Exquisitely Specific anti-KRAS Biodegraders Inform on the Cellular Prevalence of Nucleotide-Loaded States
| Autor: | Anthony W. Partridge, Simon Ng, Brian Henry, Khong Ming Peh, Jinkai Teo, Yu-Chi Juang, Pooja Gopal, Constance Yeo, Shuhui Lim, Huibin Zhang, Regina Khoo |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
chemistry.chemical_classification
Gene isoform 010405 organic chemistry General Chemical Engineering Mutant Context (language use) General Chemistry Computational biology Biology 010402 general chemistry medicine.disease_cause 01 natural sciences Fusion protein Small molecule 0104 chemical sciences Ubiquitin ligase Chemistry chemistry biology.protein medicine Nucleotide KRAS QD1-999 Research Article |
| Zdroj: | ACS Central Science ACS Central Science, Vol 7, Iss 2, Pp 274-291 (2020) |
| ISSN: | 2374-7951 2374-7943 |
| Popis: | Mutations to RAS proteins (H-, N-, and K-RAS) are among the most common oncogenic drivers, and tumors harboring these lesions are some of the most difficult to treat. Although covalent small molecules against KRASG12C have shown promising efficacy against lung cancers, traditional barriers remain for drugging the more prevalent KRASG12D and KRASG12V mutants. Targeted degradation has emerged as an attractive alternative approach, but for KRAS, identification of the required high-affinity ligands continues to be a challenge. Another significant hurdle is the discovery of a hybrid molecule that appends an E3 ligase-recruiting moiety in a manner that satisfies the precise geometries required for productive polyubiquitin transfer while maintaining favorable druglike properties. To gain insights into the advantages and feasibility of KRAS targeted degradation, we applied a protein-based degrader (biodegrader) approach. This workflow centers on the intracellular expression of a chimeric protein consisting of a high-affinity target-binding domain fused to an engineered E3 ligase adapter. A series of anti-RAS biodegraders spanning different RAS isoform/nucleotide-state specificities and leveraging different E3 ligases provided definitive evidence for RAS degradability. Further, these established that the functional consequences of KRAS degradation are context dependent. Of broader significance, using the exquisite degradation specificity that biodegraders can possess, we demonstrated how this technology can be applied to answer questions that other approaches cannot. Specifically, application of the GDP-state specific degrader uncovered the relative prevalence of the “off-state” of WT and various KRAS mutants in the cellular context. Finally, if delivery challenges can be addressed, anti-RAS biodegraders will be exciting candidates for clinical development. Biodegraders, engineered proteins that fuse a high-affinity binder (e.g., RBD, K27, R11.1.6, and NS1) to an E3 ligase (e.g., SPOP), prompt degradation of RAS and reveal novel biological insights. |
| Databáze: | OpenAIRE |
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