Precision quality check of immunotherapeutics via single-cell cytokine mapping

Adoptive transfer of autologous T cells engineered to express chimeric antigen receptorsCARshas emerged as a promising immunotherapy for patients with hematologic malignanciessuch as CDCARs in leukemias and lymphomasHoweverchallenges remain in terms of manufacturing consistency and the functional profile of the CAR T cell productsince infused cellsasliving drugscan be activated and release a variety of cytokines upon specific antigen recognitionThese secreted cytokines may result in not only therapeutic efficacybut also life threatening immunotoxicitysuch as cytokine release syndrome and neurologic toxicityIt is essential for cancer patients to have a full spectrum potency and toxicity profiling of CAR T cell products before infusionWe have developed a single cell barcode chipSCBCtechnologywhich enablesaassayingsecreted proteins per individual live cellba broad range of immune cell functions covering efficacy and safetycis designed to fit various types of immune cellsdis the first quantitative metric of these proteins per cellanderequires samples sizes totaling onlycellsall critical leaps over flow cytometry based platformsWith the collaboration of two leading CAR T pharmaceutical companiesIsoPlexissingle cell deep profiling has revealedfor the first timea pre infusion correlate to post infusion patient responseThe SCBC analysis ofpatients with lymphoma demonstrates a significant correlation of the polyfunctional strength of the CAR T cells with objective responsecomplete or partialto the CAR T cell therapypindicating a powerful metric of quality assessmentwhere the other gold standard technologies did not detect any significant correlationsBased on the clinical data and the automation progress in our SBIR Phase I grantwe are proposing the following specific aims in this two year Phase II applicationAIMDevelop automatedflow cellconsumable compartmentwhich capturesplex single cell cytokine responseto prepare for full automation required by large CAR T trialmonthsAIMProduce a fully automated cartridge analysis and workflow systemto allowsamples assayed in parallel with minimal user interactionallowing easy introduction into clinical core labsmonthsAIMProduce a comprehensive informatics suiteand fully test our automated system and informatics in twopatient cohort trialsproviding robust and predictive biomarkers for pre infusion CAR T qualityEstablish MSKCC and UCLA beta sites for post phase II transitionmonthsWith this Phase II submissionwe will deliver the first effective CAR T pre infusion quality check assay to predict objective response in patientsin an easy to useautomated system that can be used throughout all cellular immunotherapy trials