{"doi":"10.1101/2025.04.09.648007","title":"Hypoimmunogenic hPSC-derived cardiac organoids for immune evasion and heart repair","abstract":"ABSTRACT Human pluripotent stem cell (hPSC)-derived cardiac therapies hold great promise for heart regeneration but face major translational barriers due to allogeneic immune rejection. Here, we engineered hypoimmunogenic hPSCs using a two-step CRISPR-Cas9 strategy: (1) B2M knockout, eliminating HLA class I surface expression, and (2) knock-in of HLA-E or HLA-G trimer constructs in the AAVS1 safe harbor locus to confer robust immune evasion. Hypoimmunogenic hPSCs maintained pluripotency, efficiently differentiated into cardiac cell types that resisted both T and NK cell-mediated cytotoxicity in vitro , and self-assembled into engineered cardiac organoids. Comprehensive analyses of the hypoimmunogenic cells and organoids revealed preservation of transcriptomic, structural, and functional properties with minimal off-target effects from gene editing. In vivo , hypoimmunogenic cardiac organoids restored contractile function in infarcted rat hearts and demonstrated superior graft retention and immune evasion in humanized mice compared to wild-type counterparts. These findings establish the therapeutic potential of hypoimmunogenic hPSC-CMs in the cardiac organoid platform, laying the foundation for off-the-shelf cardiac cell therapies to treat cardiovascular disease, the leading cause of death worldwide.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2025,"id":554971,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":5,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9444,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1437961,"name":"Alessandro R. Howells","orcid":"0000-0001-7741-340X","position":1,"is_corresponding":false},{"id":314032,"name":"Dimitrios C. Arhontoulis","orcid":"0000-0002-7959-7692","position":2,"is_corresponding":false},{"id":320852,"name":"Lauren N. Randolph","orcid":"0000-0002-1628-8673","position":3,"is_corresponding":false},{"id":888567,"name":"Nathaniel Hyams","orcid":"0009-0007-4158-3352","position":4,"is_corresponding":false},{"id":494229,"name":"Ryan W. Barrs","orcid":"0000-0001-7747-1941","position":5,"is_corresponding":false},{"id":1028438,"name":"Mei Li","orcid":"0000-0002-4264-8505","position":6,"is_corresponding":false},{"id":228501,"name":"Charles M. Kerr","orcid":"0000-0001-8736-4795","position":7,"is_corresponding":false},{"id":361355,"name":"Rob A. Robino","orcid":"0000-0002-1645-0247","position":8,"is_corresponding":false},{"id":664749,"name":"Jordan E. Morningstar","orcid":"0000-0003-2168-9671","position":9,"is_corresponding":false},{"id":1452799,"name":"Jacelyn D. Bain","orcid":"0009-0009-5897-2711","position":10,"is_corresponding":false},{"id":447221,"name":"Martha E. Floy","orcid":"0000-0003-1992-5911","position":11,"is_corresponding":false},{"id":260341,"name":"Russell A. Norris","orcid":"0000-0001-8462-5726","position":12,"is_corresponding":false},{"id":280588,"name":"Xiaoping Bao","orcid":"0000-0003-0955-6868","position":13,"is_corresponding":false},{"id":1095110,"name":"Jean Marie Ruddy","orcid":"0000-0002-4148-9549","position":14,"is_corresponding":false},{"id":254957,"name":"Sean P. Palecek","orcid":"0000-0003-4917-5584","position":15,"is_corresponding":false},{"id":499786,"name":"Leonardo M. R. Ferreira","orcid":"0000-0003-2491-9866","position":16,"is_corresponding":false},{"id":270356,"name":"Xiaojun Lian","orcid":"0000-0002-9161-1004","position":17,"is_corresponding":false},{"id":228510,"name":"Ying Mei","orcid":"0000-0002-8508-4076","position":18,"is_corresponding":false},{"id":666720,"name":"Sophia E. Silver","orcid":"0000-0003-0718-591X","position":0,"is_corresponding":true}],"reference_count":94,"raw_metadata":null,"created_at":"2026-07-19T02:54:54.542303Z","pmid":"40291708","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}