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Accelerated DNA Methylation Alterations in Hutchinson-Gilford Progeria Syndrome

Hutchinson-Gilford 早衰综合症中 DNA 甲基化的加速改变

基本信息

批准号：
10780718
负责人：
PETER W LAIRD
金额：
$ 78.62万
依托单位：
VAN ANDEL RESEARCH INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-30 至 2028-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10780718
关键词：
Acceleration Aging Automobile Driving Biological Models Catalytic Domain Cell Aging Cell Culture Techniques Cell Nucleus Cell division Cell physiology Cells Childhood Chromatin Communities Cytosine DNA Damage DNA Methylation DNA Modification Methylases DNA analysis DNA biosynthesis Data Dioxygenases Elements Epigenetic Process Event Experimental Designs Fibroblasts Gene Expression Genes Genetic Genome Genomic Imprinting Genomics Heterochromatin Human LMNB1 gene Longevity Methods Mitosis Mitotic Modeling Monitor Morphology Mus Muscular Atrophy Mutation Nuclear Nuclear Lamin Nuclear Lamina Nuclear Protein Outcome Patients Pattern Phenotype Play Polycomb Population Premature aging syndrome Production Progeria Protein Export Pathway Protein Isoforms Publishing RNA-Directed DNA Polymerase Recording of previous events Repression Research Resolution Role Smooth Muscle Smooth Muscle Myocytes Structure Symptoms Syndrome Telomerase Testing Tissues Validation X Inactivation bisulfite sequencing cost effective design experimental study farnesylation gain of function innovation lentivirally transduced methylation pattern methylome mouse genome mouse model mutant normal aging novel therapeutic intervention overexpression oxidative damage prelamin A rare genetic disorder replication stress senescence spatiotemporal vector whole genome

项目摘要

PROJECT SUMMARY / ABSTRACT Hutchinson-Gilford progeria syndrome (HGPS) is a rare genetic disorder characterized by the rapid onset of premature aging beginning in childhood. The condition is caused by a mutation in the nuclear lamin LMNA gene, which leads to the production of a dominant gain-of-function isoform of the protein, called progerin. Progerin causes major disruptions to nuclear morphology and function, including nuclear protein export and mitosis, replication stress, and contributes to increased DNA damage as well as the loss of heterochromatin and dysregylation of other epigenetic marks, including cytosine-5 DNA methylation. We have recently provided direct experimental evidence that aging-associated loss of DNA methylation at nuclear lamina-attached regions of the genome is a direct consequence of cell division-associated DNA replication. We used this principle to develop an epigenetic mitotic clock, called RepliTali, which provides a reliable estimate of cellular replicative history. We hypothesize that altered DNA methylation patterns in patients with HGPS may contribute actively to the severely accelerated aging observed in patients with HGPS. Here, we propose to conduct high- resolution analyses of DNA methylation alterations in serially cultured HGPS cells and to investigate whether we can extend the lifespan of HGPS model systems by manipulating DNA methylation patterns. In Specific Aim 1 we will define DNA methylation dynamics in serially cultured HGPS fibroblasts from early passage through replicative senescence using cost-effective Infinium DNA methylation arrays. In Specific Aim 2 we will conduct high-resolution single-cell methylome analyses at key stages of HGPS fibroblast culture to detect arising aberrations and delineate population transitions. We have developed a single-cell whole-genome bisulfite sequencing (sc-WGBS) method that delivers genomic coverage far superior to any other published sc-WGBS methods. In Specific Aim 3 we will investigate whether DNA methylation manipulation can extend lifespan in HGPS models. In Aim 3A we will test whether overexpression of DNA methylation writers and erasers increases the replicative lifespan of HGPS fibroblasts. In Aim 3B, we will target DNA methyltransferase overexpression to arterial smooth muscle cells in an HGPS mouse model to investigate whether this reduces arterial smooth muscle loss and extends lifespan. We will monitor DNA methylation changes in this mouse model using a new cost-effective DNA methylation array. We present extensive and compelling preliminary data that demonstrates both the feasibility and relevance of the proposed aims. The outcome of this proposed research could have important impacts on our understanding of the contribution of DNA methylation alterations to HGPS phenotypes, potentially opening avenues for new therapeutic approaches to treat progeria. In addition, this study could shed light on similar mechanisms operating at a longer timescale in normal aging.

项目概要/摘要哈钦森-吉尔福德早衰综合症 (HGPS) 是一种罕见的遗传性疾病，其特点是发病迅速早衰从童年开始。这种情况是由核纤层蛋白 LMNA 突变引起的基因，它导致蛋白质的显性功能获得亚型的产生，称为早衰蛋白。早老蛋白对核形态和功能造成重大破坏，包括核蛋白输出和有丝分裂、复制应激，并导致 DNA 损伤增加以及异染色质丢失以及其他表观遗传标记的失调，包括胞嘧啶 5 DNA 甲基化。我们最近提供了直接实验证据表明，与衰老相关的核纤层附着区域 DNA 甲基化缺失基因组的变化是细胞分裂相关DNA复制的直接结果。我们利用这个原理开发了一种表观遗传有丝分裂时钟，称为 RepliTali，它提供了细胞复制的可靠估计历史。我们假设 HGPS 患者 DNA 甲基化模式的改变可能会做出积极的贡献 HGPS 患者中观察到的严重加速衰老。在此，我们建议开展高对连续培养的 HGPS 细胞中 DNA 甲基化改变进行分辨率分析，并研究是否我们可以通过操纵 DNA 甲基化模式来延长 HGPS 模型系统的寿命。在具体目标 1 中，我们将定义早期连续培养的 HGPS 成纤维细胞中的 DNA 甲基化动态。使用经济高效的 Infinium DNA 甲基化阵列穿越复制衰老。在具体目标 2 中，我们将在 HGPS 的关键阶段进行高分辨率单细胞甲基化组分析成纤维细胞培养以检测出现的畸变并描绘群体转变。我们开发了一个单细胞全基因组亚硫酸氢盐测序 (sc-WGBS) 方法可提供卓越的基因组覆盖率任何其他已发布的 sc-WGBS 方法。在特定目标 3 中，我们将研究 DNA 甲基化操作是否可以延长 HGPS 的寿命模型。在目标 3A 中，我们将测试 DNA 甲基化写入器和擦除器的过度表达是否会增加 HGPS 成纤维细胞的复制寿命。在目标 3B 中，我们将目标 DNA 甲基转移酶过度表达 HGPS 小鼠模型中的动脉平滑肌细胞，以研究这是否会降低动脉平滑度肌肉损失并延长寿命。我们将使用新的方法监测该小鼠模型中 DNA 甲基化的变化经济高效的 DNA 甲基化阵列。我们提供广泛且令人信服的初步数据，证明可行性和相关性拟议的目标。这项拟议研究的结果可能会对我们产生重要影响了解 DNA 甲基化改变对 HGPS 表型的贡献，有可能开启治疗早衰症的新治疗方法的途径。此外，这项研究可以揭示类似的情况在正常衰老过程中，机制以较长的时间尺度运行。