Targeting oncoproteins with a positive selection assay for protein degraders.
Vidyasagar KoduriLeslie DuplaquetBenjamin L LampsonAdam C WangAmin H SabetMette IshoeyJoshiawa PaulkMingxing TengIsaac S HarrisJennifer E EndressXiaoxi LiuEthan DasilvaJoao A PauloKimberly J BriggsJohn G DoenchChristopher J OttTinghu ZhangKatherine Aleisha DonovanEric S FischerSteven P GygiNathanael S GrayJames E BradnerJeffrey A MedinSara J BuhrlageMatthew G OserWilliam G KaelinPublished in: Science advances (2021)
Most intracellular proteins lack hydrophobic pockets suitable for altering their function with drug-like small molecules. Recent studies indicate that some undruggable proteins can be targeted by compounds that can degrade them. For example, thalidomide-like drugs (IMiDs) degrade the critical multiple myeloma transcription factors IKZF1 and IKZF3 by recruiting them to the cereblon E3 ubiquitin ligase. Current loss of signal ("down") assays for identifying degraders often exhibit poor signal-to-noise ratios, narrow dynamic ranges, and false positives from compounds that nonspecifically suppress transcription or translation. Here, we describe a gain of signal ("up") assay for degraders. In arrayed chemical screens, we identified novel IMiD-like IKZF1 degraders and Spautin-1, which, unlike the IMiDs, degrades IKZF1 in a cereblon-independent manner. In a pooled CRISPR-Cas9-based screen, we found that CDK2 regulates the abundance of the ASCL1 oncogenic transcription factor. This methodology should facilitate the identification of drugs that directly or indirectly degrade undruggable proteins.
Keyphrases
- transcription factor
- high throughput
- acute lymphoblastic leukemia
- crispr cas
- dna binding
- multiple myeloma
- genome editing
- cancer therapy
- genome wide identification
- single cell
- air pollution
- emergency department
- gene expression
- ionic liquid
- small molecule
- reactive oxygen species
- binding protein
- drug induced
- microbial community
- cell proliferation
- bioinformatics analysis