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

摘要 近20%的儿童肥胖或超重。据预测，一半的儿童肥胖将发生在5岁之前 超过一半的现有儿童将在35岁之前患上肥胖症。母体饮食衍生的营养信号 在围产期发育期间传播的条件，在生命后期的代谢反应。我们发现高n-6到n-3 (n6/n3)脂肪酸暴露与婴儿体内脂肪积累增加呈正相关；在小鼠中， 降低n6/n3暴露条件下的新生儿表观成脂作用，传递长期代谢 让成年人受益。我们的发现引出了中心假说： 发育条件下低n6/n3暴露通过细胞和功能成脂 干细胞样脂肪细胞前体群体内的反应。 脂肪细胞前体细胞(APC)亚型包括前体细胞、前体脂肪细胞和新发现的 “脂肪生成-调节”细胞(Areg)。Aregs减少脂肪组织扩张(ATE)的细胞数 和旁分泌方式。重要的是，我们观察到新生儿体内脂肪堆积、形态和 小鼠低n6/n3暴露所致的细胞密度。这一表型支持ATE的构型对早期敏感 Liven6/n3，然而，受低n6/n3刺激调节的APC亚型频率和分子多样性仍然存在 未知。 我们通过从高n6/n3和低n6/n3幼崽的脂肪库中分离出初级apc来解决这个问题。 曝光。初步的批量rna-seq分析表明，从低n6/n3增加了arg亚型标记。 曝光。其中包括NR2F2-一种与减弱的脂肪生成有关的关键转录调节因子。在共培养中， 低n6/n3条件下的APC在体内增殖较慢，分化较少。这些发现表明 增加ARG细胞数量(或其活性)可能会减轻新生儿中有害的ATE。 这一建议结合了最先进的单细胞RNA测序、现代流式细胞仪技术、 N6/n3比率和NR2F2的遗传操作、实时活细胞成像和体外原代APC培养 实现两个主要目标： SA1.用Areg特异的mRNA签名同时定量ARG细胞数量 出生后高、低n6/n3暴露。 SA2.为了确定NR2F2是否介导低n6/n3依赖对体内脂肪形成的影响，APC 种群和APC成脂潜力。 冲击力。确定可调节早期生命的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症的新模式。

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.