The focus of my research is to develop novel technologies to measure the proteomic response of individual patients to targeted therapies for lymphoma. I have established a strong foundation in the use of transgenic models to study cancer. I now aim to gain the skills needed to become an independent translational researcher, including rigorous knowledge of biostatistics and clinical trial design and experience as a co- investigator in ongoing studies, with the goal to successfully incorporate potential new biomarkers into patient oriented research to validate biomarkers for clinical decision making. At the bench, I have developed a novel nanofluidic proteomic immunoassay (NIA) for the analysis of oncoproteins in cells from patient specimens collected using minimally invasive techniques such as fine needle aspirate (FNA) or blood draw, in as few as 25 cells in 4 nanoliters of lysate. NIA can be used to simultaneously identify multiple phosphorylated and non- phosphorylated isoforms of individual proteins. I have developed the use of NIA to measure total oncoproteins, including BCL2, MYC, ERK and MEK in lymphoma, to measure percent phosphorylation of ERK and MEK proteins in the MAP Kinase (MAPK) pathway;and identify decreases of a specific phosphorylated ERK2 isomer early during CML treatment that could distinguish patients who would respond to tyrosine kinase inhibitors from those that were treatment refractory. Finally, I have used NIA together with flow cytometry (FACS) to identify changes in RAS/MAPK/STAT signaling in cells from patients with low grade NHL treated on a clinical trial of single agent atorvastatin. We discovered that patients with Chronic Lymphocytic Leukemia (CLL) and Marginal Zone Lymphoma (MZL) showed an increase in tumor cell apoptosis and a decrease in circulating tumor cells upon treatment with atorvastatin, indicating that atorvastatin may have a measurable clinical effect associated with apoptosis and signaling inhibition. Comparison of proteomic signatures in individual patients before and early during treatment can provide valuable information that can not be gleaned from a single timepoint. I hypothesize that nanoscale analysis of proteomic changes in blood and FNA's can reveal the molecular activity of novel therapeutics in individual patients. I also hypothesize that NIA can be used to identify when a drug preferentially effects a specific protein phospho-isoform and FACS analysis can resolve drug effects on tumor cells versus non-tumor cells. I will develop complementary NIA and FACS technologies for analysis of minimally invasive blood or fine needle aspirate samples from individual patients before and during treatment. I will determine if changes in BCL2 and RAS pathway profiles can be used to monitor the molecular activity of a BCL2 family inhibitor (ABT-263) and atorvastatin, respectively. My proposal will be amply supported by state-of-the art facilities, equipment, including a CB1000 instrument to perform NIA, and technical support. The Stanford Cancer Center and Stanford Clinical Trials Office provide resources for clinical trial support, including support for Oncore, our online research management database. Biomarker studies will be performed in the laboratory of Dr. Felsher in the Center for Clinical Sciences Research building. Finally, our new Population Health Sciences Institute (PHSI) is a newly opened patient-based research center that occupies 40,000 square feet containing a Clinical Investigation Unit to assist investigators with the development, implementation, and conduct of patient-based research studies, examination rooms and a core laboratory to perform specialized sample processing and storage in support of research protocols. The PHSI also has a Quantitative Sciences Unit and a Bioinformatics Unit to provide biostatistical support in research design and data analysis and to facilitate integration of data from multiple platforms. I will have access to all these facilities, and equipment. A key component to my career development is an ambitious plan to complete 31 units of biostatistical and clinical design coursework through the Stanford Center for Clinical and Translational Education and Research, Department of Health Research and Policy. I will gain clinical experience by becoming a co-investigator on an ongoing Phase I study of ABT-263, and as co-protocol director for our investigator-initiated study of atorvastatin in lymphoma. My instructor appointment has 100% protected time for research-related activities in order to accomplish these goals, guided my Primary Mentor, Dr. Dean Felsher and Co-Mentor, Dr. Ronald Levy. I am fortunate to have assembled this well-matched team committed to my career development. The three of us will meet together monthly. Dr. Felsher, Director of the Stanford Cancer Center Molecular Therapeutics Program, has established a superb and highly funded research program, including 5 different federal grants, studying the mechanism of "oncogene addiction". Dr. Felsher will assure that I receive the education, evaluation and guidance, financial support and resources for my studies, leadership opportunities and protected time, and provide specific guidance for career development. Dr. Levy, a world leader in translational research for lymphoma, will provide guidance in performing translational clinical trials and in achieving academic advancement. My career is dedicated to translational investigation for realizing the goal of individualized cancer treatment.
The proposed work will use innovative technologies to develop new methods to measure proteins that might predict if patients will respond to therapies designed to target specific tumor pathways. I will incorporate these new assays into two clinical trials, each evaluating the biologic and clinical response of patients with lymphoma to novel therapeutic agents.
|Eberlin, Livia S; Gabay, Meital; Fan, Alice C et al. (2014) Alteration of the lipid profile in lymphomas induced by MYC overexpression. Proc Natl Acad Sci U S A 111:10450-5|
|Beurlet, Stephanie; Omidvar, Nader; Gorombei, Petra et al. (2013) BCL-2 inhibition with ABT-737 prolongs survival in an NRAS/BCL-2 mouse model of AML by targeting primitive LSK and progenitor cells. Blood 122:2864-76|
|DeChristopher, Brian A; Fan, Alice C; Felsher, Dean W et al. (2012) "Picolog," a synthetically-available bryostatin analog, inhibits growth of MYC-induced lymphoma in vivo. Oncotarget 3:58-66|
|Seetharam, Mahesh; Fan, Alice C; Tran, Mai et al. (2012) Treatment of higher risk myelodysplastic syndrome patients unresponsive to hypomethylating agents with ON 01910.Na. Leuk Res 36:98-103|