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

肥胖者肠道微生物组的宏基因组研究

• 批准号: 7339700
• 负责人: JEFFREY I GORDON
• 金额: $ 55.66万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-16 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7339700
• 关键词: Adolescence; Adult; African American; Algorithms; Animals; Bacteroidetes; Base Sequence; Biochemical; Biological Assay; Biological Markers; Birth; Blood capillaries; Body Weight decreased; Body fat; Calorimetry; Carbohydrates; Categories; Clinical Research; Collection; Communities; Cyclotrons; DNA; DNA Sequence; Data; Data Set; Deposition; Diet; Disease; Distal; Dizygotic Twins; Ecology; Enrollment; Enzymes; European; Fatty acid glycerol esters; Feces; Female; Female Adolescents; Fermentation; Fourier Transform; Future; Genbank; Gene Expression; Gene Expression Profile; Genes; Genetic; Genome; Genotype; Germ-Free; Gnotobiotic; Harvest; Health; Human; Incidence; Individual; Inflammatory Bowel Diseases; Institutes; Ions; Irritable Bowel Syndrome; Joints; Leptin; Measurement; Mediator of activation protein; Medicine; Metabolic Pathway; Metagenomics; Metric; Missouri; Modeling; Monozygotic Twinning; Monozygotic twins; Mothers; Mus; Obese Mice; Obesity; Parents; Pathology; Pathway interactions; Patients; Physiology; Play; Predisposition; Process; Property; Prospective Studies; Psychiatry; Recruitment Activity; Relative (related person); Research Personnel; Ribosomal RNA; Role; Sampling; Sequence Analysis; Shotguns; Silicon Dioxide; Specimen; Statistically Significant; Structure; Substance abuse problem; Surveys; System; Testing; Time; Transcript; Transplantation; Twin Multiple Birth; Twin Studies; Vaginal delivery procedure; Variant; Vertical Disease Transmission; Volatile Fatty Acids; Washington; Weight; base; cDNA Library; capillary; cohort; comparative; concept; day; dietary restriction; food consumption; functional group; metagenomic sequencing; microbial; microbial community; microbial genome; microbiome; novel therapeutics; professor; prospective; rRNA Genes; research study; therapeutic target; young adult

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

我们在无菌小鼠和无关的成年肥胖和瘦人中的观察表明， 肥胖与肠道微生物生态之间的动态联系。项目1将测试假设， 一个可识别的共同的有机体和基因谱系明显的微生物群和微生物组的 肥胖的人相比，瘦的成年人，这些肥胖相关的有机体基因， 预测的属性，可能有助于提高效率的能量收获从饮食。目标1 -我们 将对从以下来源获得的粪便样本进行比较宏基因组（基于DNA）序列分析 年轻成年人（25- 35岁），阴道分娩的肥胖（BMI ≥ 35 kg/m2）的女性MZ双胞胎对，以及 偏瘦的MZ双胞胎（BMI 18.5- 25 kg/m2）：每个类别（肥胖或偏瘦）中有5对MZ双胞胎 将是欧洲血统（共10对双胞胎）。每个类别中还有五对双胞胎是非洲人- 美国人双胞胎的母亲将被用作参考对照，因为有证据表明， 肠道菌群从母体向后代的传播。两个粪便样本将首先 在一个月的时间间隔内收集每个个体。每个病人的粪便群落DNA样本 将使用“通用”细菌和古细菌引物进行基于16 S rRNA序列的计数。 将来自每个个体的两个DNA样品合并，并将产生约40 Mb的DNA序列 使用GS 20热测序仪。我们将使用UniFrac指标比较社区，该指标由 奈特实验室，并采用现有的算法，从我们的研究开发的肥胖和瘦老鼠，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（在 12，24，36，和48个月）的MZ双胞胎对和他们的母亲将允许我们检查的结构， 他们的微生物群和微生物群随着时间的推移相对于双胞胎，母亲和BMI。为了确定是否 宿主相对于共同母亲的遗传背景在选择微生物群中起主导作用， 它的微生物组，我们将进行一个比较分析的微生物组的肥胖和瘦25- 35岁 女性欧洲血统和非裔美国人双卵双胞胎对和他们的母亲。目标2 - The 在肥胖与瘦型MZ的粪便微生物组中发现的KEGG和COG官能团的代表性 双胞胎对将与粪便微生物群落转录组和代谢组相关。cDNA 从先前用于整个微生物组的相同粪便样品中分离的RNA制备的文库 鸟枪DNA测序，将进行表征。粪便中发现的基因和转录本 将肥胖个体与瘦个体的微生物组/转录组置于KEGG代谢途径上。的