CAREER: Analyzing and engineering multi-gene cooperation to generate cell lines for therapeutic protein production

Stanford University, Stanford CA

Investigators

Abstract

This award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5). 0846392 Wang Intellectual Merit: As cell biologists, geneticists, or engineers, one routinely over-expresses a gene in a cell, observes the cell, and compares its behavior to the parent. Yet multiple genes, not one, often determine cell phenotypes. One example is the sustained proliferation characteristic of immortalized mammalian cells. A mutation that changes the expression of a single gene or activity of a single protein rarely suffices to immortalize a cell; instead, multiple genes generally need to be mutated. This project investigates how multiple genes contribute to the phenotypes of immortalized cells: proliferation, apoptosis, senescence, and serum-independent growth. This project also aims to determine how different gene expression levels affect these phenotypes. In other words, the experiments address both combinatorial and dose effects. Broader Impact: This technology provides a powerful tool to generate cell lines with properties optimized for the production of therapeutic proteins. Using this experimental and analytical platform, this project will generate cell lines that exhibit high proliferation rates in low serum or serum-free media. Expression of various gene combinations will be evaluated for the ability to improve or create cell lines. Gene candidates include known oncogenes, tumor suppressor genes, and cell cycle regulators, as well as genes identified previously by retroviral mutagenesis in mice. The long-term goal is to generate human plasma B cell lines for the production of monoclonal antibodies. Faculty, graduate students, and undergraduates from the Department of Chemical Engineering at Stanford University will adopt a class of freshmen at Cupertino High School. This class is enrolled in a program named AVID (Advancement via Individual Determination). The program identifies students with poor class performance, but good attendance and minimal disciplinary problems. The program has no selection criteria based on race or gender, although the majority of students are economically disadvantaged, underrepresented minorities who will be the first in their families to attend college. The four-year program aims to help these students graduate from high school and go to college.