In this award application we outline a strategy to identify genes that regulate cancer cell invasion, an early step in cancer metastasis, using the zebrafish. We have shown that zebrafish meltdown (mlt) mutants develop a cancer-like phenotype as a result of stromal invasion of posterior intestinal epithelial cells, mlt mutants ectopically express zebrafish orthologs of human genes strongly implicated in cancer progression. Importantly, inhibition of these genes rescues the invasive mlt phenotype. Positional cloning identified zebrafish smooth muscle myosin heavy chain (MYH11) as the responsible mlt gene. Human MYH11 has also been shown to be part of a gene expression signature that is predictive of tumor metastasis. Together, these findings support mlt as an invasive cancer model. Here, we propose experiments that will allow the identification of mlt modifier genes. The human orthologs of such genes are predicted to play a role in cancer invasion and metastasis, and may be targets for anti-cancer therapy. Three interrelated sets of experiments have been designed to identify mlt modifiers. First, we will analyze in mlt mutants the expression arid function of zebrafish orthologs of human genes with an established role in cancer cell invasion. Second, we will compare transcriptional profiles of mlt and sibling wild type larvae and assess the role of differentially expressed genes using antisense knockdown and forced expression experiments. Third, we will conduct a chemical mutagenesis genetic screen to identify recessive and dominant modifiers of the mlt cancer-like phenotype. Together, these aims provide a novel approach to identify genes that regulate human cancer progression. Functional assessment of these genes in mlt mutants provides a rapid, in vivo assay of their suitability for pharmacological targeting.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
3R01DK054942-09S1
Application #
7994128
Study Section
Gastrointestinal Cell and Molecular Biology Study Section (GCMB)
Program Officer
Karp, Robert W
Project Start
2010-02-22
Project End
2011-01-31
Budget Start
2010-02-22
Budget End
2011-01-31
Support Year
9
Fiscal Year
2010
Total Cost
$101,270
Indirect Cost
Name
University of Pennsylvania
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Abrams, Joshua; Einhorn, Zev; Seiler, Christoph et al. (2016) Graded effects of unregulated smooth muscle myosin on intestinal architecture, intestinal motility and vascular function in zebrafish. Dis Model Mech 9:529-40
Seiler, Christoph; Davuluri, Gangarao; Abrams, Joshua et al. (2012) Smooth muscle tension induces invasive remodeling of the zebrafish intestine. PLoS Biol 10:e1001386
Walters, James W; Anderson, Jennifer L; Bittman, Robert et al. (2012) Visualization of lipid metabolism in the zebrafish intestine reveals a relationship between NPC1L1-mediated cholesterol uptake and dietary fatty acid. Chem Biol 19:913-25
Abrams, J; Davuluri, G; Seiler, C et al. (2012) Smooth muscle caldesmon modulates peristalsis in the wild type and non-innervated zebrafish intestine. Neurogastroenterol Motil 24:288-99
Gao, Nan; Davuluri, Gangarao; Gong, Weilong et al. (2011) The nuclear pore complex protein Elys is required for genome stability in mouse intestinal epithelial progenitor cells. Gastroenterology 140:1547-55.e10
Seiler, Christoph; Pack, Michael (2011) Transgenic labeling of the zebrafish pronephric duct and tubules using a promoter from the enpep gene. Gene Expr Patterns 11:118-21
Lorent, Kristin; Moore, John C; Siekmann, Arndt F et al. (2010) Reiterative use of the notch signal during zebrafish intrahepatic biliary development. Dev Dyn 239:855-64
Clifton, Justin D; Lucumi, Edinson; Myers, Michael C et al. (2010) Identification of novel inhibitors of dietary lipid absorption using zebrafish. PLoS One 5:e12386
Seiler, Christoph; Abrams, Joshua; Pack, Michael (2010) Characterization of zebrafish intestinal smooth muscle development using a novel sm22?-b promoter. Dev Dyn 239:2806-12
Davuluri, G; Seiler, C; Abrams, J et al. (2010) Differential effects of thin and thick filament disruption on zebrafish smooth muscle regulatory proteins. Neurogastroenterol Motil 22:1100-e285

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