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

Understanding the molecular mechanism of a protein-recycling complex in small cell lung cancer treatment resistance.

了解蛋白质回收复合物在小细胞肺癌治疗耐药中的分子机制。

基本信息

批准号：
10466892
负责人：
Benjamin H Lok
金额：
$ 33.34万
依托单位：
UNIVERSITY HEALTH NETWORK
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-25 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10466892
关键词：
Amino Acid Sequence Antibodies BRCA mutations Behavior Biochemical Biological Biological Assay Biological Markers Biotin CRISPR library CRISPR screen CRISPR/Cas technology CUL1 gene Cancer Biology Cancer Etiology Cancer Patient Cancer cell line Cell Line Cellular Assay Cessation of life Cisplatin Clinical Clinical Research Clustered Regularly Interspaced Short Palindromic Repeats Co-Immunoprecipitations Code Complementary DNA Complex Custom DNA Damage DNA Repair DNA Repair Pathway Disease Disease Resistance Enzyme Inhibitor Drugs Enzymes Etoposide Exhibits F Box Domain F-Box Proteins FDA approved Gene Targeting Genes Genetic Screening Genomics Goals In Vitro Knock-out Laboratories Lead Libraries Malignant Neoplasms Malignant neoplasm of lung Mass Spectrum Analysis Mediating Mediator of activation protein Modeling Molecular Mutagenesis Mutate Mutation Non-Small-Cell Lung Carcinoma Pathway interactions Patient-Focused Outcomes Patients Pharmaceutical Preparations Pharmacotherapy Phase Poly(ADP-ribose) Polymerases Protein Cycling Pathway Proteins Radiation therapy Recurrent disease Recycling Regulation Relapse Reporter Reporting Resistance Role Structure Tertiary Protein Structure Therapeutic Ubiquitin Up-Regulation Validation Woman Work Xenograft procedure brca gene chemotherapy chromosome 2p loss functional genomics genome-wide homologous recombination improved improved outcome in vivo inhibitor insight lung small cell carcinoma men multicatalytic endopeptidase complex novel novel therapeutics patient derived xenograft model pre-clinical recombinational repair recruit relapse patients resistance mechanism response restoration small hairpin RNA therapeutic development therapy resistant treatment response tumor ubiquitin-protein ligase vector

项目摘要

PROJECT SUMMARY Small cell lung cancer (SCLC) patients have an initial robust response to combinations of DNA damaging agents (e.g. cisplatin, etoposide, radiotherapy), however, many patients inevitably suffer from relapse and resistant disease. A clear understanding of these resistance mechanisms remains elusive. Consequently, there is a critical need to: (1) understand the mechanisms of therapeutic resistance and (2) develop novel therapeutics. Poly-(ADP)-ribose polymerase enzymes (PARP) protein levels are upregulated in SCLC relative to other lung cancers, and initial studies suggest that this upregulation is associated with increased sensitivity of SCLC to PARP inhibitors (PARPi) in vitro. PARP inhibitors are synthetic lethal with BRCA1/2 mutated homologous recombination (HR) deficient tumors and restoration of HR by BRCA reversion mutations is a known mechanism of PARPi and cisplatin resistance. However, as BRCA1/2 mutations are exceedingly rare in SCLC non-BRCA mechanisms must be operant. We performed a genome-wide CRISPR knockout screen to identify novel mechanisms of PARPi resistance. From subsequent functional validation and clinical genomic correlation, we identified deficiency in an F-box protein coding gene as a putative biomarker of resistance to PARP inhibitors and cisplatin in SCLC that may be present in up to ~20% of relapse patient tumors. Loss of this F-box protein abrogates the function of its corresponding SKP1, CUL1, F-box (SCF) E3 ubiquitin ligase complex. By proximity-dependent biotin identification (BioID) of this F-box protein, we have identified a high confidence interactor with substrate-like behavior for SCF-mediated ubiquitin-proteasomal degradation that is important for regulation of HR and DNA repair. This proposal aims to: (1) determine the mechanism of this specific SCF complex with its substrate to engage the ubiquitin-proteasome pathway; (2) elucidate the impact of this F-box protein on HR, DNA repair, and therapeutic sensitivity to PARPi/cisplatin; and (3) identify synthetic lethal interactions with deficiencies in this F-box protein to provide biologic insight and characterize immediately translatable approaches for relapsed treatment resistant SCLC.

项目概要小细胞肺癌 (SCLC) 患者对 DNA 损伤的组合有最初的强烈反应然而，许多患者不可避免地会复发并接受药物治疗（例如顺铂、依托泊苷、放射治疗）。抗病。对这些耐药机制的清晰理解仍然难以捉摸。最后，迫切需要：（1）了解治疗耐药的机制，（2）开发新的疗法。相对于其他肺组织，SCLC 中聚 (ADP)-核糖聚合酶 (PARP) 蛋白水平上调癌症，初步研究表明这种上调与 SCLC 对癌症的敏感性增加有关 PARP 抑制剂 (PARPi) 体外。 PARP抑制剂与BRCA1/2突变同源物合成致死重组（HR）缺陷型肿瘤和通过 BRCA 回复突变恢复 HR 是已知的 PARPi 和顺铂耐药机制。然而，由于 BRCA1/2 突变在 SCLC 非 BRCA 机制必须有效。我们进行了全基因组 CRISPR 敲除筛选，以确定 PARPi 抗性的新机制。根据随后的功能验证和临床基因组相关性，我们发现了 F-box 的缺陷蛋白质编码基因作为 SCLC 中 PARP 抑制剂和顺铂耐药性的假定生物标志物，可能存在于高达约 20% 的复发患者肿瘤中。该 F-box 蛋白的丢失会消除其功能相应的 SKP1、CUL1、F-box (SCF) E3 泛素连接酶复合物。通过邻近依赖性生物素对该 F-box 蛋白的鉴定（BioID），我们已经鉴定出具有底物样的高置信度相互作用子 SCF 介导的泛素蛋白酶体降解行为对于 HR 和 DNA 的调节非常重要维修。该提案的目的是：（1）确定这种特定的 SCF 复合物及其底物参与的机制泛素-蛋白酶体途径； (2) 阐明该 F-box 蛋白对 HR、DNA 修复和对 PARPi/顺铂的治疗敏感性； (3) 识别合成致死相互作用及其缺陷 F-box 蛋白可提供生物学见解并表征可立即转化的复发方法治疗难治性 SCLC。