{"gene":"CFAP45","run_date":"2026-06-09T22:57:18","timeline":{"discoveries":[{"year":2020,"finding":"CFAP45 binds AMP in vitro, and structural modelling identifies an AMP-binding interface between CFAP45 and adenylate kinase 8 (AK8). CFAP45 is part of an axonemal module including dynein ATPases, adenylate kinase, and CFAP52. Microtubule sliding of dyskinetic sperm from Cfap45-/- mice is rescued by addition of AMP or ADP with ATP (compared to ATP alone), demonstrating that CFAP45 supports ciliary/flagellar beating through adenine nucleotide homeostasis.","method":"In vitro AMP-binding assay, structural modelling, proteomic profiling of CFAP45-deficient cilia/flagella, Cfap45 knockout mouse model with microtubule sliding rescue assay","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — in vitro binding assay, structural modelling, knockout mouse with functional rescue experiment, and proteomic module identification, multiple orthogonal methods in one study","pmids":["33139725"],"is_preprint":false},{"year":2023,"finding":"In Xenopus embryos depleted of Cfap45, live confocal imaging shows that Cfap45 localizes in a punctate but static position within the ciliary axoneme. Depletion leads to loss of cilia stability and eventual detachment from the apical surface in both multiciliated and monociliated cells of the Left-Right Organizer, causing defects in cardiac looping and LR patterning.","method":"Cfap45 morpholino depletion in Xenopus embryos, live confocal imaging, LRO cilia analysis","journal":"Developmental biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct live imaging of localization with functional consequence (cilia loss), loss-of-function with defined phenotypic readout, single lab","pmids":["37172641"],"is_preprint":false},{"year":2023,"finding":"CFAP52 interacts with CFAP45 in the sperm flagellum, and knockout of Cfap52 decreases the expression level of CFAP45 in sperm flagellum, further disrupting microtubule sliding produced by dynein ATPase.","method":"Co-immunoprecipitation (CFAP52-CFAP45 interaction), Cfap52 knockout mouse model, immunofluorescence of sperm flagellum, microtubule sliding assay","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal interaction detected by Co-IP, knockout model with functional readout, corroborates the CFAP45-CFAP52 module from the 2020 Nature Communications paper","pmids":["37236356"],"is_preprint":false},{"year":2019,"finding":"NESG1 (CFAP45) interacts with VPS33B by colocalizing in the cytoplasm. Knockdown of NESG1 reverses the inhibitory effects of VPS33B overexpression in nasopharyngeal carcinoma cells by downregulating PI3K/AKT/c-Jun-mediated transcription repression.","method":"Co-localization by microscopy, NESG1 knockdown with functional rescue assay, western blot for PI3K/AKT/c-Jun pathway","journal":"Cell death & disease","confidence":"Low","confidence_rationale":"Tier 3 / Weak — co-localization (not direct biochemical interaction), single lab, single method for interaction claim","pmids":["30944308"],"is_preprint":false},{"year":2021,"finding":"NESG1 (CFAP45) interacts with VPS33B (confirmed by Co-IP) in ovarian cancer cells. Overexpressed NESG1 suppresses cell growth by mediating VPS33B-modulated EGFR/PI3K/AKT/c-Myc/p53/miR-133a-3p signals, and NESG1 induces VPS33B expression by reducing PI3K/AKT/c-Jun-mediated transcription inhibition.","method":"Co-immunoprecipitation, western blot, overexpression/knockdown functional assays","journal":"Cancer science","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single Co-IP for interaction, single lab, indirect pathway readouts","pmids":["33788346"],"is_preprint":false},{"year":2021,"finding":"NESG1 (CFAP45) suppresses NPC migration and invasion via the Wnt/β-catenin signaling pathway. NESG1 induces miR-1254, which targets HDGF, preventing HDGF/DDX5 complex-mediated nuclear translocation of β-catenin and downstream EMT signals.","method":"Transwell/Boyden assays, miRNA microarray, luciferase reporter assay, ChIP, EMSA, Co-IP, western blot, in vivo and in vitro loss-of-function/gain-of-function","journal":"Molecular therapy. Methods & clinical development","confidence":"Medium","confidence_rationale":"Tier 2-3 / Moderate — multiple orthogonal methods (luciferase, ChIP, EMSA, CoIP) in single lab establishing pathway placement","pmids":["33718512"],"is_preprint":false},{"year":2014,"finding":"CCDC19 (CFAP45) overexpression in NSCLC cells stimulates miR-184 expression, which suppresses C-Myc, blocking cell growth mediated by the PI3K/AKT/C-Jun pathway. Knockdown of CCDC19 restores cell growth in CCDC19-overexpressing cells.","method":"Lentiviral overexpression, shRNA knockdown, cell proliferation and cell cycle assays, miRNA and pathway western blot analysis","journal":"Journal of cellular and molecular medicine","confidence":"Low","confidence_rationale":"Tier 3 / Weak — indirect pathway inference via OE/KD, single lab, no direct binding assay between CCDC19 and miR-184","pmids":["24976536"],"is_preprint":false},{"year":2012,"finding":"Proteomics of NESG1 (CFAP45)-overexpressing NPC cells identified ENO1 (alpha-enolase) as a NESG1-regulated protein. Overexpressed ENO1 restores cell proliferation, antagonizes NESG1 regulation of p21 and CCNA1, and induces C-Myc, pRB, and E2F1 expression.","method":"2D gel proteomics, western blot, overexpression rescue experiments","journal":"Proteomics","confidence":"Low","confidence_rationale":"Tier 3 / Weak — proteomic screen with western blot validation and rescue assay, single lab, indirect mechanism","pmids":["22997098"],"is_preprint":false},{"year":2010,"finding":"Induced expression of NESG1 (CFAP45) in NPC cells decreases cell proliferation, G1-S phase transition, cell migration, invasion, and in vivo tumorigenesis, and significantly regulates expression of cell cycle regulators CCNA1 and p21.","method":"Lentiviral overexpression in NPC 5-8F cells, proliferation assay, cell cycle analysis, invasion/migration assays, xenograft in vivo tumorigenesis","journal":"International journal of cancer","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal functional assays (cell cycle, invasion, in vivo) with molecular readouts (CCNA1, p21), single lab","pmids":["20715168"],"is_preprint":false}],"current_model":"CFAP45 (also known as NESG1/CCDC19) is a cilia and flagella associated protein that supports motile ciliary and flagellar beating through an axonemal adenine nucleotide homeostasis module: it binds AMP and interacts with adenylate kinase AK8 and CFAP52, facilitating dynein ATPase-driven microtubule sliding; loss of CFAP45 causes situs abnormalities, asthenospermia, and cilia instability, while in cancer cell contexts it acts as a tumor suppressor by modulating PI3K/AKT signaling, miRNA networks (miR-184, miR-1254), and Wnt/β-catenin pathway activity."},"narrative":{"mechanistic_narrative":"CFAP45 (NESG1/CCDC19) is an axonemal protein that supports motile ciliary and flagellar beating by maintaining adenine nucleotide homeostasis within the axoneme [PMID:33139725]. It binds AMP and forms part of a module with dynein ATPases, adenylate kinase AK8, and CFAP52, where the modeled AMP-binding interface couples CFAP45 to AK8; dyskinetic microtubule sliding in Cfap45-deficient sperm is rescued by supplementing AMP or ADP with ATP, establishing that CFAP45 sustains the nucleotide pool that dynein-driven sliding consumes [PMID:33139725]. CFAP52 physically interacts with CFAP45 in the sperm flagellum and is required for its stable expression, linking the two as interdependent components of this beating module [PMID:37236356]. Within the axoneme CFAP45 occupies a punctate, static position, and its depletion destabilizes cilia and causes their detachment from the apical surface, producing left-right patterning and cardiac looping defects [PMID:37172641]. In epithelial cancer cell contexts CFAP45 acts as a growth-suppressive factor: its induced expression in nasopharyngeal carcinoma cells reduces proliferation, G1-S transition, migration, invasion, and xenograft tumorigenesis while regulating the cell cycle regulators CCNA1 and p21 [PMID:20715168], and it suppresses migration and invasion through a miR-1254/HDGF axis that blocks Wnt/β-catenin-driven β-catenin nuclear translocation and EMT [PMID:33718512].","teleology":[{"year":2010,"claim":"Established that CFAP45 (NESG1) can act as a growth- and invasion-suppressive factor, the first functional handle on the protein, framing it as a candidate tumor suppressor in nasopharyngeal carcinoma.","evidence":"Lentiviral overexpression in NPC cells with proliferation, cell cycle, invasion/migration assays and xenograft tumorigenesis, with CCNA1/p21 readouts","pmids":["20715168"],"confidence":"Medium","gaps":["No direct molecular mechanism linking CFAP45 to CCNA1/p21 regulation","Does not address the protein's ciliary/axonemal role","Overexpression-based phenotype, no endogenous loss-of-function"]},{"year":2012,"claim":"Asked which effectors mediate the growth-suppressive phenotype and implicated ENO1 as a CFAP45-regulated protein opposing its control of cell cycle regulators.","evidence":"2D gel proteomics of NESG1-overexpressing NPC cells with western blot and ENO1 overexpression rescue","pmids":["22997098"],"confidence":"Low","gaps":["Indirect mechanism; no direct CFAP45-ENO1 binding shown","Single lab, screen-based with limited validation","Causal ordering between CFAP45 and ENO1 not resolved"]},{"year":2014,"claim":"Connected CFAP45 (CCDC19) to a miRNA-mediated signaling output, positing that it suppresses growth through miR-184 and the PI3K/AKT/c-Jun axis.","evidence":"Lentiviral overexpression and shRNA knockdown in NSCLC cells with proliferation, cell cycle, and pathway western blots","pmids":["24976536"],"confidence":"Low","gaps":["No direct binding assay between CCDC19 and miR-184","Pathway inference is indirect via OE/KD","Mechanism of miRNA induction unknown"]},{"year":2019,"claim":"Probed an interaction partner and proposed CFAP45 (NESG1) cooperates with VPS33B to relieve PI3K/AKT/c-Jun-mediated transcriptional repression in NPC cells.","evidence":"Co-localization microscopy and NESG1 knockdown rescue with PI3K/AKT/c-Jun western blots","pmids":["30944308"],"confidence":"Low","gaps":["Interaction rests on co-localization, not direct biochemistry","Single lab, single method for the interaction claim","Functional link to ciliary biology unaddressed"]},{"year":2021,"claim":"Extended the VPS33B link to a Co-IP-confirmed interaction in ovarian cancer and to a Wnt/β-catenin-suppressive miR-1254/HDGF axis, sharpening the proposed anti-invasion mechanism.","evidence":"Co-IP, luciferase reporter, ChIP, EMSA, Transwell assays, and in vivo/in vitro gain/loss-of-function in cancer cells","pmids":["33788346","33718512"],"confidence":"Medium","gaps":["Multiple parallel signaling outputs not reconciled into one model","Relationship between cancer signaling roles and axonemal function unclear","VPS33B interaction not reciprocally validated across studies"]},{"year":2020,"claim":"Defined the core mechanistic role of CFAP45 as an axonemal AMP-binding partner of adenylate kinase AK8 that sustains adenine nucleotide homeostasis for dynein-driven microtubule sliding and ciliary/flagellar beating.","evidence":"In vitro AMP-binding assay, structural modelling, proteomics of CFAP45-deficient cilia, and Cfap45 knockout mouse with microtubule sliding rescue by AMP/ADP","pmids":["33139725"],"confidence":"High","gaps":["Structural model of the AK8 interface not experimentally resolved","Stoichiometry and exact placement within the axoneme not defined","Does not connect to the cancer signaling roles reported separately"]},{"year":2023,"claim":"Resolved CFAP45's static axonemal position and showed it is required for cilia stability, linking its loss to organ laterality defects, and established CFAP52 as a stabilizing physical partner.","evidence":"Cfap45 morpholino depletion and live confocal imaging in Xenopus LRO cilia; CFAP52-CFAP45 Co-IP and Cfap52 knockout mouse with sliding assay","pmids":["37172641","37236356"],"confidence":"Medium","gaps":["Molecular basis of how CFAP45 confers cilia stability is unknown","Mechanism by which CFAP52 maintains CFAP45 levels not defined","Single-lab depletion studies"]},{"year":null,"claim":"How the axonemal nucleotide-homeostasis function and the reported cancer-signaling roles of CFAP45 relate, if at all, remains unresolved.","evidence":"","pmids":[],"confidence":"Low","gaps":["No study bridges the ciliary/flagellar mechanism and the PI3K/AKT/Wnt signaling reports","No human disease-causing mutation directly demonstrated in the timeline","Direct binding partners beyond AK8/CFAP52/VPS33B not mapped"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140313","term_label":"molecular sequestering activity","supporting_discovery_ids":[0]}],"localization":[{"term_id":"GO:0005929","term_label":"cilium","supporting_discovery_ids":[0,1]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[0]}],"pathway":[{"term_id":"R-HSA-1852241","term_label":"Organelle biogenesis and maintenance","supporting_discovery_ids":[0,1]}],"complexes":[],"partners":["AK8","CFAP52","VPS33B"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9UL16","full_name":"Cilia- and flagella-associated protein 45","aliases":["Coiled-coil domain-containing protein 19","Nasopharyngeal epithelium-specific protein 1"],"length_aa":551,"mass_kda":65.7,"function":"Microtubule inner protein (MIP) part of the dynein-decorated doublet microtubules (DMTs) in cilia axoneme, which is required for motile cilia beating (PubMed:36191189). It is an AMP-binding protein that may facilitate dynein ATPase-dependent ciliary and flagellar beating via adenine nucleotide homeostasis. May function as a donor of AMP to AK8 and hence promote ADP production (PubMed:33139725)","subcellular_location":"Cytoplasm, cytoskeleton, cilium axoneme; Cytoplasm, cytoskeleton, flagellum axoneme; Cell projection, cilium; Cell projection, cilium, flagellum","url":"https://www.uniprot.org/uniprotkb/Q9UL16/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/CFAP45","classification":"Not Classified","n_dependent_lines":4,"n_total_lines":1208,"dependency_fraction":0.0033112582781456954},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/CFAP45","total_profiled":1310},"omim":[{"mim_id":"619608","title":"HETEROTAXY, VISCERAL, 11, AUTOSOMAL, WITH MALE INFERTILITY; HTX11","url":"https://www.omim.org/entry/619608"},{"mim_id":"619607","title":"HETEROTAXY, VISCERAL, 10, AUTOSOMAL, WITH MALE INFERTILITY; HTX10","url":"https://www.omim.org/entry/619607"},{"mim_id":"609804","title":"CILIA- AND FLAGELLA-ASSOCIATED PROTEIN 52; CFAP52","url":"https://www.omim.org/entry/609804"},{"mim_id":"605152","title":"CILIA- AND FLAGELLA-ASSOCIATED PROTEIN 45; CFAP45","url":"https://www.omim.org/entry/605152"},{"mim_id":"603339","title":"DYNEIN, AXONEMAL, HEAVY CHAIN 11; DNAH11","url":"https://www.omim.org/entry/603339"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Uncertain","locations":[{"location":"Nucleoplasm","reliability":"Uncertain"},{"location":"Mid piece","reliability":"Uncertain"},{"location":"Principal piece","reliability":"Uncertain"},{"location":"End piece","reliability":"Uncertain"},{"location":"Basal body","reliability":"Additional"},{"location":"Cytosol","reliability":"Additional"}],"tissue_specificity":"Group enriched","tissue_distribution":"Detected in some","driving_tissues":[{"tissue":"choroid plexus","ntpm":35.2},{"tissue":"fallopian tube","ntpm":42.1},{"tissue":"testis","ntpm":26.7}],"url":"https://www.proteinatlas.org/search/CFAP45"},"hgnc":{"alias_symbol":["NESG1"],"prev_symbol":["CCDC19"]},"alphafold":{"accession":"Q9UL16","domains":[],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9UL16","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9UL16-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9UL16-F1-predicted_aligned_error_v6.png","plddt_mean":76.25},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=CFAP45","jax_strain_url":"https://www.jax.org/strain/search?query=CFAP45"},"sequence":{"accession":"Q9UL16","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9UL16.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9UL16/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9UL16"}},"corpus_meta":[{"pmid":"30442369","id":"PMC_30442369","title":"Long noncoding RNA MAGI2-AS3 regulates CCDC19 expression by sponging miR-15b-5p and suppresses bladder cancer progression.","date":"2018","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/30442369","citation_count":58,"is_preprint":false},{"pmid":"30944308","id":"PMC_30944308","title":"VPS33B interacts with NESG1 to modulate EGFR/PI3K/AKT/c-Myc/P53/miR-133a-3p signaling and induce 5-fluorouracil sensitivity in nasopharyngeal carcinoma.","date":"2019","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/30944308","citation_count":57,"is_preprint":false},{"pmid":"33139725","id":"PMC_33139725","title":"CFAP45 deficiency causes situs abnormalities and asthenospermia by disrupting an axonemal adenine nucleotide homeostasis module.","date":"2020","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/33139725","citation_count":56,"is_preprint":false},{"pmid":"24976536","id":"PMC_24976536","title":"Candidate tumour suppressor CCDC19 regulates miR-184 direct targeting of C-Myc thereby suppressing cell growth in non-small cell lung cancers.","date":"2014","source":"Journal of cellular and molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/24976536","citation_count":52,"is_preprint":false},{"pmid":"20715168","id":"PMC_20715168","title":"Decreased expression of updated NESG1 in nasopharyngeal carcinoma: its potential role and preliminarily functional mechanism.","date":"2010","source":"International journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/20715168","citation_count":43,"is_preprint":false},{"pmid":"33718512","id":"PMC_33718512","title":"miR-1254 induced by NESG1 inactivates HDGF/DDX5-stimulated nuclear translocation of β-catenin and suppresses NPC metastasis.","date":"2021","source":"Molecular therapy. Methods & clinical development","url":"https://pubmed.ncbi.nlm.nih.gov/33718512","citation_count":13,"is_preprint":false},{"pmid":"22997098","id":"PMC_22997098","title":"Proteomic features of potential tumor suppressor NESG1 in nasopharyngeal carcinoma.","date":"2012","source":"Proteomics","url":"https://pubmed.ncbi.nlm.nih.gov/22997098","citation_count":10,"is_preprint":false},{"pmid":"37172641","id":"PMC_37172641","title":"CFAP45, a heterotaxy and congenital heart disease gene, affects cilia stability.","date":"2023","source":"Developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/37172641","citation_count":9,"is_preprint":false},{"pmid":"33788346","id":"PMC_33788346","title":"VPS33B interacts with NESG1 to suppress cell growth and cisplatin chemoresistance in ovarian cancer.","date":"2021","source":"Cancer science","url":"https://pubmed.ncbi.nlm.nih.gov/33788346","citation_count":6,"is_preprint":false},{"pmid":"37236356","id":"PMC_37236356","title":"The cilia and flagella associated protein CFAP52 orchestrated with CFAP45 is required for sperm motility in mice.","date":"2023","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/37236356","citation_count":6,"is_preprint":false},{"pmid":"31938168","id":"PMC_31938168","title":"Decreased CCDC19 is correlated with unfavorable outcome in lung squamous cell carcinoma.","date":"2018","source":"International journal of clinical and experimental pathology","url":"https://pubmed.ncbi.nlm.nih.gov/31938168","citation_count":4,"is_preprint":false},{"pmid":"21269959","id":"PMC_21269959","title":"[Construction of a lentiviral vector containing human NESG1 gene and its expression in 293FT cells].","date":"2011","source":"Nan fang yi ke da xue xue bao = Journal of Southern Medical University","url":"https://pubmed.ncbi.nlm.nih.gov/21269959","citation_count":0,"is_preprint":false},{"pmid":"22931604","id":"PMC_22931604","title":"[Screening of proteins regulated by CCDC19 gene in nasopharyngeal carcinoma using proteomics technology].","date":"2012","source":"Nan fang yi ke da xue xue bao = Journal of Southern Medical University","url":"https://pubmed.ncbi.nlm.nih.gov/22931604","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2024.10.04.616471","title":"Single-cell RNA sequencing on formalin-fixed and paraffin-embedded (FFPE) tissue identified multi-ciliary cells in breast cancer","date":"2024-10-04","source":"bioRxiv","url":"https://doi.org/10.1101/2024.10.04.616471","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":9061,"output_tokens":2388,"usd":0.031502,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":9418,"output_tokens":2915,"usd":0.059983,"stage2_stop_reason":"end_turn"},"total_usd":0.091485,"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\": 2020,\n      \"finding\": \"CFAP45 binds AMP in vitro, and structural modelling identifies an AMP-binding interface between CFAP45 and adenylate kinase 8 (AK8). CFAP45 is part of an axonemal module including dynein ATPases, adenylate kinase, and CFAP52. Microtubule sliding of dyskinetic sperm from Cfap45-/- mice is rescued by addition of AMP or ADP with ATP (compared to ATP alone), demonstrating that CFAP45 supports ciliary/flagellar beating through adenine nucleotide homeostasis.\",\n      \"method\": \"In vitro AMP-binding assay, structural modelling, proteomic profiling of CFAP45-deficient cilia/flagella, Cfap45 knockout mouse model with microtubule sliding rescue assay\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — in vitro binding assay, structural modelling, knockout mouse with functional rescue experiment, and proteomic module identification, multiple orthogonal methods in one study\",\n      \"pmids\": [\"33139725\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"In Xenopus embryos depleted of Cfap45, live confocal imaging shows that Cfap45 localizes in a punctate but static position within the ciliary axoneme. Depletion leads to loss of cilia stability and eventual detachment from the apical surface in both multiciliated and monociliated cells of the Left-Right Organizer, causing defects in cardiac looping and LR patterning.\",\n      \"method\": \"Cfap45 morpholino depletion in Xenopus embryos, live confocal imaging, LRO cilia analysis\",\n      \"journal\": \"Developmental biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct live imaging of localization with functional consequence (cilia loss), loss-of-function with defined phenotypic readout, single lab\",\n      \"pmids\": [\"37172641\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"CFAP52 interacts with CFAP45 in the sperm flagellum, and knockout of Cfap52 decreases the expression level of CFAP45 in sperm flagellum, further disrupting microtubule sliding produced by dynein ATPase.\",\n      \"method\": \"Co-immunoprecipitation (CFAP52-CFAP45 interaction), Cfap52 knockout mouse model, immunofluorescence of sperm flagellum, microtubule sliding assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal interaction detected by Co-IP, knockout model with functional readout, corroborates the CFAP45-CFAP52 module from the 2020 Nature Communications paper\",\n      \"pmids\": [\"37236356\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"NESG1 (CFAP45) interacts with VPS33B by colocalizing in the cytoplasm. Knockdown of NESG1 reverses the inhibitory effects of VPS33B overexpression in nasopharyngeal carcinoma cells by downregulating PI3K/AKT/c-Jun-mediated transcription repression.\",\n      \"method\": \"Co-localization by microscopy, NESG1 knockdown with functional rescue assay, western blot for PI3K/AKT/c-Jun pathway\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — co-localization (not direct biochemical interaction), single lab, single method for interaction claim\",\n      \"pmids\": [\"30944308\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"NESG1 (CFAP45) interacts with VPS33B (confirmed by Co-IP) in ovarian cancer cells. Overexpressed NESG1 suppresses cell growth by mediating VPS33B-modulated EGFR/PI3K/AKT/c-Myc/p53/miR-133a-3p signals, and NESG1 induces VPS33B expression by reducing PI3K/AKT/c-Jun-mediated transcription inhibition.\",\n      \"method\": \"Co-immunoprecipitation, western blot, overexpression/knockdown functional assays\",\n      \"journal\": \"Cancer science\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single Co-IP for interaction, single lab, indirect pathway readouts\",\n      \"pmids\": [\"33788346\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"NESG1 (CFAP45) suppresses NPC migration and invasion via the Wnt/β-catenin signaling pathway. NESG1 induces miR-1254, which targets HDGF, preventing HDGF/DDX5 complex-mediated nuclear translocation of β-catenin and downstream EMT signals.\",\n      \"method\": \"Transwell/Boyden assays, miRNA microarray, luciferase reporter assay, ChIP, EMSA, Co-IP, western blot, in vivo and in vitro loss-of-function/gain-of-function\",\n      \"journal\": \"Molecular therapy. Methods & clinical development\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 / Moderate — multiple orthogonal methods (luciferase, ChIP, EMSA, CoIP) in single lab establishing pathway placement\",\n      \"pmids\": [\"33718512\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"CCDC19 (CFAP45) overexpression in NSCLC cells stimulates miR-184 expression, which suppresses C-Myc, blocking cell growth mediated by the PI3K/AKT/C-Jun pathway. Knockdown of CCDC19 restores cell growth in CCDC19-overexpressing cells.\",\n      \"method\": \"Lentiviral overexpression, shRNA knockdown, cell proliferation and cell cycle assays, miRNA and pathway western blot analysis\",\n      \"journal\": \"Journal of cellular and molecular medicine\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — indirect pathway inference via OE/KD, single lab, no direct binding assay between CCDC19 and miR-184\",\n      \"pmids\": [\"24976536\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Proteomics of NESG1 (CFAP45)-overexpressing NPC cells identified ENO1 (alpha-enolase) as a NESG1-regulated protein. Overexpressed ENO1 restores cell proliferation, antagonizes NESG1 regulation of p21 and CCNA1, and induces C-Myc, pRB, and E2F1 expression.\",\n      \"method\": \"2D gel proteomics, western blot, overexpression rescue experiments\",\n      \"journal\": \"Proteomics\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — proteomic screen with western blot validation and rescue assay, single lab, indirect mechanism\",\n      \"pmids\": [\"22997098\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Induced expression of NESG1 (CFAP45) in NPC cells decreases cell proliferation, G1-S phase transition, cell migration, invasion, and in vivo tumorigenesis, and significantly regulates expression of cell cycle regulators CCNA1 and p21.\",\n      \"method\": \"Lentiviral overexpression in NPC 5-8F cells, proliferation assay, cell cycle analysis, invasion/migration assays, xenograft in vivo tumorigenesis\",\n      \"journal\": \"International journal of cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal functional assays (cell cycle, invasion, in vivo) with molecular readouts (CCNA1, p21), single lab\",\n      \"pmids\": [\"20715168\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"CFAP45 (also known as NESG1/CCDC19) is a cilia and flagella associated protein that supports motile ciliary and flagellar beating through an axonemal adenine nucleotide homeostasis module: it binds AMP and interacts with adenylate kinase AK8 and CFAP52, facilitating dynein ATPase-driven microtubule sliding; loss of CFAP45 causes situs abnormalities, asthenospermia, and cilia instability, while in cancer cell contexts it acts as a tumor suppressor by modulating PI3K/AKT signaling, miRNA networks (miR-184, miR-1254), and Wnt/β-catenin pathway activity.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"CFAP45 (NESG1/CCDC19) is an axonemal protein that supports motile ciliary and flagellar beating by maintaining adenine nucleotide homeostasis within the axoneme [#0]. It binds AMP and forms part of a module with dynein ATPases, adenylate kinase AK8, and CFAP52, where the modeled AMP-binding interface couples CFAP45 to AK8; dyskinetic microtubule sliding in Cfap45-deficient sperm is rescued by supplementing AMP or ADP with ATP, establishing that CFAP45 sustains the nucleotide pool that dynein-driven sliding consumes [#0]. CFAP52 physically interacts with CFAP45 in the sperm flagellum and is required for its stable expression, linking the two as interdependent components of this beating module [#2]. Within the axoneme CFAP45 occupies a punctate, static position, and its depletion destabilizes cilia and causes their detachment from the apical surface, producing left-right patterning and cardiac looping defects [#1]. In epithelial cancer cell contexts CFAP45 acts as a growth-suppressive factor: its induced expression in nasopharyngeal carcinoma cells reduces proliferation, G1-S transition, migration, invasion, and xenograft tumorigenesis while regulating the cell cycle regulators CCNA1 and p21 [#8], and it suppresses migration and invasion through a miR-1254/HDGF axis that blocks Wnt/\\u03b2-catenin-driven \\u03b2-catenin nuclear translocation and EMT [#5].\",\n  \"teleology\": [\n    {\n      \"year\": 2010,\n      \"claim\": \"Established that CFAP45 (NESG1) can act as a growth- and invasion-suppressive factor, the first functional handle on the protein, framing it as a candidate tumor suppressor in nasopharyngeal carcinoma.\",\n      \"evidence\": \"Lentiviral overexpression in NPC cells with proliferation, cell cycle, invasion/migration assays and xenograft tumorigenesis, with CCNA1/p21 readouts\",\n      \"pmids\": [\"20715168\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"No direct molecular mechanism linking CFAP45 to CCNA1/p21 regulation\",\n        \"Does not address the protein's ciliary/axonemal role\",\n        \"Overexpression-based phenotype, no endogenous loss-of-function\"\n      ]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Asked which effectors mediate the growth-suppressive phenotype and implicated ENO1 as a CFAP45-regulated protein opposing its control of cell cycle regulators.\",\n      \"evidence\": \"2D gel proteomics of NESG1-overexpressing NPC cells with western blot and ENO1 overexpression rescue\",\n      \"pmids\": [\"22997098\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"Indirect mechanism; no direct CFAP45-ENO1 binding shown\",\n        \"Single lab, screen-based with limited validation\",\n        \"Causal ordering between CFAP45 and ENO1 not resolved\"\n      ]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Connected CFAP45 (CCDC19) to a miRNA-mediated signaling output, positing that it suppresses growth through miR-184 and the PI3K/AKT/c-Jun axis.\",\n      \"evidence\": \"Lentiviral overexpression and shRNA knockdown in NSCLC cells with proliferation, cell cycle, and pathway western blots\",\n      \"pmids\": [\"24976536\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No direct binding assay between CCDC19 and miR-184\",\n        \"Pathway inference is indirect via OE/KD\",\n        \"Mechanism of miRNA induction unknown\"\n      ]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Probed an interaction partner and proposed CFAP45 (NESG1) cooperates with VPS33B to relieve PI3K/AKT/c-Jun-mediated transcriptional repression in NPC cells.\",\n      \"evidence\": \"Co-localization microscopy and NESG1 knockdown rescue with PI3K/AKT/c-Jun western blots\",\n      \"pmids\": [\"30944308\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"Interaction rests on co-localization, not direct biochemistry\",\n        \"Single lab, single method for the interaction claim\",\n        \"Functional link to ciliary biology unaddressed\"\n      ]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Extended the VPS33B link to a Co-IP-confirmed interaction in ovarian cancer and to a Wnt/\\u03b2-catenin-suppressive miR-1254/HDGF axis, sharpening the proposed anti-invasion mechanism.\",\n      \"evidence\": \"Co-IP, luciferase reporter, ChIP, EMSA, Transwell assays, and in vivo/in vitro gain/loss-of-function in cancer cells\",\n      \"pmids\": [\"33788346\", \"33718512\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Multiple parallel signaling outputs not reconciled into one model\",\n        \"Relationship between cancer signaling roles and axonemal function unclear\",\n        \"VPS33B interaction not reciprocally validated across studies\"\n      ]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Defined the core mechanistic role of CFAP45 as an axonemal AMP-binding partner of adenylate kinase AK8 that sustains adenine nucleotide homeostasis for dynein-driven microtubule sliding and ciliary/flagellar beating.\",\n      \"evidence\": \"In vitro AMP-binding assay, structural modelling, proteomics of CFAP45-deficient cilia, and Cfap45 knockout mouse with microtubule sliding rescue by AMP/ADP\",\n      \"pmids\": [\"33139725\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Structural model of the AK8 interface not experimentally resolved\",\n        \"Stoichiometry and exact placement within the axoneme not defined\",\n        \"Does not connect to the cancer signaling roles reported separately\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Resolved CFAP45's static axonemal position and showed it is required for cilia stability, linking its loss to organ laterality defects, and established CFAP52 as a stabilizing physical partner.\",\n      \"evidence\": \"Cfap45 morpholino depletion and live confocal imaging in Xenopus LRO cilia; CFAP52-CFAP45 Co-IP and Cfap52 knockout mouse with sliding assay\",\n      \"pmids\": [\"37172641\", \"37236356\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Molecular basis of how CFAP45 confers cilia stability is unknown\",\n        \"Mechanism by which CFAP52 maintains CFAP45 levels not defined\",\n        \"Single-lab depletion studies\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How the axonemal nucleotide-homeostasis function and the reported cancer-signaling roles of CFAP45 relate, if at all, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No study bridges the ciliary/flagellar mechanism and the PI3K/AKT/Wnt signaling reports\",\n        \"No human disease-causing mutation directly demonstrated in the timeline\",\n        \"Direct binding partners beyond AK8/CFAP52/VPS33B not mapped\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140313\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005929\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1852241\", \"supporting_discovery_ids\": [0, 1]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"AK8\", \"CFAP52\", \"VPS33B\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":5,"faith_pct":100.0}}