Establishing Drosophila Intestinal Tumor Models to Study Signaling interactions that regulate tumor growth.
Sydney Anderson, Matthew T. Bilotti, Elizabeth Conley, Michael K. Gruhot, Anthony Latronica, Ryann A. Mann, Kathleen McCaslin, Arushi Rai, Jibriel Saqibuddin
Colorectal cancer (CRC) is the second most common cause of cancer death in the United States, with an estimated 147,950 new cases in 2020. Some of the most common mutations found in patients with CRC are activation of oncogenic Ras, loss of function of APC, and dominant negative p53 mutations. We have developed a Drosophila melanogaster CRC model by integrating all three mutations in a single fly line. To understand the expression of each mutated gene on tumorigenesis, ‘one-’, ‘two-’, and ‘three-hit’ models were also made. Using these models we plan to establish how combinations of genetic alterations promote intestinal tumor growth, and understand the interactions between the molecular pathways of the mutated genes, understanding the expression of pathway specific target genes in the tumors, and understanding the progression of tumor formation in these CRC models.Our goal is to conduct a two part experiment using Gal4-UAS directly, where we will use esg-GAL4 to drive expression of tumor promoting genes specifically in intestinal stem cells. We will use a temperature sensitive GAL80 in combination with the esg-GAL4. This will allow temperature dependent control of GAL4 activity in the intestinal stem cells. Initially we plan to test the efficacy of GAL4 activity by testing the intestinal growth at room temperature (22℃) where the GAL4 is expected to remain inactive; and compare the phenotypes of intestinal growth with larvae grown at 29℃ where the GAL4 is active. First, we will look for survival rates in the larvae. Second, we will characterize the nature of the intestinal defects and check if the tumors are benign or malignant by dissecting the larvae, processing the intestines for antibody staining from 29℃ cultures. Our plan is to use anti-PH3 and anti- Dlg antibodies which will mark all dividing cells and cellular boundaries. This will allow us to determine if the tumors are also invasive or metastatic tumors. Based on these initial analyses, we will identify the potent combinations that cause metastatic tumors for further studies like assessment of MAPK, WNT and HIPPO pathway interactions. Here, we present our progress on the development and assessment of CRC models using the power of Drosophila.
Madhuri Kango-Singh, Amit Singh
Primary Advisor's Department
Stander Symposium project, College of Arts and Sciences
United Nations Sustainable Development Goals
Good Health and Well-Being
"Establishing Drosophila Intestinal Tumor Models to Study Signaling interactions that regulate tumor growth." (2022). Stander Symposium Projects. 2437.