Chemistry and Biology of Alkyl Phosphotriester Lesions

烷基磷酸三酯损伤的化学和生物学

• 批准号: 10307544
• 负责人: Yinsheng Wang
• 金额: $ 36.49万
• 依托单位: UNIVERSITY OF CALIFORNIA RIVERSIDE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-12-14 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10307544
• 关键词: 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol; 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone; Affect; Alkylating Agents; Alkylation; Biological; Biological Assay; Biology; Bypass; CRISPR/Cas technology; Carcinogens; Cells; Chemicals; Chemistry; Chemotherapy-Oncologic Procedure; Chromatin; DNA; DNA Adducts; DNA Alkylation; DNA Damage; DNA biosynthesis; DNA lesion; DNA-Directed DNA Polymerase; Data; Development; Dinucleoside Phosphates; Epigenetic Process; Etiology; Exposure to; Frequencies; Genetic Transcription; Health; Histones; Human; Human Genome; Induced Mutation; Investigation; Knowledge; Lead; Lesion; Liver; Lung; Malignant Neoplasms; Mammalian Cell; Measures; Metabolic Activation; Methods; Methylation; Modification; Molecular; Mutagenesis; Mutagens; N' -nitrosonornicotine; Nitrosamines; Oligodeoxyribonucleotides; Outcome; Oxygen; Rattus; Research; Risk; Rodent; Role; Shuttle Vectors; Site; Testing; Tissues; Tobacco; Tobacco smoke; Veins; Vertebral column; adduct; alkyl group; animal tissue; chemical synthesis; chemotherapeutic agent; design; drinking water; experimental study; exposed human population; genetic information; genome editing; genome integrity; human disease; improved; inorganic phosphate; insight; nucleobase; pyridine; repaired

ABSTRACT The human genome is continuously attacked by a variety of endogenous and exogenous genotoxic agents, which can lead to DNA damage and compromise genomic integrity. Alkylating species constitute a ubiquitous class of DNA damaging agents. Aside from forming nucleobase adducts, alkylating agents can also attack one of the non-carbon-bonded oxygen atoms of internucleotide phosphate group to yield backbone alkylation products, i.e. the alkyl phosphotriester (alkyl-PTE) lesions. Although the alkyl-PTE lesions are induced at relatively high frequencies, very little is known about their repair and biological consequences. We hypothesize that unrepaired alkyl-PTE lesions may lead to the development of cancer and other human diseases by compromising the flow of genetic information through inhibiting DNA replication and transcription, and inducing mutations in cells. We also posit that the effects of the alkyl-PTE lesions on DNA replication and transcription may be modulated by the size of the alkyl group conjugated with the backbone phosphate in DNA. Moreover, alkylating agents are among the earliest and most widely prescribed cancer chemotherapeutic agents. Hence, a systematic investigation about how the alkyl-PTE lesions perturb DNA replication and transcription will reveal molecular insights about how exposure to alkylating agents contributes to cancer development, and provide an important knowledge basis for designing better cancer chemotherapeutic agents. To test the above hypothesis, we propose experiments according to three specific aims: Aim #1. To synthesize oligodeoxyribonucleotides (ODNs) harboring site-specifically inserted alkyl-PTE lesions with different flanking sequences; Aim #2. To investigate how the alkyl-PTE lesions perturb the efficiency and fidelity of DNA replication in cells. Aim #3. to examine the transcriptional mutagenesis and repair of alkyl-PTE lesions. The proposed research is built upon our established expertise in the chemical syntheses of lesion-carrying ODNs and in utilizing shuttle vector methods for the assessment about how DNA lesions perturb DNA replication and transcription in cells. The outcome of the proposed studies will provide a systematic and molecule-level understanding about the repair and human health consequences of the alkyl-PTE lesions. Thus, the proposed research will lead to important knowledge for assessing the risk of human exposure toward alkylating agents, for understanding the roles of these lesions in the etiology of human diseases, and for developing better strategies for cancer chemotherapy.

抽象的 人类基因组不断受到各种内源性和外源基因毒性剂的攻击， 这可能导致DNA损伤并损害基因组完整性。烷基化物质构成无处不在的 DNA损伤代理的类别。除了形成核碱加合物外，烷基化剂还可以攻击一种 核苷酸磷酸间磷酸座间的非碳键氧原子的产生，产生骨链烷基化 产物，即烷基磷酸酯（烷基-PTE）病变。尽管诱导烷基斑的病变 频率相对较高，对它们的修复和生物学后果知之甚少。我们假设 未经修复的烷基腐病子可能会导致癌症和其他人类疾病的发展 通过抑制DNA的复制和转录来损害遗传信息的流动，并诱导 细胞中的突变。我们还认为，烷基分布病变对DNA复制和转录的影响 可以通过与DNA中的主链磷酸盐偶联的烷基大小调节。而且， 烷基化剂是最早，规定的癌症化学治疗剂之一。因此， 关于烷基-pte病变如何烧伤DNA复制和转录的系统研究 关于暴露于烷基化剂如何有助于癌症发展的分子见解，并提供 设计更好的癌症化学治疗剂的重要知识基础。为了检验上述假设， 我们根据三个特定目的提出实验：目标＃1。合成寡脱氧纤维核苷酸 （ODN）具有不同侧翼序列的特定地点插入的烷基细胞；目标＃2。到 研究烷基细胞病变如何扰动细胞中DNA复制的效率和保真度。目标＃3。到 检查转录诱变和烷基细胞病变的修复。拟议的研究建立在 我们在携带病变ODN的化学合成方面既定的专业知识，并利用班车向量 评估DNA病变如何扰动细胞中DNA复制和转录的方法。这 拟议的研究的结果将提供有关修复的系统和分子级的理解 和烷基细胞病变的人类健康后果。因此，拟议的研究将导致重要 评估人类暴露于烷基化剂的风险的知识，以了解 这些病变在人类疾病的病因学中，以及为癌症化学疗法制定更好的策略。