Using Analog-Sensitive (AS) Kinase Technology to Engineer PROTAC Targeted CDK Degraders
Faculty Mentor
Joseph Morin
Major/Area of Research
Cancer Biology, Molecular Biology
Description
We studied a chemical-genetic approach to dissect the networks of cyclin-dependent kinases (CDKs) and their targets in cell division and gene expression in human cancer cells. We focused on a human cancer cell line that expresses only a mutant version of a particular CDK. It has been engineered to be analog-sensitive (AS) or susceptible to inhibition by bulky purine analogs that do not interfere with the functions of wild-type kinases. By altering a single kinase in a cell, we can precisely determine the roles and targets of it. We can identify the sequence of pathways containing several CDKs and find genetic relationships between them by combining mutations in numerous enzymes. Analog-sensitive kinase technology enables the fast, reversible, and highly selective inhibition of engineered kinases in human cancer cells. Enhancements of both potency and selectivity for the AS kinase inhibitors are achieved with degraders, also known as proteolysis-targeting chimeras (PROTACs), which target proteins of interest for selective destruction. In this approach, a small-molecule ligand of the protein being targeted, such as a kinase, is conjugated to a functional group that binds specifically to an E3 ubiquitin ligase; when the target is engaged in cells, this causes the protein to be ubiquitinated and degraded by the proteasome. We aim to pinpoint specific CDK complexes and their precise roles and functions in controlling the cell cycle and transcription. We also aim to identify possible sensitivities of cancer cells to CDK inhibitors by using the chemical genetic study of the CDK network.
Using Analog-Sensitive (AS) Kinase Technology to Engineer PROTAC Targeted CDK Degraders
We studied a chemical-genetic approach to dissect the networks of cyclin-dependent kinases (CDKs) and their targets in cell division and gene expression in human cancer cells. We focused on a human cancer cell line that expresses only a mutant version of a particular CDK. It has been engineered to be analog-sensitive (AS) or susceptible to inhibition by bulky purine analogs that do not interfere with the functions of wild-type kinases. By altering a single kinase in a cell, we can precisely determine the roles and targets of it. We can identify the sequence of pathways containing several CDKs and find genetic relationships between them by combining mutations in numerous enzymes. Analog-sensitive kinase technology enables the fast, reversible, and highly selective inhibition of engineered kinases in human cancer cells. Enhancements of both potency and selectivity for the AS kinase inhibitors are achieved with degraders, also known as proteolysis-targeting chimeras (PROTACs), which target proteins of interest for selective destruction. In this approach, a small-molecule ligand of the protein being targeted, such as a kinase, is conjugated to a functional group that binds specifically to an E3 ubiquitin ligase; when the target is engaged in cells, this causes the protein to be ubiquitinated and degraded by the proteasome. We aim to pinpoint specific CDK complexes and their precise roles and functions in controlling the cell cycle and transcription. We also aim to identify possible sensitivities of cancer cells to CDK inhibitors by using the chemical genetic study of the CDK network.