{"gene":"CARS1","run_date":"2026-06-09T22:57:17","timeline":{"discoveries":[{"year":2015,"finding":"Loss of CARS (cysteinyl-tRNA synthetase) suppresses ferroptosis induced by cystine deprivation (erastin treatment). Knockdown of CARS prevented induction of lipid reactive oxygen species without altering iron homeostasis, and led to accumulation of cystathionine and upregulation of genes associated with serine biosynthesis and transsulfuration. Inhibition of the transsulfuration pathway resensitized CARS-knockdown cells to erastin, placing CARS upstream of the transsulfuration pathway in the ferroptosis resistance mechanism.","method":"Genome-wide siRNA screen, siRNA knockdown, cell death assays, lipid ROS measurement, iron homeostasis assays, metabolite profiling (cystathionine accumulation), gene expression analysis, pharmacological inhibition of transsulfuration pathway","journal":"Cell death and differentiation","confidence":"High","confidence_rationale":"Tier 2 / Strong — genome-wide unbiased screen confirmed by targeted knockdown, multiple orthogonal methods (ROS measurement, metabolomics, gene expression, rescue experiments), single lab but rigorous mechanistic follow-up with pathway epistasis established","pmids":["26184909"],"is_preprint":false},{"year":2021,"finding":"CARS (cysteinyl-tRNA synthetase) acts as a cysteine sensor that, under cysteine-deficient conditions, binds to AMPKγ2 and recruits CaMKK2 to activate AMPK. Cysteine itself inhibits the CARS–AMPKγ2 interaction; when cysteine is absent, this inhibition is relieved, allowing CARS to mediate binding of AMPK to CaMKK2, resulting in CaMKK2-dependent phosphorylation and activation of AMPK. Blocking AMPK activation under cysteine-deficient conditions led to cell death, establishing this cysteine-CARS-CaMKK2-AMPKγ2 axis as a cell survival mechanism under nutrient deprivation.","method":"Co-immunoprecipitation (CARS with AMPKγ2 and CaMKK2), direct binding assays, cysteine supplementation/deprivation experiments, AMPK phosphorylation assays, siRNA/genetic loss-of-function, cell viability assays","journal":"The EMBO journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal co-immunoprecipitation and binding assays with functional rescue, single lab, multiple orthogonal methods but no in vitro reconstitution or structural validation","pmids":["34472622"],"is_preprint":false}],"current_model":"CARS1 (cysteinyl-tRNA synthetase) functions beyond aminoacylation as a cysteine sensor: under cysteine deprivation it binds AMPKγ2 and recruits CaMKK2 to activate AMPK for cell survival, and its loss induces the transsulfuration pathway to suppress ferroptosis by preventing lipid ROS accumulation downstream of cystine deprivation."},"narrative":{"mechanistic_narrative":"CARS1 (cysteinyl-tRNA synthetase) operates beyond canonical aminoacylation as a cysteine-responsive node coupling cysteine availability to cell survival and ferroptotic death [PMID:26184909, PMID:34472622]. Under cysteine deprivation, CARS1 acts as a cysteine sensor: cysteine normally inhibits its interaction with AMPKγ2, and when cysteine is absent this inhibition is relieved, allowing CARS1 to bind AMPKγ2 and recruit CaMKK2, driving CaMKK2-dependent phosphorylation and activation of AMPK to sustain cell survival; blocking this axis under cysteine-deficient conditions causes cell death [PMID:34472622]. Independently, loss of CARS1 confers resistance to ferroptosis triggered by cystine deprivation, acting upstream of the transsulfuration pathway — CARS1 knockdown induces cystathionine accumulation and serine biosynthesis/transsulfuration genes and prevents lipid ROS accumulation without altering iron homeostasis, and inhibiting transsulfuration restores ferroptosis sensitivity [PMID:26184909]. No structural model or in vitro reconstitution of these activities has been characterized in the available corpus.","teleology":[{"year":2015,"claim":"Whether CARS1 contributes to cell death regulation was unknown; this work established it as a determinant of ferroptosis sensitivity acting upstream of the transsulfuration pathway.","evidence":"Genome-wide siRNA screen with targeted knockdown, lipid ROS and iron assays, metabolite profiling, and pharmacological transsulfuration inhibition in erastin-treated cells","pmids":["26184909"],"confidence":"High","gaps":["Molecular mechanism by which CARS1 loss induces transsulfuration genes not defined","Whether the effect depends on aminoacylation activity not tested","No structural or biochemical characterization of the regulatory step"]},{"year":2021,"claim":"How cysteine availability is sensed to control survival signaling was unclear; this work showed CARS1 is a cysteine sensor that, when cysteine is absent, scaffolds AMPKγ2 and CaMKK2 to activate AMPK.","evidence":"Reciprocal co-immunoprecipitation and binding assays, cysteine supplementation/deprivation, AMPK phosphorylation assays, and loss-of-function viability assays","pmids":["34472622"],"confidence":"Medium","gaps":["No in vitro reconstitution or structural validation of the cysteine-regulated CARS1–AMPKγ2 interaction","Direct cysteine-binding site on CARS1 not mapped","Relationship between the AMPK survival axis and the ferroptosis/transsulfuration role not integrated"]},{"year":null,"claim":"How CARS1's aminoacylation function relates mechanistically to its cysteine-sensing and ferroptosis-regulatory roles remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["Whether tRNA charging activity is required for either non-canonical function is untested","No structural basis for cysteine sensing","Connection between AMPK survival signaling and transsulfuration-dependent ferroptosis resistance unestablished"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140299","term_label":"molecular sensor activity","supporting_discovery_ids":[1]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[1]}],"localization":[],"pathway":[{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[0]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[1]}],"complexes":[],"partners":["PRKAG2","CAMKK2"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"P49589","full_name":"Cysteine--tRNA ligase, cytoplasmic","aliases":["Cysteinyl-tRNA synthetase","CysRS"],"length_aa":748,"mass_kda":85.5,"function":"Catalyzes the ATP-dependent ligation of cysteine to tRNA(Cys)","subcellular_location":"Cytoplasm","url":"https://www.uniprot.org/uniprotkb/P49589/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/CARS1","classification":"Common Essential","n_dependent_lines":1206,"n_total_lines":1208,"dependency_fraction":0.9983443708609272},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"EEF1G","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/CARS1","total_profiled":1310},"omim":[{"mim_id":"618891","title":"MICROCEPHALY, DEVELOPMENTAL DELAY, AND BRITTLE HAIR SYNDROME; MDBH","url":"https://www.omim.org/entry/618891"},{"mim_id":"123859","title":"CYSTEINYL-tRNA SYNTHETASE 1; CARS1","url":"https://www.omim.org/entry/123859"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Enhanced","locations":[{"location":"Cytosol","reliability":"Enhanced"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/CARS1"},"hgnc":{"alias_symbol":[],"prev_symbol":["CARS"]},"alphafold":{"accession":"P49589","domains":[{"cath_id":"3.40.50.620","chopping":"29-99_228-419","consensus_level":"high","plddt":92.5579,"start":29,"end":419},{"cath_id":"-","chopping":"105-212","consensus_level":"medium","plddt":90.4581,"start":105,"end":212},{"cath_id":"-","chopping":"580-642","consensus_level":"medium","plddt":90.133,"start":580,"end":642},{"cath_id":"-","chopping":"685-748","consensus_level":"medium","plddt":82.2514,"start":685,"end":748},{"cath_id":"1.10.730","chopping":"445-557","consensus_level":"high","plddt":93.8257,"start":445,"end":557}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P49589","model_url":"https://alphafold.ebi.ac.uk/files/AF-P49589-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P49589-F1-predicted_aligned_error_v6.png","plddt_mean":89.62},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=CARS1","jax_strain_url":"https://www.jax.org/strain/search?query=CARS1"},"sequence":{"accession":"P49589","fasta_url":"https://rest.uniprot.org/uniprotkb/P49589.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P49589/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P49589"}},"corpus_meta":[{"pmid":"25985798","id":"PMC_25985798","title":"TRUCKs: the fourth generation of CARs.","date":"2015","source":"Expert opinion on biological therapy","url":"https://pubmed.ncbi.nlm.nih.gov/25985798","citation_count":575,"is_preprint":false},{"pmid":"24578504","id":"PMC_24578504","title":"Antibody-modified T cells: CARs take the front seat for hematologic malignancies.","date":"2014","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/24578504","citation_count":501,"is_preprint":false},{"pmid":"26503962","id":"PMC_26503962","title":"Mesothelin-Targeted CARs: Driving T Cells to Solid Tumors.","date":"2015","source":"Cancer discovery","url":"https://pubmed.ncbi.nlm.nih.gov/26503962","citation_count":408,"is_preprint":false},{"pmid":"26184909","id":"PMC_26184909","title":"Loss of cysteinyl-tRNA synthetase (CARS) induces the transsulfuration pathway and inhibits ferroptosis induced by cystine deprivation.","date":"2015","source":"Cell death and differentiation","url":"https://pubmed.ncbi.nlm.nih.gov/26184909","citation_count":398,"is_preprint":false},{"pmid":"24329791","id":"PMC_24329791","title":"Of CARs and TRUCKs: chimeric antigen receptor (CAR) T cells engineered with an inducible cytokine to modulate the tumor stroma.","date":"2014","source":"Immunological reviews","url":"https://pubmed.ncbi.nlm.nih.gov/24329791","citation_count":300,"is_preprint":false},{"pmid":"30482221","id":"PMC_30482221","title":"Universal CARs, universal T cells, and universal CAR T cells.","date":"2018","source":"Journal of hematology & oncology","url":"https://pubmed.ncbi.nlm.nih.gov/30482221","citation_count":244,"is_preprint":false},{"pmid":"30012854","id":"PMC_30012854","title":"CARs versus BiTEs: A Comparison between T Cell-Redirection Strategies for Cancer Treatment.","date":"2018","source":"Cancer discovery","url":"https://pubmed.ncbi.nlm.nih.gov/30012854","citation_count":188,"is_preprint":false},{"pmid":"29288199","id":"PMC_29288199","title":"Comparison of T Cell Activities Mediated by Human TCRs and CARs That Use the Same Recognition Domains.","date":"2017","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/29288199","citation_count":159,"is_preprint":false},{"pmid":"36175679","id":"PMC_36175679","title":"KIR-based inhibitory CARs overcome CAR-NK cell trogocytosis-mediated fratricide and tumor escape.","date":"2022","source":"Nature medicine","url":"https://pubmed.ncbi.nlm.nih.gov/36175679","citation_count":150,"is_preprint":false},{"pmid":"26738472","id":"PMC_26738472","title":"Therapeutic Potential of T Cell Chimeric Antigen Receptors (CARs) in Cancer Treatment: Counteracting Off-Tumor Toxicities for Safe CAR T Cell Therapy.","date":"2016","source":"Annual review of pharmacology and toxicology","url":"https://pubmed.ncbi.nlm.nih.gov/26738472","citation_count":122,"is_preprint":false},{"pmid":"29554494","id":"PMC_29554494","title":"Driving CARs on the uneven road of antigen heterogeneity in solid tumors.","date":"2018","source":"Current opinion in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/29554494","citation_count":115,"is_preprint":false},{"pmid":"32451454","id":"PMC_32451454","title":"Signaling from T cell receptors (TCRs) and chimeric antigen receptors (CARs) on T cells.","date":"2020","source":"Cellular & molecular immunology","url":"https://pubmed.ncbi.nlm.nih.gov/32451454","citation_count":113,"is_preprint":false},{"pmid":"20810169","id":"PMC_20810169","title":"Perfluoroalkyl compounds in dust from Asian, Australian, European, and North American homes and UK cars, classrooms, and offices.","date":"2011","source":"Environment international","url":"https://pubmed.ncbi.nlm.nih.gov/20810169","citation_count":106,"is_preprint":false},{"pmid":"29385713","id":"PMC_29385713","title":"Nanobody Based Dual Specific CARs.","date":"2018","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/29385713","citation_count":104,"is_preprint":false},{"pmid":"12093923","id":"PMC_12093923","title":"Polyvalent cation receptor proteins (CaRs) are salinity sensors in fish.","date":"2002","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/12093923","citation_count":99,"is_preprint":false},{"pmid":"29127833","id":"PMC_29127833","title":"Carboxylic acid reductase enzymes (CARs).","date":"2017","source":"Current opinion in chemical biology","url":"https://pubmed.ncbi.nlm.nih.gov/29127833","citation_count":94,"is_preprint":false},{"pmid":"24667963","id":"PMC_24667963","title":"NKG2D CARs as cell therapy for cancer.","date":"2014","source":"Cancer journal (Sudbury, Mass.)","url":"https://pubmed.ncbi.nlm.nih.gov/24667963","citation_count":89,"is_preprint":false},{"pmid":"34732890","id":"PMC_34732890","title":"Cross-HLA targeting of intracellular oncoproteins with peptide-centric CARs.","date":"2021","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/34732890","citation_count":88,"is_preprint":false},{"pmid":"23162754","id":"PMC_23162754","title":"A sensitivity scale for targeting T cells with chimeric antigen receptors (CARs) and bispecific T-cell Engagers (BiTEs).","date":"2012","source":"Oncoimmunology","url":"https://pubmed.ncbi.nlm.nih.gov/23162754","citation_count":88,"is_preprint":false},{"pmid":"31272471","id":"PMC_31272471","title":"Engineering switchable and programmable universal CARs for CAR T therapy.","date":"2019","source":"Journal of hematology & oncology","url":"https://pubmed.ncbi.nlm.nih.gov/31272471","citation_count":83,"is_preprint":false},{"pmid":"28416139","id":"PMC_28416139","title":"CARs: Synthetic Immunoreceptors for Cancer Therapy and Beyond.","date":"2017","source":"Trends in molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/28416139","citation_count":82,"is_preprint":false},{"pmid":"24667958","id":"PMC_24667958","title":"CD28z CARs and armored CARs.","date":"2014","source":"Cancer journal (Sudbury, Mass.)","url":"https://pubmed.ncbi.nlm.nih.gov/24667958","citation_count":73,"is_preprint":false},{"pmid":"26963133","id":"PMC_26963133","title":"Bispecific antibodies and CARs: generalized immunotherapeutics harnessing T cell redirection.","date":"2016","source":"Current opinion in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/26963133","citation_count":73,"is_preprint":false},{"pmid":"22319014","id":"PMC_22319014","title":"Raman spectroscopy and CARS microscopy of stem cells and their derivatives.","date":"2011","source":"Journal of Raman spectroscopy : JRS","url":"https://pubmed.ncbi.nlm.nih.gov/22319014","citation_count":70,"is_preprint":false},{"pmid":"28322974","id":"PMC_28322974","title":"Current modalities in cancer immunotherapy: Immunomodulatory antibodies, CARs and vaccines.","date":"2017","source":"Pharmacology & therapeutics","url":"https://pubmed.ncbi.nlm.nih.gov/28322974","citation_count":69,"is_preprint":false},{"pmid":"24576891","id":"PMC_24576891","title":"Shedding new light on lipid functions with CARS and SRS microscopy.","date":"2014","source":"Biochimica et biophysica acta","url":"https://pubmed.ncbi.nlm.nih.gov/24576891","citation_count":67,"is_preprint":false},{"pmid":"29880908","id":"PMC_29880908","title":"Driving cars to the clinic for solid tumors.","date":"2018","source":"Gene therapy","url":"https://pubmed.ncbi.nlm.nih.gov/29880908","citation_count":64,"is_preprint":false},{"pmid":"27151947","id":"PMC_27151947","title":"Quantitative imaging of lipids in live mouse oocytes and early embryos using CARS microscopy.","date":"2016","source":"Development (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/27151947","citation_count":64,"is_preprint":false},{"pmid":"37938771","id":"PMC_37938771","title":"Targeting of intracellular oncoproteins with peptide-centric CARs.","date":"2023","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/37938771","citation_count":60,"is_preprint":false},{"pmid":"26352543","id":"PMC_26352543","title":"Smart CARs engineered for cancer immunotherapy.","date":"2015","source":"Current opinion in oncology","url":"https://pubmed.ncbi.nlm.nih.gov/26352543","citation_count":58,"is_preprint":false},{"pmid":"32013131","id":"PMC_32013131","title":"Targeting CD38-Expressing Multiple Myeloma and Burkitt Lymphoma Cells In Vitro with Nanobody-Based Chimeric Antigen Receptors (Nb-CARs).","date":"2020","source":"Cells","url":"https://pubmed.ncbi.nlm.nih.gov/32013131","citation_count":57,"is_preprint":false},{"pmid":"36732477","id":"PMC_36732477","title":"Synapse-tuned CARs enhance immune cell anti-tumor activity.","date":"2023","source":"Nature biotechnology","url":"https://pubmed.ncbi.nlm.nih.gov/36732477","citation_count":55,"is_preprint":false},{"pmid":"34274536","id":"PMC_34274536","title":"The rational development of CD5-targeting biepitopic CARs with fully human heavy-chain-only antigen recognition domains.","date":"2021","source":"Molecular therapy : the journal of the American Society of Gene Therapy","url":"https://pubmed.ncbi.nlm.nih.gov/34274536","citation_count":55,"is_preprint":false},{"pmid":"28977984","id":"PMC_28977984","title":"Driving better and safer HER2-specific CARs for cancer therapy.","date":"2017","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/28977984","citation_count":52,"is_preprint":false},{"pmid":"29861327","id":"PMC_29861327","title":"PiggyBac-Engineered T Cells Expressing CD19-Specific CARs that Lack IgG1 Fc Spacers Have Potent Activity against B-ALL Xenografts.","date":"2018","source":"Molecular therapy : the journal of the American Society of Gene Therapy","url":"https://pubmed.ncbi.nlm.nih.gov/29861327","citation_count":51,"is_preprint":false},{"pmid":"27322989","id":"PMC_27322989","title":"TriKEs and BiKEs join CARs on the cancer immunotherapy highway.","date":"2016","source":"Human vaccines & immunotherapeutics","url":"https://pubmed.ncbi.nlm.nih.gov/27322989","citation_count":50,"is_preprint":false},{"pmid":"26515978","id":"PMC_26515978","title":"Stability and activity of MCSP-specific chimeric antigen receptors (CARs) depend on the scFv antigen-binding domain and the protein backbone.","date":"2015","source":"Cancer immunology, immunotherapy : CII","url":"https://pubmed.ncbi.nlm.nih.gov/26515978","citation_count":50,"is_preprint":false},{"pmid":"33654227","id":"PMC_33654227","title":"TCR-like CARs and TCR-CARs targeting neoepitopes: an emerging potential.","date":"2021","source":"Cancer gene therapy","url":"https://pubmed.ncbi.nlm.nih.gov/33654227","citation_count":49,"is_preprint":false},{"pmid":"30827461","id":"PMC_30827461","title":"T Cell-Redirecting Strategies to 'STAb' Tumors: Beyond CARs and Bispecific Antibodies.","date":"2019","source":"Trends in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/30827461","citation_count":49,"is_preprint":false},{"pmid":"32997278","id":"PMC_32997278","title":"Programmable and multi-targeted CARs: a new breakthrough in cancer CAR-T cell therapy.","date":"2020","source":"Clinical & translational oncology : official publication of the Federation of Spanish Oncology Societies and of the National Cancer Institute of Mexico","url":"https://pubmed.ncbi.nlm.nih.gov/32997278","citation_count":49,"is_preprint":false},{"pmid":"34028069","id":"PMC_34028069","title":"The antigen-binding moiety in the driver's seat of CARs.","date":"2021","source":"Medicinal research reviews","url":"https://pubmed.ncbi.nlm.nih.gov/34028069","citation_count":48,"is_preprint":false},{"pmid":"31542798","id":"PMC_31542798","title":"Conventional CARs versus modular CARs.","date":"2019","source":"Cancer immunology, immunotherapy : CII","url":"https://pubmed.ncbi.nlm.nih.gov/31542798","citation_count":43,"is_preprint":false},{"pmid":"28688236","id":"PMC_28688236","title":"Antigen-specific regulatory T cells: are police CARs the answer?","date":"2017","source":"Translational research : the journal of laboratory and clinical medicine","url":"https://pubmed.ncbi.nlm.nih.gov/28688236","citation_count":41,"is_preprint":false},{"pmid":"36922828","id":"PMC_36922828","title":"Tuning CARs: recent advances in modulating chimeric antigen receptor (CAR) T cell activity for improved safety, efficacy, and flexibility.","date":"2023","source":"Journal of translational medicine","url":"https://pubmed.ncbi.nlm.nih.gov/36922828","citation_count":39,"is_preprint":false},{"pmid":"38493479","id":"PMC_38493479","title":"Integration of ζ-deficient CARs into the CD3ζ gene conveys potent cytotoxicity in T and NK cells.","date":"2024","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/38493479","citation_count":37,"is_preprint":false},{"pmid":"31004624","id":"PMC_31004624","title":"CAR T-cell bioengineering: Single variable domain of heavy chain antibody targeted CARs.","date":"2019","source":"Advanced drug delivery reviews","url":"https://pubmed.ncbi.nlm.nih.gov/31004624","citation_count":37,"is_preprint":false},{"pmid":"32541028","id":"PMC_32541028","title":"Structure-guided engineering of the affinity and specificity of CARs against Tn-glycopeptides.","date":"2020","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/32541028","citation_count":37,"is_preprint":false},{"pmid":"38987856","id":"PMC_38987856","title":"Fueling CARs: metabolic strategies to enhance CAR T-cell therapy.","date":"2024","source":"Experimental hematology & oncology","url":"https://pubmed.ncbi.nlm.nih.gov/38987856","citation_count":35,"is_preprint":false},{"pmid":"33789222","id":"PMC_33789222","title":"Remote controlling of CAR-T cells and toxicity management: Molecular switches and next generation CARs.","date":"2021","source":"Translational oncology","url":"https://pubmed.ncbi.nlm.nih.gov/33789222","citation_count":35,"is_preprint":false},{"pmid":"23546091","id":"PMC_23546091","title":"Simultaneous hyperspectral differential-CARS, TPF and SHG microscopy with a single 5 fs Ti:Sa laser.","date":"2013","source":"Optics express","url":"https://pubmed.ncbi.nlm.nih.gov/23546091","citation_count":35,"is_preprint":false},{"pmid":"29856949","id":"PMC_29856949","title":"Increasing T Cell Versatility with SUPRA CARs.","date":"2018","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/29856949","citation_count":34,"is_preprint":false},{"pmid":"26110321","id":"PMC_26110321","title":"Seatbelts in CAR therapy: How Safe Are CARS?","date":"2015","source":"Pharmaceuticals (Basel, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/26110321","citation_count":34,"is_preprint":false},{"pmid":"32019116","id":"PMC_32019116","title":"Rapid and Effective Generation of Nanobody Based CARs using PCR and Gibson Assembly.","date":"2020","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/32019116","citation_count":33,"is_preprint":false},{"pmid":"28233151","id":"PMC_28233151","title":"CARs in the Lead Against Multiple Myeloma.","date":"2017","source":"Current hematologic malignancy reports","url":"https://pubmed.ncbi.nlm.nih.gov/28233151","citation_count":32,"is_preprint":false},{"pmid":"32794303","id":"PMC_32794303","title":"Driving CARs with alternative navigation tools - the potential of engineered binding scaffolds.","date":"2020","source":"The FEBS journal","url":"https://pubmed.ncbi.nlm.nih.gov/32794303","citation_count":32,"is_preprint":false},{"pmid":"26065655","id":"PMC_26065655","title":"A drive through cellular therapy for CLL in 2015: allogeneic cell transplantation and CARs.","date":"2015","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/26065655","citation_count":32,"is_preprint":false},{"pmid":"34462325","id":"PMC_34462325","title":"CARs and beyond: tailoring macrophage-based cell therapeutics to combat solid malignancies.","date":"2021","source":"Journal for immunotherapy of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/34462325","citation_count":31,"is_preprint":false},{"pmid":"20431282","id":"PMC_20431282","title":"Genetic variation in innate immunity pathways and their potential contribution to the SIRS/CARS debate: evidence from human studies and animal models.","date":"2010","source":"Journal of innate immunity","url":"https://pubmed.ncbi.nlm.nih.gov/20431282","citation_count":30,"is_preprint":false},{"pmid":"24877002","id":"PMC_24877002","title":"Hyperspectral and differential CARS microscopy for quantitative chemical imaging in human adipocytes.","date":"2014","source":"Biomedical optics express","url":"https://pubmed.ncbi.nlm.nih.gov/24877002","citation_count":30,"is_preprint":false},{"pmid":"28810803","id":"PMC_28810803","title":"Driving CARs on the Highway to Solid Cancer: Some Considerations on the Adoptive Therapy with CAR T Cells.","date":"2017","source":"Human gene therapy","url":"https://pubmed.ncbi.nlm.nih.gov/28810803","citation_count":30,"is_preprint":false},{"pmid":"32429316","id":"PMC_32429316","title":"CARs: Beyond T Cells and T Cell-Derived Signaling Domains.","date":"2020","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/32429316","citation_count":29,"is_preprint":false},{"pmid":"35821636","id":"PMC_35821636","title":"Targeting glycans for CAR therapy: The advent of sweet CARs.","date":"2022","source":"Molecular therapy : the journal of the American Society of Gene Therapy","url":"https://pubmed.ncbi.nlm.nih.gov/35821636","citation_count":29,"is_preprint":false},{"pmid":"31430498","id":"PMC_31430498","title":"Carboxylic acid reductases (CARs): An industrial perspective.","date":"2019","source":"Journal of biotechnology","url":"https://pubmed.ncbi.nlm.nih.gov/31430498","citation_count":29,"is_preprint":false},{"pmid":"29909918","id":"PMC_29909918","title":"Advances in the use of natural receptor- or ligand-based chimeric antigen receptors (CARs) in haematologic malignancies.","date":"2018","source":"Best practice & research. Clinical haematology","url":"https://pubmed.ncbi.nlm.nih.gov/29909918","citation_count":29,"is_preprint":false},{"pmid":"36578789","id":"PMC_36578789","title":"Targeting TGF-β signaling in the multiple myeloma microenvironment: Steering CARs and T cells in the right direction.","date":"2022","source":"Frontiers in cell and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/36578789","citation_count":27,"is_preprint":false},{"pmid":"35618140","id":"PMC_35618140","title":"Advances in modular control of CAR-T therapy with adapter-mediated CARs.","date":"2022","source":"Advanced drug delivery reviews","url":"https://pubmed.ncbi.nlm.nih.gov/35618140","citation_count":25,"is_preprint":false},{"pmid":"32636832","id":"PMC_32636832","title":"Engineering Cytoplasmic Signaling of CD28ζ CARs for Improved Therapeutic Functions.","date":"2020","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/32636832","citation_count":25,"is_preprint":false},{"pmid":"31706506","id":"PMC_31706506","title":"Paving New Roads for CARs.","date":"2019","source":"Trends in cancer","url":"https://pubmed.ncbi.nlm.nih.gov/31706506","citation_count":24,"is_preprint":false},{"pmid":"36346550","id":"PMC_36346550","title":"CARs: a new approach for the treatment of autoimmune diseases.","date":"2022","source":"Science China. Life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/36346550","citation_count":22,"is_preprint":false},{"pmid":"32421455","id":"PMC_32421455","title":"ADCs, BiTEs, CARs, and Small Molecules: A New Era of Targeted Therapy in Non-Hodgkin Lymphoma.","date":"2020","source":"American Society of Clinical Oncology educational book. American Society of Clinical Oncology. Annual Meeting","url":"https://pubmed.ncbi.nlm.nih.gov/32421455","citation_count":22,"is_preprint":false},{"pmid":"35989138","id":"PMC_35989138","title":"Targeting CD19 for diffuse large B cell lymphoma in the era of CARs: Other modes of transportation.","date":"2022","source":"Blood reviews","url":"https://pubmed.ncbi.nlm.nih.gov/35989138","citation_count":22,"is_preprint":false},{"pmid":"29909915","id":"PMC_29909915","title":"CARs and other T cell therapies for MM: The clinical experience.","date":"2018","source":"Best practice & research. Clinical haematology","url":"https://pubmed.ncbi.nlm.nih.gov/29909915","citation_count":21,"is_preprint":false},{"pmid":"37096170","id":"PMC_37096170","title":"Determination of corn protein content using near-infrared spectroscopy combined with A-CARS-PLS.","date":"2023","source":"Food chemistry: X","url":"https://pubmed.ncbi.nlm.nih.gov/37096170","citation_count":21,"is_preprint":false},{"pmid":"35151842","id":"PMC_35151842","title":"Multipurposing CARs: Same engine, different vehicles.","date":"2022","source":"Molecular therapy : the journal of the American Society of Gene Therapy","url":"https://pubmed.ncbi.nlm.nih.gov/35151842","citation_count":20,"is_preprint":false},{"pmid":"34472622","id":"PMC_34472622","title":"CARS senses cysteine deprivation to activate AMPK for cell survival.","date":"2021","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/34472622","citation_count":20,"is_preprint":false},{"pmid":"34432728","id":"PMC_34432728","title":"Potent, Selective CARs as Potential T-Cell Therapeutics for HPV-positive Cancers.","date":"2021","source":"Journal of immunotherapy (Hagerstown, Md. : 1997)","url":"https://pubmed.ncbi.nlm.nih.gov/34432728","citation_count":20,"is_preprint":false},{"pmid":"32079207","id":"PMC_32079207","title":"Harnessing T Cells to Target Pediatric Acute Myeloid Leukemia: CARs, BiTEs, and Beyond.","date":"2020","source":"Children (Basel, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/32079207","citation_count":20,"is_preprint":false},{"pmid":"31754147","id":"PMC_31754147","title":"Single variable domains from the T cell receptor β chain function as mono- and bifunctional CARs and TCRs.","date":"2019","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/31754147","citation_count":20,"is_preprint":false},{"pmid":"32630096","id":"PMC_32630096","title":"T-Cell Gene Therapy in Cancer Immunotherapy: Why It Is No Longer Just CARs on The Road.","date":"2020","source":"Cells","url":"https://pubmed.ncbi.nlm.nih.gov/32630096","citation_count":19,"is_preprint":false},{"pmid":"15310893","id":"PMC_15310893","title":"Hybrid cars now, fuel cell cars later.","date":"2004","source":"Science (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/15310893","citation_count":19,"is_preprint":false},{"pmid":"38384128","id":"PMC_38384128","title":"Inhibitory CARs fail to protect from immediate T cell cytotoxicity.","date":"2024","source":"Molecular therapy : the journal of the American Society of Gene Therapy","url":"https://pubmed.ncbi.nlm.nih.gov/38384128","citation_count":18,"is_preprint":false},{"pmid":"29603617","id":"PMC_29603617","title":"Engineering therapeutic T cells to suppress alloimmune responses using TCRs, CARs, or BARs.","date":"2018","source":"American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons","url":"https://pubmed.ncbi.nlm.nih.gov/29603617","citation_count":18,"is_preprint":false},{"pmid":"23543145","id":"PMC_23543145","title":"Functional analysis of the carS gene of Fusarium fujikuroi.","date":"2013","source":"Molecular genetics and genomics : MGG","url":"https://pubmed.ncbi.nlm.nih.gov/23543145","citation_count":18,"is_preprint":false},{"pmid":"22131062","id":"PMC_22131062","title":"Muscle CARs and TcRs: turbo-charged technologies for the (T cell) masses.","date":"2011","source":"Cancer immunology, immunotherapy : CII","url":"https://pubmed.ncbi.nlm.nih.gov/22131062","citation_count":17,"is_preprint":false},{"pmid":"36430728","id":"PMC_36430728","title":"Learning from TCR Signaling and Immunological Synapse Assembly to Build New Chimeric Antigen Receptors (CARs).","date":"2022","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/36430728","citation_count":17,"is_preprint":false},{"pmid":"30665350","id":"PMC_30665350","title":"Comparative transcriptomic analysis unveils interactions between the regulatory CarS protein and light response in Fusarium.","date":"2019","source":"BMC genomics","url":"https://pubmed.ncbi.nlm.nih.gov/30665350","citation_count":17,"is_preprint":false},{"pmid":"32906767","id":"PMC_32906767","title":"Eosinophils and Neutrophils-Molecular Differences Revealed by Spontaneous Raman, CARS and Fluorescence Microscopy.","date":"2020","source":"Cells","url":"https://pubmed.ncbi.nlm.nih.gov/32906767","citation_count":16,"is_preprint":false},{"pmid":"34035459","id":"PMC_34035459","title":"Novel immune engagers and cellular therapies for metastatic castration-resistant prostate cancer: do we take a BiTe or ride BiKEs, TriKEs, and CARs?","date":"2021","source":"Prostate cancer and prostatic diseases","url":"https://pubmed.ncbi.nlm.nih.gov/34035459","citation_count":16,"is_preprint":false},{"pmid":"27618260","id":"PMC_27618260","title":"From therapeutic antibodies to chimeric antigen receptors (CARs): making better CARs based on antigen-binding domain.","date":"2016","source":"Expert opinion on biological therapy","url":"https://pubmed.ncbi.nlm.nih.gov/27618260","citation_count":16,"is_preprint":false},{"pmid":"35133146","id":"PMC_35133146","title":"Label-Free Live-Cell Imaging of Internalized Microplastics and Cytoplasmic Organelles with Multicolor CARS Microscopy.","date":"2022","source":"Environmental science & technology","url":"https://pubmed.ncbi.nlm.nih.gov/35133146","citation_count":15,"is_preprint":false},{"pmid":"38658805","id":"PMC_38658805","title":"Extracellular domains of CARs reprogramme T cell metabolism without antigen stimulation.","date":"2024","source":"Nature metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/38658805","citation_count":14,"is_preprint":false},{"pmid":"33768128","id":"PMC_33768128","title":"Anti-CD19 CARs displayed at the surface of lentiviral vector particles promote transduction of target-expressing cells.","date":"2021","source":"Molecular therapy. Methods & clinical development","url":"https://pubmed.ncbi.nlm.nih.gov/33768128","citation_count":14,"is_preprint":false},{"pmid":"26485445","id":"PMC_26485445","title":"Fast Cars and No Brakes: Autologous Stem Cell Transplantation as a Platform for Novel Immunotherapies.","date":"2015","source":"Biology of blood and marrow transplantation : journal of the American Society for Blood and Marrow Transplantation","url":"https://pubmed.ncbi.nlm.nih.gov/26485445","citation_count":13,"is_preprint":false},{"pmid":"36499331","id":"PMC_36499331","title":"Modern Advances in CARs Therapy and Creating a New Approach to Future Treatment.","date":"2022","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/36499331","citation_count":13,"is_preprint":false},{"pmid":"36226849","id":"PMC_36226849","title":"Do Treg Speed Up with CARs? Chimeric Antigen Receptor Treg Engineered to Induce Transplant Tolerance.","date":"2022","source":"Transplantation","url":"https://pubmed.ncbi.nlm.nih.gov/36226849","citation_count":13,"is_preprint":false},{"pmid":"25964863","id":"PMC_25964863","title":"Roadblocks to success for RNA CARs in solid tumors.","date":"2015","source":"Oncoimmunology","url":"https://pubmed.ncbi.nlm.nih.gov/25964863","citation_count":13,"is_preprint":false},{"pmid":"37460153","id":"PMC_37460153","title":"Localization of unlabeled bepirovirsen antisense oligonucleotide in murine tissues using in situ hybridization and CARS imaging.","date":"2023","source":"RNA (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/37460153","citation_count":13,"is_preprint":false},{"pmid":"31153590","id":"PMC_31153590","title":"Quantitative Mapping of Triacylglycerol Chain Length and Saturation Using Broadband CARS Microscopy.","date":"2019","source":"Biophysical journal","url":"https://pubmed.ncbi.nlm.nih.gov/31153590","citation_count":13,"is_preprint":false},{"pmid":"2129345","id":"PMC_2129345","title":"Carbohydrate and hydrophobic-carbohydrate recognition sites (CARS and HY-CARS) in solubilized glycosyltransferases.","date":"1990","source":"Indian journal of biochemistry & biophysics","url":"https://pubmed.ncbi.nlm.nih.gov/2129345","citation_count":13,"is_preprint":false},{"pmid":"38680567","id":"PMC_38680567","title":"Aptamer-Based Nongenetic Reprogramming of CARs Enables Flexible Modulation of T Cell-Mediated Tumor Immunotherapy.","date":"2024","source":"ACS central science","url":"https://pubmed.ncbi.nlm.nih.gov/38680567","citation_count":12,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":38990,"output_tokens":1034,"usd":0.06624,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":7298,"output_tokens":1471,"usd":0.036632,"stage2_stop_reason":"end_turn"},"total_usd":0.102872,"stage1_batch_id":"msgbatch_01ACAxJQabDsht39JXeWG22d","stage2_batch_id":"msgbatch_01QEU22A9U5sbPTgsMzTmvSf","note":"batch pricing = 50% of standard"},"stage1_raw":"Looking at the corpus, I need to identify papers about CARS1 (cysteinyl-tRNA synthetase, also known as CARS). The vast majority of papers are about CAR T cells (Chimeric Antigen Receptors), CARS microscopy (Coherent Anti-Stokes Raman Scattering), carboxylic acid reductases, or other unrelated topics. I need to find papers specifically about the CARS1/CARS protein (cysteinyl-tRNA synthetase).\n\nRelevant papers:\n- PMID:26184909: Loss of CARS (cysteinyl-tRNA synthetase) and ferroptosis\n- PMID:34472622: CARS senses cysteine deprivation to activate AMPK\n\nAll other papers are about CAR T cells, CARS microscopy, carboxylic acid reductases, or other unrelated topics and should be excluded.\n\n```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2015,\n      \"finding\": \"Loss of CARS (cysteinyl-tRNA synthetase) suppresses ferroptosis induced by cystine deprivation (erastin treatment). Knockdown of CARS prevented induction of lipid reactive oxygen species without altering iron homeostasis, and led to accumulation of cystathionine and upregulation of genes associated with serine biosynthesis and transsulfuration. Inhibition of the transsulfuration pathway resensitized CARS-knockdown cells to erastin, placing CARS upstream of the transsulfuration pathway in the ferroptosis resistance mechanism.\",\n      \"method\": \"Genome-wide siRNA screen, siRNA knockdown, cell death assays, lipid ROS measurement, iron homeostasis assays, metabolite profiling (cystathionine accumulation), gene expression analysis, pharmacological inhibition of transsulfuration pathway\",\n      \"journal\": \"Cell death and differentiation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genome-wide unbiased screen confirmed by targeted knockdown, multiple orthogonal methods (ROS measurement, metabolomics, gene expression, rescue experiments), single lab but rigorous mechanistic follow-up with pathway epistasis established\",\n      \"pmids\": [\"26184909\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"CARS (cysteinyl-tRNA synthetase) acts as a cysteine sensor that, under cysteine-deficient conditions, binds to AMPKγ2 and recruits CaMKK2 to activate AMPK. Cysteine itself inhibits the CARS–AMPKγ2 interaction; when cysteine is absent, this inhibition is relieved, allowing CARS to mediate binding of AMPK to CaMKK2, resulting in CaMKK2-dependent phosphorylation and activation of AMPK. Blocking AMPK activation under cysteine-deficient conditions led to cell death, establishing this cysteine-CARS-CaMKK2-AMPKγ2 axis as a cell survival mechanism under nutrient deprivation.\",\n      \"method\": \"Co-immunoprecipitation (CARS with AMPKγ2 and CaMKK2), direct binding assays, cysteine supplementation/deprivation experiments, AMPK phosphorylation assays, siRNA/genetic loss-of-function, cell viability assays\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal co-immunoprecipitation and binding assays with functional rescue, single lab, multiple orthogonal methods but no in vitro reconstitution or structural validation\",\n      \"pmids\": [\"34472622\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"CARS1 (cysteinyl-tRNA synthetase) functions beyond aminoacylation as a cysteine sensor: under cysteine deprivation it binds AMPKγ2 and recruits CaMKK2 to activate AMPK for cell survival, and its loss induces the transsulfuration pathway to suppress ferroptosis by preventing lipid ROS accumulation downstream of cystine deprivation.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"CARS1 (cysteinyl-tRNA synthetase) operates beyond canonical aminoacylation as a cysteine-responsive node coupling cysteine availability to cell survival and ferroptotic death [#0, #1]. Under cysteine deprivation, CARS1 acts as a cysteine sensor: cysteine normally inhibits its interaction with AMPKγ2, and when cysteine is absent this inhibition is relieved, allowing CARS1 to bind AMPKγ2 and recruit CaMKK2, driving CaMKK2-dependent phosphorylation and activation of AMPK to sustain cell survival; blocking this axis under cysteine-deficient conditions causes cell death [#1]. Independently, loss of CARS1 confers resistance to ferroptosis triggered by cystine deprivation, acting upstream of the transsulfuration pathway — CARS1 knockdown induces cystathionine accumulation and serine biosynthesis/transsulfuration genes and prevents lipid ROS accumulation without altering iron homeostasis, and inhibiting transsulfuration restores ferroptosis sensitivity [#0]. No structural model or in vitro reconstitution of these activities has been characterized in the available corpus.\",\n  \"teleology\": [\n    {\n      \"year\": 2015,\n      \"claim\": \"Whether CARS1 contributes to cell death regulation was unknown; this work established it as a determinant of ferroptosis sensitivity acting upstream of the transsulfuration pathway.\",\n      \"evidence\": \"Genome-wide siRNA screen with targeted knockdown, lipid ROS and iron assays, metabolite profiling, and pharmacological transsulfuration inhibition in erastin-treated cells\",\n      \"pmids\": [\"26184909\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Molecular mechanism by which CARS1 loss induces transsulfuration genes not defined\",\n        \"Whether the effect depends on aminoacylation activity not tested\",\n        \"No structural or biochemical characterization of the regulatory step\"\n      ]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"How cysteine availability is sensed to control survival signaling was unclear; this work showed CARS1 is a cysteine sensor that, when cysteine is absent, scaffolds AMPKγ2 and CaMKK2 to activate AMPK.\",\n      \"evidence\": \"Reciprocal co-immunoprecipitation and binding assays, cysteine supplementation/deprivation, AMPK phosphorylation assays, and loss-of-function viability assays\",\n      \"pmids\": [\"34472622\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"No in vitro reconstitution or structural validation of the cysteine-regulated CARS1\\u2013AMPKγ2 interaction\",\n        \"Direct cysteine-binding site on CARS1 not mapped\",\n        \"Relationship between the AMPK survival axis and the ferroptosis/transsulfuration role not integrated\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How CARS1's aminoacylation function relates mechanistically to its cysteine-sensing and ferroptosis-regulatory roles remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Whether tRNA charging activity is required for either non-canonical function is untested\",\n        \"No structural basis for cysteine sensing\",\n        \"Connection between AMPK survival signaling and transsulfuration-dependent ferroptosis resistance unestablished\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140299\", \"supporting_discovery_ids\": [1]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"localization\": [],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"PRKAG2\", \"CAMKK2\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":3,"faith_total":3,"faith_pct":100.0}}