Chemistry and Biology of Alkyl Phosphotriester Lesions

烷基磷酸三酯损伤的化学和生物学

基本信息

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
项目类别：
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&apos -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

项目摘要

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多种内源性和外源性基因毒剂的攻击，这可能会导致 DNA 损伤并损害基因组完整性。烷基化物质构成了普遍存在的 DNA 损伤剂的类别。除了形成核碱基加合物外，烷化剂还可以攻击一种核苷酸间磷酸基团的非碳键氧原子产生主链烷基化产品，即烷基磷酸三酯（烷基-PTE）病变。尽管烷基-PTE损伤是在频率相对较高，但对其修复和生物学后果知之甚少。我们假设未修复的烷基 PTE 损伤可能会导致癌症和其他人类疾病的发生通过抑制 DNA 复制和转录来损害遗传信息流，并诱导细胞突变。我们还假设烷基-PTE 损伤对 DNA 复制和转录的影响可以通过与DNA中的主链磷酸酯缀合的烷基的大小来调节。而且，烷化剂是最早和最广泛使用的癌症化疗药物之一。因此，关于烷基 PTE 损伤如何扰乱 DNA 复制和转录的系统研究将揭示关于接触烷化剂如何促进癌症发展的分子见解，并提供设计更好的癌症化疗药物的重要知识基础。为了检验上述假设，我们根据三个具体目标提出实验：目标#1。合成寡脱氧核糖核苷酸 (ODN) 包含具有不同侧翼序列的位点特异性插入的烷基-PTE 损伤；目标#2。到研究烷基-PTE 损伤如何干扰细胞中 DNA 复制的效率和保真度。目标#3。到检查烷基-PTE 损伤的转录突变和修复。拟议的研究建立在我们在携带病变的 ODN 的化学合成和利用穿梭载体方面积累了丰富的专业知识评估 DNA 损伤如何干扰细胞中 DNA 复制和转录的方法。这拟议研究的结果将提供对修复的系统和分子水平的理解以及烷基 PTE 病变对人类健康的影响。因此，拟议的研究将带来重要的评估人类接触烷化剂风险的知识，了解烷化剂的作用这些病变在人类疾病的病因学中，并用于制定更好的癌症化疗策略。