{"gene":"AMIGO3","run_date":"2026-06-09T22:02:43","timeline":{"discoveries":[{"year":2003,"finding":"AMIGO3 is a type I transmembrane protein with six leucine-rich repeats (LRRs) flanked by cysteine-rich LRR N- and C-terminal domains and one immunoglobulin domain; AMIGO family members engage in homophilic and heterophilic binding with each other, functioning as cell adhesion molecules.","method":"Molecular cloning, domain structure analysis, recombinant ectodomain binding assays, neurite outgrowth assays in hippocampal neurons","journal":"The Journal of cell biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — homophilic/heterophilic binding demonstrated with recombinant ectodomains; structural domain characterization by sequence analysis; single lab but multiple functional assays","pmids":["12629050"],"is_preprint":false},{"year":2011,"finding":"AMIGO-3 (along with AMIGO-1 and AMIGO-2) forms dimers through LRR-LRR interfaces, as inferred from the crystal structure of AMIGO-1 and small-angle X-ray scattering (SAXS) data on AMIGO-3; dimerization is necessary for proper cell-surface expression and likely stable folding in the ER.","method":"Crystal structure of AMIGO-1 at 2.0 Å resolution, SAXS data on AMIGO-3, static light-scattering, mutagenesis of dimerization interface","journal":"Journal of molecular biology","confidence":"Medium","confidence_rationale":"Tier 1 / Moderate — crystal structure plus mutagenesis for AMIGO-1 with SAXS supporting AMIGO-3 dimerization; AMIGO-3 specific data is indirect (modeled/SAXS), so confidence is moderate rather than high","pmids":["21983541"],"is_preprint":false},{"year":2013,"finding":"AMIGO3 acts as a co-receptor in the NgR1/p75-TROY inhibitory signaling complex, substituting for LINGO-1; AMIGO3 interacts physically with NgR1 and p75/TROY (demonstrated in non-neuronal cells and brain lysates), mediates RhoA activation in response to CNS myelin, and its knockdown in primary DRG and retinal cultures promotes neurite growth under myelin-inhibitory conditions.","method":"Co-immunoprecipitation from brain lysates and non-neuronal cell co-expression, RhoA activation assay, shRNA knockdown with neurite outgrowth assays in primary DRG and retinal neurons, in vivo expression profiling after dorsal column and optic nerve lesion","journal":"PloS one","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal co-IP in multiple cell contexts, functional RhoA activation assay, loss-of-function phenotype in primary neurons, replicated in vivo expression data; multiple orthogonal methods in one study","pmids":["23613963"],"is_preprint":false},{"year":2018,"finding":"In vivo shRNA-mediated knockdown of AMIGO3 in dorsal root ganglion neurons (>75% mRNA reduction) disinhibits NT3-stimulated regeneration of spinal cord dorsal column axons, restores compound action potential conduction across the lesion, and improves sensory and locomotor function, confirming AMIGO3's role in mediating axon growth inhibition through the RhoA pathway in vivo.","method":"shRNA knockdown via non-viral jetPEI delivery, in vivo dorsal column injury model, electrophysiology (compound action potential), behavioral testing, Western blot and immunofluorescence for AMIGO3 expression","journal":"Scientific reports","confidence":"High","confidence_rationale":"Tier 2 / Strong — clean in vivo loss-of-function with multiple orthogonal readouts (molecular, electrophysiological, behavioral), replicates mechanistic findings from PMID:23613963","pmids":["30013050"],"is_preprint":false},{"year":2021,"finding":"AMIGO3 expression is upregulated after status convulsion in immature mice and contributes to myelin sheath damage; downregulation of AMIGO3 alleviates myelin structural impairment (assessed by TEM and myelin basic protein levels) and inhibits the ROCK/RhoA signaling pathway, indicating AMIGO3 signals through ROCK/RhoA to regulate myelination and axon growth after seizure.","method":"Kainic acid status convulsion model in immature mice, shRNA knockdown of AMIGO3, Western blot for myelin basic protein and ROCK/RhoA pathway components, transmission electron microscopy of myelin ultrastructure","journal":"Frontiers in molecular neuroscience","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — clean loss-of-function with defined molecular and ultrastructural readouts, but single lab and single publication","pmids":["34650403"],"is_preprint":false}],"current_model":"AMIGO3 is a type I transmembrane LRR/Ig-domain protein that dimerizes via its LRR interface and functions as a co-receptor within the NgR1/p75-TROY inhibitory signaling complex—substituting for LINGO-1 acutely after CNS injury—where it transduces myelin-derived inhibitory signals through RhoA/ROCK to collapse axonal growth cones and suppress myelination, effects that are reversed by AMIGO3 knockdown both in vitro and in vivo."},"narrative":{"mechanistic_narrative":"AMIGO3 is a type I transmembrane protein composed of six leucine-rich repeats flanked by cysteine-rich LRR-N and LRR-C caps and a single immunoglobulin domain, and it acts as a cell adhesion molecule capable of homophilic and heterophilic binding with other AMIGO family members [PMID:12629050]. It dimerizes through an LRR-LRR interface, a contact required for proper cell-surface expression and stable folding [PMID:21983541]. Functionally, AMIGO3 serves as a co-receptor within the NgR1/p75-TROY inhibitory signaling complex, where it substitutes for LINGO-1, binds NgR1 and p75/TROY, and transduces CNS myelin-derived inhibitory signals through RhoA/ROCK activation to suppress axon growth [PMID:23613963, PMID:34650403]. Loss of AMIGO3 disinhibits axon regeneration: shRNA knockdown in primary DRG and retinal neurons promotes neurite outgrowth under myelin-inhibitory conditions [PMID:23613963], and in vivo knockdown in dorsal root ganglion neurons restores NT3-stimulated dorsal column axon regeneration, compound action potential conduction, and sensory-locomotor function [PMID:30013050]. AMIGO3 is also upregulated after status convulsion and contributes to myelin sheath damage, with its downregulation relieving myelin ultrastructural impairment via ROCK/RhoA suppression [PMID:34650403].","teleology":[{"year":2003,"claim":"Establishing AMIGO3's domain architecture and adhesive behavior defined it as an LRR/Ig cell adhesion molecule capable of homophilic and heterophilic interactions, framing it as a candidate neuronal surface receptor.","evidence":"Molecular cloning, domain analysis, recombinant ectodomain binding and neurite outgrowth assays in hippocampal neurons","pmids":["12629050"],"confidence":"Medium","gaps":["Did not identify the in vivo binding partners or downstream signaling","Adhesion shown for ectodomains; physiological neuronal role unresolved"]},{"year":2011,"claim":"Structural work resolved how AMIGO proteins assemble, showing AMIGO3 forms LRR-LRR dimers required for surface expression and folding, explaining a prerequisite for its receptor function.","evidence":"Crystal structure of AMIGO-1 at 2.0 Å with SAXS, light-scattering and interface mutagenesis applied to AMIGO-3","pmids":["21983541"],"confidence":"Medium","gaps":["AMIGO-3 dimerization inferred from SAXS/modeling, not a direct AMIGO3 crystal structure","Did not address heteromeric assembly with NgR1/p75/TROY"]},{"year":2013,"claim":"Identifying AMIGO3 as a LINGO-1-substituting co-receptor in the NgR1/p75-TROY complex answered how myelin inhibitory signals are transduced, placing AMIGO3 at the receptor level upstream of RhoA.","evidence":"Reciprocal co-IP in brain lysates and non-neuronal cells, RhoA activation assay, shRNA knockdown with neurite outgrowth in primary DRG and retinal neurons, in vivo lesion expression profiling","pmids":["23613963"],"confidence":"High","gaps":["Stoichiometry and direct binding interface within the complex not defined","Whether AMIGO3 binds myelin ligands directly was not established"]},{"year":2018,"claim":"In vivo knockdown demonstrated that removing AMIGO3 disinhibits axon regeneration with functional recovery, confirming it is a causally required mediator of growth inhibition rather than a correlate.","evidence":"Non-viral shRNA knockdown in DRG neurons, dorsal column injury model, compound action potential electrophysiology, behavioral testing, Western blot/immunofluorescence","pmids":["30013050"],"confidence":"High","gaps":["Did not dissect the RhoA/ROCK step at molecular resolution in vivo","Durability and clinical translatability of the regenerative effect not addressed"]},{"year":2021,"claim":"Extending AMIGO3 to a seizure context showed it drives myelin sheath damage via ROCK/RhoA, generalizing its inhibitory signaling role beyond traumatic axon injury to demyelinating pathology.","evidence":"Kainic acid status convulsion model in immature mice, shRNA knockdown, Western blot for MBP and ROCK/RhoA components, TEM of myelin ultrastructure","pmids":["34650403"],"confidence":"Medium","gaps":["Single lab, single publication","Cell type driving the effect (neuron vs oligodendrocyte) not resolved"]},{"year":null,"claim":"The direct molecular ligand sensed by the AMIGO3-containing receptor complex and the structural basis of AMIGO3 incorporation into the NgR1/p75-TROY assembly remain undefined.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No direct AMIGO3-myelin ligand interaction characterized","No structure of the AMIGO3-containing receptor complex","Whether AMIGO3 substitution for LINGO-1 is regulated transcriptionally or post-translationally is unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098631","term_label":"cell adhesion mediator activity","supporting_discovery_ids":[0]},{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[2]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[2]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[1,2]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[2,4]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[2,3]}],"complexes":["NgR1/p75-TROY inhibitory receptor complex"],"partners":["RTN4R","NGFR","TROY","AMIGO1","AMIGO2"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q86WK7","full_name":"Amphoterin-induced protein 3","aliases":["AMIGO-3","Alivin-3"],"length_aa":504,"mass_kda":55.2,"function":"May mediate heterophilic cell-cell interaction. May contribute to signal transduction through its intracellular domain (By similarity)","subcellular_location":"Membrane","url":"https://www.uniprot.org/uniprotkb/Q86WK7/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/AMIGO3","classification":"Not Classified","n_dependent_lines":1,"n_total_lines":1208,"dependency_fraction":0.0008278145695364238},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/AMIGO3","total_profiled":1310},"omim":[{"mim_id":"615691","title":"ADHESION MOLECULE WITH Ig-LIKE DOMAIN 3; AMIGO3","url":"https://www.omim.org/entry/615691"},{"mim_id":"615690","title":"ADHESION MOLECULE WITH Ig-LIKE DOMAIN 2; AMIGO2","url":"https://www.omim.org/entry/615690"},{"mim_id":"615689","title":"ADHESION MOLECULE WITH Ig-LIKE DOMAIN 1; AMIGO1","url":"https://www.omim.org/entry/615689"},{"mim_id":"615320","title":"GDP-MANNOSE PYROPHOSPHORYLASE B; GMPPB","url":"https://www.omim.org/entry/615320"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Actin filaments","reliability":"Approved"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in many","driving_tissues":[],"url":"https://www.proteinatlas.org/search/AMIGO3"},"hgnc":{"alias_symbol":[],"prev_symbol":[]},"alphafold":{"accession":"Q86WK7","domains":[{"cath_id":"2.60.40.10","chopping":"287-373","consensus_level":"high","plddt":83.3967,"start":287,"end":373}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q86WK7","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q86WK7-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q86WK7-F1-predicted_aligned_error_v6.png","plddt_mean":73.5},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=AMIGO3","jax_strain_url":"https://www.jax.org/strain/search?query=AMIGO3"},"sequence":{"accession":"Q86WK7","fasta_url":"https://rest.uniprot.org/uniprotkb/Q86WK7.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q86WK7/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q86WK7"}},"corpus_meta":[{"pmid":"12629050","id":"PMC_12629050","title":"AMIGO, a transmembrane protein implicated in axon tract development, defines a novel protein family with leucine-rich repeats.","date":"2003","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/12629050","citation_count":135,"is_preprint":false},{"pmid":"23613963","id":"PMC_23613963","title":"AMIGO3 is an NgR1/p75 co-receptor signalling axon growth inhibition in the acute phase of adult central nervous system injury.","date":"2013","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/23613963","citation_count":31,"is_preprint":false},{"pmid":"21983541","id":"PMC_21983541","title":"Crystal structure and role of glycans and dimerization in folding of neuronal leucine-rich repeat protein AMIGO-1.","date":"2011","source":"Journal of molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/21983541","citation_count":25,"is_preprint":false},{"pmid":"28966634","id":"PMC_28966634","title":"LINGO-1 and AMIGO3, potential therapeutic targets for neurological and dysmyelinating disorders?","date":"2017","source":"Neural regeneration research","url":"https://pubmed.ncbi.nlm.nih.gov/28966634","citation_count":21,"is_preprint":false},{"pmid":"30013050","id":"PMC_30013050","title":"Non-viral-mediated suppression of AMIGO3 promotes disinhibited NT3-mediated regeneration of spinal cord dorsal column axons.","date":"2018","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/30013050","citation_count":16,"is_preprint":false},{"pmid":"35094710","id":"PMC_35094710","title":"Establishment of an antibody specific for AMIGO2 improves immunohistochemical evaluation of liver metastases and clinical outcomes in patients with colorectal cancer.","date":"2022","source":"Diagnostic pathology","url":"https://pubmed.ncbi.nlm.nih.gov/35094710","citation_count":13,"is_preprint":false},{"pmid":"37279148","id":"PMC_37279148","title":"Comparative and integrative analysis of transcriptomic and epigenomic-wide DNA methylation changes in African American prostate cancer.","date":"2023","source":"Epigenetics","url":"https://pubmed.ncbi.nlm.nih.gov/37279148","citation_count":11,"is_preprint":false},{"pmid":"34650403","id":"PMC_34650403","title":"The Role and Mechanism of AMIGO3 in the Formation of Aberrant Neural Circuits After Status Convulsion in Immature Mice.","date":"2021","source":"Frontiers in molecular neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/34650403","citation_count":1,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":6294,"output_tokens":1517,"usd":0.020819,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":8267,"output_tokens":2343,"usd":0.049955,"stage2_stop_reason":"end_turn"},"total_usd":0.070774,"stage1_batch_id":"msgbatch_0179Z2pxRYBsXi2xADneafRw","stage2_batch_id":"msgbatch_01LaY9ZaVdgjWArcZo9eqQE5","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2003,\n      \"finding\": \"AMIGO3 is a type I transmembrane protein with six leucine-rich repeats (LRRs) flanked by cysteine-rich LRR N- and C-terminal domains and one immunoglobulin domain; AMIGO family members engage in homophilic and heterophilic binding with each other, functioning as cell adhesion molecules.\",\n      \"method\": \"Molecular cloning, domain structure analysis, recombinant ectodomain binding assays, neurite outgrowth assays in hippocampal neurons\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — homophilic/heterophilic binding demonstrated with recombinant ectodomains; structural domain characterization by sequence analysis; single lab but multiple functional assays\",\n      \"pmids\": [\"12629050\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"AMIGO-3 (along with AMIGO-1 and AMIGO-2) forms dimers through LRR-LRR interfaces, as inferred from the crystal structure of AMIGO-1 and small-angle X-ray scattering (SAXS) data on AMIGO-3; dimerization is necessary for proper cell-surface expression and likely stable folding in the ER.\",\n      \"method\": \"Crystal structure of AMIGO-1 at 2.0 Å resolution, SAXS data on AMIGO-3, static light-scattering, mutagenesis of dimerization interface\",\n      \"journal\": \"Journal of molecular biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — crystal structure plus mutagenesis for AMIGO-1 with SAXS supporting AMIGO-3 dimerization; AMIGO-3 specific data is indirect (modeled/SAXS), so confidence is moderate rather than high\",\n      \"pmids\": [\"21983541\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"AMIGO3 acts as a co-receptor in the NgR1/p75-TROY inhibitory signaling complex, substituting for LINGO-1; AMIGO3 interacts physically with NgR1 and p75/TROY (demonstrated in non-neuronal cells and brain lysates), mediates RhoA activation in response to CNS myelin, and its knockdown in primary DRG and retinal cultures promotes neurite growth under myelin-inhibitory conditions.\",\n      \"method\": \"Co-immunoprecipitation from brain lysates and non-neuronal cell co-expression, RhoA activation assay, shRNA knockdown with neurite outgrowth assays in primary DRG and retinal neurons, in vivo expression profiling after dorsal column and optic nerve lesion\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal co-IP in multiple cell contexts, functional RhoA activation assay, loss-of-function phenotype in primary neurons, replicated in vivo expression data; multiple orthogonal methods in one study\",\n      \"pmids\": [\"23613963\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"In vivo shRNA-mediated knockdown of AMIGO3 in dorsal root ganglion neurons (>75% mRNA reduction) disinhibits NT3-stimulated regeneration of spinal cord dorsal column axons, restores compound action potential conduction across the lesion, and improves sensory and locomotor function, confirming AMIGO3's role in mediating axon growth inhibition through the RhoA pathway in vivo.\",\n      \"method\": \"shRNA knockdown via non-viral jetPEI delivery, in vivo dorsal column injury model, electrophysiology (compound action potential), behavioral testing, Western blot and immunofluorescence for AMIGO3 expression\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — clean in vivo loss-of-function with multiple orthogonal readouts (molecular, electrophysiological, behavioral), replicates mechanistic findings from PMID:23613963\",\n      \"pmids\": [\"30013050\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"AMIGO3 expression is upregulated after status convulsion in immature mice and contributes to myelin sheath damage; downregulation of AMIGO3 alleviates myelin structural impairment (assessed by TEM and myelin basic protein levels) and inhibits the ROCK/RhoA signaling pathway, indicating AMIGO3 signals through ROCK/RhoA to regulate myelination and axon growth after seizure.\",\n      \"method\": \"Kainic acid status convulsion model in immature mice, shRNA knockdown of AMIGO3, Western blot for myelin basic protein and ROCK/RhoA pathway components, transmission electron microscopy of myelin ultrastructure\",\n      \"journal\": \"Frontiers in molecular neuroscience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — clean loss-of-function with defined molecular and ultrastructural readouts, but single lab and single publication\",\n      \"pmids\": [\"34650403\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"AMIGO3 is a type I transmembrane LRR/Ig-domain protein that dimerizes via its LRR interface and functions as a co-receptor within the NgR1/p75-TROY inhibitory signaling complex—substituting for LINGO-1 acutely after CNS injury—where it transduces myelin-derived inhibitory signals through RhoA/ROCK to collapse axonal growth cones and suppress myelination, effects that are reversed by AMIGO3 knockdown both in vitro and in vivo.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"AMIGO3 is a type I transmembrane protein composed of six leucine-rich repeats flanked by cysteine-rich LRR-N and LRR-C caps and a single immunoglobulin domain, and it acts as a cell adhesion molecule capable of homophilic and heterophilic binding with other AMIGO family members [#0]. It dimerizes through an LRR-LRR interface, a contact required for proper cell-surface expression and stable folding [#1]. Functionally, AMIGO3 serves as a co-receptor within the NgR1/p75-TROY inhibitory signaling complex, where it substitutes for LINGO-1, binds NgR1 and p75/TROY, and transduces CNS myelin-derived inhibitory signals through RhoA/ROCK activation to suppress axon growth [#2, #4]. Loss of AMIGO3 disinhibits axon regeneration: shRNA knockdown in primary DRG and retinal neurons promotes neurite outgrowth under myelin-inhibitory conditions [#2], and in vivo knockdown in dorsal root ganglion neurons restores NT3-stimulated dorsal column axon regeneration, compound action potential conduction, and sensory-locomotor function [#3]. AMIGO3 is also upregulated after status convulsion and contributes to myelin sheath damage, with its downregulation relieving myelin ultrastructural impairment via ROCK/RhoA suppression [#4].\",\n  \"teleology\": [\n    {\n      \"year\": 2003,\n      \"claim\": \"Establishing AMIGO3's domain architecture and adhesive behavior defined it as an LRR/Ig cell adhesion molecule capable of homophilic and heterophilic interactions, framing it as a candidate neuronal surface receptor.\",\n      \"evidence\": \"Molecular cloning, domain analysis, recombinant ectodomain binding and neurite outgrowth assays in hippocampal neurons\",\n      \"pmids\": [\"12629050\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Did not identify the in vivo binding partners or downstream signaling\", \"Adhesion shown for ectodomains; physiological neuronal role unresolved\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Structural work resolved how AMIGO proteins assemble, showing AMIGO3 forms LRR-LRR dimers required for surface expression and folding, explaining a prerequisite for its receptor function.\",\n      \"evidence\": \"Crystal structure of AMIGO-1 at 2.0 Å with SAXS, light-scattering and interface mutagenesis applied to AMIGO-3\",\n      \"pmids\": [\"21983541\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"AMIGO-3 dimerization inferred from SAXS/modeling, not a direct AMIGO3 crystal structure\", \"Did not address heteromeric assembly with NgR1/p75/TROY\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Identifying AMIGO3 as a LINGO-1-substituting co-receptor in the NgR1/p75-TROY complex answered how myelin inhibitory signals are transduced, placing AMIGO3 at the receptor level upstream of RhoA.\",\n      \"evidence\": \"Reciprocal co-IP in brain lysates and non-neuronal cells, RhoA activation assay, shRNA knockdown with neurite outgrowth in primary DRG and retinal neurons, in vivo lesion expression profiling\",\n      \"pmids\": [\"23613963\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Stoichiometry and direct binding interface within the complex not defined\", \"Whether AMIGO3 binds myelin ligands directly was not established\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"In vivo knockdown demonstrated that removing AMIGO3 disinhibits axon regeneration with functional recovery, confirming it is a causally required mediator of growth inhibition rather than a correlate.\",\n      \"evidence\": \"Non-viral shRNA knockdown in DRG neurons, dorsal column injury model, compound action potential electrophysiology, behavioral testing, Western blot/immunofluorescence\",\n      \"pmids\": [\"30013050\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not dissect the RhoA/ROCK step at molecular resolution in vivo\", \"Durability and clinical translatability of the regenerative effect not addressed\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Extending AMIGO3 to a seizure context showed it drives myelin sheath damage via ROCK/RhoA, generalizing its inhibitory signaling role beyond traumatic axon injury to demyelinating pathology.\",\n      \"evidence\": \"Kainic acid status convulsion model in immature mice, shRNA knockdown, Western blot for MBP and ROCK/RhoA components, TEM of myelin ultrastructure\",\n      \"pmids\": [\"34650403\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab, single publication\", \"Cell type driving the effect (neuron vs oligodendrocyte) not resolved\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The direct molecular ligand sensed by the AMIGO3-containing receptor complex and the structural basis of AMIGO3 incorporation into the NgR1/p75-TROY assembly remain undefined.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No direct AMIGO3-myelin ligand interaction characterized\", \"No structure of the AMIGO3-containing receptor complex\", \"Whether AMIGO3 substitution for LINGO-1 is regulated transcriptionally or post-translationally is unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098631\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [2]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [2]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [1, 2]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [2, 4]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [2, 3]}\n    ],\n    \"complexes\": [\"NgR1/p75-TROY inhibitory receptor complex\"],\n    \"partners\": [\"RTN4R\", \"NGFR\", \"TROY\", \"AMIGO1\", \"AMIGO2\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":4,"faith_total":5,"faith_pct":80.0}}