Early life n-3 fatty acids increase novel Adipogenesis-regulatory cells to condition adipogenesis in a NR2F2 dependent manner

生命早期 n-3 脂肪酸增加新型脂肪生成调节细胞，以 NR2F2 依赖性方式调节脂肪生成

• 批准号: 9807608
• 负责人: Michael C. Rudolph
• 金额: $ 11.66万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2019-12-23
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9807608
• 关键词: Address; Adipocytes; Adipose tissue; Adolescent; Adult; Affect; Age; Attenuated; Biological Assay; Blood; Body fat; Breastfed infant; CCAAT-Enhancer-Binding Proteins; Caenorhabditis elegans; Cancer Burden; Cell Count; Cell Culture Techniques; Cells; Cellular Morphology; Cellularity; Child; Coculture Techniques; DNA Methylation; Data; Development; Diet; Disease; Epigenetic Process; Exposure to; F3 gene; FASN gene; Fatty Acids; Fatty acid glycerol esters; Flow Cytometry; Fostering; Frequencies; Genetic Transcription; Growth; Heart Diseases; Histology; In Vitro; Incidence; Individual; Infant; Insurance; Leptin; Life; Link; Lipids; LoxP-flanked allele; Magnetic Resonance; Measures; Mediating; Mediator of activation protein; Medical; Mesenchymal Stem Cells; Messenger RNA; Metabolic; Modality; Modernization; Molecular; Morphology; Mothers; Mus; Neonatal; Non-Insulin-Dependent Diabetes Mellitus; Nutritional; Obesity; Omega-3 Fatty Acids; Overweight; Pattern; Peroxisome Proliferator-Activated Receptors; Phenotype; Play; Polyunsaturated Fatty Acids; Population; Pregnancy; Proliferating; Proteins; Quality of life; RNA; Research; Resolution; Role; Signal Transduction; Stem cells; Stimulus; System; Techniques; Testing; Therapeutic; Time; Tissue Expansion; Well in self; Youth; apoAI regulatory protein-1; cancer type; genetic manipulation; in vivo; insulin sensitivity; lipid biosynthesis; live cell imaging; mother nutrition; neonate; novel; obesity in children; obesity prevention; offspring; paracrine; perinatal development; postnatal; precursor cell; progenitor; public health relevance; pup; response; single-cell RNA sequencing; stem; transcriptome sequencing

ABSTRACT Nearly 20% of children are obese or overweight. It is anticipated half of childhood obesity will occur by age 5 and more than half of existing children will develop obesity by age 35. Maternal diet-derived nutritional signals transmitted during perinatal development condition metabolic responses later in life. We found high n-6 to n-3 (n6/n3) fatty acid exposures positively associate with increased infant body fat accumulation; and in mice, lowering n6/n3 exposure conditioned neonatal adipogenesis epigenetically, imparting long-lasting metabolic benefit to adults. Our findings led to the central hypothesis: Low n6/n3 exposure during development conditions adipogenesis via cellular and functional responses within stem-like adipocyte precursor populations. Adipocyte precursor cell (APC) subtypes include progenitors, preadipocytes, and newly discovered “Adipogenesis-regulatory” cells (Areg). Aregs attenuate adipose tissue expansion (ATE) in a cell number and paracrine way. Importantly, we observe attenuated neonatal body fat accumulation, morphology, and cellularity from low n6/n3 exposure in mice. This phenotype supports that patterning of ATE is sensitive to early life n6/n3, yet, the APC subtype frequencies and molecular diversity regulated by low n6/n3 stimuli remain unknown. We began addressing this gap by isolating primary APCs from fat depots of pups with high and low n6/n3 exposure. Preliminary bulk RNA-seq analysis identified increased Areg subtype markers from low n6/n3 exposure. Included is NR2F2 -a key transcriptional regulator linked to attenuated adipogenesis. In co-culture, APCs conditioned in vivo by low n6/n3 proliferated slower and differentiated less. These findings suggest increases Areg cell number (or their activity) might attenuate detrimental ATE in neonates. This proposal combines state-of-the-art single cell RNA-sequencing, modern flow cytometry techniques, genetic manipulation of n6/n3 ratios and NR2F2, real-time live cell imaging, and in vitro primary APC culture to achieve two primary objectives: SA1. To quantify Areg cell number simultaneously with Areg specific mRNA signatures triggered by high and low n6/n3 postnatal exposures. SA2. To determine if NR2F2 mediates the low n6/n3 dependent effect on in vivo adipogenesis, APC populations, and APC adipogenic potential. Impact. Defining molecular responses and cellular diversity of APC subtypes that can condition early life ATE in vivo could help inform new modalities of obesity prevention.

抽象的 近 20% 的儿童肥胖或超重，预计一半的儿童肥胖将在 5 岁时发生。 超过一半的现有儿童将在 35 岁时患上肥胖症。 母亲饮食来源的营养信号 我们发现，n-6 至 n-3 的代谢反应在围产期发育期间传播。 (n6/n3) 脂肪酸暴露与婴儿体内脂肪积累的增加呈正相关；并且在小鼠中， 降低 n6/n3 暴露，通过表观遗传调节新生儿脂肪形成，赋予持久的代谢能力 我们的研究结果得出了中心假设： 发育条件下 n6/n3 暴露量低 通过细胞和功能形成脂肪 干样脂肪细胞前体群体内的反应。 脂肪细胞前体细胞（APC）亚型包括祖细胞、前脂肪细胞和新发现的 “脂肪生成调节”细胞 (Areg) 会减弱一定数量的脂肪组织扩张 (ATE)。 重要的是，我们观察了新生儿体内脂肪积累、形态和减弱的情况。 这种表型支持 ATE 的模式对早期敏感。 生命 n6/n3，然而，由低 n6/n3 刺激调节的 APC 亚型频率和分子多样性仍然存在 未知。 我们开始通过从具有高和低 n6/n3 的幼崽的脂肪库中分离初级 APC 来解决这一差距 初步的批量 RNA 测序分析发现，低 n6/n3 的 Areg 亚型标记物有所增加。 其中包括 NR2F2——一种与脂肪生成减弱相关的关键转录调节因子。 在体内，通过低 n6/n3 调节的 APC 增殖较慢且分化程度较低。 增加 Areg 细胞数量（或其活性）可能会减轻新生儿的痛苦 ATE。 该提案结合了最先进的单细胞 RNA 测序、现代流式细胞术技术、 n6/n3 比率和 NR2F2 的基因操作、实时活细胞成像以及体外原代 APC 培养 实现两个主要目标： SA1 与 Areg 特异性 mRNA 签名同时量化 Areg 细胞数量。 高和低 n6/n3 产后暴露。 SA2。确定 NR2F2 是否介导对体内脂肪生成的低 n6/n3 依赖性作用，APC 人群和 APC 脂肪形成潜力。 影响。定义可以调节早期生命 ATE 的 APC 亚型的分子反应和细胞多样性。 vivo 可以帮助提供预防肥胖的新方式。