"Mechanisms of Damage-Induced Homologous Recombination"

“损伤诱导的同源重组机制”

• 批准号: 8014926
• 负责人: Bevin P. Engelward
• 金额: $ 23.24万
• 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 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8014926
• 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; Endogenous Factors; Environmental Risk Factor; Event; Excision Repair; Exposure to; 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); 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; 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.

每次细胞分裂时，必须准确地复制数十亿个碱基对的信息 DNA损伤的猛烈攻击。同源重组(HR)为耐受和 修复受损的DNA。虽然人力资源总体上是有益的，但人力资源期间的不协调可能会导致 致瘤序列重排。尽管它很重要，但人们对体内的HR知之甚少。 主要是因为与检测HR相关的技术困难。我们最近创造了转基因 在完整的胰腺组织中可以直接检测到荧光重组细胞的动物 第一次。这一技术突破使评估基因和环境如何 随着时间的推移，各种因素会调节重组细胞的积累。胰腺癌排在第四位 癌症死亡的原因，胰腺癌的危险因素包括慢性炎症和暴露于 烷基化试剂，这两种试剂都已知会导致碱基受损，然后通过碱基切除来修复 修复(BER)途径。在之前的资助周期中，我们发现未修复的损伤和下游 BER中间体可在体外诱导HR。在这里，我们建议进行有史以来第一次关于 BER对活体胰腺组织心率的影响特别是，我们将重点介绍AAG DNA糖基酶， 它移除了广泛的损伤，包括许多由烷基化损伤和 发炎的化学物质。我们的假设是，BER底物和中间体诱导HR，并且 慢性炎症或激素诱导的损害会加剧损害的影响。 有丝分裂刺激。在DNA损伤方面，我们将重点关注AAG通常修复的DNA损伤，以及 在下游误码率中间体上。我们的具体目标是：i)揭示AAG基板或下行误码率 中间体调节体内自发或烷基化诱导的重组；ii)研究 体内慢性胰腺炎症和激素诱导的细胞分裂的重组效应；以及 三)确定炎症或有丝分裂刺激是否调节DNA损伤对HR的影响或改变 重组细胞体内克隆扩增的程度。这项工作的长期目标是 为了阐明影响人的自发易感性的分子和细胞过程， 环境诱导和癌症治疗诱导的DNA序列重排。

项目成果

Tissue-specific differences in the accumulation of sequence rearrangements with age.

随着年龄的增长，序列重排累积的组织特异性差异。

• DOI: 10.1016/j.dnarep.2008.01.012
• 发表时间: 2008
• 期刊: DNA repair
• 影响因子: 3.8
• 作者: Wiktor-Brown,DominikaM;Olipitz,Werner;Hendricks,CarrieA;Rugo,RebeccaE;Engelward,BevinP
• 通讯作者: Engelward,BevinP

Recombinational repair is critical for survival of Escherichia coli exposed to nitric oxide.

重组修复对于暴露于一氧化氮的大肠杆菌的生存至关重要。

• DOI: 10.1128/jb.183.1.131-138.2001
• 发表时间: 2001
• 期刊: Journal of bacteriology
• 影响因子: 3.2
• 作者: Spek,EJ;Wright,TL;Stitt,MS;Taghizadeh,NR;Tannenbaum,SR;Marinus,MG;Engelward,BP
• 通讯作者: Engelward,BP

Contributions of DNA repair and damage response pathways to the non-linear genotoxic responses of alkylating agents.

• DOI: 10.1016/j.mrrev.2015.11.001
• 发表时间: 2016-01
• 期刊: Mutation research. Reviews in mutation research
• 作者: Klapacz J;Pottenger LH;Engelward BP;Heinen CD;Johnson GE;Clewell RA;Carmichael PL;Adeleye Y;Andersen ME
• 通讯作者: Andersen ME

In vivo recombination after chronic damage exposure falls to below spontaneous levels in "recombomice".

• DOI: 10.1158/1541-7786.567.2.10
• 发表时间: 2004-10
• 期刊: Molecular cancer research : MCR
• 作者: O. Kovalchuk;Carrie A. Hendricks;S. Cassie;Andrew J Engelward;B. Engelward

p53 null fluorescent yellow direct repeat (FYDR) mice have normal levels of homologous recombination.

• DOI: 10.1016/j.dnarep.2011.09.009
• 发表时间: 2011-12-10
• 期刊: DNA repair
• 影响因子: 3.8
• 作者: Wiktor-Brown DM;Sukup-Jackson MR;Fakhraldeen SA;Hendricks CA;Engelward BP
• 通讯作者: Engelward BP