The contribution of DNA interstrand crosslinks to aging

DNA链间交联对衰老的贡献

• 批准号: 8097589
• 负责人: Laura Jane Niedernhofer
• 金额: $ 37.78万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-10 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8097589
• 关键词: 8-MOP; Affect; Age; Aging; Apoptosis; Biological; Biological Models; Body Weight decreased; Bone Marrow; CCL4 gene; Cadmium; Cardiovascular Diseases; Cell Aging; Cell Survival; Cells; Cessation of life; Chromosome abnormality; Complex; Coupled; DNA; DNA Crosslinking; DNA Crosslinking Agent; DNA Damage; DNA Interstrand Crosslinking; DNA lesion; Disease; Dose; ERCC1 gene; Elderly; Engineering; Environment; Environmental Exposure; Epidermis; Exposure to; Ficusin; Fracture; Frequencies; Genomic Instability; Genotoxic Stress; Health; Human; In Vitro; Individual; Kidney; Lesion; Life; Lipid Peroxidation; Liver; Malignant Bone Neoplasm; Malignant Neoplasms; Measures; Mechlorethamine; Membrane; Methoxsalen; Molecular; Mus; Mutagens; Mutation; Nerve Degeneration; Nucleotide Excision Repair; Osteopenia; Pathology; Pharmaceutical Preparations; Phenotype; Plants; Predisposition; Prevention; Progeria; Psoralens; Reactive Oxygen Species; Research; Research Personnel; Risk; Severities; Skin; Skin Aging; Source; Symptoms; Syndrome; Testing; Wild Type Mouse; Work; aging population; angelicin; body system; cohort; crosslink; cytotoxic; cytotoxicity; demographics; disorder risk; driving force; early onset; endonuclease; environmental agent; functional loss; in vivo; photoactivation; programs; repaired; research study; response; sarcopenia

DESCRIPTION (provided by applicant): Aging is characterized by loss of functional reserve, placing the elderly at increased risk of numerous diseases. Identifying the driving force behind this functional loss is essential for maintaining a healthy populace. Recent evidence from our lab and others implicates DNA damage as a cause of aging. This implies a strong environmental component to aging. The long term objective of this research is to understand the molecular mechanism by which DNA damage promotes aging. This coupled with identifying environmental causes of genotoxic stress will greatly facilitate prevention of age-associated diseases. ERCC1-XPF is an endonuclease required for repair of bulky monoadducts via nucleotide excision repair (NER) and DNA interstrand crosslinks (ICLs) via a distinct mechanism. Deletion of ERCC1-XPF in the mouse causes early onset aging. These mice therefore offer a unique, rapid and sensitive model system for discovering which genotoxins promote aging and how they do so. The phenotype of the Erccl mice cannot be attributed to loss of NER. Thus our working hypothesis is that rapid aging in ERCC1-XPF deficient mice is caused by their inability to repair ICLs and therefore a consequence of endogenous ICLs which are cytotoxic. To test this, the investigators engineered mice hypomorphic for ERCC1-XPF which age over the course of months, permitting interventional studies. These mice will be exposed to DNA crosslinking drugs and environmental agents that promote lipid peroxidation (LPO), a likely source of endogenous ICLs, to determine if these exposures exacerbate the progeroid symptoms of the mice. The investigators discovered a human progeria caused by mutation of XPF. Thus identifying the cause of rapid aging in ERCC1-XPF-deficient mice will have direct implications for human health. The specific aims of this project are: Aim I: To define the cellular response of ERCCl-XPF-deficient cells to DNA ICLs and LPO. ERCCl-XPF-deficient cells will be exposed to 8-MOP or angelicin, plant-derived psoralens. Photoactivation of 8-MOP induces ICLs and monoadducts, whereas angelicin produces only monoadducts. Cell survival, cellular senescence, apoptosis, mutation frequency and chromosomal aberrations will be measured. If our hypothesis is correct then, 8-MOP will be significantly more cytotoxic than angelicin under conditions where an equal number of DNA lesions are induced. ERCCl-XPF-deficient cells will also be exposed to cadmium, an environmental agent that promotes LPO, to determine if LPO elicits the same cellular response as ICLs. Aim II: To directly test the hypothesis that unrepaired DNA ICLs promote aging. ERCC1-XPF hypomorphic mice will be chronically exposed to the crosslinking agent mechlorethamine. A second cohort will be exposed to 2-chloroethylamine (which induces structurally related monoadducts but not ICLs) using a dose that induces the same number of lesions as mechlorethamine. If our hypothesis is correct, mechlorethamine, but not 2-chloroethylamine, will exacerbate the progeria in these mice. Results will be confirmed by comparing skin aging in response to topical 8-MOP versus angelicin plus UV-A in mice genetically deleted for ERCC1-XPF in the skin only. Aim III: To determine if lipid peroxidation (LPO) promotes aging in mice with defective ICL repair. LPO is caused by oxygen radical damage to membranes and yields products able to crosslink DNA. We hypothesize that LPO is a source of ICLs that contribute to the phenotype of the Erccl mice. LPO will be induced in ERCCl-XPF-deficient mice via exposure to CCL4 or cadmium. If our hypothesis is correct, Erccl mice will be hypersensitive to LPO compared to wild type mice and LPO will exacerbate their progeria. Results from these experiments will indicate if LPO promotes aging and if so, whether it does so by inducing DNA damage. These experiments will also reveal if two common industrial exposures promote aging.

描述（由申请人提供）： 衰老的特征是功能储备的丧失，使老年人的风险增加了许多疾病。确定这种功能损失背后的驱动力对于维持健康的民众至关重要。我们实验室和其他实验室的最新证据暗示DNA损害是衰老的原因。这意味着衰老的强大环境成分。这项研究的长期目标是了解DNA损伤促进衰老的分子机制。结合确定遗传毒性应激的环境原因将极大地促进预防与年龄相关的疾病。 ERCC1-XPF是通过核苷酸切除修复（NER）和DNA Interstrand Interstrand交叉链接（ICL）通过独特的机制修复笨重的单核酶。小鼠中ERCC1-XPF的删除会导致早期衰老。因此，这些小鼠提供了一种独特，快速和敏感的模型系统，用于发现哪种基因毒素促进衰老以及如何进行衰老。 ERCCL小鼠的表型不能归因于NER的丧失。因此，我们的工作假设是，ERCC1-XPF缺乏小鼠的快速衰老是由于它们无法修复ICL的原因，因此是内源性ICL的结果，后者是细胞毒性的。为了测试这一点，研究人员为MICE的ERCC1-XPF设计了小鼠，该年龄在几个月内，允许介入研究。这些小鼠将暴露于DNA交联的药物和环境药物，这些药物促进脂质过氧化（LPO）（LPO）是内源性ICL的可能来源，以确定这些暴露是否会加剧小鼠的孕前症状。研究人员发现了由XPF突变引起的人类后代。因此，确定ERCC1-XPF缺陷小鼠的快速衰老原因将对人类健康产生直接影响。该项目的具体目的是：目标I：定义ERCCL-XPF缺陷型细胞对DNA ICL和LPO的细胞反应。 ERCCL-XPF缺陷型细胞将暴露于8-MOP或Angelicin，植物衍生的薄膜。 8-MOP的光活化诱导ICL和单载合，而Angelicin仅产生单核。将测量细胞存活，细胞衰老，凋亡，突变频率和染色体畸变。如果我们的假设是正确的，那么在诱导相等数量的DNA病变的条件下，8-MOP将比安吉林蛋白更明显地比天然气毒素。 ERCCL-XPF缺陷型细胞也将暴露于促进LPO的环境剂镉，以确定LPO是否引起与ICL相同的细胞反应。 AIM II：直接检验未修复的DNA ICL促进衰老的假设。 ERCC1-XPF肌型小鼠将长期暴露于交联剂机电甲胺中。第二个队列将使用与诱导与机电甲胺相同数量的病变数量的剂量暴露于2-氯乙胺（诱导与结构相关的单载合）。如果我们的假设是正确的，则机电甲胺，而不是2-氯乙胺，将加剧这些小鼠的后代。通过对局部8-MOP与Angelicin和Angelicin Plus UV-A进行比较，仅在皮肤中遗传上删除的ERCC1-XPF的小鼠中的局部性8-MOP与Angelicin Plus UV-A进行比较，可以证实结果。 AIM III：确定脂质过氧化（LPO）是否促进ICL修复有缺陷的小鼠的衰老。 LPO是由对膜的氧自由基损害引起的，并产生能够交叉链接DNA的产品。我们假设LPO是有助于ERCCL小鼠表型的ICL的来源。通过暴露于CCL4或镉，将在ERCCL-XPF缺乏的小鼠中诱导LPO。如果我们的假设正确，与野生型小鼠相比，ERCCL小鼠对LPO高度敏感，而LPO将加剧其早期。这些实验的结果将指示LPO是否促进衰老，如果是这样，是否通过诱导DNA损伤来促进衰老。这些实验还将揭示是否有两个常见的工业暴露促进衰老。

项目成果

Spine degeneration in a murine model of chronic human tobacco smokers.

• DOI: 10.1016/j.joca.2012.04.010
• 发表时间: 2012-08
• 期刊: OSTEOARTHRITIS AND CARTILAGE
• 影响因子: 7
• 作者: Wang, D.;Nasto, L. A.;Roughley, P.;Leme, A. S.;Houghton, A. M.;Usas, A.;Sowa, G.;Lee, J.;Niedernhofer, L.;Shapiro, S.;Kang, J.;Vo, N.
• 通讯作者: Vo, N.

Genotoxic stress accelerates age-associated degenerative changes in intervertebral discs.

• DOI: 10.1016/j.mad.2012.11.002
• 发表时间: 2013-01
• 期刊: MECHANISMS OF AGEING AND DEVELOPMENT
• 影响因子: 5.3
• 作者: Nasto, Luigi A.;Wang, Dong;Robinson, Andria R.;Clauson, Cheryl L.;Ngo, Kevin;Dong, Qing;Roughley, Peter;Epperly, Michael;Huq, Saiful M.;Pola, Enrico;Sowa, Gwendolyn;Robbins, Paul D.;Kang, James;Niedernhofer, Laura J.;Vo, Nam V.
• 通讯作者: Vo, Nam V.

DNA damage triggers a chronic autoinflammatory response, leading to fat depletion in NER progeria.

• DOI: 10.1016/j.cmet.2013.08.011
• 发表时间: 2013-09-03
• 期刊: Cell metabolism
• 影响因子: 29
• 作者: Karakasilioti I;Kamileri I;Chatzinikolaou G;Kosteas T;Vergadi E;Robinson AR;Tsamardinos I;Rozgaja TA;Siakouli S;Tsatsanis C;Niedernhofer LJ;Garinis GA
• 通讯作者: Garinis GA

ISSLS prize winner: inhibition of NF-κB activity ameliorates age-associated disc degeneration in a mouse model of accelerated aging.

• DOI: 10.1097/brs.0b013e31824ee8f7
• 发表时间: 2012-10-01
• 期刊: Spine
• 影响因子: 3
• 作者: Nasto LA;Seo HY;Robinson AR;Tilstra JS;Clauson CL;Sowa GA;Ngo K;Dong Q;Pola E;Lee JY;Niedernhofer LJ;Kang JD;Robbins PD;Vo NV
• 通讯作者: Vo NV

Isolation of muscle-derived stem/progenitor cells based on adhesion characteristics to collagen-coated surfaces.

• DOI: 10.1007/978-1-62703-317-6_5
• 发表时间: 2013
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Lavasani, Mitra;Lu, Aiping;Thompson, Seth D;Robbins, Paul D;Huard, Johnny;Niedernhofer, Laura J
• 通讯作者: Niedernhofer, Laura J