"Mechanisms of Damage-Induced Homologous Recombination"

“损伤诱导的同源重组机制”

• 批准号: 7554137
• 负责人: Bevin P. Engelward
• 金额: $ 24.06万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-02-25 至 2012-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7554137
• 关键词: Adenine; Alkylating Agents; Alkylation; Animals; Base Excision Repairs; Base Pairing; Cancer Etiology; Cell division; Cell physiology; Cells; Cessation of life; Chemicals; Chronic; Clonal Expansion; Code; DNA Damage; DNA Repair; DNA Sequence Rearrangement; DNA glycosylase; DNA lesion; Direct Repeats; Double Strand Break Repair; Environmental Risk Factor; Event; Excision Repair; Exposure to; Face; Frequencies; Genetic; Genetic Recombination; Genomic Instability; Goals; Grant; In Vitro; Inflammation; Inflammatory; Knowledge; Large-Scale Sequencing; Lead; Learning; Length; Lesion; Light; Loss of Heterozygosity; Malignant Neoplasms; Malignant neoplasm of pancreas; Mammalian Cell; Methylnitrosourea; Mitogens; Modeling; Molecular; Mus; Pancreas; Pathway interactions; Persons; Predisposition; Process; Proteins; Recombinants; Relative (related person); Research Personnel; Risk Factors; Signal Transduction; Site; Testing; Time; Tissues; Transgenic Animals; Work; base; cancer prevention; cancer risk; cancer therapy; chronic pancreatitis; homologous recombination; in vivo; interest; mutant; overexpression; programs; reconstitution; repair enzyme; repaired; tumorigenic

Every time a cell divides, billions of base pairs of information must be accurately copied in the face of an onslaught of DNA damage. Homologous recombination (HR) provides a critical mechanism for tolerating and repairing damaged DNA. Although HR is generally beneficial, misalignments during HR can lead to tumorigenic sequence rearrangements. Despite its fundamental importance, little is known about HR in vivo, primarily because of technical difficulties associated with detecting HR. We have recently created transgenic animals in which fluorescent recombinant cells can be directly detected within intact pancreatic tissue for the first time. This technological breakthrough makes it possible to assess how genetic and environmental factors modulate the accumulation of recombinant cells over time. Pancreatic cancer is the fourth leading cause of cancer death, and risk factors for pancreatic cancer include chronic inflammation and exposure to alkylating agents, both of which are known to induce damaged bases that are repaired by the base excision repair (BER) pathway. In the previous grant cycle, we found that both unrepaired lesions and downstream BER intermediates can induce HR in vitro. Here, we propose to undertake the first ever studies of the influence of BER on HR in pancreatic tissue in vivo. In particular, we will focus on the Aag DNA glycosylase, which removes a broad range of lesions, including many that are created by alkylation damage and inflammatory chemicals. Our hypothesis is that BER substrates and intermediates induce HR, and that the effects of damage are exacerbated by conditions of chronic inflammation or by hormonally-induced mitogenic stimulation. In terms of DNA damage, we will focus on DNA lesions normally repaired by Aag, and on downstream BER intermediates. Our Specific Aims are I) Reveal if Aag substrates or downstream BER intermediates modulate spontaneous or alkylation-induced recombination in vivo; II) Study the potential recombinogenic effects of chronic pancreatic inflammation and hormonally induced cell division in vivo; and III) Determine if inflammation or mitogenic stimulation modulates the effects of DNA damage on HR or alters the extent of clonal expansion of recombinant cells in vivo. The broad long term objectives of this work are to shed light on the molecular and cellular processes that influence a person's susceptibility to spontaneous, environmentally-induced, and cancer therapy-induced DNA sequence rearrangements.

每当一个细胞分裂时，数十亿个碱基对的信息必须被精确地复制