{"gene":"RSPH4A","run_date":"2026-06-10T07:46:28","timeline":{"discoveries":[{"year":2009,"finding":"RSPH4A encodes a protein component of the axonemal radial spoke head; loss-of-function mutations cause primary ciliary dyskinesia with central-microtubular-pair abnormalities (intermittent or complete absence of the central pair), establishing its role in maintaining normal motile cilia movement in 9+2-structure cilia and flagella.","method":"SNP-based linkage analysis, positional candidate gene identification, in situ hybridization (murine Rsph9), zebrafish knockdown/rescue experiments for RSPH9 ortholog (functional context for radial spoke head proteins), human mutation analysis","journal":"American journal of human genetics","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic mapping, mutation identification, and zebrafish knockdown/rescue replicated across multiple families and species; independently supported by subsequent structural studies","pmids":["19200523"],"is_preprint":false},{"year":2020,"finding":"Rsph4a is required for assembly of all three radial spoke heads (RS1, RS2, and RS3) in mouse tracheal cilia; Rsph4a-deficient mice lack all triplet spoke heads as revealed by cryo-electron tomography, and Rsph4a contributes to planar beating of motile cilia in trachea, ependymal tissue, and oviduct by building the distal architecture of radial spokes.","method":"Cryo-electron tomography of Rsph4a-knockout mouse tracheal cilia, ciliary movement observation, immunofluorescence analysis","journal":"PLoS genetics","confidence":"High","confidence_rationale":"Tier 1 / Moderate — cryo-electron tomography structural analysis combined with functional (ciliary movement) and immunofluorescence readouts in a genetic knockout model, single lab but multiple orthogonal methods","pmids":["32203505"],"is_preprint":false},{"year":2021,"finding":"In RSPH4A-/- human respiratory cilia, the radial spoke heads of RS1 and RS2 (but not RS3) are missing, and there are additional defects in the arch domains adjacent to RS1 and RS2 heads (distinct from RSPH1-/- defects); secondary heterogeneous defects in the central pair complex are also observed, establishing the specific structural role of RSPH4A at RS1 and RS2 spoke heads and adjacent arch domains.","method":"Cryo-electron tomography (cryo-ET) and subtomogram averaging of noninvasively collected human PCD patient respiratory cilia compared to controls","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 1 / Moderate — cryo-ET with subtomogram averaging directly on patient-derived human cilia, structural comparison with controls and RSPH1-/- cilia, single lab but highly rigorous structural method","pmids":["33852348"],"is_preprint":false},{"year":2013,"finding":"A splice-site mutation (c.921+3_6delAAGT) in RSPH4A leads to a premature translation termination signal and loss of function, confirmed by quantitative ciliary ultrastructural analysis showing central apparatus defects, reduced ciliary beat frequency and abnormal waveform, and transcript analysis.","method":"Genetic sequencing, transcript analysis, ciliary ultrastructural analysis by electron microscopy, measurement of ciliary beat frequency and waveform","journal":"Human mutation","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function confirmed by multiple orthogonal methods (ultrastructure, beat frequency/waveform, transcript analysis) in a single study","pmids":["23798057"],"is_preprint":false},{"year":2026,"finding":"AAV-mediated adenosine base editing targeting RSPH4A mutations in patient-derived cells partially restored normal ciliary motion pattern (~30.4% increase in normal motion pattern with reduction in circular motions) demonstrating that correction of RSPH4A sequence restores ciliary function.","method":"AAV-mediated base editing (nickase Cas9 fused to adenosine deaminase), Western blot, digital PCR for editing efficiency, high-speed video and confocal microscopy for ciliary beat frequency and motion pattern in patient cells","journal":"Human gene therapy","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — functional restoration of ciliary motility after RSPH4A correction demonstrated with multiple readouts in patient cells, but single lab proof-of-concept study with limited editing efficiency","pmids":["42050778"],"is_preprint":false}],"current_model":"RSPH4A encodes a radial spoke head protein required for assembly of the distal heads of radial spokes RS1 and RS2 (and RS3 in mice) within the 9+2 axonemal repeat unit of motile cilia; loss of RSPH4A causes absence of RS1/RS2 spoke heads and arch domain defects with secondary central pair complex abnormalities, leading to abnormal (circling) ciliary beat pattern and primary ciliary dyskinesia without laterality defects."},"narrative":{"mechanistic_narrative":"RSPH4A encodes a structural component of the axonemal radial spoke head required for normal motility of 9+2 motile cilia, and its loss causes primary ciliary dyskinesia [PMID:19200523]. RSPH4A builds the distal head architecture of radial spokes: in human respiratory cilia its loss eliminates the radial spoke heads of RS1 and RS2 (but spares RS3) and produces additional defects in the arch domains adjacent to those heads, with secondary heterogeneous abnormalities of the central pair complex [PMID:33852348]. In mouse tracheal cilia Rsph4a is required for assembly of all three radial spoke heads (RS1, RS2, RS3), and it supports planar ciliary beating across tracheal, ependymal, and oviduct epithelia [PMID:32203505]. Loss-of-function mutations, including a splice-site allele generating premature termination, abolish this function and yield central-apparatus ultrastructural defects, reduced beat frequency, and abnormal waveform [PMID:19200523, PMID:23798057]; correction of the RSPH4A sequence in patient-derived cells partially restores normal ciliary motion [PMID:42050778].","teleology":[{"year":2009,"claim":"Established RSPH4A as a radial spoke head gene whose loss-of-function mutations cause primary ciliary dyskinesia, defining a genetic basis for central-pair-associated ciliary dysmotility.","evidence":"SNP linkage, positional candidate identification, and human mutation analysis with ortholog functional context","pmids":["19200523"],"confidence":"High","gaps":["Did not resolve which radial spoke heads RSPH4A builds at structural resolution","Mechanism linking radial spoke loss to central pair abnormality not defined"]},{"year":2013,"claim":"Showed that a specific splice-site mutation causes premature termination and loss of function, linking RSPH4A sequence disruption to measurable ciliary ultrastructural and motility defects.","evidence":"Genetic sequencing, transcript analysis, electron microscopy, beat frequency and waveform measurement in patient cilia","pmids":["23798057"],"confidence":"Medium","gaps":["Single study; structural localization of the defect within radial spokes not resolved","Protein-level consequence inferred from transcript, not directly visualized"]},{"year":2020,"claim":"Defined the structural role of Rsph4a in vivo by showing it is required to assemble all three radial spoke heads and to support planar beating across multiple ciliated epithelia in mouse.","evidence":"Cryo-electron tomography of knockout mouse tracheal cilia with ciliary movement and immunofluorescence readouts","pmids":["32203505"],"confidence":"High","gaps":["Mouse three-head requirement differs from later human findings; species-specific architecture unresolved","Does not establish direct protein-protein contacts within the spoke head"]},{"year":2021,"claim":"Pinpointed the human structural defect to absence of RS1 and RS2 spoke heads (sparing RS3) plus adjacent arch domain defects, distinguishing RSPH4A loss from RSPH1 loss and revealing secondary central pair abnormalities.","evidence":"Cryo-ET with subtomogram averaging of patient-derived human respiratory cilia versus controls and RSPH1-/- cilia","pmids":["33852348"],"confidence":"High","gaps":["Mechanism by which spoke head loss secondarily perturbs the central pair not defined","Atomic-resolution position of RSPH4A within the spoke head head not determined"]},{"year":2026,"claim":"Demonstrated that correcting the RSPH4A sequence is sufficient to restore ciliary function, providing causal confirmation that the motility defect stems from the gene lesion.","evidence":"AAV-mediated adenosine base editing of patient-derived cells with Western blot, digital PCR, and high-speed/confocal microscopy of ciliary motion","pmids":["42050778"],"confidence":"Medium","gaps":["Single proof-of-concept study with limited editing efficiency and partial (~30%) functional rescue","Does not show restoration of radial spoke head ultrastructure"]},{"year":null,"claim":"The mechanism by which loss of RS1/RS2 spoke heads secondarily destabilizes the central pair complex, and the molecular contacts RSPH4A makes within the spoke head, remain unresolved.","evidence":"","pmids":[],"confidence":"High","gaps":["No atomic-resolution model of RSPH4A within the spoke head","Direct binding partners of RSPH4A in the spoke head not biochemically defined","Basis of species difference (three heads in mouse vs RS1/RS2 in human) unexplained"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0005198","term_label":"structural molecule activity","supporting_discovery_ids":[0,1,2]}],"localization":[{"term_id":"GO:0005929","term_label":"cilium","supporting_discovery_ids":[0,1,2]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[1,2]}],"pathway":[{"term_id":"R-HSA-1852241","term_label":"Organelle biogenesis and maintenance","supporting_discovery_ids":[1,2]}],"complexes":["radial spoke head"],"partners":[],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q5TD94","full_name":"Radial spoke head protein 4 homolog A","aliases":["Radial spoke head-like protein 3"],"length_aa":716,"mass_kda":80.7,"function":"Component of the axonemal radial spoke head which plays an important role in ciliary motility (PubMed:19200523). Essential for triplet radial spokes (RS1, RS2 and RS3) head assembly in the motile cilia (By similarity)","subcellular_location":"Cytoplasm, cytoskeleton, cilium axoneme; Cell projection, cilium","url":"https://www.uniprot.org/uniprotkb/Q5TD94/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/RSPH4A","classification":"Not Classified","n_dependent_lines":2,"n_total_lines":1208,"dependency_fraction":0.0016556291390728477},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/RSPH4A","total_profiled":1310},"omim":[{"mim_id":"616481","title":"CILIARY DYSKINESIA, PRIMARY, 32; CILD32","url":"https://www.omim.org/entry/616481"},{"mim_id":"615876","title":"RADIAL SPOKE HEAD 3; RSPH3","url":"https://www.omim.org/entry/615876"},{"mim_id":"612650","title":"CILIARY DYSKINESIA, PRIMARY, 12; CILD12","url":"https://www.omim.org/entry/612650"},{"mim_id":"612649","title":"CILIARY DYSKINESIA, PRIMARY, 11; CILD11","url":"https://www.omim.org/entry/612649"},{"mim_id":"612648","title":"RADIAL SPOKE HEAD COMPONENT 9; RSPH9","url":"https://www.omim.org/entry/612648"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Connecting piece","reliability":"Approved"},{"location":"Flagellar centriole","reliability":"Approved"},{"location":"Annulus","reliability":"Additional"}],"tissue_specificity":"Group enriched","tissue_distribution":"Detected in some","driving_tissues":[{"tissue":"choroid plexus","ntpm":22.0},{"tissue":"fallopian tube","ntpm":35.0}],"url":"https://www.proteinatlas.org/search/RSPH4A"},"hgnc":{"alias_symbol":["dJ412I7.1","FLJ37974","RSPH6B","CILD11"],"prev_symbol":["RSHL3"]},"alphafold":{"accession":"Q5TD94","domains":[{"cath_id":"-","chopping":"208-264","consensus_level":"high","plddt":86.596,"start":208,"end":264},{"cath_id":"-","chopping":"301-380_415-507_522-560_587-648","consensus_level":"medium","plddt":88.5861,"start":301,"end":648}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q5TD94","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q5TD94-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q5TD94-F1-predicted_aligned_error_v6.png","plddt_mean":67.44},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=RSPH4A","jax_strain_url":"https://www.jax.org/strain/search?query=RSPH4A"},"sequence":{"accession":"Q5TD94","fasta_url":"https://rest.uniprot.org/uniprotkb/Q5TD94.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q5TD94/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q5TD94"}},"corpus_meta":[{"pmid":"19200523","id":"PMC_19200523","title":"Mutations in radial spoke head protein genes RSPH9 and RSPH4A cause primary ciliary dyskinesia with central-microtubular-pair abnormalities.","date":"2009","source":"American journal of human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/19200523","citation_count":271,"is_preprint":false},{"pmid":"23798057","id":"PMC_23798057","title":"Founder mutation in RSPH4A identified in patients of Hispanic descent with primary ciliary dyskinesia.","date":"2013","source":"Human mutation","url":"https://pubmed.ncbi.nlm.nih.gov/23798057","citation_count":52,"is_preprint":false},{"pmid":"32203505","id":"PMC_32203505","title":"Rsph4a is essential for the triplet radial spoke head assembly of the mouse motile cilia.","date":"2020","source":"PLoS genetics","url":"https://pubmed.ncbi.nlm.nih.gov/32203505","citation_count":31,"is_preprint":false},{"pmid":"36768259","id":"PMC_36768259","title":"The RSPH4A Gene in Primary Ciliary Dyskinesia.","date":"2023","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/36768259","citation_count":18,"is_preprint":false},{"pmid":"33670432","id":"PMC_33670432","title":"Primary Ciliary Dyskinesia Diagnostic Challenges: Understanding the Clinical Phenotype of the Puerto Rican RSPH4A Founder Mutation.","date":"2021","source":"Diagnostics (Basel, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/33670432","citation_count":17,"is_preprint":false},{"pmid":"33852348","id":"PMC_33852348","title":"Structural insights into the cause of human RSPH4A primary ciliary dyskinesia.","date":"2021","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/33852348","citation_count":16,"is_preprint":false},{"pmid":"34513534","id":"PMC_34513534","title":"Primary Ciliary Dyskinesia: Ancestral Haplotypes Analysis of the RSPH4A Founder Mutation in Puerto Rico.","date":"2021","source":"Cureus","url":"https://pubmed.ncbi.nlm.nih.gov/34513534","citation_count":4,"is_preprint":false},{"pmid":"40429607","id":"PMC_40429607","title":"Assessing Olfactory Acuity in Primary Ciliary Dyskinesia with the RSPH4A Founder Mutation.","date":"2025","source":"Journal of clinical medicine","url":"https://pubmed.ncbi.nlm.nih.gov/40429607","citation_count":2,"is_preprint":false},{"pmid":"38818043","id":"PMC_38818043","title":"A novel homozygous RSPH4A variant in a family with primary ciliary dyskinesia and literature review.","date":"2024","source":"Frontiers in genetics","url":"https://pubmed.ncbi.nlm.nih.gov/38818043","citation_count":1,"is_preprint":false},{"pmid":"40262389","id":"PMC_40262389","title":"Primary ciliary dyskinesia in a Japanese woman caused by a novel RSPH4A variant.","date":"2025","source":"Respiratory investigation","url":"https://pubmed.ncbi.nlm.nih.gov/40262389","citation_count":1,"is_preprint":false},{"pmid":"40852414","id":"PMC_40852414","title":"Analysis of clinical and genetic features in an adolescent patient with primary ciliary dyskinesia induced by homozygous mutation in the RSPH4A gene: a case report.","date":"2025","source":"Frontiers in pediatrics","url":"https://pubmed.ncbi.nlm.nih.gov/40852414","citation_count":0,"is_preprint":false},{"pmid":"42050778","id":"PMC_42050778","title":"AAV-Mediated Base Editing for Correction of RSPH4A Mutations in Primary Ciliary Dyskinesia: A Proof-of-Concept Study.","date":"2026","source":"Human gene therapy","url":"https://pubmed.ncbi.nlm.nih.gov/42050778","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":7759,"output_tokens":1384,"usd":0.022018,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":8135,"output_tokens":2010,"usd":0.045463,"stage2_stop_reason":"end_turn"},"total_usd":0.067481,"stage1_batch_id":"msgbatch_013LWZZT6MWaqB9qyTSeQBmU","stage2_batch_id":"msgbatch_0194ZzQaTGNpruoNJBbEMbt3","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2009,\n      \"finding\": \"RSPH4A encodes a protein component of the axonemal radial spoke head; loss-of-function mutations cause primary ciliary dyskinesia with central-microtubular-pair abnormalities (intermittent or complete absence of the central pair), establishing its role in maintaining normal motile cilia movement in 9+2-structure cilia and flagella.\",\n      \"method\": \"SNP-based linkage analysis, positional candidate gene identification, in situ hybridization (murine Rsph9), zebrafish knockdown/rescue experiments for RSPH9 ortholog (functional context for radial spoke head proteins), human mutation analysis\",\n      \"journal\": \"American journal of human genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic mapping, mutation identification, and zebrafish knockdown/rescue replicated across multiple families and species; independently supported by subsequent structural studies\",\n      \"pmids\": [\"19200523\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Rsph4a is required for assembly of all three radial spoke heads (RS1, RS2, and RS3) in mouse tracheal cilia; Rsph4a-deficient mice lack all triplet spoke heads as revealed by cryo-electron tomography, and Rsph4a contributes to planar beating of motile cilia in trachea, ependymal tissue, and oviduct by building the distal architecture of radial spokes.\",\n      \"method\": \"Cryo-electron tomography of Rsph4a-knockout mouse tracheal cilia, ciliary movement observation, immunofluorescence analysis\",\n      \"journal\": \"PLoS genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — cryo-electron tomography structural analysis combined with functional (ciliary movement) and immunofluorescence readouts in a genetic knockout model, single lab but multiple orthogonal methods\",\n      \"pmids\": [\"32203505\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"In RSPH4A-/- human respiratory cilia, the radial spoke heads of RS1 and RS2 (but not RS3) are missing, and there are additional defects in the arch domains adjacent to RS1 and RS2 heads (distinct from RSPH1-/- defects); secondary heterogeneous defects in the central pair complex are also observed, establishing the specific structural role of RSPH4A at RS1 and RS2 spoke heads and adjacent arch domains.\",\n      \"method\": \"Cryo-electron tomography (cryo-ET) and subtomogram averaging of noninvasively collected human PCD patient respiratory cilia compared to controls\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — cryo-ET with subtomogram averaging directly on patient-derived human cilia, structural comparison with controls and RSPH1-/- cilia, single lab but highly rigorous structural method\",\n      \"pmids\": [\"33852348\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"A splice-site mutation (c.921+3_6delAAGT) in RSPH4A leads to a premature translation termination signal and loss of function, confirmed by quantitative ciliary ultrastructural analysis showing central apparatus defects, reduced ciliary beat frequency and abnormal waveform, and transcript analysis.\",\n      \"method\": \"Genetic sequencing, transcript analysis, ciliary ultrastructural analysis by electron microscopy, measurement of ciliary beat frequency and waveform\",\n      \"journal\": \"Human mutation\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function confirmed by multiple orthogonal methods (ultrastructure, beat frequency/waveform, transcript analysis) in a single study\",\n      \"pmids\": [\"23798057\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"AAV-mediated adenosine base editing targeting RSPH4A mutations in patient-derived cells partially restored normal ciliary motion pattern (~30.4% increase in normal motion pattern with reduction in circular motions) demonstrating that correction of RSPH4A sequence restores ciliary function.\",\n      \"method\": \"AAV-mediated base editing (nickase Cas9 fused to adenosine deaminase), Western blot, digital PCR for editing efficiency, high-speed video and confocal microscopy for ciliary beat frequency and motion pattern in patient cells\",\n      \"journal\": \"Human gene therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — functional restoration of ciliary motility after RSPH4A correction demonstrated with multiple readouts in patient cells, but single lab proof-of-concept study with limited editing efficiency\",\n      \"pmids\": [\"42050778\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"RSPH4A encodes a radial spoke head protein required for assembly of the distal heads of radial spokes RS1 and RS2 (and RS3 in mice) within the 9+2 axonemal repeat unit of motile cilia; loss of RSPH4A causes absence of RS1/RS2 spoke heads and arch domain defects with secondary central pair complex abnormalities, leading to abnormal (circling) ciliary beat pattern and primary ciliary dyskinesia without laterality defects.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"RSPH4A encodes a structural component of the axonemal radial spoke head required for normal motility of 9+2 motile cilia, and its loss causes primary ciliary dyskinesia [#0]. RSPH4A builds the distal head architecture of radial spokes: in human respiratory cilia its loss eliminates the radial spoke heads of RS1 and RS2 (but spares RS3) and produces additional defects in the arch domains adjacent to those heads, with secondary heterogeneous abnormalities of the central pair complex [#2]. In mouse tracheal cilia Rsph4a is required for assembly of all three radial spoke heads (RS1, RS2, RS3), and it supports planar ciliary beating across tracheal, ependymal, and oviduct epithelia [#1]. Loss-of-function mutations, including a splice-site allele generating premature termination, abolish this function and yield central-apparatus ultrastructural defects, reduced beat frequency, and abnormal waveform [#0, #3]; correction of the RSPH4A sequence in patient-derived cells partially restores normal ciliary motion [#4].\",\n  \"teleology\": [\n    {\n      \"year\": 2009,\n      \"claim\": \"Established RSPH4A as a radial spoke head gene whose loss-of-function mutations cause primary ciliary dyskinesia, defining a genetic basis for central-pair-associated ciliary dysmotility.\",\n      \"evidence\": \"SNP linkage, positional candidate identification, and human mutation analysis with ortholog functional context\",\n      \"pmids\": [\"19200523\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not resolve which radial spoke heads RSPH4A builds at structural resolution\", \"Mechanism linking radial spoke loss to central pair abnormality not defined\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Showed that a specific splice-site mutation causes premature termination and loss of function, linking RSPH4A sequence disruption to measurable ciliary ultrastructural and motility defects.\",\n      \"evidence\": \"Genetic sequencing, transcript analysis, electron microscopy, beat frequency and waveform measurement in patient cilia\",\n      \"pmids\": [\"23798057\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single study; structural localization of the defect within radial spokes not resolved\", \"Protein-level consequence inferred from transcript, not directly visualized\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Defined the structural role of Rsph4a in vivo by showing it is required to assemble all three radial spoke heads and to support planar beating across multiple ciliated epithelia in mouse.\",\n      \"evidence\": \"Cryo-electron tomography of knockout mouse tracheal cilia with ciliary movement and immunofluorescence readouts\",\n      \"pmids\": [\"32203505\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mouse three-head requirement differs from later human findings; species-specific architecture unresolved\", \"Does not establish direct protein-protein contacts within the spoke head\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Pinpointed the human structural defect to absence of RS1 and RS2 spoke heads (sparing RS3) plus adjacent arch domain defects, distinguishing RSPH4A loss from RSPH1 loss and revealing secondary central pair abnormalities.\",\n      \"evidence\": \"Cryo-ET with subtomogram averaging of patient-derived human respiratory cilia versus controls and RSPH1-/- cilia\",\n      \"pmids\": [\"33852348\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism by which spoke head loss secondarily perturbs the central pair not defined\", \"Atomic-resolution position of RSPH4A within the spoke head head not determined\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Demonstrated that correcting the RSPH4A sequence is sufficient to restore ciliary function, providing causal confirmation that the motility defect stems from the gene lesion.\",\n      \"evidence\": \"AAV-mediated adenosine base editing of patient-derived cells with Western blot, digital PCR, and high-speed/confocal microscopy of ciliary motion\",\n      \"pmids\": [\"42050778\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single proof-of-concept study with limited editing efficiency and partial (~30%) functional rescue\", \"Does not show restoration of radial spoke head ultrastructure\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The mechanism by which loss of RS1/RS2 spoke heads secondarily destabilizes the central pair complex, and the molecular contacts RSPH4A makes within the spoke head, remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No atomic-resolution model of RSPH4A within the spoke head\", \"Direct binding partners of RSPH4A in the spoke head not biochemically defined\", \"Basis of species difference (three heads in mouse vs RS1/RS2 in human) unexplained\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0005198\", \"supporting_discovery_ids\": [0, 1, 2]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005929\", \"supporting_discovery_ids\": [0, 1, 2]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [1, 2]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1852241\", \"supporting_discovery_ids\": [1, 2]}\n    ],\n    \"complexes\": [\"radial spoke head\"],\n    \"partners\": [],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":4,"faith_total":4,"faith_pct":100.0}}