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Understanding Kinase Target Engagement in Live Cells

This webinar is Session 3 of a 4-Part Discover Glo Webinar Series

Discover Glo Webinar Series

Series Summary:

Over the last 30 years, assays and technologies utilizing the natural phenomenon of bioluminescence have gone from novel research tools to foundational elements of the modern research toolbox. Innovation utilizing bioluminescent approaches continues, further expanding both the available methods and the scientific advancements made by life science and drug discovery researchers. In this webinar series you will hear from industry experts and Promega scientists who will share research updates on important research topics included Kinases, GPCRs, and Targeted Protein Degradation. You will also learn about recent technology advancements that can be applied to these as well as other drug targets and research areas.

Session 3 - Understanding Kinase Target Engagement in Live Cells

Summary

Kinases are among the most successful targets of small molecule therapeutics.  Despite growing success and interest in drugging this broad enzyme family, selective engagement of individual kinases in cells represents a significant challenge. Kinase inhibition and selectivity is routinely quantified using purified kinase domains in cell-free assay formats, which unfortunately often fail to reflect the complex environment where kinase engagement would naturally occur.  In cells, the composite effects of dynamic multiprotein complexes and target activation state can have a pronounced impact on kinase engagement pharmacology.  The dynamics present in the cellular milieu can be challenging to simulate in a cell free system. To more accurately query kinase target engagement, we have developed panels of NanoBRET assays that measure compound binding against this protein family in live cells.  This method can be adapted to conditionally monitor target engagement at selected protein-protein interactions.  Here we explore the landscape of target engagement to a number of kinase subfamilies that function in multiprotein complexes, including cyclin dependent kinases (CDKs) as well as RAF (BRAF/CRAF/ARAF) kinases.  We uncover a surprising spectrum of engagement against some dark (lesser studied) family members. Our intracellular selectivity analysis reveals both liabilities as well as key opportunities for drug repurposing. 


Other webinars in this series:

Bioluminescence: Past, Present, Future

Applications of CRISPR Knock-in Tagging for Studying Endogenous Protein Dynamics

Advances in Targeted Protein Degradation


Speakers

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Dr. Susanne Mueller-Knapp
COO, Director of Operations of the Chemical Probes Portal
Structural Genomics Consortium, Frankfurt

Susanne Müller-Knapp studied Human Biology in Marburg Germany followed by a PhD in molecular biology at the Karolinska Institute in Stockholm, Sweden (1997). She then had more than 6 years of postdoctoral training in the area of inflammation and gene regulation at the Karolinska Institute and at the DIBIT San Raffaele Scientific Institute in Milan, Italy.

In 2004 Susanne joined the Structural Genomics Consortium, SGC, in Oxford. The SGC is an international public private partnership that currently comprises 8 international pharmaceutical companies and a large network of academic and industrial collaborators. Susanne worked at the SGC first as External Research Manager and then Scientific Coordinator. She has been the Project Manager of the Epigenetic Probe Project, which generates tool compounds with defined specificity and selectivity for epigenetic targets and the cell based assay group at the SGC in Oxford testing the cellular activity of the in vitro characterised tool compounds. In her role as Chief Operating Officer at the SGC Frankfurt Susanne is now coordinating several probe programs including the global SGC kinase chemical probe program and the donated probe program, which makes probes available from the pharmaceutical partners of the SGC. She is also Director of Operations of the Chemical Probes Portal, an online platform, providing e recommendations for the right choice and use of chemical probes.

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Dr. Benedict-Tilman Berger
Team leader Cellular Assays
Structural Genomics Consortium, Frankfurt

Benedict-Tilman Berger studied Biochemistry at the Freie Universität Berlin. Both his Bachelor thesis (Bayer HealthCare, Department of Screening) and his Master thesis (SGC Oxford, Department of Protein Kinases and Chemical/Structural Biology) focused on binding kinetics of protein kinase inhibitors and possible structural explanations for a slow binding of compounds, which might be a parameter for drug optimization in terms of selectivity, safety or dosing intervals. In 2016, he joined the group of Prof. Stefan Knapp at the SGC site in Frankfurt am Main to work on his PhD thesis that he finished in 2020. He was following up on his previous work on kinases, using more advanced techniques, e.g., the NanoBRET cellular target engagement assay, to explain in vitro and in vivo binding kinetics of kinase-ligand interactions on a structural level. Currently, he is contributing to a consortium (SFB 1399) that aims to identify new epigenetic targets in small cell lung cancer with cellular screening and helps setting up a kinome-wide NanoBRET screening platform.

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Dr. Matthew Robers
Senior Research Scientist
Promega Corporation

Matthew Robers is a Senior Research Scientist and Group Leader at Promega Corporation.  Matthew received post-graduate training at University of Wisconsin-Madison, studying iron-sulfur cluster enzymes in bacteria. Following graduate school, Matthew emphasized technology development for cellular pathway analysis at Life Technologies (Invitrogen), where he led a platform to quantify post-translational modifications in live cells.  Since joining Promega, Matthew has built a team focused on the development of new technologies to assess intracellular target occupancy, and has developed biophysical techniques for quantifying compound affinity and engagement kinetics within intact cells.  Matthew’s target engagement team is aggressively expanding the NanoBRET platform for complex intracellular targets in oncology, inflammation, and CNS disease.