{"gene":"DNAH10","run_date":"2026-04-28T17:46:02","timeline":{"discoveries":[{"year":2021,"finding":"DNAH10 encodes an inner dynein arm (IDA) heavy chain protein in sperm flagella; bi-allelic loss-of-function variants in humans cause significant IDA deficiency in sperm flagella along with loss of DNAH1 protein, resulting in MMAF and asthenoteratozoospermia","method":"Whole-exome sequencing, immunofluorescence, transmission electron microscopy, scanning electron microscopy","journal":"Journal of assisted reproduction and genetics","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods (TEM ultrastructure, IF protein localization, genetics) in human patients with clear phenotypic readout","pmids":["34657236"],"is_preprint":false},{"year":2023,"finding":"DNAH10 deficiency causes primary ciliary dyskinesia (PCD) in humans and mice; loss of DNAH10 results in absence of DNAH10 and DNALI1 in respiratory cilia, disordered 9+2 axoneme architecture, and IDA defects in both respiratory cilia and sperm flagella; Dnah10-knockout and knockin mice recapitulate PCD phenotypes including chronic respiratory infection, male infertility, and hydrocephalus","method":"Exome sequencing, immunostaining, transmission electron microscopy, Dnah10-knockout and knockin mouse models","journal":"Frontiers of medicine","confidence":"High","confidence_rationale":"Tier 2 — orthogonal methods including animal models, ultrastructural analysis, and immunostaining, replicated in human and mouse","pmids":["37314648"],"is_preprint":false},{"year":2025,"finding":"DNAH10 interacts with CFAP57, DYNLL1, and CCDC73 to form a double-headed inner dynein arm f (IDAf) complex in cilia; loss of DNAH10 leads to reduced expression of CFAP57, DYNLL1, and CCDC73, improper IDAf assembly, ciliary dysfunction, and pulmonary fibrosis","method":"Co-immunoprecipitation, immunostaining, Dnah10-knockout mouse model, proteomic analysis, scanning electron microscopy","journal":"Orphanet journal of rare diseases","confidence":"High","confidence_rationale":"Tier 1-2 — Co-IP confirmed interactions, KO mouse model, proteomic and structural validation","pmids":["40898283"],"is_preprint":false},{"year":2025,"finding":"DNAH10 acts as a bridging protein that enhances the interaction between UCHL3 and PACRG; UCHL3 stabilizes PACRG via deubiquitination, and the DNAH10-UCHL3-PACRG complex facilitates intra-manchette transport during spermiogenesis; DNAH10 deficiency causes abnormal sperm head and flagella morphology, impaired manchette function, and aberrant localization of axonemal proteins","method":"Co-immunoprecipitation, immunofluorescence, protein interaction assays, loss-of-function studies in mice","journal":"Development (Cambridge, England)","confidence":"High","confidence_rationale":"Tier 1-2 — mechanistic Co-IP, deubiquitination pathway placement, and defined cellular phenotype with multiple methods","pmids":["41058558"],"is_preprint":false},{"year":2021,"finding":"Morpholino knockdown and CRISPR/Cas9-mediated mutation of dnah10 in zebrafish disrupts cardiac looping and causes abnormal expression of left-right patterning markers lefty2 and pitx2, implicating DNAH10 in left-right body patterning","method":"Zebrafish morpholino knockdown, CRISPR/Cas9 mutation, in situ hybridization for lefty2/pitx2","journal":"Genome medicine","confidence":"Medium","confidence_rationale":"Tier 2 — genetic loss-of-function in zebrafish with molecular marker readout, single study","pmids":["29843777"],"is_preprint":false},{"year":2021,"finding":"Disruption of dnah10 in zebrafish causes ependymal monocilia dysfunction (reduced beat frequency), disassembly of the Reissner fiber, and adult-viable scoliosis; monocilia function driven by dnah10 contributes to Reissner fiber polymerization and spine straightening","method":"F0 CRISPR screening, stable dnah10 mutant zebrafish, monocilia beat frequency measurement, Reissner fiber imaging","journal":"Developmental biology","confidence":"High","confidence_rationale":"Tier 2 — stable genetic mutant with direct functional measurement of cilia beat frequency, Reissner fiber assembly, and spine phenotype","pmids":["34915022"],"is_preprint":false},{"year":2019,"finding":"RNAi knockdown of the DNAH10 Drosophila ortholog results in a significant increase in total body triglyceride levels, implicating DNAH10 in lipid/triglyceride homeostasis","method":"Drosophila RNAi knockdown, triglyceride quantification","journal":"Nature genetics","confidence":"Medium","confidence_rationale":"Tier 2 — direct functional assay in model organism with defined metabolic phenotype, single study","pmids":["30778226"],"is_preprint":false},{"year":2024,"finding":"Repeated freeze-thaw cycles in bovine sperm significantly decrease DNAH10 protein levels in the axoneme (9+2 structure disrupted), correlating with reduced sperm motility, establishing DNAH10 as a functional axonemal component required for sperm motility","method":"Transmission electron microscopy, western blot/protein quantification, motility analysis","journal":"Cryobiology","confidence":"Medium","confidence_rationale":"Tier 3 — correlative protein loss with functional motility readout, single study in bovine model","pmids":["38593909"],"is_preprint":false}],"current_model":"DNAH10 is an inner dynein arm (IDA) heavy chain protein that assembles into the double-headed IDAf complex (together with CFAP57, DYNLL1, and CCDC73) in motile cilia and sperm flagella, where it is required for ciliary and flagellar beat; additionally, DNAH10 acts as a bridging factor enhancing the UCHL3-PACRG deubiquitination complex to support intra-manchette transport during spermiogenesis, and its loss in humans and mice causes primary ciliary dyskinesia, male infertility (MMAF/asthenoteratozoospermia), and scoliosis via monocilia dysfunction and Reissner fiber disassembly."},"narrative":{"teleology":[{"year":2019,"claim":"Before any ciliary role was defined, a Drosophila screen linked the DNAH10 ortholog to triglyceride homeostasis, raising the possibility of non-canonical functions for this dynein heavy chain.","evidence":"RNAi knockdown in Drosophila with triglyceride quantification","pmids":["30778226"],"confidence":"Medium","gaps":["Mechanism connecting a ciliary dynein to lipid metabolism is unknown","Not replicated in vertebrate systems","Single phenotypic readout without mechanistic dissection"]},{"year":2021,"claim":"Genetic and ultrastructural studies in humans and zebrafish established DNAH10 as an IDA heavy chain required for sperm flagellar motility, ciliary-driven left-right patterning, and ependymal monocilia function, revealing that its loss causes MMAF/asthenoteratozoospermia, cardiac laterality defects, Reissner fiber disassembly, and scoliosis.","evidence":"Whole-exome sequencing in infertile men with TEM/SEM/IF; zebrafish CRISPR mutants with monocilia beat frequency measurement, Reissner fiber imaging, and in situ hybridization for laterality markers","pmids":["34657236","29843777","34915022"],"confidence":"High","gaps":["Identity of the specific IDA sub-complex containing DNAH10 was unknown","Whether DNAH10 functions beyond the axoneme (e.g. during spermiogenesis) was not addressed","Human respiratory cilia phenotype had not yet been characterized"]},{"year":2023,"claim":"Extension to respiratory cilia and mouse genetics demonstrated that DNAH10 deficiency causes primary ciliary dyskinesia with IDA loss in both respiratory cilia and sperm, confirmed by knockout and knockin mouse models recapitulating chronic infection, hydrocephalus, and infertility.","evidence":"Exome sequencing in PCD patients; Dnah10-KO and knockin mice with immunostaining and TEM","pmids":["37314648"],"confidence":"High","gaps":["Subunit composition of the DNAH10-containing IDA complex remained unresolved","Whether DNAH10 has roles outside the axonemal beat machinery was not explored"]},{"year":2024,"claim":"Correlative evidence in bovine sperm showed that axonemal DNAH10 protein levels decline under cryodamage in parallel with motility loss, supporting its functional requirement for flagellar beating.","evidence":"Western blot quantification and TEM in freeze-thawed bovine sperm with motility analysis","pmids":["38593909"],"confidence":"Medium","gaps":["Correlative design; no causal manipulation of DNAH10 in bovine sperm","Does not distinguish whether DNAH10 loss is cause or consequence of structural collapse"]},{"year":2025,"claim":"Biochemical and proteomic work defined DNAH10 as a subunit of the double-headed IDAf complex (with CFAP57, DYNLL1, CCDC73) and revealed a second, non-axonemal function: DNAH10 bridges UCHL3 to PACRG, stabilizing PACRG via deubiquitination to support intra-manchette transport during spermiogenesis.","evidence":"Co-IP, proteomic analysis, immunostaining, and Dnah10-KO mice for IDAf complex; Co-IP and protein interaction assays with deubiquitination readouts in manchette studies","pmids":["40898283","41058558"],"confidence":"High","gaps":["Structural basis of IDAf assembly and DNAH10 motor domain architecture remain unresolved","Whether the UCHL3-PACRG bridging function is independent of or integrated with axonemal assembly is unclear","Downstream cargo of DNAH10-dependent manchette transport not fully cataloged"]},{"year":null,"claim":"Key open questions include the atomic-resolution structure of the IDAf complex, the full catalog of DNAH10-dependent manchette cargo, and whether the triglyceride phenotype observed in Drosophila reflects a conserved metabolic role or a secondary consequence of ciliary defects.","evidence":"","pmids":[],"confidence":"Low","gaps":["No cryo-EM or crystal structure of DNAH10 or the IDAf complex","Lipid metabolism link lacks vertebrate replication or mechanistic explanation","Genotype-phenotype correlations across different DNAH10 mutations (missense vs. null) not systematically defined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0003774","term_label":"cytoskeletal motor activity","supporting_discovery_ids":[0,1,2,5,7]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[3]}],"localization":[{"term_id":"GO:0005929","term_label":"cilium","supporting_discovery_ids":[0,1,2,5]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[0,2,7]}],"pathway":[{"term_id":"R-HSA-1852241","term_label":"Organelle biogenesis and maintenance","supporting_discovery_ids":[0,1,2,5]},{"term_id":"R-HSA-1474165","term_label":"Reproduction","supporting_discovery_ids":[0,3]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[4,5]}],"complexes":["IDAf (inner dynein arm f complex)","DNAH10-UCHL3-PACRG deubiquitination/bridging complex"],"partners":["CFAP57","DYNLL1","CCDC73","UCHL3","PACRG","DNAH1","DNALI1"],"other_free_text":[]},"mechanistic_narrative":"DNAH10 is an inner dynein arm heavy chain essential for motile cilia and flagellar beating, left-right body patterning, and spermiogenesis. DNAH10 assembles with CFAP57, DYNLL1, and CCDC73 into the double-headed inner dynein arm f (IDAf) complex; loss of DNAH10 destabilizes these subunits, disrupts IDA ultrastructure, and causes ciliary dysfunction in respiratory epithelia, ependymal cells, and sperm flagella [PMID:40898283, PMID:34657236, PMID:37314648]. Beyond its structural role in the axoneme, DNAH10 bridges the deubiquitinase UCHL3 to PACRG, stabilizing PACRG via deubiquitination and facilitating intra-manchette transport during sperm head and flagellum shaping [PMID:41058558]. Bi-allelic loss-of-function mutations in DNAH10 cause primary ciliary dyskinesia with chronic respiratory disease, male infertility (MMAF/asthenoteratozoospermia), hydrocephalus, and situs defects, while ependymal monocilia dysfunction leads to Reissner fiber disassembly and scoliosis [PMID:37314648, PMID:34915022, PMID:29843777]."},"prefetch_data":{"uniprot":{"accession":"Q8IVF4","full_name":"Dynein axonemal heavy chain 10","aliases":["Axonemal beta dynein heavy chain 10","Ciliary dynein heavy chain 10"],"length_aa":4471,"mass_kda":514.8,"function":"Force generating protein of respiratory cilia. Produces force towards the minus ends of microtubules. Dynein has ATPase activity; the force-producing power stroke is thought to occur on release of ADP. Involved in sperm motility; implicated in sperm flagellar assembly (PubMed:34237282). Probable inner arm dynein heavy chain","subcellular_location":"Cytoplasm, cytoskeleton, cilium axoneme","url":"https://www.uniprot.org/uniprotkb/Q8IVF4/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/DNAH10","classification":"Not Classified","n_dependent_lines":5,"n_total_lines":1208,"dependency_fraction":0.0041390728476821195},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/DNAH10","total_profiled":1310},"omim":[{"mim_id":"619515","title":"SPERMATOGENIC FAILURE 56; SPGF56","url":"https://www.omim.org/entry/619515"},{"mim_id":"605884","title":"DYNEIN, AXONEMAL, HEAVY CHAIN 10; DNAH10","url":"https://www.omim.org/entry/605884"},{"mim_id":"258150","title":"SPERMATOGENIC FAILURE 1; SPGF1","url":"https://www.omim.org/entry/258150"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Principal piece","reliability":"Supported"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in some","driving_tissues":[{"tissue":"fallopian tube","ntpm":3.2},{"tissue":"testis","ntpm":2.8}],"url":"https://www.proteinatlas.org/search/DNAH10"},"hgnc":{"alias_symbol":["FLJ43808"],"prev_symbol":[]},"alphafold":{"accession":"Q8IVF4","domains":[],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8IVF4","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q8IVF4-2-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q8IVF4-2-F1-predicted_aligned_error_v6.png","plddt_mean":79.75},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=DNAH10","jax_strain_url":"https://www.jax.org/strain/search?query=DNAH10"},"sequence":{"accession":"Q8IVF4","fasta_url":"https://rest.uniprot.org/uniprotkb/Q8IVF4.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q8IVF4/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8IVF4"}},"corpus_meta":[{"pmid":"30778226","id":"PMC_30778226","title":"Protein-coding variants implicate novel genes related to lipid homeostasis contributing to body-fat distribution.","date":"2019","source":"Nature genetics","url":"https://pubmed.ncbi.nlm.nih.gov/30778226","citation_count":83,"is_preprint":false},{"pmid":"36792588","id":"PMC_36792588","title":"DNALI1 deficiency causes male infertility with severe asthenozoospermia in humans and mice by disrupting the assembly of the flagellar inner dynein arms and fibrous sheath.","date":"2023","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/36792588","citation_count":42,"is_preprint":false},{"pmid":"26061684","id":"PMC_26061684","title":"Alterations of the spindle checkpoint pathway in clinicopathologically aggressive CpG island methylator phenotype clear cell renal cell carcinomas.","date":"2015","source":"International journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/26061684","citation_count":37,"is_preprint":false},{"pmid":"34791246","id":"PMC_34791246","title":"Whole-exome sequencing of a cohort of infertile men reveals novel causative genes in teratozoospermia that are chiefly related to sperm head defects.","date":"2021","source":"Human reproduction (Oxford, England)","url":"https://pubmed.ncbi.nlm.nih.gov/34791246","citation_count":37,"is_preprint":false},{"pmid":"29843777","id":"PMC_29843777","title":"Rare copy number variants analysis identifies novel candidate genes in heterotaxy syndrome patients with congenital heart defects.","date":"2018","source":"Genome medicine","url":"https://pubmed.ncbi.nlm.nih.gov/29843777","citation_count":34,"is_preprint":false},{"pmid":"34657236","id":"PMC_34657236","title":"Bi-allelic variants in DNAH10 cause asthenoteratozoospermia and male infertility.","date":"2021","source":"Journal of assisted reproduction and genetics","url":"https://pubmed.ncbi.nlm.nih.gov/34657236","citation_count":29,"is_preprint":false},{"pmid":"31959735","id":"PMC_31959735","title":"DNAH10 mutation correlates with cisplatin sensitivity and tumor mutation burden in small-cell lung cancer.","date":"2020","source":"Aging","url":"https://pubmed.ncbi.nlm.nih.gov/31959735","citation_count":27,"is_preprint":false},{"pmid":"24891332","id":"PMC_24891332","title":"Identification of four novel genes contributing to familial elevated plasma HDL cholesterol in humans.","date":"2014","source":"Journal of lipid research","url":"https://pubmed.ncbi.nlm.nih.gov/24891332","citation_count":21,"is_preprint":false},{"pmid":"32128068","id":"PMC_32128068","title":"Identification of rare variants in novel candidate genes in pulmonary atresia patients by next generation sequencing.","date":"2020","source":"Computational and structural biotechnology journal","url":"https://pubmed.ncbi.nlm.nih.gov/32128068","citation_count":17,"is_preprint":false},{"pmid":"34915022","id":"PMC_34915022","title":"The axonemal dynein heavy chain 10 gene is essential for monocilia motility and spine alignment in zebrafish.","date":"2021","source":"Developmental 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Genetics","url":"https://pubmed.ncbi.nlm.nih.gov/35812163","citation_count":10,"is_preprint":false},{"pmid":"36881978","id":"PMC_36881978","title":"Comparative transcriptome analysis identified crucial genes and pathways affecting sperm motility in the reproductive tract of drakes with different libido.","date":"2023","source":"Poultry science","url":"https://pubmed.ncbi.nlm.nih.gov/36881978","citation_count":7,"is_preprint":false},{"pmid":"38986426","id":"PMC_38986426","title":"Prenatal EDC exposure, DNA Methylation, and early childhood growth: A prospective birth cohort study.","date":"2024","source":"Environment international","url":"https://pubmed.ncbi.nlm.nih.gov/38986426","citation_count":5,"is_preprint":false},{"pmid":"38593909","id":"PMC_38593909","title":"The effects of repeated freezing and thawing on bovine sperm morphometry and function.","date":"2024","source":"Cryobiology","url":"https://pubmed.ncbi.nlm.nih.gov/38593909","citation_count":5,"is_preprint":false},{"pmid":"31836722","id":"PMC_31836722","title":"Discovery of increased epidermal DNAH10 expression after regeneration of dermis in a randomized with-in person trial - reflections on psoriatic inflammation.","date":"2019","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/31836722","citation_count":4,"is_preprint":false},{"pmid":"39996363","id":"PMC_39996363","title":"Novel bi-allelic variants in DNAH10 lead to multiple morphological abnormalities of sperm flagella and male infertility.","date":"2025","source":"Asian journal of andrology","url":"https://pubmed.ncbi.nlm.nih.gov/39996363","citation_count":4,"is_preprint":false},{"pmid":"26420031","id":"PMC_26420031","title":"Exome sequencing in a patient with Catel-Manzke-like syndrome excludes the involvement of the known genes and reveals a possible candidate.","date":"2015","source":"European journal of medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/26420031","citation_count":3,"is_preprint":false},{"pmid":"39523437","id":"PMC_39523437","title":"Novel variants in DNAH9 are present in two infertile patients with severe asthenospermia.","date":"2024","source":"Journal of human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/39523437","citation_count":3,"is_preprint":false},{"pmid":"39270811","id":"PMC_39270811","title":"Identification of competing endogenous RNA networks associated with circRNA and lncRNA in TCDD-induced cleft palate development.","date":"2024","source":"Toxicology letters","url":"https://pubmed.ncbi.nlm.nih.gov/39270811","citation_count":1,"is_preprint":false},{"pmid":"40898283","id":"PMC_40898283","title":"DNAH10 mutation cause primary ciliary dyskinesia with defects of IDAf complex assembly and lung fibrosis manifestation.","date":"2025","source":"Orphanet journal of rare diseases","url":"https://pubmed.ncbi.nlm.nih.gov/40898283","citation_count":0,"is_preprint":false},{"pmid":"41058558","id":"PMC_41058558","title":"DNAH10 interacts with UCHL3-PACRG complex to coordinate sperm head and flagella development during spermiogenesis.","date":"2025","source":"Development (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/41058558","citation_count":0,"is_preprint":false},{"pmid":"40376536","id":"PMC_40376536","title":"Establishment and clinical significance of genetic factor screening method for patients with nonobstructive azoospermia based on whole exon sequencing technology.","date":"2025","source":"Translational andrology and urology","url":"https://pubmed.ncbi.nlm.nih.gov/40376536","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2024.06.28.24309634","title":"Transcriptomics-Driven Machine Learning Models Accurately Predict Chemotherapy Response in Muscle-invasive Bladder Cancer","date":"2024-06-28","source":"bioRxiv","url":"https://doi.org/10.1101/2024.06.28.24309634","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":15562,"output_tokens":1907,"usd":0.037645},"stage2":{"model":"claude-opus-4-6","input_tokens":5178,"output_tokens":2033,"usd":0.115072},"total_usd":0.152717,"stage1_batch_id":"msgbatch_0118RScg3WGaGkaZRtWZbtce","stage2_batch_id":"msgbatch_01Cpfzvtbj42aqp5oYZ5Te2s","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2021,\n      \"finding\": \"DNAH10 encodes an inner dynein arm (IDA) heavy chain protein in sperm flagella; bi-allelic loss-of-function variants in humans cause significant IDA deficiency in sperm flagella along with loss of DNAH1 protein, resulting in MMAF and asthenoteratozoospermia\",\n      \"method\": \"Whole-exome sequencing, immunofluorescence, transmission electron microscopy, scanning electron microscopy\",\n      \"journal\": \"Journal of assisted reproduction and genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods (TEM ultrastructure, IF protein localization, genetics) in human patients with clear phenotypic readout\",\n      \"pmids\": [\"34657236\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"DNAH10 deficiency causes primary ciliary dyskinesia (PCD) in humans and mice; loss of DNAH10 results in absence of DNAH10 and DNALI1 in respiratory cilia, disordered 9+2 axoneme architecture, and IDA defects in both respiratory cilia and sperm flagella; Dnah10-knockout and knockin mice recapitulate PCD phenotypes including chronic respiratory infection, male infertility, and hydrocephalus\",\n      \"method\": \"Exome sequencing, immunostaining, transmission electron microscopy, Dnah10-knockout and knockin mouse models\",\n      \"journal\": \"Frontiers of medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — orthogonal methods including animal models, ultrastructural analysis, and immunostaining, replicated in human and mouse\",\n      \"pmids\": [\"37314648\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"DNAH10 interacts with CFAP57, DYNLL1, and CCDC73 to form a double-headed inner dynein arm f (IDAf) complex in cilia; loss of DNAH10 leads to reduced expression of CFAP57, DYNLL1, and CCDC73, improper IDAf assembly, ciliary dysfunction, and pulmonary fibrosis\",\n      \"method\": \"Co-immunoprecipitation, immunostaining, Dnah10-knockout mouse model, proteomic analysis, scanning electron microscopy\",\n      \"journal\": \"Orphanet journal of rare diseases\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — Co-IP confirmed interactions, KO mouse model, proteomic and structural validation\",\n      \"pmids\": [\"40898283\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"DNAH10 acts as a bridging protein that enhances the interaction between UCHL3 and PACRG; UCHL3 stabilizes PACRG via deubiquitination, and the DNAH10-UCHL3-PACRG complex facilitates intra-manchette transport during spermiogenesis; DNAH10 deficiency causes abnormal sperm head and flagella morphology, impaired manchette function, and aberrant localization of axonemal proteins\",\n      \"method\": \"Co-immunoprecipitation, immunofluorescence, protein interaction assays, loss-of-function studies in mice\",\n      \"journal\": \"Development (Cambridge, England)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — mechanistic Co-IP, deubiquitination pathway placement, and defined cellular phenotype with multiple methods\",\n      \"pmids\": [\"41058558\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Morpholino knockdown and CRISPR/Cas9-mediated mutation of dnah10 in zebrafish disrupts cardiac looping and causes abnormal expression of left-right patterning markers lefty2 and pitx2, implicating DNAH10 in left-right body patterning\",\n      \"method\": \"Zebrafish morpholino knockdown, CRISPR/Cas9 mutation, in situ hybridization for lefty2/pitx2\",\n      \"journal\": \"Genome medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic loss-of-function in zebrafish with molecular marker readout, single study\",\n      \"pmids\": [\"29843777\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Disruption of dnah10 in zebrafish causes ependymal monocilia dysfunction (reduced beat frequency), disassembly of the Reissner fiber, and adult-viable scoliosis; monocilia function driven by dnah10 contributes to Reissner fiber polymerization and spine straightening\",\n      \"method\": \"F0 CRISPR screening, stable dnah10 mutant zebrafish, monocilia beat frequency measurement, Reissner fiber imaging\",\n      \"journal\": \"Developmental biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — stable genetic mutant with direct functional measurement of cilia beat frequency, Reissner fiber assembly, and spine phenotype\",\n      \"pmids\": [\"34915022\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"RNAi knockdown of the DNAH10 Drosophila ortholog results in a significant increase in total body triglyceride levels, implicating DNAH10 in lipid/triglyceride homeostasis\",\n      \"method\": \"Drosophila RNAi knockdown, triglyceride quantification\",\n      \"journal\": \"Nature genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct functional assay in model organism with defined metabolic phenotype, single study\",\n      \"pmids\": [\"30778226\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Repeated freeze-thaw cycles in bovine sperm significantly decrease DNAH10 protein levels in the axoneme (9+2 structure disrupted), correlating with reduced sperm motility, establishing DNAH10 as a functional axonemal component required for sperm motility\",\n      \"method\": \"Transmission electron microscopy, western blot/protein quantification, motility analysis\",\n      \"journal\": \"Cryobiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — correlative protein loss with functional motility readout, single study in bovine model\",\n      \"pmids\": [\"38593909\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"DNAH10 is an inner dynein arm (IDA) heavy chain protein that assembles into the double-headed IDAf complex (together with CFAP57, DYNLL1, and CCDC73) in motile cilia and sperm flagella, where it is required for ciliary and flagellar beat; additionally, DNAH10 acts as a bridging factor enhancing the UCHL3-PACRG deubiquitination complex to support intra-manchette transport during spermiogenesis, and its loss in humans and mice causes primary ciliary dyskinesia, male infertility (MMAF/asthenoteratozoospermia), and scoliosis via monocilia dysfunction and Reissner fiber disassembly.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"DNAH10 is an inner dynein arm heavy chain essential for motile cilia and flagellar beating, left-right body patterning, and spermiogenesis. DNAH10 assembles with CFAP57, DYNLL1, and CCDC73 into the double-headed inner dynein arm f (IDAf) complex; loss of DNAH10 destabilizes these subunits, disrupts IDA ultrastructure, and causes ciliary dysfunction in respiratory epithelia, ependymal cells, and sperm flagella [PMID:40898283, PMID:34657236, PMID:37314648]. Beyond its structural role in the axoneme, DNAH10 bridges the deubiquitinase UCHL3 to PACRG, stabilizing PACRG via deubiquitination and facilitating intra-manchette transport during sperm head and flagellum shaping [PMID:41058558]. Bi-allelic loss-of-function mutations in DNAH10 cause primary ciliary dyskinesia with chronic respiratory disease, male infertility (MMAF/asthenoteratozoospermia), hydrocephalus, and situs defects, while ependymal monocilia dysfunction leads to Reissner fiber disassembly and scoliosis [PMID:37314648, PMID:34915022, PMID:29843777].\",\n  \"teleology\": [\n    {\n      \"year\": 2019,\n      \"claim\": \"Before any ciliary role was defined, a Drosophila screen linked the DNAH10 ortholog to triglyceride homeostasis, raising the possibility of non-canonical functions for this dynein heavy chain.\",\n      \"evidence\": \"RNAi knockdown in Drosophila with triglyceride quantification\",\n      \"pmids\": [\"30778226\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Mechanism connecting a ciliary dynein to lipid metabolism is unknown\",\n        \"Not replicated in vertebrate systems\",\n        \"Single phenotypic readout without mechanistic dissection\"\n      ]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Genetic and ultrastructural studies in humans and zebrafish established DNAH10 as an IDA heavy chain required for sperm flagellar motility, ciliary-driven left-right patterning, and ependymal monocilia function, revealing that its loss causes MMAF/asthenoteratozoospermia, cardiac laterality defects, Reissner fiber disassembly, and scoliosis.\",\n      \"evidence\": \"Whole-exome sequencing in infertile men with TEM/SEM/IF; zebrafish CRISPR mutants with monocilia beat frequency measurement, Reissner fiber imaging, and in situ hybridization for laterality markers\",\n      \"pmids\": [\"34657236\", \"29843777\", \"34915022\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Identity of the specific IDA sub-complex containing DNAH10 was unknown\",\n        \"Whether DNAH10 functions beyond the axoneme (e.g. during spermiogenesis) was not addressed\",\n        \"Human respiratory cilia phenotype had not yet been characterized\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Extension to respiratory cilia and mouse genetics demonstrated that DNAH10 deficiency causes primary ciliary dyskinesia with IDA loss in both respiratory cilia and sperm, confirmed by knockout and knockin mouse models recapitulating chronic infection, hydrocephalus, and infertility.\",\n      \"evidence\": \"Exome sequencing in PCD patients; Dnah10-KO and knockin mice with immunostaining and TEM\",\n      \"pmids\": [\"37314648\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Subunit composition of the DNAH10-containing IDA complex remained unresolved\",\n        \"Whether DNAH10 has roles outside the axonemal beat machinery was not explored\"\n      ]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Correlative evidence in bovine sperm showed that axonemal DNAH10 protein levels decline under cryodamage in parallel with motility loss, supporting its functional requirement for flagellar beating.\",\n      \"evidence\": \"Western blot quantification and TEM in freeze-thawed bovine sperm with motility analysis\",\n      \"pmids\": [\"38593909\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Correlative design; no causal manipulation of DNAH10 in bovine sperm\",\n        \"Does not distinguish whether DNAH10 loss is cause or consequence of structural collapse\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Biochemical and proteomic work defined DNAH10 as a subunit of the double-headed IDAf complex (with CFAP57, DYNLL1, CCDC73) and revealed a second, non-axonemal function: DNAH10 bridges UCHL3 to PACRG, stabilizing PACRG via deubiquitination to support intra-manchette transport during spermiogenesis.\",\n      \"evidence\": \"Co-IP, proteomic analysis, immunostaining, and Dnah10-KO mice for IDAf complex; Co-IP and protein interaction assays with deubiquitination readouts in manchette studies\",\n      \"pmids\": [\"40898283\", \"41058558\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Structural basis of IDAf assembly and DNAH10 motor domain architecture remain unresolved\",\n        \"Whether the UCHL3-PACRG bridging function is independent of or integrated with axonemal assembly is unclear\",\n        \"Downstream cargo of DNAH10-dependent manchette transport not fully cataloged\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Key open questions include the atomic-resolution structure of the IDAf complex, the full catalog of DNAH10-dependent manchette cargo, and whether the triglyceride phenotype observed in Drosophila reflects a conserved metabolic role or a secondary consequence of ciliary defects.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No cryo-EM or crystal structure of DNAH10 or the IDAf complex\",\n        \"Lipid metabolism link lacks vertebrate replication or mechanistic explanation\",\n        \"Genotype-phenotype correlations across different DNAH10 mutations (missense vs. null) not systematically defined\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0003774\", \"supporting_discovery_ids\": [0, 1, 2, 5, 7]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [3]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005929\", \"supporting_discovery_ids\": [0, 1, 2, 5]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [0, 2, 7]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1852241\", \"supporting_discovery_ids\": [0, 1, 2, 5]},\n      {\"term_id\": \"R-HSA-1474165\", \"supporting_discovery_ids\": [0, 3]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [4, 5]}\n    ],\n    \"complexes\": [\n      \"IDAf (inner dynein arm f complex)\",\n      \"DNAH10-UCHL3-PACRG deubiquitination/bridging complex\"\n    ],\n    \"partners\": [\n      \"CFAP57\",\n      \"DYNLL1\",\n      \"CCDC73\",\n      \"UCHL3\",\n      \"PACRG\",\n      \"DNAH1\",\n      \"DNALI1\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}