Aim of the work
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CHAPTER 2: AIM OF THE WORK
Hereditary breast cancer is associated with specific mutations in BRCA1 tumor suppressor gene. In BRCA1 mutation carriers, cancer is generally triggered by full inactivation of BRCA activity throughout subsequent somatic mutations of the wild-type allele. Requirement of total inactivation of BRCA for insurgence of the tumor justifies the design of preventive strategies to restore functional activity or replace the inactive gene. From this ground the aim of this thesis was to develop a novel approach to insert BRCA1 into mutated cells as a preventative strategy against breast cancer. On this purpose BRCA1 was transduced in target cells using a lentiviral vector derived from FIV, one of the most efficient tools for gene therapy. Indeed, integration of proviral genome in host cell DNA ensures long-term expression of the heterologous gene (transgene), an essential feature for permanent restoration of gene effectiveness. In this work BRCA1 gene was cloned into a FIV vector previously produced in our laboratory and optimized for heterologous cell transduction of different species. The FIV-BRCA vector was evaluated for its ability to transduce HCC1937, a tumor cell line with a mutated BRCA. The BRCA1 was successfully expressed in transduced cells leading to full restoration of gene activity.
