Creation of AlkB Mutants for Protection of Bone Marrow

创建用于保护骨髓的 AlkB 突变体

基本信息

批准号：
7463139
负责人：
Manel Camps
金额：
$ 13.25万
依托单位：
UNIVERSITY OF CALIFORNIA SANTA CRUZ
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-07-01 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7463139
关键词：
Address Adenine Adverse effects Alkylating Agents Alkylation Alleles Animal Model Bacteriophage M13 Base Excision Repairs Base Pairing Biological Assay Blood Blood Cells Bone Marrow Bone Marrow Suppression Brain Neoplasms Bypass Camping Carmustine Catalysis Cell Line Cells Charge Clinic Clinical Clinical Trials Collaborations Coupled DNA DNA Adducts DNA Modification Process Dacarbazine Dose Dose-Limiting Enzymes Escherichia coli Evolution Excision Figs - dietary Generations Genes Genetic Materials Genomics Goals Guanine Hematopoietic Hemorrhage Homologous Gene Human Human Cell Line Infection Insecta Lesion Libraries Lymphoma Mammalian Cell Marrow Mediating Methylation Modeling Modification Mus Mutagenesis Mutation Nitrogen O(6)-Methylguanine-DNA Methyltransferase Performance Pharmaceutical Preparations Positioning Attribute Procarbazine Property Proteins Purines Pyrimidine Pyrimidines Reaction Relative (related person)Resistance Single-Stranded DNA Site Soft Tissue Neoplasms Solid Stem cells Substrate Specificity Suicide System Testing Toxic effect Training Translations Universities Variant Vascular blood supply Veterinarians Washington Work adduct base cancer therapy clinically relevant comparative cytotoxic cytotoxicity design directed evolution ds-DNA enzyme pathway gene therapy improved in vivo killings lexitropsin melanoma methyl group methyl lexitropsin methylpurine mouse model mutant neoplastic cell preference protective effect purine repaired temozolomide tissue culture

项目摘要

DESCRIPTION (provided by applicant): Methylating agents are mainstay drugs for the treatment of cancer and are amongst the few agents available against brain tumors. Cells responsible for keeping the blood cell supply, "blood progenitor cells", are highly sensitive to these agents, which results in severe side-effects, including bleeding and infection. Blood progenitor cells can be protected from the toxic effects of methylation by resistance genes. The most effective one is AGT, which directly removes toxic modifications caused by methylation of the cell's genetic material (DNA). However, AGT does not recognize certain toxic DNA modifications. Here I propose the use of a different gene (AlkB), which targets some of these alternative lesions, to improve protection against the toxic effects of methylation. This gene, which has only recently been characterized, should therefore enhance AGT protection. Moreover, unlike AGT, AlkB remains active after mediating a repair reaction, which implies that AlkB repair should be more amenable to modulation. Here I propose a powerful approach to identify variants of one human homologue of AlkB (ABH2) with improved repair performance. Specifically, I want to enhance the enzyme's ability to recognize lesions generated by agents used in the clinic. Selected variants will be tested in human progenitor cells in culture and in a mouse animal model for protection against treatment with high doses of these agents. Ultimately, the mutants identified in this work should improve the management of side effects of methylating agents and would also allow the use of higher doses for more effective treatment.

描述（由申请人提供）：甲基化剂是用于治疗癌症的主要药物，并且是针对脑肿瘤的少数药物之一。负责保持血细胞供应的细胞“血祖细胞”对这些药物高度敏感，这会导致严重的副作用，包括出血和感染。可以保护血祖细胞免受耐药基因的毒性作用。最有效的是AGT，它直接去除由细胞遗传物质（DNA）甲基化引起的毒性修饰。但是，AGT不能识别某些有毒的DNA修饰。在这里，我提出了针对其中一些替代病变的不同基因（ALKB）的使用，以改善对甲基化毒性作用的保护。因此，该基因直到最近才被表征，因此应增强AGT保护。此外，与AGT不同，ALKB介导了修复反应后保持活跃，这意味着ALKB修复应该更适合调节。在这里，我提出了一种强大的方法，以识别一个人类ALKB（ABH2）的一种人类同源物的变体，并改善了维修性能。具体而言，我想增强酶识别诊所使用药物产生的病变的能力。选定的变体将在人类祖细胞中的培养物和小鼠动物模型中进行测试，以防止使用这些药物高剂量的治疗。最终，这项工作中确定的突变体应改善甲基化剂的副作用的管理，还可以使用更高剂量来进行更有效的治疗。