Mitochondrial Aging and Reprogramming

线粒体老化和重编程

• 批准号: 9142455
• 负责人: SHOUKHRAT M MITALIPOV
• 金额: $ 52.31万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-30 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9142455
• 关键词: Affect; Age; Aging; Aging-Related Process; Alopecia; Animals; Biochemical; Biological Assay; Biological Process; Blood; Body Weight decreased; Cell Aging; Cell Differentiation process; Cell Line; Cell physiology; Cells; Chimera organism; Competence; Complement; Complex; Cytogenetics; Cytoplasmic Organelle; DNA; Derivation procedure; Development; Disease; Embryo; Epigenetic Process; Etiology; Fatty acid glycerol esters; Female; Fertility; Fibroblasts; Generations; Genetic; Genome; Germ Lines; Growth and Development function; Health; Heart; Human; In Vitro; Inherited; Karyotype; Kyphosis deformity of spine; Liver; Maintenance; Malignant Neoplasms; Measures; Metabolic; Metabolism; Mitochondria; Mitochondrial DNA; Molecular; Mus; Mutagenesis; Mutate; Mutation; Nuclear; Oocytes; Organ; Osteoporosis; Oxidative Phosphorylation; Patients; Phenotype; Physiological; Play; Pluripotent Stem Cells; Premature aging syndrome; Production; Replacement Therapy; Role; Skin; Somatic Cell; Spleen; Staging; Stem cells; Techniques; Teratoma; Testing; Therapeutic; Tissues; age effect; age related; aged; cell age; cell type; design; egg; fetal; genome integrity; in vivo; induced pluripotent stem cell; insight; meetings; mitochondrial DNA mutation; mitochondrial genome; mouse model; offspring; pluripotency; postnatal; programs; somatic cell nuclear transfer; subcutaneous

 DESCRIPTION (provided by applicant): Aging is a complex biological process characterized by a gradual but persistent decline in biochemical, physiological and metabolic function of many cells, tissues and organs. Mitochondrial abnormalities have been implicated in the etiology of age-related diseases. Mitochondria contain their own genome (mtDNA) and it has been proposed that mtDNA mutations in somatic tissues accumulate with age, causing a decline in mitochondrial function, and subsequently, age-related diseases and aging itself. In contrast, female germ line (oocyte) mtDNA largely evades mutagenesis and is preserved and replicated with minimal damage. The objective of this proposal is to test the hypothesis that age-related mitochondrial erosion in somatic cells is associated with mtDNA mutations, which in turn adversely affects the developmental competence of iPSCs derived from these cells. Therefore, complete replacement of mutated somatic cell mtDNA is critical for resetting the full developmental competence and therapeutic potential of human iPSCs. We propose to evaluate the effect of aged mitochondria and mutated mtDNA in iPSCs on development and aging in a mouse model, using the definitive chimera assay. To meet our objective, three Specific Aims have been designed. In Aim 1, we will generate genetically identical iPSC lines carrying old and ntESCs with pristine young oocyte mitochondria. In Aim 2, we will define stem cells by genetic, cytogenetic, epigenetic, transcriptional and metabolic profiling. In Aim 3, we will study the development and aging of chimeric offspring generated from stem cells. When completed, this project will provide the direct experimental proof that accumulation of damage and loss of mitochondrial genome integrity plays a central role in the aging process. We will also determine whether the age related mitochondrial erosion in somatic cells affects the developmental potential of experimental PSCs.

 描述（由申请人提供）：衰老是一个复杂的生物学过程，其特征是许多细胞、组织和器官的生化、生理和代谢功能逐渐但持续下降。线粒体异常与年龄相关疾病的病因有关。线粒体包含其自身的基因组（mtDNA），并且已经提出，体细胞组织中的mtDNA突变随着年龄的增长而积累，导致线粒体功能下降，随后导致与年龄相关的疾病和衰老本身。相反，雌性生殖系（卵母细胞）mtDNA在很大程度上避免了诱变，并以最小的损伤保存和复制。 该提案的目的是检验体细胞中与年龄相关的线粒体侵蚀与mtDNA突变相关的假设，这反过来又对来自这些细胞的iPSC的发育能力产生不利影响。因此，突变体细胞mtDNA的完全替换对于重置人类iPSC的完全发育能力和治疗潜力至关重要。我们建议使用确定的嵌合体测定来评估iPSCs中老化的线粒体和突变的mtDNA对小鼠模型中发育和衰老的影响。为了实现我们的目标，我们制定了三个具体目标。在目标1中，我们将产生携带具有原始年轻卵母细胞线粒体的老年和ntESC的遗传上相同的iPSC系。在目标2中，我们将通过遗传学、细胞遗传学、表观遗传学、转录和代谢谱来定义干细胞。在目标3中，我们将研究由干细胞产生的嵌合后代的发育和衰老。 完成后，该项目将提供直接的实验证据，证明线粒体基因组完整性的损伤和损失的积累在衰老过程中起着核心作用。我们还将确定体细胞中与年龄相关的线粒体侵蚀是否影响实验PSC的发育潜力。