Integrative metabolism of oocyte development and its modulation by maternal diet

卵母细胞发育的综合代谢及其母亲饮食的调节

• 批准号: 9792269
• 负责人: ADAM J CHICCO
• 金额: $ 21.96万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-24 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9792269
• 关键词: Affect; Age; Aging; Animal Model; Assisted Reproductive Technology; Biology; Biomedical Engineering; Cattle; Cell physiology; Cellular Metabolic Process; Chromosome abnormality; Clinical; Comparative Study; Competence; Custom; Development; Diabetes Mellitus; Diet; Dietary Intervention; Embryo; Embryonic Development; Energy Metabolism; Environment; Equus caballus; Experimental Models; Failure; Fertilization; Fluorometry; Formulation; Future; Glucose; Goals; Human; Hyperglycemia; Impairment; In Vitro; Incidence; Infertility; Intervention; Life Style; Link; Maternal Age; Metabolic; Metabolic syndrome; Metabolism; Methods; Mitochondria; Mitochondrial DNA; Modeling; Monitor; Nutrient; Obesity; Omega-3 Fatty Acids; Oocytes; Outcome; Oxidative Phosphorylation; Population; Process; Protocols documentation; Reactive Oxygen Species; Reference Values; Research Project Grants; Resolution; Respiration; Role; Sampling; Secondary to; Supplementation; System; Testing; Therapeutic; Time; Woman; aged; assisted reproduction; base; blastocyst; diabetic; enzyme activity; extracellular; fatty acid supplementation; granulosa cell; human model; impaired capacity; improved; in vivo; insight; instrumentation; metabolic phenotype; miniaturize; mitochondrial metabolism; mother nutrition; new technology; oocyte maturation; real time monitoring; reproductive fitness; respiratory; sensor

Project Summary AAccumulating evidence indicates that an inability of oocyte mitochondria to meet the energy demands of maturation, fertilization, and embryo development contributes to infertility, chromosomal abnormalities and poor assisted reproduction outcomes. This is particularly relevant in the context of maternal aging, where a loss of oocyte mitochondrial content is thought to limit oxidative phosphorylation (OXPHOS) capacity secondary to reductions in mtDNA content or replication. However, our understanding of oocyte and embryo energy metabolism has been limited by inherent technical challenges, and so is currently based largely on indirect estimates of mitochondrial capacity/content, marker enzyme activities and metabolite analyses. To overcome this limitation, our team has developed the first miniaturized metabolic multi-sensor capable of real-time monitoring of mitochondrial respiration, glycolytic flux and extracellular acidification in single oocytes and embryos. Using this new technology in an equine model, we have found that oocyte OXPHOS rate indeed declines with maternal age, but its mitochondrial respiratory capacity actually increases, arguing against the hypothesis that a loss of mitochondrial content/capacity impairs oocyte metabolic potential with age. In parallel studies of granulosa cell metabolism using high-resolution respirometry and fluorometry, we found that maternal aging does not impair OXPHOS capacity, but dramatically increases release of mitochondria-derived reactive oxygen species. This could be reversed by modulating maternal dietary composition for 8 weeks, consistent with potential links between maternal lifestyle, the follicular environment, and oocyte metabolic competence. The aim of this proposal is to apply state-of-the-art methods toward characterizing the integrative metabolic phenotype of mammalian oocytes and embryos during development, and evaluate the impact of nutritional interventions applied both in vivo (maternal) and in vitro (media composition) on these parameters. In Aim 1, we will establish the developmental time-course of metabolic changes that occur from oocyte maturation to the blastocyst stage, and evaluate the influence of incubation media composition, maternal aging and obesity/hyperglycemia on these processes. Studies are proposed in bovine, equine and human oocyte samples for feasibility testing and to establish the value of animal modeling for future applications. In Aim 2, we will test the hypothesis that maternal dietary omega-3 fatty acid supplementation alters oocyte metabolism and improves embryo development, based on preliminary studies in an equine model. Our long-term goal is to establish methods, instrumentation and reference values for integrative metabolic monitoring of oocyte and embryo development to facilitate scientific and therapeutic advances that optimize human reproductive fitness and assisted reproduction outcomes. Our investigative team combines expertise in oocyte biology and assisted reproduction technology, mitochondrial metabolism and respirometry, and biomedical engineering that are uniquely suited to achieving the aims and long-term goals of this project.

项目概要 A越来越多的证据表明，卵母细胞线粒体无法满足成熟、受精和胚胎发育的能量需求，会导致不孕、染色体异常和辅助生殖结果不佳。这在母亲衰老的背景下尤其重要，卵母细胞线粒体含量的损失被认为会限制继发于线粒体 DNA 含量或复制减少的氧化磷酸化 (OXPHOS) 能力。然而，我们对卵母细胞和胚胎能量代谢的理解受到固有的技术挑战的限制，因此目前主要基于线粒体容量/含量、标记酶活性和代谢物分析的间接估计。为了克服这一限制，我们的团队开发了第一个小型化代谢多传感器，能够实时监测单个卵母细胞和胚胎中的线粒体呼吸、糖酵解通量和细胞外酸化。在马模型中使用这项新技术，我们发现卵母细胞 OXPHOS 率确实会随着母亲年龄的增长而下降，但其线粒体呼吸能力实际上会增加，这反驳了线粒体内容/能力的损失会随着年龄的增长而损害卵母细胞代谢潜力的假设。在使用高分辨率呼​​吸测定法和荧光测定法对颗粒细胞代谢进行的平行研究中，我们发现母亲衰老不会损害 OXPHOS 能力，但会显着增加线粒体衍生的活性氧的释放。这可以通过调整母亲饮食成分8周来逆转，这与母亲生活方式、卵泡环境和卵母细胞代谢能力之间的潜在联系一致。该提案的目的是应用最先进的方法来表征哺乳动物卵母细胞和胚胎在发育过程中的综合代谢表型，并评估体内（母体）和体外（培养基成分）应用营养干预措施的影响）关于这些参数。在目标 1 中，我们将建立从卵母细胞成熟到囊胚阶段发生的代谢变化的发育时间过程，并评估孵化培养基成分、母亲衰老和肥胖/高血糖对这些过程的影响。建议对牛、马和人类卵母细胞样本进行研究，以进行可行性测试并确定动物模型对未来应用的价值。在目标 2 中，我们将根据马模型的初步研究，检验母亲膳食 omega-3 脂肪酸补充剂可改变卵母细胞代谢并改善胚胎发育的假设。我们的长期目标是建立卵母细胞和胚胎发育综合代谢监测的方法、仪器和参考值，以促进科学和治疗的进步，优化人类生殖健康和辅助生殖结果。我们的研究团队结合了卵母细胞生物学和辅助生殖技术、线粒体代谢和呼吸测量以及生物医学工程方面的专业知识，非常适合实现该项目的目标和长期目标。

项目成果

A multi-sensor system for measuring bovine embryo metabolism.

• DOI: 10.1016/j.bios.2018.09.071
• 发表时间: 2019-02
• 期刊: Biosensors & bioelectronics
• 影响因子: 12.6
• 作者: Yusra Obeidat;G. Catandi;E. Carnevale;A. Chicco;A. Demann;S. Field;Tom Chen

Equine maternal aging affects oocyte lipid content, metabolic function and developmental potential.

• DOI: 10.1530/rep-20-0494
• 发表时间: 2021-04
• 期刊: Reproduction (Cambridge, England)
• 作者: Catandi GD;Obeidat YM;Broeckling CD;Chen TW;Chicco AJ;Carnevale EM
• 通讯作者: Carnevale EM

Oocyte metabolic function, lipid composition, and developmental potential are altered by diet in older mares.

• DOI: 10.1530/rep-21-0351
• 发表时间: 2022-04-01
• 期刊: Reproduction (Cambridge, England)
• 作者: Catandi GD;LiPuma L;Obeidat YM;Maclellan LJ;Broeckling CD;Chen T;Chicco AJ;Carnevale EM
• 通讯作者: Carnevale EM