Mitochondrial mechanisms of maternal age effects on offspring health and lifespan

母亲年龄影响后代健康和寿命的线粒体机制

• 批准号: 10889850
• 负责人: Kristin Gribble
• 金额: $ 12.2万
• 依托单位: MARINE BIOLOGICAL LABORATORY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-15 至 2027-03-01
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10889850
• 关键词: Acceleration; Administrative Supplement; Aging; Autophagocytosis; Biogenesis; CRISPR/Cas technology; Clustered Regularly Interspaced Short Palindromic Repeats; Communication; DNA cassette; Development; Future; Generations; Genes; Goals; Health; Heritability; Human; Induced Mutation; Inherited; Knock-in; Knock-out; Longevity; Maternal Age; Mediating; Metabolic Pathway; Methods; Mitochondria; Mitochondrial DNA; Molecular; Mothers; Mutagenesis; Mutation; Nuclear; Parents; Phenotype; Protocols documentation; Public Health; Reporter; Reproduction; Role; Testing; Time; Work; advanced maternal age; age effect; genetic testing; healthspan; improved; inter-individual variation; mitochondrial dysfunction; mutant; offspring; parent project; targeted treatment; tool

PROJECT SUMMARY Advanced maternal age at the time of reproduction decreases offspring lifespan and health in a range of species, including humans. The mechanisms controlling maternal age effects on offspring are unknown, however. In our parent R01 project, we hypothesize that mitochondrial dysfunction in advanced maternal age causes accumulation of dysfunctional mitochondria in offspring through compensatory biogenesis and decreased autophagy during development. This disrupts offspring mitochondrial function and mitochondrial-nuclear communication, leading to accelerated offspring aging. The objectives of the parent project are to understand the mitochondrial mechanisms by which maternal age determines offspring aging. In the parent R01, we proposed to identify the maternal mitochondrial mechanisms that trigger maternal age effects; identify the developmental mechanisms causing accumulation of mtDNA and damaged mitochondria in old-mother offspring; and identify the offspring mitochondrial mechanisms involved in negative maternal age effects and determine if these mechanisms are ROS- dependent. When the parent project proposal was submitted, a method for CRISPR gene editing in Brachionus rotifers was still in development. We have now successfully created a protocol for high-efficiency CRISPR-mediated mutagenesis in Brachionus manjavacas. As proof-of-principle for this new method, we created knockout mutations of VASA and MLH3, and a knock-in mutation of a stop cassette in MLH3. Mutations were inherited over many generations, demonstrating our ability to establish stable mutant lines. The availability of heritable CRISPR induced mutation in Brachionus rotifers now makes it possible to test mitochondrial mechanisms of maternal age effects more directly. The goal of this small-scale, short-term Administrative Supplement project is to establish 5-7 stable knockout mutants and knock-in fluorescent reporter strains for mitochondria-related genes in Brachionus manjavacas. Establishing mutants for genes related to mitochondrial function and dynamics will allow us to test the role of specific mitochondrial dynamics and metabolic pathways in maternal age effects on lifespan and healthspan in the parent R01 and future studies.

项目摘要 高龄产妇在生育时会减少后代的寿命和健康， 包括人类在内的各种物种。控制母亲年龄影响的机制 然而，后代是未知的。在我们的父R 01项目中，我们假设线粒体 高龄产妇的功能障碍导致功能障碍的线粒体在 后代通过补偿性生物合成和减少自噬在发展过程中。这 破坏后代线粒体功能和细胞核通讯，导致 加速后代衰老父项目的目标是了解 母亲年龄决定后代衰老的线粒体机制。在母 R 01，我们建议确定触发母体年龄的母体线粒体机制 影响;确定导致mtDNA积累和受损的发育机制 老年母亲后代中的线粒体;并确定后代线粒体机制 参与负面的母体年龄效应，并确定这些机制是否是ROS- 依赖。当提交母项目提案时，一种CRISPR基因的方法 臂尾轮虫中的基因编辑仍在发展中。我们现在已经成功地创建了一个 在Manjavacas臂尾轮虫中进行高效CRISPR介导的诱变的方案。作为 为了验证这种新方法的原理，我们创建了VASA和MLH 3的敲除突变， MLH 3中终止盒的敲入突变。突变遗传了许多 这证明了我们建立稳定突变株系的能力。的可用性 臂尾轮虫中可遗传的CRISPR诱导突变现在使得测试 线粒体机制的母亲年龄的影响更直接。这个小规模的， 短期行政补充项目是建立5-7个稳定的敲除突变体， 敲入Manjavacas臂尾轮虫相关基因的荧光报告菌株。 建立与线粒体功能和动力学相关的基因突变体将使我们能够 测试特定线粒体动力学和代谢途径在母体年龄效应中的作用 在母体R 01和未来研究中的寿命和健康寿命。