Development of 'enhanced' potency immunotherapy products using nonviral approaches. (November 2015)

• Record Type: Journal Article
• Authors: Brady, James; Li, Linhong; Viley, Angelia; Natarajan, Pachai; Allen, Cornell; Shivakumar, Rama; Duskin, Meg; Peshwa, Madhusudan V
• Is Part Of: Pharmaceutical bioprocessing. Volume 3:Number 7(2015)
• Page Start: 463
• Page End: 470
• Publication Date: 2015-11
• Subjects: cancer immunotherapy -- cell therapy -- chimeric antigen receptors -- dendritic cells -- electroporation -- GMP-compliant -- mRNA -- NK cells -- T cells; Drugs -- Testing -- Periodicals; Drug development -- Periodicals; Clinical trials -- Periodicals; Biopharmaceutics -- Periodicals; 615.19
• DOI: 10.4155/pbp.15.24
• Languages: English
• ISSNs: 2048-9153

Abstract

In the next 5–10 years we could see cellular-based pharmaceuticals, or cell therapy, meeting the unmet medical needs of thousands of people. How this therapy will meet these needs depends on the ability of researchers and manufacturers to successfully and cost effectively manufacture and deliver engineered cell-based therapeutic products that are safe and exhibit enhanced potency with resulting durable, meaningful clinical efficacy. The ability to engineer such enhanced potency using nonviral, cGMP-compliant, automated and closed system manufacturing processes will represent a significant advantage. To outline how such a process might work, we have summarized the application of a scalable, cGMP-compliant, electroporation platform for engineering dendritic cells (DCs), NK cells and T cells for development of cellular immunotherapies targeting hematological malignancies and solid tumors. Autologous cellular immunotherapy refers to a class of therapies that are designed to stimulate a specific immune response against cancer cells or other disease targets. Many autologous cellular immunotherapy protocols involve ex vivo modification of a patient's immune cells to enhance biological potency. Cellular modification strategies include pulsing DCs with tumor antigens/lysate and expression of chimeric antigen receptors or modified T-cell receptors (TCRs) on T cells and NK cells. DC pulsing is a process that stimulates uptake of tumor antigens or tumor cell lysate by DCs for the purpose of presenting tumor antigens on the DC surface in complex with major histocompatibility class molecules. DC pulsing is used in autologous cellular therapies for treating cancer. A chimeric antigen receptor (CAR) is a recombinant protein that typically consists of an extracellular antigen recognition domain (usually derived from an immunoglobulin variable region or TCR), a transmembrane domain, and one or more intracellular signaling domains (typically derived from TCRs and other proteins that are involved in T-cell signal transduction). Expression of CARs on the surfaces of NK cells or T cells following viral gene delivery or transfection with mRNA allows specific targeting of autologous immune cells to cancer cells. Scalable electroporation is a method for loading molecules into large numbers of cells based on the application of an electric field that produces temporary permeabilization of cell membranes. In contrast to conventional electroporation, which typically takes place in small cuvettes, scalable electroporation enables the processing of larger numbers of cells by flowing cells and loading agents between a pair of electrodes. mRNA transfection involves introducing mRNA molecules into cells for the purpose of expressing exogenous proteins. Compared to transfection of cells with plasmid DNA, mRNA transfection typically results in higher levels of cell viability and higher percentages of cells that express the encoded proteins. Transfection with mRNA also eliminates the potential for undesirable genomic insertion events that are associated with viral gene delivery methods.