While mechanisms regulating oncogenesis have been elucidated substantially in recent years, comparatively little is yet known about the mechanisms that underlie metastasis of tumor cells, the ultimate cause of death in cancer. Metastasis suppressor genes play pivotal roles in controlling this process, but only a handful have been identified. NM23 (""""""""negative in metastasis"""""""") genes represent the first described metastasis suppressors and are recognized for their antimetastatic activity in melanoma, breast and gastric carcinoma. The overall goal of this project to systematically dissect the molecular mechanisms and genomic pathways underlying the metastasis suppressor activity of NM23. We recently found that the NM23-H1 isoform is a 3'-5' exonuclease (3'5' EXO), an activity often associated with genome integrity, apoptosis and chromatin remodeling. Although there is clear potential relevance of these processes to malignant progression, we will also measure the extent to which the nucleoside diphosphate kinase (NDPK) and histidine kinase (hisK) activities of NM23-H1 contribute to its metastasis suppressor properties.
In Specific Aim 1, we will complete a detailed structure-function analysis to delineate the active site of the 3'5 EXO, and characterize a panel of mutants harboring lesions that selectively disrupt the 3'5' EXO, NDPK and hisK functions of the NM23-H1 molecule. The mutants will be used as molecular tools to determine the relative contributions of each to suppression of the metastatic phenotype (Specific Aim 2) in human melanoma cells, using cell culture and in vivo models to measure tumorigenic and metastatic potential. Gene expression changes elicited by expression of NM23-H1 variants will be determined by microarray analysis and real-time PCR (Specific Aim 3). Correlational analysis will identify NM23-regulated genes whose expression tracks with metastatic phenotype and metastatic progression. Functional relevance of identified genes will be validated by viral-mediated overexpression and knockdown approaches.
In Specific Aim 4, we will directly test the DMA repair activity of NM23-H1 in mammalian and yeast cell models, and identify the relevant enzymatic function. These studies should provide the first systematic analysis of pathways through which NM23 proteins oppose metastasis in multiple cancer types and, therefore could suggest novel strategies to combat this final and intractable stage of the disease. ? ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA083237-08
Application #
7472332
Study Section
Cancer Genetics Study Section (CG)
Program Officer
Woodhouse, Elizabeth
Project Start
1999-08-01
Project End
2011-07-31
Budget Start
2008-08-01
Budget End
2009-07-31
Support Year
8
Fiscal Year
2008
Total Cost
$203,946
Indirect Cost
Name
University of Kentucky
Department
Pharmacology
Type
Schools of Medicine
DUNS #
939017877
City
Lexington
State
KY
Country
United States
Zip Code
40506
Puts, Gemma S; Leonard, M Kathryn; Pamidimukkala, Nidhi V et al. (2018) Nuclear functions of NME proteins. Lab Invest 98:211-218
Leonard, M Kathryn; McCorkle, Joseph R; Snyder, Devin E et al. (2018) Identification of a gene expression signature associated with the metastasis suppressor function of NME1: prognostic value in human melanoma. Lab Invest 98:327-338
Leonard, M Kathryn; Pamidimukkala, Nidhi; Puts, Gemma S et al. (2017) The HGF/SF Mouse Model of UV-Induced Melanoma as an In Vivo Sensor for Metastasis-Regulating Gene. Int J Mol Sci 18:
Novak, Marián; Leonard, Mary Kathryn; Yang, Xiuwei H et al. (2015) Metastasis suppressor NME1 regulates melanoma cell morphology, self-adhesion and motility via induction of fibronectin expression. Exp Dermatol 24:455-61
Kaetzel, David M; Leonard, Mary K; Cook, Gemma S et al. (2015) Dual functions of NME1 in suppression of cell motility and enhancement of genomic stability in melanoma. Naunyn Schmiedebergs Arch Pharmacol 388:199-206
McCorkle, Joseph R; Leonard, Mary K; Kraner, Susan D et al. (2014) The metastasis suppressor NME1 regulates expression of genes linked to metastasis and patient outcome in melanoma and breast carcinoma. Cancer Genomics Proteomics 11:175-94
Jarrett, Stuart G; Novak, Marian; Harris, Nathan et al. (2013) NM23 deficiency promotes metastasis in a UV radiation-induced mouse model of human melanoma. Clin Exp Metastasis 30:25-36
Jarrett, Stuart G; Novak, Marian; Dabernat, Sandrine et al. (2012) Metastasis suppressor NM23-H1 promotes repair of UV-induced DNA damage and suppresses UV-induced melanomagenesis. Cancer Res 72:133-43
Ganguly, S S; Fiore, L S; Sims, J T et al. (2012) c-Abl and Arg are activated in human primary melanomas, promote melanoma cell invasion via distinct pathways, and drive metastatic progression. Oncogene 31:1804-16
Zhang, Qingbei; McCorkle, Joseph R; Novak, Marian et al. (2011) Metastasis suppressor function of NM23-H1 requires its 3'-5' exonuclease activity. Int J Cancer 128:40-50

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