De Novo Lipogenesis and Insulin Sensitivity in Obese

Study Overview
=================
Brief Summary
-----------------
Disturbances of de novo lipogenesis (DNL) are one of the features of dysfunction of adipose tissue (AT). Disturbances of DNL play a role in development of metabolic complications of obesity. The goal of this project is to investigate novel pathways of DNL regulation. DNL will be studied during nutritional interventions in healthy and obese subjects in exposure to 2-days high carbohydrate diet preceded by a) 2-days fasting b) several weeks´ ketogenic diet. This nutritional protocol creates conditions for the study of prominent changes in DNL: suppression of DNL during fasting or ketogenic diet followed by stimulation during high-carbohydrate diet. Systemic phenotypic features and molecular indices of DNL regulation in AT will be followed during the protocols. Specific attention will be paid to newly reported pathway- hormone sensitive lipase and transcription factor ChREBP. The results will contribute to development of pharmacological approaches in the treatment of metabolic complications of obesity, targeted selectively to AT, without side effects in other tissues.

Detailed Description
-----------------
The main goal of the proposed project is to characterize the regulation of de novo lipogenesis in AT, a pathway strongly associated with insulin sensitivity in humans. The project should provide information that will bring proof-of-concept for the development of AT DNL-targeting therapeutic strategies to decrease the metabolic risk in obese individuals. Two protocols in lean and obese women to modulate (inhibit/induce) DNL will be implemented: 1) two days of fasting followed by two days of high-carbohydrate diet refeeding (FAST/RF) in lean and obese women; 2) fasting-mimicking intervention in obese women with high fat low carbohydrate ketogenic diet followed by two days of high-carbohydrate diet refeeding (KETO/RF). KETO diet should provide long lasting AT DNL inhibition, and as such it should further highlight the processes necessary for DNL activation in the refeeding phase. The unique protocols proposed in the application will allow to investigate in humans the relationship between AT DNL and whole body insulin sensitivity and glucose tolerance. Moreover, the state of the art experimental methodologies applied for the analyses of AT samples should uncover the possible mechanisms of regulation of DNL by ChREBP, HSL and other factors as well as the related AT secretory capacity in humans. The findings obtained in lean subjects will be compared to obese subjects, as the deregulated response of these pathways might be expected. The project will provide a proof-of-principle for the role of AT DNL in the regulation of insulin sensitivity in lean and obese individuals. The results will indicate novel pathways for future development of drugs targeting the relevant sites in AT in the context of treatment of obesity-induced insulin resistance and associated disorders.

Official Title
-----------------
The Role of De Novo Lipogenesis in Regulation of Insulin Sensitivity in Adipose Tissue in Obese

Conditions
-----------------
Insulin Resistance, Obesity

Intervention / Treatment
-----------------
* Behavioral: Fasting/refeeding
* Behavioral: Fasting/refeeding
* Behavioral: Ketogenic diet/ fasting


Participation Criteria
=================
Eligibility Criteria
-----------------
Inclusion Criteria: sedentary premenopausal women lean (n=20-25, age 25-40 years, BMI 20-25 kg/m2) obese (n=20-25, age 25-40 years, BMI 30-40 kg/m) Exclusion Criteria: diagnosed cancer diabetes (T1DM and T2DM) liver and renal diseases major cardiovascular event bariatric surgery allergy to lidocaine positive serology for hepatitis (B and C) and HIV smoking above 10 cigarettes/day, alcohol consumption above 66g/day sleep apnea poor venous status weight-change more than 3kg in last 3 months untreated hyper- or hypo-thyroidism long term use of medication and/or steroids shift workers and individuals with abnormal sleep/wake pattern

Ages Eligible for Study
-----------------
Minimum Age: 25 Years
Maximum Age: 45 Years

Sexes Eligible for Study
-----------------
Female

Accepts Healthy Volunteers
-----------------
Accepts Healthy Volunteers

Study Plan
=================
How is the study designed?
-----------------

Design Details

Primary Purpose: Basic Science
Allocation: Randomized
Intervention Model: Parallel Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Active Comparator: FAST Lean<br>Short-term fasting (FAST), comparator group of lean subjects | Behavioral: Fasting/refeeding<br>* 2-days fasting followed by 2-days of refeeding. During the fasting period the subjects will be hospitalized to control their state of health and ensure fasting compliance. During refeeding the standardized meals will be provided to cover individual daily requirements (60% carbohydrates, 15% protein, 25 % fat).<br>* Other names: FAST lean;|
| Experimental: FAST Obese<br>Short-term fasting (FAST), experimental group of obese subjects | Behavioral: Fasting/refeeding<br>* 2-days fasting followed by 2-days of refeeding. During the fasting period the subjects will be hospitalized to control their state of health and ensure fasting compliance. During refeeding the standardized meals will be provided to cover individual daily requirements (60% carbohydrates, 15% protein, 25 % fat).<br>* Other names: FAST obese;|
| Experimental: KETO Obese<br>Medium-term ketogenic diet intervention (KETO), experimental group of obese subjects | Behavioral: Ketogenic diet/ fasting<br>* 1 month of fasting-mimicking ketogenic diet and subsequent 2-days of refeeding. The subjects will be instructed by nutritional specialist to follow isocaloric ketogenic diet consisting of 6% carbohydrates, 17 % of proteins and 77% fat to cover individual energy demand. During refeeding the standardized meals will be provided to cover individual daily requirements (60% carbohydrates, 15% protein, 25 % fat).<br>* Other names: KETO obese;|

What is the study measuring?
-----------------
Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Insulin sensitivity | Change in insulin sensitivity index (ΔiAUC insulin/ΔiAUC glucose from OGTT) | through study completion, an average of 1 year | 
| De Novo Lipogenesis in adipose tissue | Change in mRNA expression of lipogenic genes and by targeted and non-targeted lipid analysis (ΔCT) | through study completion, an average of 1 year | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| gene expression profiling of adipose tissue | Change in long non-coding RNA analysis by microarrays (ΔCT) | through study completion, an average of 1 year | 

 Terms related to the study
=================
Keywords Provided by Centre Hospitalier Valida
-----------------
adipose tissue, insulin resistance, obesity, ketogenic diet, de novo lipogenesis