce oncogene-induced senescence with nuclear G-actin accumulation [PMID:41888102], and CFL1 promotes PHGDH transcription to enhance serine synthesis and antioxidant output, conferring sorafenib resistance in hepatocellular carcinoma [PMID:37203277].","teleology":[{"year":1996,"claim":"Establishing the molecular identity and chromosomal locus of human non-muscle cofilin distinguished CFL1 from its muscle paralog and framed it as an actin-binding protein capable of nuclear translocation.","evidence":"PCR in somatic cell hybrids, FISH, and radiation hybrid mapping","pmids":["8800436"],"confidence":"High","gaps":["Did not define the biochemical regulation of severing activity","Functional consequences of cytoplasm-to-nucleus translocation left unresolved"]},{"year":2017,"claim":"Identifying LIMK1 as a direct upstream kinase that phosphorylates and inactivates CFL1 connected cofilin regulation to cancer cell motility.","evidence":"LIMK1 knockdown with pCFL1/CFL1 Western blot and migration/invasion assays in TNBC cells","pmids":["29156719"],"confidence":"Medium","gaps":["No in vitro kinase reconstitution","Single cell-line context"]},{"year":2019,"claim":"Linking CFL1 to mechanotransduction showed that vinculin recruits the SSH1 phosphatase to activate cofilin for F-actin rearrangement during cardiomyocyte myofilament maturation.","evidence":"Interactome profiling, Co-IP, genetic loss-of-function and epistasis with live imaging in zebrafish cardiomyocytes","pmids":["31495694"],"confidence":"High","gaps":["Direct binding stoichiometry of the VCL-SSH1-CFL axis not resolved","Mammalian generalization not tested in this study"]},{"year":2020,"claim":"Demonstrating that SSH1 dephosphorylates p62/SQSTM1 independently of cofilin established that the cofilin phosphatase has CFL-independent substrates, refining how SSH1 activity relates to CFL1.","evidence":"RNAi, overexpression of defined mutants, fluorescent reporters in cell lines and primary neurons","pmids":["33044112"],"confidence":"High","gaps":["Does not directly address CFL1 catalytic mechanism","Relative flux through CFL-dependent vs independent SSH1 actions in vivo unknown"]},{"year":2021,"claim":"Placing CFL1 downstream of RHOA/CDC42-LIMK1 and under Hhex/RHOGDIA control clarified how upstream small-GTPase regulation tunes cofilin-mediated severing and migration.","evidence":"Co-IP, scratch assays, confocal imaging, and pathway epistasis in NSCLC cells","pmids":["34321041"],"confidence":"Medium","gaps":["Single lab and cell-type","Direct effect on CFL1 severing kinetics not measured"]},{"year":2023,"claim":"Identifying HUNK-GEF-H1-RhoA as an upstream input into the LIMK1/CFL1 axis extended the regulatory map of cofilin phosphorylation to suppression of EMT and metastasis.","evidence":"In vitro kinase assay on GEF-H1, phospho-CFL1 Western blot, and metastasis assays in colorectal cancer cells","pmids":["37193711"],"confidence":"Medium","gaps":["CFL1 phosphorylation inferred from cascade, not directly reconstituted","Single lab"]},{"year":2023,"claim":"Revealing that CFL1 promotes PHGDH transcription assigned cofilin a non-cytoskeletal role in serine/antioxidant metabolism that drives chemoresistance.","evidence":"Transcriptome sequencing, siRNA knockdown, metabolic assays, and in vivo nanoparticle co-delivery in HCC","pmids":["37203277"],"confidence":"Medium","gaps":["Mechanism by which CFL1 reaches/regulates the PHGDH promoter not defined","Whether this requires nuclear actin binding unknown"]},{"year":2024,"claim":"Genetic knock-in of non-functional Cfl1 in connective tissue mast cells proved that cofilin-dependent actin depolymerization is required for mast cell formation and anaphylaxis.","evidence":"Cell-type-specific knock-in (Mcpt5-Cre-nf-Cfl1), flow cytometry, passive systemic anaphylaxis","pmids":["41684538"],"confidence":"High","gaps":["Developmental step at which CTMCs are lost not pinpointed","Upstream regulators of CFL1 in mast cells not identified"]},{"year":2024,"claim":"Conditional loss of ADF/Cfl1 in microglia established cofilin as required for F-actin/microtubule balance, process motility, and lesion-directed migration with cognitive consequences.","evidence":"Conditional KO, in vivo two-photon imaging, cytoskeletal staining, behavioral tests (preprint)","pmids":["bio_10.1101_2024.09.27.615114"],"confidence":"Medium","gaps":["Preprint, not yet peer-reviewed","ADF and Cfl1 contributions not separated"]},{"year":2024,"claim":"Linking CFL1 to chemokine signaling and to perineural invasion broadened its migratory roles to MSC homing (CCR1 axis) and tumor-nerve interactions.","evidence":"Lentiviral CFL1 overexpression with CCR1 inhibitor rescue and NOD model; conditional neuronal CFL1 KO with tumor-nerve co-culture and multiplex IHC","pmids":["38183912","38353363"],"confidence":"Medium","gaps":["Mechanism connecting CFL1 to CCR1 transcription unclear","Cell-intrinsic vs microenvironmental contributions only partly dissected"]},{"year":2025,"claim":"Bidirectional manipulation of hippocampal Cfl1 demonstrated a role in regulating synaptic transmission and alcohol-seeking behavior, extending cofilin function to neural circuits.","evidence":"Viral overexpression/knockdown in dentate gyrus subregions, two-photon synaptic imaging, IntelliCage behavior, RNA-seq","pmids":["40931167"],"confidence":"Medium","gaps":["Molecular link between cofilin activity and synaptic change not resolved","Single lab"]},{"year":2026,"claim":"Showing c-Myc directly transactivates CFL1 via promoter E-boxes positioned cofilin as a transcriptional effector driving oncogene-induced senescence and nuclear G-actin accumulation.","evidence":"ChIP-qPCR, cofilin-1 knockdown, c-Myc mutants, Co-IP, F-actin/G-actin assays in lung cancer cells","pmids":["41888102"],"confidence":"Medium","gaps":["Functional significance of the c-Myc-cofilin physical interaction unclear","How nuclear G-actin accumulation enforces senescence not defined"]},{"year":null,"claim":"How the phosphorylation switch, nuclear actin shuttling, and transcriptional/metabolic functions of CFL1 are coordinated within a single cell remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model integrating regulatory inputs","Direct vs indirect mechanism of CFL1 transcriptional effects unknown","Tissue-specific regulators upstream of LIMK/SSH1 not mapped"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[12,1,8]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[1,4]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[12]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[12,10]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[0,11]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[1,3,4]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[0,8]}],"complexes":[],"partners":["LIMK1","SSH1","VCL","MYC"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"P23528","full_name":"Cofilin-1","aliases":["18 kDa phosphoprotein","p18","Cofilin, non-muscle isoform"],"length_aa":166,"mass_kda":18.5,"function":"Binds to F-actin and exhibits pH-sensitive F-actin depolymerizing activity (PubMed:11812157). In conjunction with the subcortical maternal complex (SCMC), plays an essential role for zygotes to progress beyond the first embryonic cell divisions via regulation of actin dynamics (PubMed:15580268). Required for the centralization of the mitotic spindle and symmetric division of zygotes (By similarity). Plays a role in the regulation of cell morphology and cytoskeletal organization in epithelial cells (PubMed:21834987). Required for the up-regulation of atypical chemokine receptor ACKR2 from endosomal compartment to cell membrane, increasing its efficiency in chemokine uptake and degradation (PubMed:23633677). Required for neural tube morphogenesis and neural crest cell migration (By similarity)","subcellular_location":"Nucleus matrix; Cytoplasm, cytoskeleton; Cell projection, ruffle membrane; Cell projection, lamellipodium membrane; Cell projection, lamellipodium; Cell projection, growth cone; Cell projection, axon","url":"https://www.uniprot.org/uniprotkb/P23528/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/CFL1","classification":"Common Essential","n_dependent_lines":745,"n_total_lines":1208,"dependency_fraction":0.6167218543046358},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"ACTB","stoichiometry":10.0},{"gene":"ACTG1","stoichiometry":0.2},{"gene":"CALM3","stoichiometry":0.2},{"gene":"CAPZB","stoichiometry":0.2},{"gene":"CTTN","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/CFL1","total_profiled":1310},"omim":[{"mim_id":"621464","title":"LIMB- AND CNS-EXPRESSED GENE 1-LIKE; LIX1L","url":"https://www.omim.org/entry/621464"},{"mim_id":"621425","title":"ABRA C-TERMINAL-LIKE PROTEIN; ABRACL","url":"https://www.omim.org/entry/621425"},{"mim_id":"616450","title":"EF-HAND DOMAIN FAMILY, MEMBER D2; EFHD2","url":"https://www.omim.org/entry/616450"},{"mim_id":"616128","title":"FAMILY WITH SEQUENCE SIMILARITY 89, MEMBER B; FAM89B","url":"https://www.omim.org/entry/616128"},{"mim_id":"610932","title":"TWINFILIN ACTIN-BINDING PROTEIN 1; TWF1","url":"https://www.omim.org/entry/610932"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Plasma membrane","reliability":"Approved"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/CFL1"},"hgnc":{"alias_symbol":[],"prev_symbol":["CFL"]},"alphafold":{"accession":"P23528","domains":[{"cath_id":"3.40.20.10","chopping":"5-164","consensus_level":"high","plddt":88.0897,"start":5,"end":164}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P23528","model_url":"https://alphafold.ebi.ac.uk/files/AF-P23528-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P23528-F1-predicted_aligned_error_v6.png","plddt_mean":87.56},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=CFL1","jax_strain_url":"https://www.jax.org/strain/search?query=CFL1"},"sequence":{"accession":"P23528","fasta_url":"https://rest.uniprot.org/uniprotkb/P23528.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P23528/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P23528"}},"corpus_meta":[{"pmid":"29156719","id":"PMC_29156719","title":"The microRNAs miR-200b-3p and miR-429-5p target the LIMK1/CFL1 pathway to inhibit growth and motility of breast cancer cells.","date":"2017","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/29156719","citation_count":78,"is_preprint":false},{"pmid":"20095995","id":"PMC_20095995","title":"The expression of CFL1 and N-WASP in esophageal squamous cell carcinoma and its correlation with clinicopathological features.","date":"2010","source":"Diseases of the esophagus : official journal of the International Society for Diseases of the Esophagus","url":"https://pubmed.ncbi.nlm.nih.gov/20095995","citation_count":59,"is_preprint":false},{"pmid":"10784045","id":"PMC_10784045","title":"Candida albicans CFL1 encodes a functional ferric reductase activity that can rescue a Saccharomyces cerevisiae fre1 mutant.","date":"2000","source":"Microbiology (Reading, England)","url":"https://pubmed.ncbi.nlm.nih.gov/10784045","citation_count":56,"is_preprint":false},{"pmid":"31495694","id":"PMC_31495694","title":"Mechanical Forces Regulate Cardiomyocyte Myofilament Maturation via the VCL-SSH1-CFL Axis.","date":"2019","source":"Developmental cell","url":"https://pubmed.ncbi.nlm.nih.gov/31495694","citation_count":44,"is_preprint":false},{"pmid":"23810590","id":"PMC_23810590","title":"A low membrane lipid phase transition temperature is associated with a high cryotolerance of Lactobacillus delbrueckii subspecies bulgaricus CFL1.","date":"2013","source":"Journal of dairy science","url":"https://pubmed.ncbi.nlm.nih.gov/23810590","citation_count":40,"is_preprint":false},{"pmid":"37203277","id":"PMC_37203277","title":"Remodeling Serine Synthesis and Metabolism via Nanoparticles (NPs)-Mediated CFL1 Silencing to Enhance the Sensitivity of Hepatocellular Carcinoma to Sorafenib.","date":"2023","source":"Advanced science (Weinheim, Baden-Wurttemberg, Germany)","url":"https://pubmed.ncbi.nlm.nih.gov/37203277","citation_count":33,"is_preprint":false},{"pmid":"8800436","id":"PMC_8800436","title":"Mapping of human non-muscle type cofilin (CFL1) to chromosome 11q13 and muscle-type cofilin (CFL2) to chromosome 14.","date":"1996","source":"Annals of human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/8800436","citation_count":33,"is_preprint":false},{"pmid":"24631590","id":"PMC_24631590","title":"Novel role of the Candida albicans ferric reductase gene CFL1 in iron acquisition, oxidative stress tolerance, morphogenesis and virulence.","date":"2014","source":"Research in microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/24631590","citation_count":32,"is_preprint":false},{"pmid":"29563135","id":"PMC_29563135","title":"Identification of CRKII, CFL1, CNTN1, NME2, and TKT as Novel and Frequent T-Cell Targets in Human IDH-Mutant Glioma.","date":"2018","source":"Clinical cancer research : an official journal of the American Association for Cancer Research","url":"https://pubmed.ncbi.nlm.nih.gov/29563135","citation_count":26,"is_preprint":false},{"pmid":"28190471","id":"PMC_28190471","title":"Plant lectins ConBr and CFL modulate expression toll-like receptors, pro-inflammatory cytokines and reduce the bacterial burden in macrophages infected with Salmonella enterica serovar Typhimurium.","date":"2016","source":"Phytomedicine : international journal of phytotherapy and phytopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/28190471","citation_count":24,"is_preprint":false},{"pmid":"25130162","id":"PMC_25130162","title":"A novel role of the ferric reductase Cfl1 in cell wall integrity, mitochondrial function, and invasion to host cells in Candida albicans.","date":"2014","source":"FEMS yeast research","url":"https://pubmed.ncbi.nlm.nih.gov/25130162","citation_count":22,"is_preprint":false},{"pmid":"34321041","id":"PMC_34321041","title":"Hhex inhibits cell migration via regulating RHOA/CDC42-CFL1 axis in human lung cancer cells.","date":"2021","source":"Cell communication and signaling : CCS","url":"https://pubmed.ncbi.nlm.nih.gov/34321041","citation_count":18,"is_preprint":false},{"pmid":"17352815","id":"PMC_17352815","title":"Association between CFL1 gene polymorphisms and spina bifida risk in a California population.","date":"2007","source":"BMC medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/17352815","citation_count":18,"is_preprint":false},{"pmid":"33044112","id":"PMC_33044112","title":"SSH1 impedes SQSTM1/p62 flux and MAPT/Tau clearance independent of CFL (cofilin) activation.","date":"2020","source":"Autophagy","url":"https://pubmed.ncbi.nlm.nih.gov/33044112","citation_count":18,"is_preprint":false},{"pmid":"34838479","id":"PMC_34838479","title":"E2F4-induced AGAP2-AS1 up-regulation accelerates the progression of colorectal cancer via miR-182-5p/CFL1 axis.","date":"2021","source":"Digestive and liver disease : official journal of the Italian Society of Gastroenterology and the Italian Association for the Study of the Liver","url":"https://pubmed.ncbi.nlm.nih.gov/34838479","citation_count":17,"is_preprint":false},{"pmid":"18392697","id":"PMC_18392697","title":"Overexpression of the cucumber LEAFY homolog CFL and hormone treatments alter flower development in gloxinia (Sinningia speciosa).","date":"2008","source":"Plant molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/18392697","citation_count":17,"is_preprint":false},{"pmid":"37193711","id":"PMC_37193711","title":"HUNK inhibits epithelial-mesenchymal transition of CRC via direct phosphorylation of GEF-H1 and activating RhoA/LIMK-1/CFL-1.","date":"2023","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/37193711","citation_count":16,"is_preprint":false},{"pmid":"39114368","id":"PMC_39114368","title":"The role of actin cytoskeleton CFL1 and ADF/cofilin superfamily in inflammatory response.","date":"2024","source":"Frontiers in molecular biosciences","url":"https://pubmed.ncbi.nlm.nih.gov/39114368","citation_count":14,"is_preprint":false},{"pmid":"35092861","id":"PMC_35092861","title":"Identification of the most damaging nsSNPs in the human CFL1 gene and their functional and structural impacts on cofilin-1 protein.","date":"2022","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/35092861","citation_count":14,"is_preprint":false},{"pmid":"26717562","id":"PMC_26717562","title":"Phylogenetic Patterns of Codon Evolution in the ACTIN-DEPOLYMERIZING FACTOR/COFILIN (ADF/CFL) Gene Family.","date":"2015","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/26717562","citation_count":14,"is_preprint":false},{"pmid":"31990066","id":"PMC_31990066","title":"Overexpression of CFL1 in gastric cancer and the effects of its silencing by siRNA with a nanoparticle delivery system in the gastric cancer cell line.","date":"2020","source":"Journal of cellular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/31990066","citation_count":10,"is_preprint":false},{"pmid":"24567775","id":"PMC_24567775","title":"Matricellular protein Cfl1 regulates cell differentiation.","date":"2013","source":"Communicative & integrative biology","url":"https://pubmed.ncbi.nlm.nih.gov/24567775","citation_count":8,"is_preprint":false},{"pmid":"36860487","id":"PMC_36860487","title":"Bio-removal of rare earth elements from hazardous industrial waste of CFL bulbs by the extremophile red alga Galdieria sulphuraria.","date":"2023","source":"Frontiers in microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/36860487","citation_count":7,"is_preprint":false},{"pmid":"38183912","id":"PMC_38183912","title":"CFL1 restores the migratory capacity of bone marrow mesenchymal stem cells in primary Sjögren's syndrome by regulating CCR1 expression.","date":"2024","source":"International immunopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/38183912","citation_count":5,"is_preprint":false},{"pmid":"38353363","id":"PMC_38353363","title":"Neuronal CFL1 upregulation in head and neck squamous cell carcinoma enhances tumor-nerve crosstalk and promotes tumor growth.","date":"2024","source":"Molecular carcinogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/38353363","citation_count":4,"is_preprint":false},{"pmid":"37406176","id":"PMC_37406176","title":"Identification cloning and functional analysis of novel natural antisense lncRNA CFL1-AS1 in cattle.","date":"2023","source":"Epigenetics","url":"https://pubmed.ncbi.nlm.nih.gov/37406176","citation_count":3,"is_preprint":false},{"pmid":"35418661","id":"PMC_35418661","title":"Estradiol inhibits HIV-1BaL infection and induces CFL1 expression in peripheral blood mononuclear cells and endocervical mucosa.","date":"2022","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/35418661","citation_count":3,"is_preprint":false},{"pmid":"15643084","id":"PMC_15643084","title":"[Expression of CFL gene during differentiation of floral and vegetative buds in cucumber cotyledonary nodes cultured in vitro].","date":"2004","source":"Zhi wu sheng li yu fen zi sheng wu xue xue bao = Journal of plant physiology and molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/15643084","citation_count":3,"is_preprint":false},{"pmid":"40146350","id":"PMC_40146350","title":"In vitro chrysene degradation by purified cell free laccase (P-CFL) from Cochliobolus lunatus strain CHR4D in the presence of various redox mediator systems (RMSs) and computational evaluation of their laccase-ligand interactions.","date":"2025","source":"Environmental science and pollution research international","url":"https://pubmed.ncbi.nlm.nih.gov/40146350","citation_count":3,"is_preprint":false},{"pmid":"39152428","id":"PMC_39152428","title":"Sensitization of hepatocellular carcinoma cells to HDACi is regulated through hsa-miR-342-5p/CFL1.","date":"2024","source":"Cancer cell international","url":"https://pubmed.ncbi.nlm.nih.gov/39152428","citation_count":2,"is_preprint":false},{"pmid":"17537423","id":"PMC_17537423","title":"Cryotolerance of Lactobacillus delbrueckii subsp. bulgaricus CFL1 is modified by acquisition of antibiotic resistance.","date":"2007","source":"Cryobiology","url":"https://pubmed.ncbi.nlm.nih.gov/17537423","citation_count":2,"is_preprint":false},{"pmid":"34781231","id":"PMC_34781231","title":"Anti-infective activity of Cratylia argentea lectin (CFL) against experimental infection with virulent Listeria monocytogenes in Swiss mice.","date":"2021","source":"Phytomedicine : international journal of phytotherapy and phytopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/34781231","citation_count":2,"is_preprint":false},{"pmid":"40931167","id":"PMC_40931167","title":"Hippocampal Cofilin and CFL1 gene variants are linked to Alcohol Use Disorder phenotypes.","date":"2025","source":"Molecular psychiatry","url":"https://pubmed.ncbi.nlm.nih.gov/40931167","citation_count":1,"is_preprint":false},{"pmid":"38684875","id":"PMC_38684875","title":"Serum anti-CFL1, anti-EZR, and anti-CYPA autoantibody as diagnostic markers in ovarian cancer.","date":"2024","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/38684875","citation_count":1,"is_preprint":false},{"pmid":"36547626","id":"PMC_36547626","title":"Heterologous Expression of CFL1 Confers Flocculating Ability to Cutaneotrichosporon oleaginosus Lipid-Rich Cells.","date":"2022","source":"Journal of fungi (Basel, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/36547626","citation_count":1,"is_preprint":false},{"pmid":"36639125","id":"PMC_36639125","title":"Multiobjective optimization of frozen and freeze-dried Lactobacillus delbrueckii subsp. bulgaricus CFL1 production via the modification of fermentation conditions.","date":"2023","source":"Journal of applied microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/36639125","citation_count":1,"is_preprint":false},{"pmid":"30460068","id":"PMC_30460068","title":"CFL-1, a novel F-box protein with leucine-rich repeat may interact with UNC-10 for the regulation of defecation and daumone response in Caenorhabditis elegans.","date":"2017","source":"Animal cells and systems","url":"https://pubmed.ncbi.nlm.nih.gov/30460068","citation_count":1,"is_preprint":false},{"pmid":"38495482","id":"PMC_38495482","title":"CFL1 is Implicated in Chronic Myeloid Leukemia Response during Imatinib Therapy.","date":"2024","source":"Journal of Cancer","url":"https://pubmed.ncbi.nlm.nih.gov/38495482","citation_count":0,"is_preprint":false},{"pmid":"40179112","id":"PMC_40179112","title":"TENS improves CFL injury rat and regulates the intestinal microbiota.","date":"2025","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/40179112","citation_count":0,"is_preprint":false},{"pmid":"41888102","id":"PMC_41888102","title":"c-Myc transactivates CFL1 to induce senescence-like phenotype and potentiate the bystander effects for the migration and proliferation in lung cancer cells.","date":"2026","source":"Cell death discovery","url":"https://pubmed.ncbi.nlm.nih.gov/41888102","citation_count":0,"is_preprint":false},{"pmid":"25414520","id":"PMC_25414520","title":"A New Runge-Kutta Discontinuous Galerkin Method with Conservation Constraint to Improve CFL Condition for Solving Conservation Laws.","date":"2014","source":"Journal of computational physics","url":"https://pubmed.ncbi.nlm.nih.gov/25414520","citation_count":0,"is_preprint":false},{"pmid":"41684538","id":"PMC_41684538","title":"Inactivation of cofilin-1 in Mcpt5-Cre-nf-Cfl1fl/fl mice prevents the formation of connective tissue mast cells without affecting basophils: a new tool to investigate the specific role of CTMCs in disease.","date":"2026","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/41684538","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.09.11.25335447","title":"Anterior Talofibular Ligament and Calcaneofibular Ligament Repair Reinforced by Bio-inductive Implants","date":"2025-09-12","source":"bioRxiv","url":"https://doi.org/10.1101/2025.09.11.25335447","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.06.25.661657","title":"A Two-Phase Core-Plasma Model for Microvascular Blood Flow: Comparative Analysis of Hemodynamic Models","date":"2025-06-28","source":"bioRxiv","url":"https://doi.org/10.1101/2025.06.25.661657","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.09.22.677682","title":"High-Fidelity Simulation of Cell Adhesion and Flow with Coarse Grain Modelling: A Numerical and Experimental Approach","date":"2025-09-23","source":"bioRxiv","url":"https://doi.org/10.1101/2025.09.22.677682","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.09.27.615114","title":"Deficiency of actin depolymerizing factors ADF/Cfl1 in microglia decreases motility and impairs memory","date":"2024-09-29","source":"bioRxiv","url":"https://doi.org/10.1101/2024.09.27.615114","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.07.13.603408","title":"Androgen receptor plays critical role in regulating cervical cancer cell migration","date":"2024-07-17","source":"bioRxiv","url":"https://doi.org/10.1101/2024.07.13.603408","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.08.27.609917","title":"Extracellular Vesicles from Stem cells Rescue Cellular Phenotypes and Behavioral Deficits in SHANK3-Associated ASD Neuronal and Mouse Models","date":"2024-08-27","source":"bioRxiv","url":"https://doi.org/10.1101/2024.08.27.609917","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":25561,"output_tokens":3886,"usd":0.067487,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11656,"output_tokens":3917,"usd":0.078103,"stage2_stop_reason":"end_turn"},"total_usd":0.14559,"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\": 2019,\n      \"finding\": \"Mechanical forces from cardiac contractility regulate cardiomyocyte myofilament maturation through the VCL-SSH1-CFL axis: vinculin (VCL) recruits the phosphatase SSH1 and its effector cofilin (CFL) to regulate F-actin rearrangement and promote myofilament maturation in zebrafish cardiomyocytes.\",\n      \"method\": \"Interactome profiling (contracting vs. non-contracting cardiomyocytes), Co-IP, genetic loss-of-function, live imaging, epistasis analysis\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal interactome, Co-IP, genetic epistasis, and functional phenotype across multiple orthogonal methods in a single rigorous study\",\n      \"pmids\": [\"31495694\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"LIMK1 directly phosphorylates and inactivates CFL1; inhibition of LIMK1 reduces CFL1 expression and phosphorylation, thereby impairing CFL1-dependent F-actin/G-actin remodeling and suppressing breast cancer cell migration and invasion.\",\n      \"method\": \"Western blot for pCFL1/CFL1 following LIMK1 knockdown, migration/invasion assays, miRNA mimic transfection in TNBC cells\",\n      \"journal\": \"Oncotarget\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — multiple cell-based assays in single lab, consistent with known LIMK1-cofilin pathway but no in vitro kinase reconstitution\",\n      \"pmids\": [\"29156719\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"SSH1, the canonical CFL (cofilin) phosphatase, dephosphorylates phospho-Ser403-SQSTM1/p62, impairing autophagic cargo clearance; this action is separable from SSH1-mediated CFL activation, demonstrating that SSH1 has a CFL-independent substrate.\",\n      \"method\": \"RNAi knockdown, overexpression with defined mutant constructs, genetically encoded fluorescent reporters, cell lines and primary neurons\",\n      \"journal\": \"Autophagy\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (RNAi, OE, mutant constructs, reporters) in cell lines and primary neurons, with genetic dissection of CFL-dependent vs. independent actions\",\n      \"pmids\": [\"33044112\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Hhex enhances the interaction of RHOGDIA with RHOA/CDC42, maintaining them in inactive forms, which blocks the RHOA/CDC42–LIMK1–pCFL1 signaling cascade, preserving CFL1 F-actin-severing activity and suppressing filopodium/lamellipodium formation and cell migration in lung cancer cells.\",\n      \"method\": \"Western blot, co-immunoprecipitation, wound-healing scratch assay, laser confocal imaging, overexpression/knockdown in NSCLC cells\",\n      \"journal\": \"Cell communication and signaling : CCS\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 / Moderate — Co-IP, functional migration assays, and pathway epistasis in single lab with multiple orthogonal methods\",\n      \"pmids\": [\"34321041\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"HUNK kinase directly phosphorylates GEF-H1 at serine 645, which activates RhoA and triggers a cascade of LIMK-1/CFL-1 phosphorylation that stabilizes F-actin and inhibits EMT and metastasis in colorectal cancer cells.\",\n      \"method\": \"In vitro kinase assay (direct phosphorylation), Western blot for phospho-CFL-1, overexpression/knockdown, metastasis assays\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct in vitro kinase phosphorylation assay plus cell-based epistasis, single lab\",\n      \"pmids\": [\"37193711\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"CFL1 promotes phosphoglycerate dehydrogenase (PHGDH) transcription to enhance serine synthesis and metabolism, increasing antioxidant production that scavenges sorafenib-induced ROS, thereby impairing sorafenib sensitivity in hepatocellular carcinoma.\",\n      \"method\": \"Transcriptome sequencing of sorafenib-sensitive vs. insensitive HCC patients, siRNA knockdown, metabolic assays, nanoparticle co-delivery experiments in vivo\",\n      \"journal\": \"Advanced science (Weinheim, Baden-Wurttemberg, Germany)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 / Moderate — RNA-seq, siRNA loss-of-function, metabolic readouts, and in vivo validation in single lab\",\n      \"pmids\": [\"37203277\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Estradiol (E2) dose-dependently increases total CFL1 and phosphorylated CFL1 (pCFL1) expression in PBMCs and endocervical mucosa, and this induction is abrogated by LIMK1/2 inhibitor LIMKi3; CFL1 knockdown partially restores HIV-1 infection suppressed by E2, linking E2-mediated CFL1 phosphorylation to inhibition of HIV-1BaL infection.\",\n      \"method\": \"LIMK inhibitor treatment, CFL1 siRNA knockdown, protein expression by Western blot, HIV-1 infectivity assay in PBMCs and endocervical explants\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 / Moderate — pharmacological inhibition, siRNA knockdown, and functional infection assay in two tissue models, single lab\",\n      \"pmids\": [\"35418661\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CFL1 overexpression in pSS bone marrow MSCs restores their migratory capacity by upregulating CCR1 expression; pharmacological inhibition of CCR1 suppresses the CFL1-induced migration and proliferation, placing CFL1 upstream of the CCL5/CCR1 axis.\",\n      \"method\": \"Lentivirus-mediated CFL1 overexpression, RNA-seq, Transwell migration assay, wound healing assay, CCR1 inhibitor rescue experiment, NOD mouse model\",\n      \"journal\": \"International immunopharmacology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 / Moderate — RNA-seq pathway identification, pharmacological rescue epistasis, and in vivo validation, single lab\",\n      \"pmids\": [\"38183912\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Expression of a non-functional form of Cfl1 (nf-Cfl1) in connective tissue mast cells (CTMCs) via Mcpt5-Cre results in complete absence of CTMCs, demonstrating that Cfl1-dependent actin depolymerization is essential for CTMC formation; CTMCs lacking Cfl1 activity show impaired systemic anaphylaxis.\",\n      \"method\": \"Conditional knock-in of non-functional Cfl1 in CTMCs (Mcpt5-Cre-nf-Cfl1fl/fl), flow cytometry, passive systemic anaphylaxis model\",\n      \"journal\": \"Frontiers in immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — genetic knock-in of non-functional protein with cell-type-specific Cre, clear phenotypic readout (complete CTMC absence), functional anaphylaxis assay\",\n      \"pmids\": [\"41684538\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Overexpression of Cfl1 in the polymorphic layer of the hippocampal dentate gyrus (PoDG) inhibits ML-DG synapses, increases motivation to seek alcohol, impairs extinction of alcohol-seeking, and decreases reward consumption during relapse; reducing Cfl1 levels has opposite effects, establishing hippocampal Cfl1 as a regulator of synaptic transmission and AUD-relevant behavior.\",\n      \"method\": \"Local viral overexpression/knockdown of Cfl in hippocampal DG subregions, in vivo two-photon imaging of synaptic activity, automated IntelliCage behavioral testing, RNA sequencing\",\n      \"journal\": \"Molecular psychiatry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — bidirectional (OE and KD) viral manipulation with synaptic electrophysiology and behavioral phenotype, single lab\",\n      \"pmids\": [\"40931167\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"c-Myc transcriptionally activates CFL1 by binding E-boxes in the CFL1 promoter (confirmed by ChIP-qPCR); cofilin-1 upregulation mediates c-Myc-induced oncogene-induced senescence (OIS) and F-actin/nuclear G-actin accumulation in lung cancer cells; knockdown of cofilin-1 suppresses cMIS, and a physical interaction between c-Myc and cofilin-1 was detected by co-immunoprecipitation.\",\n      \"method\": \"ChIP-qPCR, cofilin-1 knockdown, c-Myc overexpression/truncation mutants, Co-IP, F-actin/G-actin assays, conditioned medium bystander assays\",\n      \"journal\": \"Cell death discovery\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP-qPCR identifies direct promoter binding, Co-IP shows physical interaction, loss-of-function rescues phenotype; multiple orthogonal methods, single lab\",\n      \"pmids\": [\"41888102\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Conditional knockout of ADF and Cfl1 in microglia (ADF/Cfl1-KO) causes increased accumulation of stabilized F-actin and altered microtubule dynamics, reduces microglial fine process motility, and impairs migration toward laser-induced lesions in vivo; microglial ADF/Cfl1-deficiency also decreases learning and memory in mice.\",\n      \"method\": \"Conditional KO mouse model, in vivo two-photon imaging, F-actin staining, microtubule dynamics assays, behavioral memory tests\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO with in vivo imaging and behavioral readouts; preprint, not yet peer-reviewed\",\n      \"pmids\": [\"bio_10.1101_2024.09.27.615114\"],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 1996,\n      \"finding\": \"Human non-muscle cofilin (CFL1) was mapped to chromosome 11q13 by PCR in somatic cell hybrid panels and confirmed by FISH, while muscle-type cofilin (CFL2) was mapped to chromosome 14; CFL1 was identified as an actin-binding protein involved in actin-cofilin complex translocation from cytoplasm to nucleus.\",\n      \"method\": \"PCR in rodent-human somatic cell hybrids, FISH of genomic cosmid clones, radiation hybrid mapping\",\n      \"journal\": \"Annals of human genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — multiple independent mapping methods (somatic cell hybrids, FISH, radiation hybrids) converging on same chromosomal locus\",\n      \"pmids\": [\"8800436\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CFL1 expression in nerves (rather than in tumor cells) within the head and neck squamous cell carcinoma (HNSCC) tumor microenvironment is associated with perineural invasion; HNSCC cells induce neuronal CFL1 expression, and conditional knockout of neuronal CFL1 impedes tumor-nerve interactions.\",\n      \"method\": \"Multiplex fluorescent immunohistochemistry, conditional neuronal CFL1 knockout, tumor-nerve co-culture models\",\n      \"journal\": \"Molecular carcinogenesis\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 / Moderate — conditional KO with functional tumor-nerve interaction readout and localization by multiplex IHC, single lab\",\n      \"pmids\": [\"38353363\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Imatinib therapy in CML upregulates CFL1 expression and activity (cofilin, P-cofilin) and increases F-actin (including branched F-actin), indicating that imatinib affects F-actin remodeling in CML cells by regulating CFL1.\",\n      \"method\": \"Western blot for CFL1 and P-cofilin in CML patient samples and cell lines pre/post imatinib, F-actin staining\",\n      \"journal\": \"Journal of Cancer\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single-lab observational protein measurements with limited mechanistic dissection\",\n      \"pmids\": [\"38495482\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"CFL1 (non-muscle cofilin-1) is an actin-severing and depolymerizing protein whose activity is tightly regulated by phosphorylation: LIMK1/2 phosphorylates and inactivates CFL1, while SSH1 dephosphorylates and activates it; mechanical forces transmitted through vinculin (VCL) recruit SSH1 to activate CFL1 for F-actin rearrangement during cardiomyocyte myofilament maturation; CFL1 is also transcriptionally activated by c-Myc (via direct E-box binding) to drive oncogene-induced senescence, promotes serine synthesis metabolism to confer chemoresistance, regulates cell migration via the RHOA/CDC42–LIMK1 axis downstream of Hhex and upstream of CCR1, and is essential for connective tissue mast cell formation and microglial cytoskeletal dynamics and motility.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"CFL1 (non-muscle cofilin-1) is an actin-binding protein that severs and depolymerizes F-actin to drive cytoskeletal remodeling underlying cell migration, tissue morphogenesis, and synaptic function [#12, #8]. Its activity is gated by reversible phosphorylation: LIMK1 directly phosphorylates and inactivates CFL1, blocking its F-actin/G-actin remodeling activity [#1], while the phosphatase SSH1 dephosphorylates and reactivates it [#2]. This kinase-phosphatase switch sits downstream of RHOA/CDC42 signaling, where upstream regulators such as Hhex (via RHOGDIA sequestration of RHOA/CDC42) and HUNK (via GEF-H1/RhoA activation) tune the RHOA/CDC42-LIMK1-pCFL1 cascade to control filopodium/lamellipodium formation, EMT, and metastasis [#3, #4]. Mechanical forces transmitted through vinculin recruit SSH1 and cofilin to remodel F-actin during cardiomyocyte myofilament maturation, embedding CFL1 in mechanotransduction [#0]. CFL1-dependent actin depolymerization is genetically essential for connective tissue mast cell formation and systemic anaphylaxis [#8] and, together with ADF, for microglial process motility, migration, and learning-related behavior [#11]. Beyond its cytoskeletal role, CFL1 acts as a transcriptional driver: c-Myc binds E-boxes in the CFL1 promoter to upregulate cofilin-1 and induce oncogene-induced senescence with nuclear G-actin accumulation [#10], and CFL1 promotes PHGDH transcription to enhance serine synthesis and antioxidant output, conferring sorafenib resistance in hepatocellular carcinoma [#5].\",\n  \"teleology\": [\n    {\n      \"year\": 1996,\n      \"claim\": \"Establishing the molecular identity and chromosomal locus of human non-muscle cofilin distinguished CFL1 from its muscle paralog and framed it as an actin-binding protein capable of nuclear translocation.\",\n      \"evidence\": \"PCR in somatic cell hybrids, FISH, and radiation hybrid mapping\",\n      \"pmids\": [\"8800436\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not define the biochemical regulation of severing activity\", \"Functional consequences of cytoplasm-to-nucleus translocation left unresolved\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Identifying LIMK1 as a direct upstream kinase that phosphorylates and inactivates CFL1 connected cofilin regulation to cancer cell motility.\",\n      \"evidence\": \"LIMK1 knockdown with pCFL1/CFL1 Western blot and migration/invasion assays in TNBC cells\",\n      \"pmids\": [\"29156719\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No in vitro kinase reconstitution\", \"Single cell-line context\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Linking CFL1 to mechanotransduction showed that vinculin recruits the SSH1 phosphatase to activate cofilin for F-actin rearrangement during cardiomyocyte myofilament maturation.\",\n      \"evidence\": \"Interactome profiling, Co-IP, genetic loss-of-function and epistasis with live imaging in zebrafish cardiomyocytes\",\n      \"pmids\": [\"31495694\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct binding stoichiometry of the VCL-SSH1-CFL axis not resolved\", \"Mammalian generalization not tested in this study\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Demonstrating that SSH1 dephosphorylates p62/SQSTM1 independently of cofilin established that the cofilin phosphatase has CFL-independent substrates, refining how SSH1 activity relates to CFL1.\",\n      \"evidence\": \"RNAi, overexpression of defined mutants, fluorescent reporters in cell lines and primary neurons\",\n      \"pmids\": [\"33044112\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Does not directly address CFL1 catalytic mechanism\", \"Relative flux through CFL-dependent vs independent SSH1 actions in vivo unknown\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Placing CFL1 downstream of RHOA/CDC42-LIMK1 and under Hhex/RHOGDIA control clarified how upstream small-GTPase regulation tunes cofilin-mediated severing and migration.\",\n      \"evidence\": \"Co-IP, scratch assays, confocal imaging, and pathway epistasis in NSCLC cells\",\n      \"pmids\": [\"34321041\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab and cell-type\", \"Direct effect on CFL1 severing kinetics not measured\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Identifying HUNK-GEF-H1-RhoA as an upstream input into the LIMK1/CFL1 axis extended the regulatory map of cofilin phosphorylation to suppression of EMT and metastasis.\",\n      \"evidence\": \"In vitro kinase assay on GEF-H1, phospho-CFL1 Western blot, and metastasis assays in colorectal cancer cells\",\n      \"pmids\": [\"37193711\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"CFL1 phosphorylation inferred from cascade, not directly reconstituted\", \"Single lab\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Revealing that CFL1 promotes PHGDH transcription assigned cofilin a non-cytoskeletal role in serine/antioxidant metabolism that drives chemoresistance.\",\n      \"evidence\": \"Transcriptome sequencing, siRNA knockdown, metabolic assays, and in vivo nanoparticle co-delivery in HCC\",\n      \"pmids\": [\"37203277\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which CFL1 reaches/regulates the PHGDH promoter not defined\", \"Whether this requires nuclear actin binding unknown\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Genetic knock-in of non-functional Cfl1 in connective tissue mast cells proved that cofilin-dependent actin depolymerization is required for mast cell formation and anaphylaxis.\",\n      \"evidence\": \"Cell-type-specific knock-in (Mcpt5-Cre-nf-Cfl1), flow cytometry, passive systemic anaphylaxis\",\n      \"pmids\": [\"41684538\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Developmental step at which CTMCs are lost not pinpointed\", \"Upstream regulators of CFL1 in mast cells not identified\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Conditional loss of ADF/Cfl1 in microglia established cofilin as required for F-actin/microtubule balance, process motility, and lesion-directed migration with cognitive consequences.\",\n      \"evidence\": \"Conditional KO, in vivo two-photon imaging, cytoskeletal staining, behavioral tests (preprint)\",\n      \"pmids\": [\"bio_10.1101_2024.09.27.615114\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Preprint, not yet peer-reviewed\", \"ADF and Cfl1 contributions not separated\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Linking CFL1 to chemokine signaling and to perineural invasion broadened its migratory roles to MSC homing (CCR1 axis) and tumor-nerve interactions.\",\n      \"evidence\": \"Lentiviral CFL1 overexpression with CCR1 inhibitor rescue and NOD model; conditional neuronal CFL1 KO with tumor-nerve co-culture and multiplex IHC\",\n      \"pmids\": [\"38183912\", \"38353363\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism connecting CFL1 to CCR1 transcription unclear\", \"Cell-intrinsic vs microenvironmental contributions only partly dissected\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Bidirectional manipulation of hippocampal Cfl1 demonstrated a role in regulating synaptic transmission and alcohol-seeking behavior, extending cofilin function to neural circuits.\",\n      \"evidence\": \"Viral overexpression/knockdown in dentate gyrus subregions, two-photon synaptic imaging, IntelliCage behavior, RNA-seq\",\n      \"pmids\": [\"40931167\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular link between cofilin activity and synaptic change not resolved\", \"Single lab\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Showing c-Myc directly transactivates CFL1 via promoter E-boxes positioned cofilin as a transcriptional effector driving oncogene-induced senescence and nuclear G-actin accumulation.\",\n      \"evidence\": \"ChIP-qPCR, cofilin-1 knockdown, c-Myc mutants, Co-IP, F-actin/G-actin assays in lung cancer cells\",\n      \"pmids\": [\"41888102\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional significance of the c-Myc-cofilin physical interaction unclear\", \"How nuclear G-actin accumulation enforces senescence not defined\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How the phosphorylation switch, nuclear actin shuttling, and transcriptional/metabolic functions of CFL1 are coordinated within a single cell remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model integrating regulatory inputs\", \"Direct vs indirect mechanism of CFL1 transcriptional effects unknown\", \"Tissue-specific regulators upstream of LIMK/SSH1 not mapped\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [12, 1, 8]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [1, 4]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [12]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [12, 10]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [0, 11]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [1, 3, 4]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [0, 8]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"LIMK1\", \"SSH1\", \"VCL\", \"MYC\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}