In this application, we propose studies that will enable us to retrieve cancer cells that are hypoxic and/or cycling from tumor xenografts using our newly developed pair of fluorescent protein reporter system that we engineered and termed HypoxCR (Hypoxia - cell Cycle Reporters). With HypoxCR, we will study a panel of human cancer cell lines in vitro and in vivo to characterize their genomic and metabolic reprogramming upon exposure to hypoxia in vitro or in tumor xenografts. The delineation of the genomic and metabolic programs of the subpopulations of cancer cells in a tumor will instruct us about the commensal relations amongst cancer cells in a tumor tissue and lead us to novel therapeutic targets in addition to candidates that we identified from Myc and/or HIF targets, such as lactate dehydrogenase A (LDHA) and glutatminase (GLS). Our long term goal is to identify molecular targets from the reprogramming of cancer cells in the tumor tissue and find potential drug-like inhibitors of these targets as experimental molecular probes and potential therapeutic agents. To reach our goals, we set the following Aims:
Aim 1. To characterize cancer cells in the tumor microenvironment with a novel dual fluorescent protein reporter system for hypoxia and cell cycling (HypoxCR). Using this double reporter system, we propose to retrieve cancer cells from different subpopulations of the tumor xenograft to characterize their genomic changes and phenotypes and compare these changes with those arising in vitro under hypoxia.
Aim 2. To delineate the genomic reprogramming of cancer cells by MYC and HIF-1 in vitro and in vivo. Using in vitro models of tumor hypoxia and cells retrieved from xenografts marked by HypoxCR, we will use high throughput microarray-based methods to delineate the genomic reprogramming of cells by MYC and HIF-1 in the tumor microenvironment.
Aim3. To determine the role of key nodal points in metabolic reprogramming of cancer cells in hypoxia and in the tumor microenvironment. We propose to study miR-23a/b (which is suppressed by Myc and regulates glutaminase protein levels) and MCT1 (monocarboxylate/lactate transporter) in the reprogramming of cancer cells in the tumor microenvironment.
Aim 4. To determine the effects of inhibitors of key metabolic nodes in cancer cells in vitro and in vivo. We propose to use fluorescent HypoxCR reporters to determine the sensitivity of subpopulations of tumor cells to therapy in vivo and establish complementation between drug-like molecules (eg., LDHA and GLS inhibitors) that target different tumor cell subpopulations.

Public Health Relevance

Tumors are a mixture of cancer cells with variable characteristics that resist therapies. We will use a new way of retrieving subpopulations of cancer cells from tumors and determine their sensitivities to specific therapies, informing strategic combination therapy targeting metabolism.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA057341-23
Application #
8597540
Study Section
Tumor Cell Biology Study Section (TCB)
Program Officer
Mietz, Judy
Project Start
1992-09-01
Project End
2014-12-31
Budget Start
2014-01-01
Budget End
2014-12-31
Support Year
23
Fiscal Year
2014
Total Cost
$384,216
Indirect Cost
$127,460
Name
University of Pennsylvania
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Stine, Zachary E; Walton, Zandra E; Altman, Brian J et al. (2015) MYC, Metabolism, and Cancer. Cancer Discov 5:1024-39
Hsieh, Annie L; Walton, Zandra E; Altman, Brian J et al. (2015) MYC and metabolism on the path to cancer. Semin Cell Dev Biol 43:11-21
Altman, Brian J; Hsieh, Annie L; Sengupta, Arjun et al. (2015) MYC Disrupts the Circadian Clock and Metabolism in Cancer Cells. Cell Metab 22:1009-19
Xiang, Yan; Stine, Zachary E; Xia, Jinsong et al. (2015) Targeted inhibition of tumor-specific glutaminase diminishes cell-autonomous tumorigenesis. J Clin Invest 125:2293-306
Yan, Xiaohui; Hu, Zhongyi; Feng, Yi et al. (2015) Comprehensive Genomic Characterization of Long Non-coding RNAs across Human Cancers. Cancer Cell 28:529-40
Rajeshkumar, N V; Dutta, Prasanta; Yabuuchi, Shinichi et al. (2015) Therapeutic Targeting of the Warburg Effect in Pancreatic Cancer Relies on an Absence of p53 Function. Cancer Res 75:3355-64
Doherty, Joanne R; Yang, Chunying; Scott, Kristen E N et al. (2014) Blocking lactate export by inhibiting the Myc target MCT1 Disables glycolysis and glutathione synthesis. Cancer Res 74:908-20
Le, Anne; Stine, Zachary E; Nguyen, Christopher et al. (2014) Tumorigenicity of hypoxic respiring cancer cells revealed by a hypoxia-cell cycle dual reporter. Proc Natl Acad Sci U S A 111:12486-91
Dang, Chi V (2013) MYC, metabolism, cell growth, and tumorigenesis. Cold Spring Harb Perspect Med 3:
Dutta, Prasanta; Le, Anne; Vander Jagt, David L et al. (2013) Evaluation of LDH-A and glutaminase inhibition in vivo by hyperpolarized 13C-pyruvate magnetic resonance spectroscopy of tumors. Cancer Res 73:4190-5

Showing the most recent 10 out of 75 publications