Evaluation of Intestinal Microbiome in Obese Kids

The goal of this study is to investigate and compare the composition of the intestinal
microbiome in patients with obesity only to patients with obesity and Non-Alcoholic Fatty
Liver Disease (NAFLD). In addition, we will investigate the relationship between alterations
in the intestinal microbiome, immune activation, and the progression of NAFLD to
Non-Alcoholic Steatohepatitis (NASH). We hypothesize that alterations in the intestinal
microbiome are associated with increased immune activation and progression of obesity
associated NAFLD. Based on this hypothesis we propose the following aims:

Aim 1. Identify and enroll pediatric cohort with obesity or obesity/NAFLD to study the role
of the intestinal microbiome on the development of NAFLD.

1. Enroll participants through the NEW Kids program for treatment of pediatric obesity at
Children's Hospital of Wisconsin (CHW). Identify and classify participants through
initial clinical evaluation, collect clinical metadata, and obtain and process blood
and stool samples for analysis. Demonstrate feasibility by showing that participants
can be recruited and participate in the specific study protocol.

2. Follow study population through nutritional/exercise intervention, with follow up
collection of clinical data, stool, and blood samples.

Aim 2. Characterize the intestinal microbiome through quantitative PCR and high throughput
sequencing analysis of stool specimens in participants with obesity and obesity/NAFLD.

1. Demonstrate feasibility by showing that sampling aliquots from patient fecal samples
can be successfully analyzed by proposed methods and yield consistent results for
duplicate samples.

2. Compare sequencing results and metagenomic analysis for study groups with particular
attention to bacterial composition and metabolic capacity associated with energy
harvest, lipid and carbohydrate transport, enhancement of epithelial barrier integrity,
and choline metabolism.

3. Investigate whether intervention (nutritional/exercise) results in longitudinal
alterations in the intestinal microbiome.

Aim 3. Characterize evidence of systemic inflammation by C-reactive protein (CRP), Tumor
necrosis factor alpha (TNF-alpha)Transforming growth factor beta 1 (TGF-beta and LPS Binding
Protein (LBP) levels, and analyze results in relationship to the intestinal microbiome and
the presence of NAFLD.

1. Compare levels of systemic inflammatory markers of participants with obesity vs.
obesity/NAFLD

2. Investigate whether intervention (nutritional/exercise) results in longitudinal
alterations in systemic inflammation.

Inclusion Criteria:

- Age 5-18 Years old

- Willing to consent/undergo necessary procedures

- Obese (BMI>30)

- Speak English or Spanish

Exclusion Criteria:

- any other causes of liver disease

- any chronic illnesses or life threatening conditions