{"doi":"10.1126/sciimmunol.ade3369","title":"In vitro modeling of CD8 <sup>+</sup> T cell exhaustion enables CRISPR screening to reveal a role for BHLHE40","abstract":"Identifying molecular mechanisms of exhausted CD8 T cells (T ex ) is a key goal of improving immunotherapy of cancer and other diseases. However, high-throughput interrogation of in vivo T ex can be costly and inefficient. In vitro models of T ex are easily customizable and quickly generate high cellular yield, enabling CRISPR screening and other high-throughput assays. We established an in vitro model of chronic stimulation and benchmarked key phenotypic, functional, transcriptional, and epigenetic features against bona fide in vivo T ex . We leveraged this model of in vitro chronic stimulation in combination with CRISPR screening to identify transcriptional regulators of T cell exhaustion. This approach identified several transcription factors, including BHLHE40. In vitro and in vivo validation defined a role for BHLHE40 in regulating a key differentiation checkpoint between progenitor and intermediate T ex subsets. By developing and benchmarking an in vitro model of T ex , then applying high-throughput CRISPR screening, we demonstrate the utility of mechanistically annotated in vitro models of T ex .","journal":"Science Immunology","year":2023,"id":316727,"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":94,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9421,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2023-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":105455,"name":"Sasikanth Manne","orcid":"0000-0001-9856-0871","position":1,"is_corresponding":false},{"id":265861,"name":"Shin Foong Ngiow","orcid":"0000-0002-5738-2834","position":2,"is_corresponding":false},{"id":105448,"name":"Amy E. Baxter","orcid":"0000-0002-1555-0713","position":3,"is_corresponding":false},{"id":567066,"name":"Hua Huang","orcid":"0000-0002-8936-3371","position":4,"is_corresponding":false},{"id":260597,"name":"Elizabeth Freilich","orcid":"0009-0009-6866-6398","position":5,"is_corresponding":false},{"id":915567,"name":"Megan L. Clark","orcid":"0000-0002-2631-3721","position":6,"is_corresponding":false},{"id":1021015,"name":"Joanna H. Lee","orcid":"0000-0002-7032-9261","position":7,"is_corresponding":false},{"id":52789,"name":"Zeyu Chen","orcid":"0000-0001-8534-0829","position":8,"is_corresponding":false},{"id":567064,"name":"Omar Khan","orcid":"0000-0002-7641-6163","position":9,"is_corresponding":false},{"id":632216,"name":"Ryan P. Staupe","orcid":"0000-0003-0793-524X","position":10,"is_corresponding":false},{"id":550295,"name":"Yinghui Huang","orcid":"0000-0003-0974-9539","position":11,"is_corresponding":false},{"id":260600,"name":"Junwei Shi","orcid":"0000-0002-8427-6316","position":12,"is_corresponding":false},{"id":105447,"name":"Josephine R. Giles","orcid":"0000-0002-3713-338X","position":13,"is_corresponding":false},{"id":36451,"name":"E. John Wherry","orcid":"0000-0003-0477-1956","position":14,"is_corresponding":false},{"id":105451,"name":"Jennifer E. Wu","orcid":"0000-0002-9302-8904","position":0,"is_corresponding":true}],"reference_count":81,"raw_metadata":null,"created_at":"2026-07-19T01:06:44.698773Z","pmid":"37595022","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":[]}