Dissecting Mechanism of Neurofibromin Regulation in the Context of Sporadic Cancers and Neurofibromatosis Type 1

University Of California, San Francisco, San Francisco CA

Investigators

Abstract

Mutations affecting proteins in the RAS pathway have been found in 46% of patient samples, more than any other signaling pathway. The RAS GTPase activating protein neurofibromin is a crucial component of this pathway, with mutations in this protein leading to both sporadic cancers and the tumor predisposition syndrome Neurofibromatosis Type I (NF1 ). Despite its crucial role in modulating RAS activity, little is known about how NF1 gene expression or its function as a regulator of RAS is controlled by the cell. To investigate this crucial mechanism, we propose using high-throughput genetic screens to identify novel regulators of the neurofibromin-KRAS interaction in both healthy and cancerous cells, which could serve as therapeutic targets for both sporadic cancers and patients with NF1. To do so, we will utilize a novel protein-protein interaction assay to directly measure neurofibromin-KRAS binding in the context of genetic perturbations created through CRISPRi and CRISPRa technologies. We will then use biochemical, pharmacological, and physiological assays to determine the exact mechanism behind these regulators. This work represents the first example of high-throughput, unbiased genetic screens being applied to neurofibromin regulation and thus will fill a crucial gap in our understanding of neurofibromin and its role in controlling RAS signaling. Furthermore, this project has been tailored to provide me the training, knowledge, and resources I need to establish my independent research career at the interface of cancer biology and gene therapy.