权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Screening for Cys-Reactive Ligands to Target PAX3-FOXO1

筛选针对 PAX3-FOXO1 的 Cys 反应性配体

基本信息

批准号：
10611002
负责人：
CHRISTOPHER M COUNTER
金额：
$ 36.94万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-01 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10611002
关键词：
Alveolar Rhabdomyosarcoma Binding Biochemical Biological Assay Biology Cell Line Cells Chemicals Chimeric Proteins Chromosomal translocation Collaborations Critical Pathways DNA Dana-Farber Cancer Institute Data Development Disease Drug Targeting Epitopes FOXO1A gene Family member Fire - disasters Future Genetic Genetic Transcription Goals Grant Individual Laboratories Length Libraries Ligands Luciferases Malignant Childhood Neoplasm Mass Spectrum Analysis Mediator of activation protein Modification Monitor Oncogenic Oxidation-Reduction PAX3 gene Palate Parents Patient-Focused Outcomes Pharmaceutical Preparations Production Proteins Reporter Scientist Secondary to Site Structure Transactivation Transcript Translational Research Trypsin Validation Work base candidate identification childhood sarcoma cofactor design drug candidate drug discovery experience genetic approach innovation member multidisciplinary mutant novel therapeutics protein complex screening sensor small molecule stoichiometry therapeutic target transcription factor

项目摘要

Supplemental application U54 CA231630 PI: Angela Koehler, Co-PIs: Sara Buhrlage, Jarrod Marto Abstract Alveolar rhabdomyosarcoma (ARMS) is a lethal pediatric cancer driven by the oncogenic transcription factor fusion protein PAX3-FOXO1 (P3F). While genetic data demonstrate P3F is a potential therapeutic target, transcription factors are often considered challenging pharmalogical targets. The goal of this U54 supplement is to expand drug identification strategies past our current small-molecule microarrays (SMMs) and implement covalent ligand screening against P3F protein Cys residues, thus increasing the pool of potential drug candidates. To achieve this goal, team will collaborate to pursue three objectives. First, Dr. Koehler’s team will increase production of purified full-length P3F as well as truncated forms of the protein which contain Cys-793, a residue identified as a critical mediator of cofactor CBP. Second, Dr. Koehler’s team will work with Drs. Buhrlage and Marto to optimize screening conditions for a full cys-reactive library utilizing a pooling-approach to increase throughput. Individual hits will be validated through LC-MS/MS to confirm biochemical results, such as covalent modifications and stoichiometry. Lastly, covalent ligand screen hits will be subjected to secondary assays, such as cellular thermal shift or reporter assays, outlined in the parent U45 for SMM drug discovery. This collaboration between Dr. Koehler and Drs. Buhrlage and Marto will expand the U54 team’s drug discovery capabilities, and enable a more thorough and efficient assessment of candidate drugs to target P3F. In the future, this technological approach could be extended to other transcription factors for drug candidate identification.

补充申请U54 CA231630 PI: Angela Koehler， co -PI: Sara Buhrlage, Jarrod Marto