This proposal addresses (I) the epigenetic control of cancer and (II) the relation between stem cells and cancer stem cells. I. Genome-wide hypomethylation, leading to genomic instability and increased rates of LOH, and regional hypermethylation, leading to silencing of tumor suppressor genes, are both hallmarks of cancer. The focus of Aim 1 is to establish a causal relation between genomic methylation and tumor formation in multistage cancers of the intestine, pancreas and prostate. Two complementary approaches will be pursued that either cause inhibition or ectopic activation of Dnmtl and Dnmt3b, two methyltransferases (MTases) that are crucial for maintaining and establishing genomic methylation. Because epigenetic changes - in contrast to genetic alterations - are reversible, the DNA methyltransferases represent promising targets for drug therapy. This proposal seeks to define the role these enzymes play in the development of cancers in * different tissues. II. Most cancers comprise a heterogeneous population of cells with marked differences in their proliferative potential. In the cancer stem cell concept, minor populations of tumor cells that possess the stem cell property of self-renewal are the only cells that are immortal and can sustain tumor growth. It is the second focus of this proposal to define epigenetic and genetic differences that distinguish normal stem cells from cancer stem cells. Because cancer stem cells are thought to be resistant to conventional cancer therapy it is vital to understand the differences between the tumor-initiating stem cells and the bulk tumor cells on a molecular level. One of the key issues for our understanding of the relation between adult stem cells and cancer stem cells is defining the molecular circuitry that drives self renewal in the normal stem cell and its malignant counterpart. We have shown that activation of Oct4 in cells of the adult mouse leads to a rapid and fully reversible dysplastic proliferation of the intestinal and skin stem/progenitor cell compartment. Oct4 is a pluripotency gene that in combination with Nanog and Sox2 controls the self-renewal of embryonic stem cells.
In Aim 2 we will investigate whether these key genes of ES cell renewal function also in self-renewal of adult stem cells and assess their possible role in cancer. We have used nuclear cloning as a novel and unbiased experimental tool to reprogram the genome of cancer cells and to distinguish between genetic and epigenetic changes that determine the malignant cell phenotype.
Aim 3 will use nuclear transplantation as a criterion to define the genetic and epigenetic alterations of the cancer cell genome that interfere with reversion to pluripotency.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA087869-10
Application #
7789607
Study Section
Cancer Genetics Study Section (CG)
Program Officer
Okano, Paul
Project Start
2001-05-01
Project End
2011-03-31
Budget Start
2010-04-01
Budget End
2011-03-31
Support Year
10
Fiscal Year
2010
Total Cost
$900,709
Indirect Cost
Name
Whitehead Institute for Biomedical Research
Department
Type
DUNS #
120989983
City
Cambridge
State
MA
Country
United States
Zip Code
02142
Forster, Ryan; Chiba, Kunitoshi; Schaeffer, Lorian et al. (2014) Human intestinal tissue with adult stem cell properties derived from pluripotent stem cells. Stem Cell Reports 2:838-52
Sarkar, Sovan; Maetzel, Dorothea; Korolchuk, Viktor I et al. (2014) Restarting stalled autophagy a potential therapeutic approach for the lipid storage disorder, Niemann-Pick type C1 disease. Autophagy 10:1137-40
Sexton, Alec N; Regalado, Samuel G; Lai, Christine S et al. (2014) Genetic and molecular identification of three human TPP1 functions in telomerase action: recruitment, activation, and homeostasis set point regulation. Genes Dev 28:1885-99
Torikai, Hiroki; Reik, Andreas; Soldner, Frank et al. (2013) Toward eliminating HLA class I expression to generate universal cells from allogeneic donors. Blood 122:1341-9
Buganim, Yosef; Faddah, Dina A; Jaenisch, Rudolf (2013) Mechanisms and models of somatic cell reprogramming. Nat Rev Genet 14:427-39
Dougan, Stephanie K; Dougan, Michael; Kim, Jun et al. (2013) Transnuclear TRP1-specific CD8 T cells with high or low affinity TCRs show equivalent antitumor activity. Cancer Immunol Res 1:99-111
Sarkar, Sovan; Carroll, Bernadette; Buganim, Yosef et al. (2013) Impaired autophagy in the lipid-storage disorder Niemann-Pick type C1 disease. Cell Rep 5:1302-15
Steine, Eveline J; Ehrich, Mathias; Bell, George W et al. (2011) Genes methylated by DNA methyltransferase 3b are similar in mouse intestine and human colon cancer. J Clin Invest 121:1748-52
Pawlak, Mathias; Jaenisch, Rudolf (2011) De novo DNA methylation by Dnmt3a and Dnmt3b is dispensable for nuclear reprogramming of somatic cells to a pluripotent state. Genes Dev 25:1035-40
Dawlaty, Meelad M; Ganz, Kibibi; Powell, Benjamin E et al. (2011) Tet1 is dispensable for maintaining pluripotency and its loss is compatible with embryonic and postnatal development. Cell Stem Cell 9:166-75

Showing the most recent 10 out of 59 publications