Microfluidic Engineering of Artificial-Antigen Presenting Cells for the Biomanufacturing of Tumor-Specific T-Cells

University Of California-Irvine, Irvine CA

Investigators

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Abstract

Principal Investigator/Program Director (Last, First, Middle): Lee, Abraham, Phillip 7. Project Summary A major limitation of artificial antigen presenting cells (aAPCs) for adoptive T-cell therapy is the absence of a fluid membrane. This is a major concern since once a T-cell binds to an antigen-presenting cell (APC), the T- cell rearranges its membrane and cytoskeleton to form what is known as the ?immune synapse?. The synapse consists of a cluster of T-cell receptors surrounded by costimulatory and adhesion molecules. The formation of this synapse is essential to trigger a signaling cascade that activates the T-cell to divide and differentiate, and also causes it to produce cytokines. This project will focus on engineering cell-sized unilamellar vesicles (CUVs) as aAPCs to generate antigen-specific CD4+ T-cell responses. These aAPCs are converted from a storable (>8 months) double emulsion precursor generated by a novel microfluidic process. Aim 1 will focus on optimizing the process to covert double emulsion droplets (DEDs) to cell-sized unilamellar vesicles (CUVs), both in size and membrane formation and stability. The microfluidic generated cell-sized unilamellar vesicles (CUVs) will be characterized for stability and membrane properties. The expected outcome would be a robust process to generate artificial cells for a wide range of cell-based therapies. Aim 2 will focus on the functionalization of the lipid bilayer membranes of CUVs with MHC-II protein complexes to form immune synapses between the aAPCs and the T-cells. The formation of immune synapse synergizes enhancement of aAPC/T-cell binding through an increase in avidity. Membrane-bound adhesive proteins, such as the T-cell bound integrin LFA-1, surrounding these MHC-T clusters stabilize and prolong aAPC/T-cell interactions. The expected outcome is that a limited number of activating MHC?TCR complexes on the aAPCs will be sufficient to trigger T-cells. Aim 3 will focus on investigating the conditions for optimal T-cell expansion and differentiation. By modifying the peptide sequences presented in the MHC-II complex of the aAPC it is possible to direct T-cells to attack different types of cancer cells in a specified manner. The expected outcome is significant increase in IFN-? production in CUVs coupled with NY-ESO as well as TT tetramer both by ELISA and flow cytometry. Enhanced T cell proliferation is also expected in this group.