A Study of Intracellular Signaling in Muscle and Fat Cells During Ketosis

Study Overview
=================
Brief Summary
-----------------
Hypothesis To define whether stimulation of ATGL and suppression of G0/G1 switch gene occur in the initial phases of diabetic ketoacidosis and thus can be identified as the primary mechanisms behind this life threatening condition. Make a human model for studying ketoacidosis. The investigators plan to reduce in their regular insulin over night. In the morning we administer endotoxin, which together with a relative lack of insulin will initiate ketogenesis - a state of ketoacidosis. On another occasion strict glycemic control is imposed by means of intravenous insulin. The testing is done two separate days with at least 3 weeks in between and patients are admitted to hospital the evening before the day of testing. The investigators use isotopic tracers to determine metabolic fluxes and analyse fat (ATGL, G0/G1 switch gene) and muscle biopsies.

Official Title
-----------------
The Role of ATGL and G0/G1 Switch Gene Complex in Lipopolysaccaride (LPS) Induced Ketosis - a Controlled, Randomised, Clinical Experimental Study

Conditions
-----------------
Ketoacidosis, Diabetes Mellitus Type 1

Intervention / Treatment
-----------------
* Biological: LPS


Participation Criteria
=================
Eligibility Criteria
-----------------
Inclusion Criteria: Diabetes type 1 19 < BMI < 26 minimal or negative C-peptide written consent Exclusion Criteria: Severe comorbidity regular medication apart from insulin

Ages Eligible for Study
-----------------
Minimum Age: 18 Years
Maximum Age: 45 Years

Sexes Eligible for Study
-----------------
Male

Accepts Healthy Volunteers
-----------------
No

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

Design Details

Primary Purpose: Basic Science
Allocation: Randomized
Intervention Model: Crossover Assignment
Masking: Single


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Intervention<br>Insulin reduction and mimic infection with LPS | Biological: LPS<br>* LPS is endotoxin from gram negative bacteria. It is used scientifically to mimic infection lasting 4-8 hours.<br>|
| No Intervention: Control<br>Normal insulin and no LPS | |

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

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Insulin signaling expressed as a CHANGE in phosphorylation of intracellular target proteins and CHANGE in mRNA expression of target genes in muscle- and fat-tissue. | Change in phosphorylation of target proteins and messenger RNA (mRNA) expression of target genes assessed with western blotting technique. | Muscle and fat biopsies obtained on each study day (arm): t1= 6.45 (-75min) am t2=11.15 (195min) am t3= 12.30 pm (270min) | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Change in Intracellular markers of lipid metabolism in muscle- and fat tissue biopsies | Muscle and fat at t1 and t2. Muscle biopsy at t3. Intracellular markers are assessed by western blotting. | Muscle and fat biopsies obtained on each study day (arm): t1= 6.45 am (-75min) t2=11.15 (195min) am t3= 12.30 pm (270min) | 
| Metabolism | Change in glucose, fat and protein metabolism assessed by tracer kinetics on every study day (specific times below) and by indirect calorimetry. [3H 3]Glucose tracer from t=0 - 360min. Palmitic acid tracer from t=165min - 360min. Urea tracer from 0min - 240min. amino acid tracer from 60 min - 360 min. | Change in glucose, fat and protein metabolism between study days and during each study day | 
| Cytokines and stress hormones | Measurement of immune response to endotoxin and hypoinsulinaemia. Estimating the whole body stress during ketoacidosis and pre ketoacidosis. | In basal period t=0-240 minutes and in clamp period t=240-390 minutes |