Genetic Analysis of the Putative Oncogene INT6 in Zebrafish
Lead Research Organisation:
University of Edinburgh
Department Name: MRC Human Genetics Unit-Cancer
Abstract
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Technical Summary
Significant insight into cancer development and treatment is found, in part, by studying cancer genes in model systems, such as the zebrafish. Many genes are highly conserved between zebrafish and humans, and mutations in zebrafish cancer genes can induce tumours that strongly resemble human cancers. Here, I propose to use the zebrafish system to rigorously probe the function of the putative cancer gene, INT6, in multi-drug resistance and cancer development.
INT6 is highly conserved with a clear role in cancer development, although the mechanisms behind this involvement remain mysterious. In fission yeast, Int6 maintains genome integrity, and regulates the multi-drug exposure response. In mice, integration of a virus into the INT6 gene results in truncated alleles of INT6 that appear to act in a dominant manner to promote breast cancer formation. Interestingly, in human cancers, loss of INT6 gene expression is frequently detected suggesting Int6 may also participate in suppressing tumour formation.
For my first Aim, I will use gene knockdown and overexpression methods in zebrafish embryos to rapidly establish Int6 function in genome stability and multi-drug resistance. Genomic instability is a hallmark of cancer, and these studies will be greatly enhanced by collaborating with Dr. Charles Lee, Assistant Director, Harvard Cancer Center Cytogenetic Core.
For my second Aim, I will create transgenic and mutant zebrafish lines that mimic Int6 mutations found in mammalian cancers, and yeast multi-drug resistance strains. In collaboration with Dr. Leonard Zon (Children‘s Hospital Boston) I will identify Int6 loss-of-function alleles using a new method called TILLING that involves screening over 1500 F1 mutagenized fish for INT6 mutations. With an array of Int6 lines, I will ultimately determine the contribution of Int6 to tumourigenesis. As well, I will identify Int6 genetic interacting partners, focusing on those that reveal novel insights into Int6 mechanisms that lead to multi-drug resistance and increased rates of tumour development. Zebrafish tumour pathology will be in consultation with Dr. James Amatruda (Children‘s Hospital), who has established a carcinogenesis assay in zebrafish. Finally, I will correlate Int6 tumour promoting mutations found in the zebrafish to those found in human cancers.
These Aims will be the first to establish INT6 gene networks in vertebrates, as well as address the effects of multiple Int6 mutations in tumourigenesis. Many tumours acquire resistance to chemotherapy drugs, and pursuing the function of Int6 in multi-drug resistance has direct implications for a clinically important question in cancer treatment.
INT6 is highly conserved with a clear role in cancer development, although the mechanisms behind this involvement remain mysterious. In fission yeast, Int6 maintains genome integrity, and regulates the multi-drug exposure response. In mice, integration of a virus into the INT6 gene results in truncated alleles of INT6 that appear to act in a dominant manner to promote breast cancer formation. Interestingly, in human cancers, loss of INT6 gene expression is frequently detected suggesting Int6 may also participate in suppressing tumour formation.
For my first Aim, I will use gene knockdown and overexpression methods in zebrafish embryos to rapidly establish Int6 function in genome stability and multi-drug resistance. Genomic instability is a hallmark of cancer, and these studies will be greatly enhanced by collaborating with Dr. Charles Lee, Assistant Director, Harvard Cancer Center Cytogenetic Core.
For my second Aim, I will create transgenic and mutant zebrafish lines that mimic Int6 mutations found in mammalian cancers, and yeast multi-drug resistance strains. In collaboration with Dr. Leonard Zon (Children‘s Hospital Boston) I will identify Int6 loss-of-function alleles using a new method called TILLING that involves screening over 1500 F1 mutagenized fish for INT6 mutations. With an array of Int6 lines, I will ultimately determine the contribution of Int6 to tumourigenesis. As well, I will identify Int6 genetic interacting partners, focusing on those that reveal novel insights into Int6 mechanisms that lead to multi-drug resistance and increased rates of tumour development. Zebrafish tumour pathology will be in consultation with Dr. James Amatruda (Children‘s Hospital), who has established a carcinogenesis assay in zebrafish. Finally, I will correlate Int6 tumour promoting mutations found in the zebrafish to those found in human cancers.
These Aims will be the first to establish INT6 gene networks in vertebrates, as well as address the effects of multiple Int6 mutations in tumourigenesis. Many tumours acquire resistance to chemotherapy drugs, and pursuing the function of Int6 in multi-drug resistance has direct implications for a clinically important question in cancer treatment.