التفاصيل البيبلوغرافية
العنوان:
The SMARCA2/4 ATPase Domain Surpasses the Bromodomain as a Drug Target in SWI/SNF-Mutant Cancers: Insights from cDNA Rescue and PFI-3 Inhibitor Studies
المؤلفون:
Xi Shi , Alessia Petrocchi , Shikhar Sharma , Lisa Nottebaum , Jannik N. Andersen , Elisabetta Leo , Maria Kost-Alimova , Timothy P. Heffernan , Philip Jones , Bhavatarini Vangamudi , Trang N. Tieu , Dominique Verhelle , Harshad S. Mahadeshwar , Parantu K. Shah , Dafydd R. Owen , Andrew Futreal , Mike Peoples , Giulio Draetta , Thomas A Paul , Yanai Zhan , Wylie S. Palmer , Joseph R. Marszalek , Carlo Toniatti , Alexei Protopopov
المصدر:
Cancer research . 75(18)
سنة النشر:
2015
مصطلحات موضوعية:
Cancer Research , DNA, Complementary , Lung Neoplasms , Chromosomal Proteins, Non-Histone , Pyridines , Protein domain , Biology , Binding, Competitive , Catalysis , Article , Gene Knockout Techniques , Sarcoma, Synovial , RNA interference , Cell Line, Tumor , Neoplasms , Humans , Molecular Targeted Therapy , RNA, Small Interfering , Rhabdoid Tumor , Gene knockdown , Genetic Complementation Test , DNA Helicases , Nuclear Proteins , Chromatin Assembly and Disassembly , Microarray Analysis , SWI/SNF , Chromatin , Bromodomain , Neoplasm Proteins , Protein Structure, Tertiary , Oncology , SMARCA4 , Cancer research , RNA Interference , Chromatin immunoprecipitation , Azabicyclo Compounds , Transcription Factors
الوصف:
The SWI/SNF multisubunit complex modulates chromatin structure through the activity of two mutually exclusive catalytic subunits, SMARCA2 and SMARCA4, which both contain a bromodomain and an ATPase domain. Using RNAi, cancer-specific vulnerabilities have been identified in SWI/SNF-mutant tumors, including SMARCA4-deficient lung cancer; however, the contribution of conserved, druggable protein domains to this anticancer phenotype is unknown. Here, we functionally deconstruct the SMARCA2/4 paralog dependence of cancer cells using bioinformatics, genetic, and pharmacologic tools. We evaluate a selective SMARCA2/4 bromodomain inhibitor (PFI-3) and characterize its activity in chromatin-binding and cell-functional assays focusing on cells with altered SWI/SNF complex (e.g., lung, synovial sarcoma, leukemia, and rhabdoid tumors). We demonstrate that PFI-3 is a potent, cell-permeable probe capable of displacing ectopically expressed, GFP-tagged SMARCA2-bromodomain from chromatin, yet contrary to target knockdown, the inhibitor fails to display an antiproliferative phenotype. Mechanistically, the lack of pharmacologic efficacy is reconciled by the failure of bromodomain inhibition to displace endogenous, full-length SMARCA2 from chromatin as determined by in situ cell extraction, chromatin immunoprecipitation, and target gene expression studies. Furthermore, using inducible RNAi and cDNA complementation (bromodomain- and ATPase-dead constructs), we unequivocally identify the ATPase domain, and not the bromodomain of SMARCA2, as the relevant therapeutic target with the catalytic activity suppressing defined transcriptional programs. Taken together, our complementary genetic and pharmacologic studies exemplify a general strategy for multidomain protein drug-target validation and in case of SMARCA2/4 highlight the potential for drugging the more challenging helicase/ATPase domain to deliver on the promise of synthetic-lethality therapy. Cancer Res; 75(18); 3865–78. ©2015 AACR.
تدمد:
1538-7445
URL الوصول:
https://explore.openaire.eu/search/publication?articleId=doi_dedup___::b46a37cc723ee28ec2ca78b006ca9829 https://pubmed.ncbi.nlm.nih.gov/26139243
حقوق:
OPEN
رقم الأكسشن:
edsair.doi.dedup.....b46a37cc723ee28ec2ca78b006ca9829
قاعدة البيانات:
OpenAIRE
