Rapid Genetic Manipulation of Patient-derived Breast Tumors for Metastatic Study (2013)

Undergraduates: Nicole Zalles, Dr. Chuck Harrell

Faculty Advisor: Charles Perou
Department: Biology

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While surgery, chemotherapy and radiation therapy remove or destroy the primary tumor in cancer patients, the spread and growth of cancer cells in secondary sites contribute to subsequent morbidity and mortality. There are five genetically distinct types of human breast cancer, each with a distinguishing set of genes defining their inherent biology, response to therapy, and metastatic behavior. Our goal is to identify and target specific genes that may be responsible for metastatic behavior of each subtype. Our study is focused on a basal-like tumor derived from a patient and maintained in-vivo through xenografting in mice (WHIM2); we aim to identify the genetic determinants of the metastatic abilities of this human basal-like tumor. We are developing a procedure for the labeling of this tumor so that it can be tracked/imaged in-vivo, to determine its pattern of metastasis. We established a protocol where cells were rapidly labeled with lentivirus that expresses GFP and Luciferase. Preliminary results found that the rapid labeling protocol successfully incorporated into the genome in as little as one-hour. Subsequent FACS for GFP-positive cells further purified the cell population. Future studies will incorporate metastasis promoting/inhibiting genes into the rapid labeling protocol. These will establish a baseline from which we can rapidly modify patient derived samples for further metastatic research to better understand how to improve the prognoses of patients.