{"artifact":{"id":"experiment-exp-wiki-experiments-c9orf72-hexanucleotide-repeat-mechanism","artifact_type":"experiment","entity_ids":null,"title":"Mechanism: C9orf72 Hexanucleotide Repeat Expansion in ALS/FTD","quality_score":0.675,"created_by":"crosslink-v2","provenance_chain":null,"content_hash":null,"metadata":{"source":{"type":"manual","source_name":"wiki","extracted_by":"backfill_v1","extraction_date":"2026-04-16T01:00:16.897119Z"},"summary":"# Mechanism: C9orf72 Hexanucleotide Repeat Expansion in ALS/FTD\n## Background and Rationale\nThis comprehensive validation study addresses one of the most significant questions in neurodegeneration: how a single genetic mutation in C9orf72 can manifest as distinct diseases (ALS, FTD, or combined phenotypes). The C9orf72 hexanucleotide repeat expansion represents the most common genetic cause of both familial ALS (~40%) and familial FTD (~25%), yet the mechanisms underlying phenotypic diversity re","entities":{"genes":["FTD"],"diseases":["ALS"]},"model_system":"human","_schema_version":1,"experiment_type":"validation","primary_outcome":"Validate Mechanism: C9orf72 Hexanucleotide Repeat Expansion in ALS/FTD","methodology_notes":"**Phase 1: Patient Cohort Assembly and Characterization (Months 1-3)**\n\nRecruit 150 participants: 50 C9orf72+ ALS patients, 50 C9orf72+ FTD patients, 25 C9orf72+ ALS/FTD patients, and 25 healthy controls with C9orf72 mutations but no symptoms. Perform comprehensive clinical phenotyping including ALSFRS-R, CDR-FTLD, and neuropsychological testing. Extract genomic DNA and perform Southern blot analysis to quantify hexanucleotide repeat length. Establish iPSC lines from all participants using episomal reprogramming.\n\n**Phase 2: Molecular Pathology Analysis (Months 4-8)**\n\nGenerate motor neurons and cortical neurons from iPSCs using dual SMAD inhibition protocol. Perform RNA-seq on differentiated neurons at days 21, 35, and 50 to identify disease-specific transcriptional signatures. Quantify RNA foci formation using FISH with (GGGGCC)4 and (CCCCGG)4 probes. Measure dipeptide repeat protein (DPR) accumulation via immunofluorescence and Western blot for poly-GA, poly-GP, poly-GR, poly-PA, an","replication_status":"replicated","extraction_metadata":{"backfill_at":"2026-04-16T01:00:16.897126","needs_review":true,"extraction_notes":"Backfilled from wiki source (no PMID available)","extraction_confidence":0.4}},"created_at":"2026-04-02T10:01:41.825940-07:00","updated_at":"2026-04-16T01:00:16-07:00","version_number":4,"parent_version_id":null,"version_tag":null,"changelog":null,"is_latest":1,"lifecycle_state":"active","superseded_by":null,"deprecated_at":null,"deprecated_reason":null,"dependencies":null,"market_price":0.5,"origin_type":"internal","origin_url":null,"lifecycle_changed_at":"2026-04-02T10:01:41.825940-07:00","citation_count":0,"embed_count":0,"derivation_count":0,"support_count":0,"contradiction_count":0,"total_usage":0.0,"usage_score":0.5,"usage_computed_at":null,"quality_status":"ok","contributors":[],"answers_question_ids":null,"deprecated_reason_detail":null,"deprecated_reason_code":null,"commit_sha":null,"commit_submodule":null,"last_mutated_at":"2026-05-16T14:51:34.657673-07:00","disputed_at":null,"gap_id":null,"mission_id":null,"intrinsic_priority":null,"effective_priority":null,"artifact_id":"3f56f432-7a66-4082-b1d2-688583923b42","artifact_dir":null,"primary_filename":null,"accessory_filenames":null,"folder_layout_version":1,"migrated_to_folder_at":null,"hypothesis_id":null,"authorship":{"kind":"human","contributors":[{"role":"author","actor_ref":"crosslink-v2"}]},"epistemic_tier":"T3_provisional","created_by_agent_id":null},"outgoing_links":[{"target_artifact_id":"wiki-hypotheses","link_type":"related","strength":0.54,"evidence":"Reverse: 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