Functional Analysis of Structure Specific Nucleases in Genome Stability and Cancers

结构特异性核酸酶在基因组稳定性和癌症中的功能分析

• 批准号: 10491113
• 负责人: LI ZHENG
• 金额: $ 37.74万
• 依托单位: BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-20 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10491113
• 关键词: Androgen Receptor; Award; Biochemical; Cells; Cellular Assay; Centromere; Chemoprevention; Cities; Collaborations; Complex; DNA; DNA Ligation; DNA Repair; DNA Structure; DNA biosynthesis; Deposition; Development; Etiology; Event; Funding; Future; Gene Mutation; Genes; Genome Stability; Genomic Instability; Goals; Histones; Human; Institution; Knockout Mice; Knowledge; Laboratories; Lead; Malignant Neoplasms; Malignant neoplasm of prostate; Mediating; Metabolism; Molecular; Mutation; Okazaki fragments; Peer Review; Pharmaceutical Preparations; Phosphodiesterase I; Play; Post-Translational Protein Processing; Process; Prostate; Publications; RTH-1 Nuclease; Regulation; Research; Research Institute; Resources; Role; Science; Scientist; Specialist; Structure; Technology; Time; chemotherapy; cohesion; experience; gene repair; genome integrity; helicase; mouse genetics; mutant mouse model; next generation sequencing; nuclease; programs; repaired; replication stress; telomere

Abstract The proposed R50 award will support my critical contributions to three cohesive research programs conducted in the laboratory of Dr. Binghui Shen at the Beckman Research Institute of City of Hope: NCI R01 CA073764 (1997-2022), NCI R01 CA085344 (1999-2024), and NCI R01 CA233664 (2019-2024). Various DNA intermediates are formed during DNA metabolic processes, and these structures must be properly and efficiently processed in a timely manner to avoid severe genomic instability and cancer development. This process requires structure-specific nucleases to specifically recognize and cleave certain forms of DNA structures for further processing and eventual DNA ligation into an intact DNA duplex. Over the last 20 years, I have supported the Shen laboratory in investigating the structure, function, and regulation of structure-specific nucleases typified by flap endonuclease 1 (FEN1) and DNA2 nuclease/helicase, as well as how their functional deficiency causes genome instabilities and cancer development. More recently, our research has expanded to characterizing the role of exonuclease 5 (EXO5) in resecting androgen receptor-induced double-strand breaks in prostate cancer. As the senior research scientist in the Shen laboratory, I have leveraged my expertise in biochemical and cellular assays of DNA replication and repair, mouse genetics, and cutting-edge next-generation sequencing technologies to continuously contribute to breakthrough findings that advance Dr. Shen’s research programs. Since I joined Dr. Shen’s group in 2001, I have co-authored 50 peer-reviewed publications, including 17 peer- reviewed articles resulting from my R50-sponsored research in the previous funding period (2016-2021). Currently, I am supporting the CA073764 program by defining post-translational modifications (PTMs) of FEN1 and the dynamic interaction between Okazaki fragment maturation (OFM) and histone deposition machinery as key regulatory mechanisms for efficient and accurate FEN1-mediated OFM. The current goal of the CA085344 program is to define the roles of human DNA2 in counteracting replication stress. In support of this, I will lead a team to determine how human DNA2 complexes process G4 and other non-B-form DNA structures to facilitate DNA replication at telomeres and centromeres, using DNA2 mutant mouse models. I am also directing a portion of the proposed research in the CA233664 program, which aims to define the etiological role of homology- directed repair (HDR) gene mutations in prostate cancer. I created knockout mice for the EXO5 gene, which we recently discovered to play a crucial role in HDR in prostate cells. I will lead the effort to define the causal role of EXO5 in prostate cancer and reveal the underlying molecular events. These studies will leverage my knowledge and experience accumulated during my previous research in FEN1 and DNA2. As an NCI-sponsored research specialist, I have and will continue to maximize my contribution to science by leveraging the resources and close intellectual collaborations available in an established laboratory within a resource-rich institution.

摘要 拟议的R50奖将支持我对三个有凝聚力的研究项目的关键贡献， 在希望之城贝克曼研究所沈炳辉博士的实验室中：NCI R 01 CA 073764 （1997-2022）、NCI R 01 CA 085344（1999-2024）和NCI R 01 CA 233664（2019-2024）。各种DNA 中间体在DNA代谢过程中形成，这些结构必须正确有效地 及时处理，以避免严重的基因组不稳定性和癌症发展。这个过程需要 结构特异性核酸酶特异性识别和切割某些形式的DNA结构， 加工并最终连接成完整的DNA双链体。在过去的20年里，我一直支持 在研究结构特异性核酸酶的结构、功能和调节方面， 瓣状核酸内切酶1（FEN 1）和DNA 2核酸酶/解旋酶，以及它们的功能缺陷如何导致 基因组不稳定性和癌症发展。最近，我们的研究已经扩展到表征 外切核酸酶5（EXO 5）在切除前列腺癌雄激素受体诱导的双链断裂中的作用。 作为沈实验室的高级研究科学家，我利用我在生物化学和细胞学方面的专业知识， DNA复制和修复、小鼠遗传学和尖端的下一代测序分析 技术不断为推动沈博士研究计划的突破性发现做出贡献。 自从我2001年加入沈博士的团队以来，我已经合著了50篇同行评议的出版物，其中包括17篇同行评议的出版物。 回顾了我在上一个资助期（2016-2021年）的R50赞助研究的文章。 目前，我正在通过定义FEN 1的翻译后修饰（PTM）来支持CA 073764计划 冈崎片段成熟（OFM）和组蛋白沉积机制之间的动态相互作用， FEN 1介导的OFM的有效和准确的关键调控机制。CA 085344的当前目标 该项目的目的是确定人类DNA 2在对抗复制应激中的作用。为了支持这一点，我将领导一个 研究小组确定人类DNA 2复合物如何处理G4和其他非B型DNA结构，以促进 端粒和着丝粒的DNA复制，使用DNA 2突变小鼠模型。我也在导演一部 CA 233664计划中拟议的研究，旨在定义同源性的病因学作用- 定向修复（HDR）基因突变在前列腺癌。我创造了EXO 5基因敲除小鼠， 最近发现在前列腺细胞的HDR中起关键作用。我将领导努力定义的因果作用， EXO 5在前列腺癌中的作用，并揭示潜在的分子事件。这些研究将利用我的知识 和我以前在FEN 1和DNA 2研究中积累的经验。作为一项国家癌症研究所赞助的研究， 作为一名专家，我已经并将继续通过利用资源和密切合作来最大限度地发挥我对科学的贡献。 在资源丰富的机构内建立的实验室中提供智力合作。