The zebrafish is a powerful vertebrate genetic model and has recently been used as a system to understand cancer. We have isolated 19 cell cycle mutants and several mutated genes are associated with cancer in our model system. Our studies of B-myb identified a novel pathway that affects cyclin B and participates in cancer, and our recent study of a separase mutant demonstrated that genome instability predisposes epithelial cells to cancer. For this proposal, we plan to investigate two independent cancer models based on ras pathway activation. RAS family members and the downstream targets are mutated in a number of human cancers, including melanoma and rhabdomyosarcoma. In the first model, BRAF and p53 deficiency induced melanoma. In a second model, a constitutive KRAS allele leads to embryonal rhabdomyosarcoma at 10 days of age. This is the fastest tumor model in any vertebrate. In both cases, we have demonstrated that the cells that initiate the tumor are transplantable and represent early stages of melanocytic and myogenic differentiation, respectively. We hypothesize that RAS induction will activate similar self-renewal pathways within tumors of different tissue types. Using transgenic fish, we plan to label the cancer cells with a number of reporter constructs and determine the subpopulation of tumor cells that are transplantable. The models will be further used to understand the signaling pathways operative in the cancer-initiating cell population. We plan to determine if the wnt-prostaglandin pathway can modify the ability of the cancer-initiating cell to function. We also plan to undertake a chemical screen defining molecules that will inhibit cancer stem cells. Two novel chemicals have already been found that block an expansion of early neural crest progenitors in the BRAF;p53 deficiency embryos. These chemicals are now being tested in our zebrafish model for the ability to block adult melanoma. Our studies will lead to a better understanding of cell heterogeneity within a cancer and will also lead to potential therapies for rhabdomyosarcoma and melanoma.

Public Health Relevance

Cancer is a difficult disease to treat because cells remaining after chemotherapy can recapitulate the tumor, leading to relapse. Here we plan to examine the differentiation of these rare cells within a tumor, and find pathways that disrupt them. By defining such pathways, new therapies can be designed to use in conjunction with conventional chemotherapy in the treatment of cancer.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA103846-10
Application #
8233548
Study Section
Molecular Oncogenesis Study Section (MONC)
Program Officer
Mietz, Judy
Project Start
2003-07-03
Project End
2013-02-28
Budget Start
2012-03-01
Budget End
2013-02-28
Support Year
10
Fiscal Year
2012
Total Cost
$366,757
Indirect Cost
$155,977
Name
Children's Hospital Boston
Department
Type
DUNS #
076593722
City
Boston
State
MA
Country
United States
Zip Code
02115
McConnell, Alicia M; Mito, Jeffrey K; Ablain, Julien et al. (2018) Neural crest state activation in NRAS driven melanoma, but not in NRAS-driven melanocyte expansion. Dev Biol :
Yu, Yong; Schleich, Kolja; Yue, Bin et al. (2018) Targeting the Senescence-Overriding Cooperative Activity of Structurally Unrelated H3K9 Demethylases in Melanoma. Cancer Cell 33:322-336.e8
Ablain, Julien; Xu, Mengshu; Rothschild, Harriet et al. (2018) Human tumor genomics and zebrafish modeling identify SPRED1 loss as a driver of mucosal melanoma. Science 362:1055-1060
Wattrus, Samuel J; Zon, Leonard I (2018) Stem cell safe harbor: the hematopoietic stem cell niche in zebrafish. Blood Adv 2:3063-3069
van Rooijen, Ellen; Fazio, Maurizio; Zon, Leonard I (2017) From fish bowl to bedside: The power of zebrafish to unravel melanoma pathogenesis and discover new therapeutics. Pigment Cell Melanoma Res 30:402-412
Fazio, Maurizio; Zon, Leonard I (2017) Fishing for answers in precision cancer medicine. Proc Natl Acad Sci U S A 114:10306-10308
Ciarlo, Christie; Kaufman, Charles K; Kinikoglu, Beste et al. (2017) A chemical screen in zebrafish embryonic cells establishes that Akt activation is required for neural crest development. Elife 6:
Fazio, Maurizio; Avagyan, Serine; van Rooijen, Ellen et al. (2017) Efficient Transduction of Zebrafish Melanoma Cell Lines and Embryos Using Lentiviral Vectors. Zebrafish 14:379-382
Ablain, J; Zon, L I (2016) Tissue-specific gene targeting using CRISPR/Cas9. Methods Cell Biol 135:189-202
Dang, Michelle; Henderson, Rachel E; Garraway, Levi A et al. (2016) Long-term drug administration in the adult zebrafish using oral gavage for cancer preclinical studies. Dis Model Mech 9:811-20

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