Metagenomic studies of the gut microbiomes of obese & lean Twin Pairs

我们在灵知生菌小鼠和无关的成年肥胖和瘦人身上的观察表明，有一个 肥胖与肠道微生物生态之间的动态联系。项目1将检验以下假设： 一组可识别的共享的生物和基因谱系，在微生物群和微生物群中很明显 肥胖与瘦弱的成年人相比，这些与肥胖相关的有机基因具有 预测可能有助于提高从饮食中获取能量的效率的特性。目标1-我们 将对从以下地点获得的粪便样本进行比较元基因组(基于DNA)序列分析 青壮年(25-35岁)，经阴道分娩的肥胖女性MZ双胞胎(BMI^35公斤/平方米)，以及 苗条的MZ双胞胎(BMI 18.5-25 kg/m2)：每类MZ双胞胎中的5对(肥胖或苗条) 将是欧洲血统(总共10对双胞胎)。每个类别的另外五对双胞胎将是非洲人- 美国人。双胞胎的母亲将被用作参照对照，因为有证据表明 肠道微生物群从母亲传给后代的过程。两个粪便样本最初将是 在一个月的时间间隔内按个人收集。每个患者的每个粪便群落DNA样本 将接受基于16S rRNA序列的计数，使用“通用”细菌和古细菌引物。 每个个体的两个DNA样本将被合并，并将生成-40Mb的DNA序列 使用GS20型焦炭测序仪。我们将使用由 奈特实验室，并使用现有的算法，从我们对肥胖和瘦小鼠的研究中开发出来，以进行比较 他们的微生物群。收集短期(1个月)和长期随访粪便样本(见 12个月、24个月、36个月和48个月)来自MZ双胞胎和他们的母亲将允许我们检查 他们的微生物群和微生物群随着时间的推移相对于同卵双胞胎、母亲和BMI。为了确定 宿主与共同母亲的遗传背景在微生物区系的选择中起主导作用 它的微生物群，我们将对25-35岁的肥胖者和消瘦的人进行微生物群的对比分析 欧洲女性血统和非裔美国异卵双胞胎及其母亲。目标2-- 肥胖与瘦MZ患者粪便微生物群中KEGG和COG官能团的表达 双胞胎对将与粪便微生物群落转录本和代谢体相关。Cdna 从以前用于整个微生物组的相同粪便样本中分离的RNA制备的文库 猎枪DNA测序，将被定性。在粪便中发现的基因和转录本 肥胖与瘦个体的微生物组/转录组将被置于KEGG代谢途径上。这个

Our observations in gnotobiotic mice and unrelated adult obese and lean humans indicate that there is a dynamic linkage between adiposity and gut microbial ecology. Project 1will test the hypothesis that thereis an identifiable shared set of organismal and gene lineages evident in the microbiota and microbiomes of obese compared to lean adult individuals, and that these obesity-associated organismal genes have predicted properties that could contribute to increased efficiency of energy harvest from a diet. Aim 1 -We will perform comparative metagenomic (DNA-based) sequence analysis of fecal samples obtained from young adult (25-35year old), vaginally delivered female MZtwin pairs who are obese (BMI^35 kg/m2), and MZ twin pairs who are lean (BMI 18.5-25kg/m2): five of the MZ twin pairs in each category (obese or lean) will be of European ancestry (total of 10 twin pairs). Five more twin pairs in each category will be African- American. Mothers of twin pairs will be used as reference controls since there is evidence for vertical transmission of the gut microbiota from a mother to her offspring. Two fecal samples will initially be collected per individual over a one-month interval. Each fecal community DNA sample from each patient will be subjected to 16S rRNA sequence-based enumeration using 'universal' bacterial, and archaeal primers. The two DNA samples from each individual will be pooled, and -40 Mb of DNA sequence will be generated using the GS20 pyrosequencer. We will compare communities using the UniFrac metric developed by the Knight lab, and employ existing algorithms, developed from our studies of obese and lean mice, to compare their microbiomes. The collection of both short-term (1-month) and longer-term followup fecal samples (at 12, 24, 36, and 48 months) from MZ twin pairs and their mothers will allow us to examine the structureof their microbiotas and microbiomes over time relative to co-twin, mother, and BMI.To determine whether genetic background of the host versus a shared mother plays a dominant role in selecting a microbiota and its microbiome, we will perform a comparable analysis of the microbiomes of obese and lean 25-35year-old female European ancestry and African-American dizygotic twin pairs and their mothers. Aim 2 - The representation of KEGG and COG functional groups found in the fecal microbiomes of obese vs. lean MZ twin pairs will be correlated with fecal microbial community transcriptomes and metabolomes. cDNA libraries prepared from RNAs isolated from the same fecal samples previously used for whole microbiome shotgun DNA sequencing, will be characterized. Genes and transcripts identified in the fecal microbiome/transcriptome of obese vs. lean individuals will be placed onto KEGG metabolic pathways. The