Characterisation of energy metabolism during lactation in cellular and mouse models

细胞和小鼠模型哺乳期间能量代谢的表征

• 批准号: 2484095
• 金额: --
• 依托单位: University of Oxford
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2484095
• 关键词: Characterisation; energy; metabolism; during; lactation

Maternal metabolic diseases such as diabetes and obesity are associated with lactation insufficiency and breastfeeding failure. The goal of this project is to undertake discovery science to characterise how molecules and tissues involved in energy metabolism contribute to the optimal supply of breast milk. The first part of this project will investigate the adiponectin receptor 1 (AdiporR1), a highly expressed hormone receptor in human mammary epithelial cells during pregnancy and lactation. AdiporR1 detects alterations in an adipocyte-derived hormone known as adiponectin, which has been reported to upregulate mitochondrial biogenesis in tissues such as skeletal muscle. The role of AdiporR1 in the mammary gland is uncertain, and this will be investigated using the MCF-10A human mammary epithelial cell line, which expresses AdiporR1. Cultured MCF-10A cells will be stimulated with recombinant adiponectin, and mitochondrial abundance and function assessed using mitochondrial DNA copy number and enzyme activity assays, respectively. RNA silencing of AdiporR1 in MCF-10A cells will be performed to determine whether this abrogates any observed mitochondrial effects of adiponectin. If AdipoR1 is shown to influence mammary epithelial cell function, then AdipoRon, a commercially available orally active adiponectin receptor agonist will be administered to female wild-type mice during pregnancy and the early post-partum period to assess the effect on lactation, as measured by pup growth prior to weaning. The second part of this project will identify which hormones are involved in regulating tissues involved in energy metabolism during lactation. Age-matched pregnant and lactating adult female mice will undergo metabolic profiling by assessment of glucose tolerance and insulin sensitivity, plasma lipids and body fat composition. Plasma will be obtained for measurement of key lactogenic and metabolic hormones. Relevant tissues e.g. adipose tissue, skeletal muscle, pancreatic islets, and mammary glands will be dissected from these mice for RNA-sequencing. A global survey of hormone receptor and signalling protein expression will be undertaken in these tissues, and correlated with hormonal and metabolic changes occurring during the transition from pregnancy to lactation. Thus, these studies aim to determine how maternal tissues adapt to the metabolic demands of breastfeeding, and also to provide a basis for understanding how diseases such as diabetes and obesity lead to lactation insufficiency.

糖尿病和肥胖症等母体代谢疾病与泌乳不足和母乳喂养失败有关。该项目的目标是进行科学发现，以揭示参与能量代谢的分子和组织如何有助于母乳的最佳供应。本项目的第一部分将研究脂联素受体1（AdiporR 1），一种在妊娠和哺乳期人类乳腺上皮细胞中高度表达的激素受体。AdiporR 1检测脂肪细胞衍生的激素脂联素的改变，据报道，脂联素可上调骨骼肌等组织中的线粒体生物合成。AdiporR 1在乳腺中的作用尚不确定，这将使用表达AdiporR 1的MCF-10A人乳腺上皮细胞系进行研究。将用重组脂联素刺激培养的MCF-10A细胞，并分别使用线粒体DNA拷贝数和酶活性测定评估线粒体丰度和功能。将在MCF-10A细胞中进行AdiporR 1的RNA沉默，以确定这是否消除了脂联素的任何观察到的线粒体效应。如果AdipoR 1被证明影响乳腺上皮细胞功能，那么AdipoRon，一种市售的口服活性脂联素受体激动剂，将在妊娠期间和产后早期给予雌性野生型小鼠，以评估对哺乳的影响，通过断奶前幼鼠的生长来测量。该项目的第二部分将确定哪些激素参与调节哺乳期能量代谢的组织。通过评估葡萄糖耐量和胰岛素敏感性、血浆脂质和体脂组成，对经配对的妊娠和哺乳期成年雌性小鼠进行代谢分析。将采集血浆用于测量关键催乳激素和代谢激素。将从这些小鼠中解剖相关组织，例如脂肪组织、骨骼肌、胰岛和乳腺，用于RNA测序。将在这些组织中进行激素受体和信号蛋白表达的全球调查，并与从怀孕到哺乳过渡期间发生的激素和代谢变化相关。因此，这些研究旨在确定母体组织如何适应母乳喂养的代谢需求，并为理解糖尿病和肥胖等疾病如何导致泌乳不足提供基础。