Numeric Expansion of CD-19 Specific T cells in Magnetic 3D Culture

Project Summary Adoptive immunotherapy trials using genetically modified natural killerNKand T cells with redirected specificity against B cell malignancies have demonstrated therapeutic efficacyTwo early phase clinical trials have been initiated at The University of Texas MDAnderson Cancer Center to evaluate the safety and efficacy of adoptive transfer of autologous and allogeneic T cells expressing a CDspecific chimeric antigen receptorCARinto patients with advanced B cell malignanciesThese CDspecific CART cells are selectively propagated in vitro in the presence of irradiated artificial antigen presenting cellsaAPCexpressing co stimulatory molecules that sustain their proliferation to clinically sufficient numbers in compliance with current good manufacturing practicecGMPfor Phase I II trialsThis conventional culturing process requiresdays to generate the required andgtCART cells for each infusionReduction of the time required to generate clinically sufficient numbers in tissue culture will reduce manufacturing costs significantly and will yield a clinical product in which the CART cells have increased replicative potential and are less differentiatedTo enhance the production processwe propose to develop novel co culture protocol that utilizes magnetic co levitation to increase the contact interface between CARNK or T cells and the stimulating aAPCs to significantly improve the kinetics of numeric expansion of CARlymphocytes with desired specificity and preservation of memory phenotypeOur long term goal is to develop a cGMP compliant protocol based on three dimensionalDmagnetic levitation co cultures for the rapid and specific production of CART and NK cells to support ongoing immunotherapy trialsOur novel expansion method is based on the central hypothesis that increasing the number and duration of contact points between CART or NK cells and aAPCs will accelerate the proliferation of the targeting cellsyielding clinically sufficient number of cells in less timeIn this Phase I SBIR proposalwe aim to develop and optimizeD cell magnetic levitation co culture conditions to improve the propagation of CART cells by magnetic nanoparticle labeling of aAPCsNKand T cells and co culturing inD to maximize expansion of the genetically modified CDspecific NK and T cellsDevelopment of this expansion methodology will significantly impact the treatmentmanagementand cost of adoptive cell therapies of B cell malignancies Narrative Adoptive T cell therapy offers a promising approach for treating B cell malignanciesThe process of preparing these targeting T cells for transplantation involves isolation of antigen specific cells and expansion in vitro in the presence of target antigenThe process of preparing these cells is lengthytaking up todaysIt is recognized that reducing the preparation time in culture will save manufacturing costs will yield less differentiated T cells that have increased targeting potentialThe proposed work will implement and optimize T cell expansion methods that are based on magnetically levitated cell cultures to produce clinically sufficient number of T cells in one third of the time required to manufacture by conventional methods