TRAINING IN PHARMACOLOGY, TOXICOLOGY & PHARMACEUTICAL SCIENCES 2008 ASPET SUMMER UNDERGRADUATE RESEARCH FELLOWSHIP OPPORTUNITY at the UNIVERSITY OF CINCINNATI
The Department of Genome Sciences, College of Medicine is pleased to offer this research project as part of the 20078 summer ASPET SURF Training Program offered by the Department of Pharmacology & Cell Biophysics.  Students interested in this project are advised to contact Professor Plas to discover more about the project, learn what your responsibilities will be during the ten-week research training program.
2008 ASPET SURF Project #:  08 - 010 Faculty Supervisor/Mentor: David R. Plas, Ph.D. Assistant Professor Genome Sciences Genome Research Institute, University of Cincinnati Email:  plasdr@uc.edu
Project Title: Identifying Selective Inhibitors of the AKT and Bcl-xL survival pathways
Research Program Description: Cancer incidence has reached 1.4M in the US.  Prevalence of Akt activation in cancer varies by tissue, but is greater than 25% for leukemia, prostate carcinoma, breast carcinoma, and glioblastoma.  These malignancies will account for approximately 500,000 new cancer diagnoses in 2006.  To achieve curative chemotherapy for cancer, compounds must induce programmed cell death (apoptosis) in cancer cells without significant toxicity towards normal cells.  This can be especially difficult in Akt-expressing cells, which have increased resistance to multiple apoptotic stimuli, including chemotherapeutic drugs (Wendel, Nature 2004).  We have shown that by manipulating cellular metabolism, apoptosis can be specifically induced in Akt-expressing cells without affecting cells expressing other pro-survival genes such as Bcl-xL (review: Plas and Thompson, Oncogene 2005).  We therefore propose screening for compounds that mimic the effects of metabolic intervention in Akt survival: successful compounds will induce apoptosis specifically in Akt-expressing cells, with low toxicity in Bcl-xL-expressing cells.  Moving forward, experiments will test compounds alone and in combination with standard chemotherapeutics for effects on metabolism and apoptosis in cells expressing oncogenes.  By targeting the metabolic program required for Akt-dependent apoptosis resistance, compounds identified in this project will contribute a novel approach for overcoming apoptosis resistance in cancer cells.
ASPET SURF Project Description:  The ASPET SURF project will screen a set of small molecules for viability in FL5.12 cells cultured without IL‑3, expressing either Bcl-xL or myristoylated Akt.  ‘Hits’ will be those compounds that show selective toxicity for Akt-expressing cells with little impact on Bcl-xL survival.  ‘Hits’ will be further tested in cells expressing other anti-apoptotic genes, such as BCR-ABL. A secondary screen to confirm the activity of the ‘hits’ from the primary screen will be used to test for those ‘hits’ that specifically inhibit Akt-dependent increases in mitochondrial membrane potential as assessed by fluorescence in live cells stained with tetramethylrhodamine ethyl ester (TMRE). Phosphorylation of the Akt pathway components, as well as several aspects of cell metabolism, and molecular markers of apoptosis and autophagy will be measured using in a tissue culture system.  The ASPET SURF student will use cell culture, an apoptosis assay developed for high-throughput screening (HTS), and high-throughput screening (Evotec-Technologies Plate::Explorer ultra HTS system) of a library of small molecules and assembling a chemically-structured focused set of small molecules for HTS. The project seeks to identify small molecules that can selectively inhibit two survival pathways used by cancer cells, the AKT pathway and Bcl-xL pathways. Identification of small molecules that interact with these two survival pathways are the first set of steps in the discovery of new drugs for the treatment of cancer.  The compounds screened will be from a recently acquired large compound repository.  The student will be co-mentored by a GRI member (e.g., Dr. Sandra Nelson or Dr. Bill Seibel) who is knowledgeable in this research field.