Optimizing murine cells for in vitro modeling of high-grade serous ovarian cancer
Mentor: Daniela Dinulescu, PhD (Brigham and Women's Hospital)
High-grade serous ovarian carcinoma (HGSC) is known to be the most aggressive and lethal subtype of ovarian cancer. Recent research indicates that improved, well-characterized HGSC cell lines are needed in order to perform more effective in vitro pre-clinical studies that accurately represent HGSC. The culturing of high-grade serous murine/human ovarian cancer cells from tumors, fallopian tissues or ascites has been challenging, as the establishment of primary lines most often requires the use of specialized medias and immortalization of cells. These techniques frequently lead to alterations in the cell pathways that may not be representative of the actual tumor. Our study aims to highlight a new method of generating novel HGSC cell lines from fallopian tube secretory cells, which have recently been acknowledged in literature as a significant initiation or origin site for HGSC. We will be isolating fallopian tube cells directly from a genetically engineered murine model generated by the Dinulescu laboratory. Using this model, we plan to generate a normal murine FTSEC cell line using cells that harbor conditional Brca;TP53;PTEN mutational signatures which closely recapitulate those of human HGSC. We will then treat the normal cells with doxycycline in order to induce the Brca;TP53;PTEN mutations, establishing a cancerous subset of cells. Using these cells directly derived from the mouse model and induced using doxycycline, we hypothesize that we will be able to accurately mimic human HGSC in vitro by engrafting the cancerous cells into immunodeficient mice intraperitoneally. We hypothesize that these cells will develop in the immunocompromised mice and model HGSC tumor growth in vivo. We will confirm the development of the transplanted cells using luciferase imaging in living mice and through necropsy of euthanized mice. These cells will serve as a unique tool for future research into understanding “ovarian” cancer etiology, oncogenic pathway aberrations, and allow reliable in vitro trials for novel chemotherapeutics such as nanoparticles, before performing in vivo studies.
Murine fallopian tube cells grown in vitro as a spheroid that was originally isolated from the BRCA1 (fl/+) p53 (fl/mut) PTEN (fl/fl) murine cohort. The micrograph reveals red fluorescence from the presence of mCherry, indicating a successful transfection of our mCherry/luc plasmid (Image taken at 40X). Source: Unpublished original research, Morgan Krush, Dinulescu Laboratory. Source: