1411-B - Confluence Discovery Technologies: Combination of In Vitro and In Vivo Systems to Study and Identify Novel Immuno-Oncology Drugs

Monday, February 5, 2018
5:00 PM - 6:00 PM

Confluence Discovery Technologies (CDT) is a full service CRO that specializes in supporting drug discovery and development projects from idea to IND. Fully equipped laboratories staffed with extensive drug discovery experience allow CDT to support entire programs (screening, computational and medicinal chemistry, biochemistry/mechanistic enzymology, cell biology, biomarker and in vivo pharmacology) or developing and executing on specific assays. A major focus of recent efforts has been to build assays interrogating the impact of drugs on immune cell function.

CDT has expertise in setting up in vitro lymphocyte systems to monitor drug effects on cytokine production, signal transduction, membrane protein expression, mRNA levels and cytotoxicity. Both CD4+ and CD8+ lymphocytes can be studied separately, and murine and human systems are available. Furthermore, methods have been validated for lineage specific differentiation of murine CD4+ T helper cells (Th1, Th2, Th17). Data will be shown that characterizes the kinetics of Th17 cell differentiation at the level of transcription factor analysis. In addition to generic activation of lymphocytes with CD3, murine transgenic T cells selective for a defined peptide:MHC complex can be studied as well (DO11.10 for CD4 cells, OT-1 for CD8 cells). Global activation of human T cells via soluble antibodies or bead-bound antibodies to CD3 and CD28 can be used. We have identified peripheral blood mononuclear cell (PBMC) donors, reactive to either lysates of CMV-infected cells or specific CMV peptides. With numerous frozen aliquots of each PBMC donor available, once identified as CMV-reactive, the same cells can be used for multiple rounds of compound or antibody assay. Further, allo-activation of PBMC or purified subsets of T cells (e.g., CD4 or CD8 cells) have been studied in co-culture with immature dendritic cells. Data will be shown indicating the range of responses to CMV across multiple donors, the ability of those responses to be amplified by checkpoint antibody blockade, and finally, the response of whole blood to CMV. This latter assay can be useful for clinical trials in which the effect of compounds on the memory T cell response is important to document.

In parallel with these in vitro cellular systems, CDT scientists are able to translate these observations to related in vivo systems. Studies can involve global activation of lymphocytes with injection of CD3 antibodies, or immunization with peptides specific for certain peptide:MHC complexes. Finally, with the implantation of tumor cells that express model antigens (e.g., EL4 thymoma cells or epithelial tumor cells engineered to express the OVA peptide recognized by the OT-1 transgenic T cell), effects on the anti-tumor immune response can be measured at both the gross level (data for the CT26 and MC38 tumors to be shown) of tumor growth, or infiltrating T cell. With respect to the innate immune response, data will be shown with NK cell activation and tumor killing. For a copy of the poster and to request additional information on these immunology assays please visit CDT scientists at Booth #1350.

Paul Changelian

Director of Biology
Confluence Discovery Technologies
St. Louis, MO

My laboratory has focused on the evaluation of small molecules in models of autoimmune disease, organ transplant and oncology, combining readouts of efficacy, pharmacodynamics and PK. Models include orthotopic models of pancreatic, breast and colon cancer, murine/rat collagen-induced arthritis, murine heterotopic heart transplant and cynomolgus kidney transplant. I have held positions of increasing responsibility beginning as a staff scientist at Pfizer in 1989 and currently serving as Director of Biology at Confluence Discovery Technologies. My education involved a BS in chemistry from the University of Michigan, a PhD in Immunology and Harvard Medical School and a postdoctoral fellowship in neurobiology at Washington University School of Medicine.