A single-cell device to capture and analyze rare T-cells from solid tumors.

Checkpoint inhibitors towards targets such as PDand PD Lare being developed to activate quiescent T cells embedded in tumors and enable them to recruit the rest of the immune system to fight a cancer massHow these new cures reactivate the relatively few Tumor Infiltrating LymphocytesTILsenabling the release of a complex message of cytokines to recruit the immune systemthus impacting treatment efficacyDetecting these T cells and their cytokines provide critically needed predictive biomarkers for defining responders versus nonrespondersIsoPlexis technology provides both the ability to rapidly isolate TILs from tumor samples and to detect their full functional cytokine excretion response to treatmentIn Phase I of our IMAT SBIR ProjectIsoPlexis developed a dual function device that can capture a specific cell type from a complex mixture of cells via DNA tetheredDEALantibody technology developed inSAB memberCal Tech ProfessorJim Heath s labUpon removal of the non targeted cellsthe captured T cells are then immediately released via site specific DNA cleavage of the DEAL tether and transferred into a field of single cell capture micro troughsThen these microtroughs are encapsulated by an antibody barcoded slideSCBCTMOnce isolated in each microchamberthe barcode of antibodies collectsdifferent clinically relevant proteins being sent from each T cellFor Phase II of this projectwe will further develop the duo cell capture technology integrating IsoPlexis automation technology to isolate melanoma TILsAimDevelop TILs capture and release system for immunotherapy and highly multiplexed SCBC protein capture panel customized for TILs functional response profilingAimDevelop robustfully automated assay processing for seamless incorporation into the clinical laboratoryDemonstrate the utility of the automation system for easy to use enrichment and analysis protocol validating withmelanoma TILs samplesAimImplement fully automated workflow at Yale trial site to profile responders vsnon responders acrosspatientsCombining technological improvements of AimsandIsoPlexis will demonstrate pre and post monitoring of responder differencesusing multi functional and phenotypic profile of cytotoxic T cells monitoring a mid scale cohort of patientsover amonth period