Title and Speaker Bio for Eurofins Discovery Chat: Connecting with Protein Degradation
Biosensor Development Using CRISPR to Quantify Endogenous Protein Modulated by Targeted Protein Degraders
Jane E. Lamerdin, Ph.D. Director, R&D, Eurofins DiscoverX
A cell-based assay platform that robustly and sensitively quantifies the kinetics of endogenous protein turnover is crucial for discovery of disease-relevant therapeutic agents. This need is particularly relevant for a new class of therapeutics known as targeted protein degraders. One example of such a molecule is the PROTAC, a bifunctional molecule that targets a specific disease-relevant protein for degradation by the cellular ubiquitin-proteasome system. Current technologies employed to evaluate target modulation with PROTACs include low throughput Westerns (for on-target activity) and cell proliferation assays. The ideal discovery platform should be sensitive, easy to use, and amenable to medium to high throughput screens.
In this presentation, we will describe how Eurofins DiscoverX has combined CRISPR/Cas9 technology with our well-established Enzyme Fragment Complementation (EFC) system in physiologically relevant cell backgrounds to quantify modulation of endogenous protein levels in response to therapeutic agents. To illustrate the approach, we will share a case study for development of biosensor cell lines in several cancer cell lines, including a blood cancer line, to characterize molecules that target BRD4.? Inhibition of BRD4 with small molecules has been shown to significantly reduce c-Myc expression in multiple cancer models (MM and CML), and PROTACs targeting BRD4 have also shown promise in pre-clinical models. We will present results from several biosensor lines generated with a panel of PROTACs and small molecules that target BRD4. Rank order data were consistent with previous reports for using other physiological endpoints, such as proliferation, but were achieved with a much shorter incubation time. ?Due to the simplicity of engineering and prosecuting the assays, as well as the physiological relevance of the cell models that can be used, this assay format should be amenable to discovery of new molecular entities that modulate protein levels of a diverse array of therapeutic targets.
(PROTAC is a registered trademark of Arvinas)
Dr. Jane Lamerdin is the Director of R&D at Eurofins DiscoverX where she currently oversees the development of novel cell-based assays and tools to support portfolio expansion as well as client-focused projects. She has nearly 20 years of industry experience developing and prosecuting diverse cell-based assays to support client drug discovery campaigns, and high throughput, molecular and systems biology research. Prior to joining Eurofins DiscoverX, Jane was Executive Director of Research at Odyssey Thera, where she utilized her expertise in cell signaling pathways, oncology and DNA repair to guide the development of a broad panel of high content cell-based assays for compound safety and selectivity screening, which supported programs in major pharma companies as well as the ToxCast program at the EPA. ?Jane received her B.S. and Ph.D. in Genetics from the University of California at Davis; she is con-inventor on over 5 patents and has co-authored over 50 peer-reviewed articles.