The applicants previously showed that chromosome 6 suppresses human melanoma metastasis without affecting tumorigenicity, suggesting that a melanoma metastasis-suppressor gene is encoded on chromosome 6. Using differential display and subtractive hybridization, seven differentially expressed transcripts were identified and full-length cDNAs obtained. One clone with novel nucleotide and protein sequences, KiSS-1, was expressed only in nonmetastatic neo6/melanoma hybrids, normal melanocytes and RGP melanomas by Northern blot and by RT-PCR. The KiSS-1 gene mapped to the chromosome 1q32-q41 by FISH. Transfection of KiSS-1 suppressed human melanoma C8161 metastasis by 50-99%. Tumorigenicity was not affected in the transfectants. KiSS-1, therefore, met the criteria of a melanoma metastasis-suppressor gene. Three fundamental questions arise based upon the preliminary data--(1) Is KiSS-1 a universal melanoma metastasis-suppressor gene? (2) What regulates KiSS-1 expression? (3) How does KiSS-1 suppress metastasis? Specific Aim 1: will address the first question by transfection of KiSS-1 cDNA into other metastatic human melanoma cell lines and assessment of tumorigenicity and metastasis in athymic nude mice.
Specific Aim 2 : since KiSS-1 maps to chromosome 1, it is hypothesized that a gene(s) on chromosome 6 fragments, having already mapped a locus to 6q21-q23. For this aim, the applicants will continue using this strategy to further refine the locus to <5 Mb.
Specific Aim 3 : will test the hypothesis that KiSS-1 functions in a signaling pathway, mediating its metastasis-suppressor effects through interaction with an SH3-containing protein. This hypothesis is based upon the predicted amino acid sequence of the KiSS-1 protein. The most notable feature of the Mre18 kDa, largely-hydrophilic protein is a proline-rich domain with conserved PXP motif and flanking regions indicative of an SH3 domain ligand. In addition, KiSS-1 has multiple Ser, Thr and Tyr residues as well as a consensus PKC-alpha phosphorylation site, suggesting that it could also be phosphorylated. First, they will describe basic biochemical properties of KiSS-1 such as post-translational modification and cellular localization. Second, to directly test the hypothesis, we will prepare site-directed and deletion-mutant KiSS-1 and assess their abilities to suppress melanoma metastasis. Third, they will identify critical KiSS-1 protein/macromolecule interactions using plaque library screening an immune co-precipitation experiments.
| Cook, Leah M; Hurst, Douglas R; Welch, Danny R (2011) Metastasis suppressors and the tumor microenvironment. Semin Cancer Biol 21:113-22 |
| Hurst, Douglas R; Welch, Danny R (2011) Unraveling the enigmatic complexities of BRMS1-mediated metastasis suppression. FEBS Lett 585:3185-90 |
| Hurst, Douglas R; Welch, Danny R (2011) Metastasis suppressor genes at the interface between the environment and tumor cell growth. Int Rev Cell Mol Biol 286:107-80 |
| Beck, Benjamin H; Welch, Danny R (2010) The KISS1 metastasis suppressor: a good night kiss for disseminated cancer cells. Eur J Cancer 46:1283-9 |
| Stafford, Lewis J; Vaidya, Kedar S; Welch, Danny R (2008) Metastasis suppressors genes in cancer. Int J Biochem Cell Biol 40:874-91 |
| Hurst, Douglas R; Welch, Danny R (2007) A MSC-ing link in metastasis? Nat Med 13:1289-91 |
| Nash, Kevin T; Phadke, Pushkar A; Navenot, Jean-Marc et al. (2007) Requirement of KISS1 secretion for multiple organ metastasis suppression and maintenance of tumor dormancy. J Natl Cancer Inst 99:309-21 |
| Eccles, Suzanne A; Welch, Danny R (2007) Metastasis: recent discoveries and novel treatment strategies. Lancet 369:1742-57 |
| Nash, Kevin T; Welch, Danny R (2006) The KISS1 metastasis suppressor: mechanistic insights and clinical utility. Front Biosci 11:647-59 |
| Welch, Danny R (2006) Do we need to redefine a cancer metastasis and staging definitions? Breast Dis 26:3-12 |
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