Characterizing the role of RNF25 in repair of DNA alkylation in blood cancers

表征 RNF25 在血癌 DNA 烷基化修复中的作用

基本信息

批准号：
10580070
负责人：
David Paul Toczyski
金额：
$ 8.08万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
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：
2022-03-01 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10580070
关键词：
Acute Adenine Affect Alkylating Agents Alkylation Base Excision Repairs Biological Biological Process Busulfan CRISPR screen Carbon Cell Cycle Cell Line Cells Chemicals Chlorambucil Clinical Combined Modality Therapy Cyclophosphamide Cytoskeleton DNA DNA Alkylation DNA Damage DNA Methylation DNA Repair DNA lesion Deubiquitinating Enzyme Disease Dose Drug Targeting Excision Genes Glioblastoma Goals Guanine Hematopoietic Neoplasms In Vitro Lesion Ligase Lymphoma MGMT gene Malignant Neoplasms Mass Spectrum Analysis Mechlorethamine Mediating Melphalan Mesylates Methylation Mitochondria Mitosis Molecular Multiple Myeloma Mutation Pathway interactions Patients Pharmaceutical Preparations Phosphotransferases Poly(ADP-ribose) Polymerase Inhibitor Positioning Attribute Proteins Resistance Role Sampling Signal Pathway Signal Transduction Testing Therapeutic Therapeutic Uses Translations Ubiquitin Ultraviolet Rays Up-Regulation base chemotherapy conditioning crosslink fighting gene repair inhibitor member mutant prognostic indicator repaired response small molecule small molecule inhibitor targeted treatment temozolomide therapeutic DNA therapeutic target tumor ubiquitin isopeptidase ubiquitin ligase

Acute Adenine Affect Alkylating Agents Alkylation Base Excision Repairs Biological Biological Process Busulfan CRISPR screen Carbon Cell Cycle Cell Line Cells Chemicals Chlorambucil Clinical Combined Modality Therapy Cyclophosphamide Cytoskeleton DNA DNA Alkylation DNA Damage DNA Methylation DNA Repair DNA lesion Deubiquitinating Enzyme Disease Dose Drug Targeting Excision Genes Glioblastoma Goals Guanine Hematopoietic Neoplasms In Vitro Lesion Ligase Lymphoma MGMT gene Malignant Neoplasms Mass Spectrum Analysis Mechlorethamine Mediating Melphalan Mesylates Methylation Mitochondria Mitosis Molecular Multiple Myeloma Mutation Pathway interactions Patients Pharmaceutical Preparations Phosphotransferases Poly(ADP-ribose) Polymerase Inhibitor Positioning Attribute Proteins Resistance Role Sampling Signal Pathway Signal Transduction Testing Therapeutic Therapeutic Uses Translations Ubiquitin Ultraviolet Rays Up-Regulation base chemotherapy conditioning crosslink fighting gene repair inhibitor member mutant prognostic indicator repaired response small molecule small molecule inhibitor targeted treatment temozolomide therapeutic DNA therapeutic target tumor ubiquitin isopeptidase ubiquitin ligase

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项目摘要

Abstract Nitrogen mustards were the first chemotherapeutics used to fight cancer. These drugs, some of which are still in use today, are members of a large group of therapeutic DNA alkylating agents. Alkylating agents such as Melphalan, Busulfan, and Cyclophosphamide are currently employed for aggressive lymphoma, multiple myeloma and AML. While some of these agents modify bases with longer carbon chains, others generate simple methylations. For example, temozolomide, which is widely used for glioblastoma, methylates both adenine and guanine at several positions. The methylating agent MMS is a methane sulfonate chemically very similar to Busulfan. We have carried out an unbiased CRISPR screen to examine the sensitivity of deletions of each ubiquitin ligase and DUB to forty different small molecule inhibitors of a wide array of biological pathways, ranging from the cytoskeleton, to mitochondrial function to mitosis. We found that deletion of the poorly-studied ubiquitin ligase RNF25 rendered cells extremely sensitive to the DNA alkylating agent MMS, but not at all sensitive to forty other drugs, including other DNA damaging agents. Given the specific role of RNF25 in resistance to MMS, it is likely that RNF25 has a direct role in the response to, or repair of, DNA alkylation. We propose to explore the role of RNF25 in the repair of DNA alkylation. We will extend our findings, obtained in a CML-derived cell line, to other blood cancers. We will determine the rate at which alkylated bases are removed from DNA in RNF25 mutants and examine the spectrum of alkylation agents to which RNF25 mutants are sensitive. Finally, we will identify substrates of RNF25 to determine the mechanism by which it promotes resistance to these agents.

抽象的氮芥末是第一个用于抵抗癌症的化学治疗剂。这些药物，其中一些仍然是如今，是大量治疗性DNA烷基化剂的成员。烷基化剂，例如 Melphalan，Busulfan和环磷酰胺目前用于攻击性淋巴瘤，多个骨髓瘤和AML。这些代理中的一些用更长的碳链修饰基地，而另一些则产生简单的甲基化。例如，已广泛用于胶质母细胞瘤的替莫唑胺甲基化腺嘌呤和鸟嘌呤在多个位置。甲基化剂MMS是一种化学上的甲烷磺酸盐类似于Busulfan。我们进行了一个公正的CRISPR屏幕，以检查缺失的敏感性每个泛素连接酶和将多种生物途径的四十个不同的小分子抑制剂配音，从细胞骨架到线粒体功能到有丝分裂。我们发现删除不佳的人泛素连接酶RNF25使细胞对DNA烷基化剂MM极为敏感，但根本没有对其他四十种药物敏感，包括其他DNA损伤药物。鉴于RNF25在对MMS的抗性，RNF25可能在对DNA烷基化的反应或修复中具有直接作用。我们建议探索RNF25在DNA烷基化修复中的作用。我们将扩展我们的发现，以 CML衍生的细胞系，其他血液癌。我们将确定去除烷基化碱的速率从RNF25突变体中的DNA，检查RNF25突变体的烷基化剂光谱敏感的。最后，我们将确定RNF25的底物以确定其促进的机制对这些药物的抵抗力。