Meal-regulated Substrate Metabolism, Influence of Obesity and IL-6

• ClinicalTrials.gov Identifier: NCT04687540
• Recruitment Status: Recruiting
• First Posted: December 29, 2020
• Last Update Posted: April 19, 2021
• Sponsor: Rigshospitalet, Denmark
• Information provided by (Responsible Party): Helga Ellingsgaard, Rigshospitalet, Denmark

Study Description

Brief Summary:

The overall purpose of this explorative yet quantitative study project is to understand how blocking IL-6 signaling leads to the expansion of adipose tissue mass in humans in vivo. The aim is to gain in depth knowledge about how IL-6 receptor blockade affects human lipid, glucose and protein metabolism, specifically the uptake and storage of substrates from a meal vs. their utilization, hence the balance determining whether one gains or loses fat mass.

Condition or disease	Intervention/treatment	Phase
Obesity; Healthy	Drug: Tocilizumab; Drug: Saline 0.9%	Not Applicable

Detailed Description:

Lacking IL-6 signaling leads to an expansion of adipose tissue mass in rodents and humans. However, the underlying mechanisms have not been identified.This project aims to investigate the overall hypothesis that IL-6 receptor blockade changes substrate metabolism during postabsorptive and postprandial states to favor storage over mobilization of fat and to favor glucose over fat as a source for energy production. This hypothesis finds some support in the literature: Infusion of recombinant IL-6 into humans, leading to high concentrations of IL-6 in the circulation, stimulates lipolysis and free fatty acid oxidation.

Therefore, the investigators hypothesize that IL-6 receptor blockade impairs the mobilization of FFA from adipose tissue and impairs fat oxidation in skeletal muscle in the postabsorptive state. In the postprandial, state the investigators hypothesize that IL-6 receptor blockade reduces the insulin-induced uptake and deposition of fat by adipose tissue and skeletal muscle, therefore contributing to ectopic fat deposition in the liver.

In this study 12 lean and 12 obese male participants will be included. The participants will attend one screening visit and two study visits. The IL-6 receptor antibody tocilizumab will be infused on study visit 1.

Isotope dilution techniques, blood flow measurements, arterio-venous differences across adipose tissue and skeletal muscle, fat and skeletal muscle biopsies will be used to assess lipid, glucose and protein kinetics on a whole-body as well as fat and skeletal muscle level in the fasting state and after the ingestion of a liquid mixed-meal. Respiratory exchange ratio will be measured by indirect calorimetry.

Study Design

• Study Type: Interventional (Clinical Trial)
• Estimated Enrollment: 24 participants
• Allocation: Non-Randomized
• Intervention Model: Crossover Assignment
• Intervention Model Description: The study is designed in a placebo-controlled crossover manner, consisting of a screening visit and two study visits. Due to the 4-week wash out period of the IL-6 receptor antibody tocilizumab, the order of study visits will be identical in all subjects; hence, study visits will not be randomized. Subjects will be infused with tocilizumab at the end of study visit 1. This will allow us to study the effect of tocilizumab on study visit 2.
• Masking: Single (Participant)
• Masking Description: Only subjects will be masked regarding order of saline and tocilizumab infusion
• Primary Purpose: Basic Science
• Official Title: Meal-regulated Substrate Metabolism, Influence of Obesity and IL-6
• Actual Study Start Date: April 9, 2021
• Estimated Primary Completion Date: September 1, 2021
• Estimated Study Completion Date: December 1, 2021

Arms and Interventions

Arm	Intervention/treatment
Placebo Comparator: Study day 1 (study visit 1); Baseline measurements (pre-intervention) are obtained on study visit 1.	Drug: Tocilizumab; Baseline: Tocilizumab (infusion of 8 mg/kg bodyweight or a maximum of 800 mg) will be infused over 60 minutes at the end for the study day, therefore study visit 1 (study day 1) measurements are baseline.; Other Name: RoActemra
Active Comparator: Study day 21 (study visit 2); Post-intervention measurements: Participants will be under the influence of tocilizumab, which was injected at the end of study visit 1.	Drug: Saline 0.9%; Participants are under influence of tocilizumab since the effect of the drug will last for 4 weeks. Participants will be infused with saline at study visit 2 (study day 21). Placebo to tocilizumab will be saline (NaCl 0.9%) as tocilizumab is a colorless solution and has to be diluted with NaCl 0.9% prior to administration

Outcome Measures

Primary Outcome Measures :

1. Whole-body, fat and skeletal muscle fat turnover [ Time Frame: 0-21 days ]
   Rate of appearance and disappearance of glycerol and palmitate, fatty acid oxidation and re-esterification, arterio-venous differences of glycerol, palmitate, triglycerides across adipose tissue and skeletal muscle, triglycerides fractional synthesis rate in the postabsorptive and postprandial state in the presence of tocilizumab as compared to placebo
2. Whole-body, fat and skeletal muscle glucose turnover [ Time Frame: 0-21 days ]
   Rate of appearance and disappearance of glucose, arterio-venous differences of glucose across adipose tissue and skeletal muscle, glycogen fractional synthesis rate in the postabsorptive and postprandial state in the presence of tocilizumab as compared to placebo
3. Whole-body, fat and skeletal muscle amino acid and protein turnover [ Time Frame: 0-21 days ]
   Rate of appearance and disappearance of amino acids, arterio-venous differences of amino acids across adipose tissue and skeletal muscle, protein fractional synthesis rate in the postabsorptive and postprandial state, in the presence of tocilizumab as compared to placebo
4. Nutrient uptake [ Time Frame: 0-21 days ]
   Uptake of fatty acids, glucose and amino acids from a meal in the presence of tocilizumab as compared to placebo

Secondary Outcome Measures :

1. Free fatty acids (FFA) (plasma concentration) [ Time Frame: 0-21 days ]
   Change in postabsorptive and postprandial FFA levels in the presence of tocilizumab as compared to placebo
2. Triglycerides (plasma concentration) [ Time Frame: 0-21 days ]
   Change in postabsorptive and postprandial triglycerides levels in the presence of tocilizumab as compared to placebo
3. Subjective feeling of hunger and fullness [ Time Frame: 0-21 days ]
   Hunger and fullness will be assessed on a VAS scale.
4. Insulin (plasma concentration) [ Time Frame: 0-21 days ]
   Change in postabsorptive and postprandial insulin levels in the presence of tocilizumab as compared to placebo
5. C-peptide (plasma concentration) [ Time Frame: 0-21 days ]
   Change in postabsorptive and postprandial c-peptide levels in the presence of tocilizumab as compared to placebo
6. Glucagon (plasma concentration) [ Time Frame: 0-21 days ]
   Change in postabsorptive and postprandial glucagon levels in the presence of tocilizumab as compared to placebo
7. Cortisol (plasma concentration) [ Time Frame: 0-21 days ]
   Change in postabsorptive and postprandial cortisol levels in the presence of tocilizumab as compared to placebo
8. Adrenaline (plasma concentration) [ Time Frame: 0-21 days ]
   Change in postabsorptive and postprandial adrenaline levels in the presence of tocilizumab as compared to placebo
9. Noradrenaline (plasma concentration) [ Time Frame: 0-21 days ]
   Change in postabsorptive and postprandial noradrenaline levels in the presence of tocilizumab as compared to placebo
10. Cytokines, incl. interleukin-6 (IL-6) (plasma concentration) [ Time Frame: 0-21 days ]
    Change in postabsorptive and postprandial cytokine levels in the presence of tocilizumab as compared to placebo
11. Total and active GLP-1 (plasma concentration) [ Time Frame: 0-21 days ]
    Change in postabsorptive and postprandial total and active GLP-1 levels in the presence of tocilizumab as compared to placebo
12. GIP (plasma concentration) [ Time Frame: 0-21 days ]
    Change in postabsorptive and postprandial GIP levels in the presence of tocilizumab as compared to placebo
13. PYY (plasma concentration) [ Time Frame: 0-21 days ]
    Change in postabsorptive and postprandial PYY postabsorptive and postprandial in the presence of tocilizumab as compared to placebo
14. Leptin (plasma concentration) [ Time Frame: 0-21 days ]
    Change in postabsorptive and postprandial leptin levels in the presence of tocilizumab as compared to placebo
15. Testosterone (plasma concentration) [ Time Frame: 0-21 days ]
    Change in postabsorptive testosterone levels in the presence of tocilizumab as compared to placebo
16. TSH (plasma concentration) [ Time Frame: 0-21 days ]
    Change in postabsorptive TSH levels in the presence of tocilizumab as compared to placebo
17. GH (plasma concentration) [ Time Frame: 0-21 days ]
    Change in postabsorptive and postprandial GH levels in the presence of tocilizumab as compared to placebo
18. Respiratory exchange ratio (RER) [ Time Frame: 0-21 days ]
    Indirect calorimetry measured in post-absorptive and postprandial states
19. Femoral artery blood flow [ Time Frame: 0-21 days ]
    Change in femoral artery blood flow in the presence of tocilizumab as compared to placebo
20. RNA sequencing [ Time Frame: 0-21 days ]
    RNA sequencing on adipose tissue and skeletal muscle biopsies, monocytes with or without the influence of tocilizumab
21. Mitochondrial respiration (Oroboros) [ Time Frame: 0-21 days ]
    Mitochondrial respiration in skeletal muscle biopsies with or without the influence of tocilizumab
22. Gastric emptying rate [ Time Frame: 0-21 days ]
    Gastric emptying rate in the presence of tocilizumab as compared to placebo
23. Plasma metabolome [ Time Frame: 0-21 days ]
    Plasma metabolome with or without the influence of tocilizumab
24. Plasma, lipidome [ Time Frame: 0-21 days ]
    Plasma lipidome with or without the influence of tocilizumab
25. Adipose tissue proteome [ Time Frame: 0-21 days ]
    Adipose tissue proteome with or without the influence of tocilizumab
26. Skeletal muscle proteome [ Time Frame: 0-21 days ]
    Skeletal muscle proteome with or without the influence of tocilizumab
27. Monocyte secretome [ Time Frame: 0-21 days ]
    Change in the postabsorptive and postprandial secretory profile of monocytes in the presence of tocilizumab as compared to placebo
28. IL-6 signaling activation in monocytes [ Time Frame: 0 days ]
    IL-6 signaling pathway activation in monocytes from lean participants compared to from obese participants

Eligibility Criteria

• Ages Eligible for Study: 18 Years to 40 Years (Adult)
• Sexes Eligible for Study: Male
• Accepts Healthy Volunteers: Yes

Criteria

Inclusion Criteria:

Healthy males:

• Age ≥ 18 years and ≤ 40 years
• BMI < 18 and > 25 kg/m2
• Healthy (based on screening)
• Stable body weight for 6 months

Obese males:

• Age ≥ 18 years and ≤ 40 years
• BMI ≥ 30 and ≤ 40 kg/m2
• Healthy (based on screening)
• Stable body weight for 6 months

Exclusion Criteria:

• Smoking
• Evidence of severe thyroid or heart disease, inflammatory diseases, current infection, liver disease (transaminases >2x upper normal range), kidney disease (creatinine >1.5 mg/dl), known immunosuppressive disease, corticosteroid use, regular NSAID or paracetamol usage, aspirin use >100 mg/d, history of carcinoma, history of tuberculosis, anemia (hematocrit <33%), WBC <2 x 10^3/ul, platelets <100 x 10^3/ul, bleeding disorders, obstructive pulmonary disease
• Femoral hernia, vascular prosthesis, vascular thrombosis
• Previous nerve damage, many previous femoral catheter installations

Contacts and Locations

Contacts

Contact: Helga Ellingsgaard; (+45) 3545 6550; Helga.Ellingsgaard@regionh.dk

Locations

Denmark

Rigshospitalet, Centre of Inflammation and Metabolism (CIM) Centre for Physical Activity Research (CFAS); Recruiting

Copenhagen, Denmark, 2100

Contact: Helga Ellingsgaard, Ph.D. (+45) 3545 6550 Helga.Ellingsgaard@regionh.dk

Sub-Investigator: Beckey Trinh, MD

Sponsors and Collaborators

Rigshospitalet, Denmark

Investigators

• Principal Investigator: Helga Ellingsgaard, PhD; CFAS, Rigshospitalet

More Information

Publications:

Wallenius V, Wallenius K, Ahrén B, Rudling M, Carlsten H, Dickson SL, Ohlsson C, Jansson JO. Interleukin-6-deficient mice develop mature-onset obesity. Nat Med. 2002 Jan;8(1):75-9.

Wueest S, Item F, Boyle CN, Jirkof P, Cesarovic N, Ellingsgaard H, Böni-Schnetzler M, Timper K, Arras M, Donath MY, Lutz TA, Schoenle EJ, Konrad D. Interleukin-6 contributes to early fasting-induced free fatty acid mobilization in mice. Am J Physiol Regul Integr Comp Physiol. 2014 Jun 1;306(11):R861-7. doi: 10.1152/ajpregu.00533.2013. Epub 2014 Apr 2.

Wedell-Neergaard AS, Lang Lehrskov L, Christensen RH, Legaard GE, Dorph E, Larsen MK, Launbo N, Fagerlind SR, Seide SK, Nymand S, Ball M, Vinum N, Dahl CN, Henneberg M, Ried-Larsen M, Nybing JD, Christensen R, Rosenmeier JB, Karstoft K, Pedersen BK, Ellingsgaard H, Krogh-Madsen R. Exercise-Induced Changes in Visceral Adipose Tissue Mass Are Regulated by IL-6 Signaling: A Randomized Controlled Trial. Cell Metab. 2019 Apr 2;29(4):844-855.e3. doi: 10.1016/j.cmet.2018.12.007. Epub 2018 Dec 27.

Christensen RH, Lehrskov LL, Wedell-Neergaard AS, Legaard GE, Ried-Larsen M, Karstoft K, Krogh-Madsen R, Pedersen BK, Ellingsgaard H, Rosenmeier JB. Aerobic Exercise Induces Cardiac Fat Loss and Alters Cardiac Muscle Mass Through an Interleukin-6 Receptor-Dependent Mechanism: Cardiac Analysis of a Double-Blind Randomized Controlled Clinical Trial in Abdominally Obese Humans. Circulation. 2019 Nov 12;140(20):1684-1686. doi: 10.1161/CIRCULATIONAHA.119.042287. Epub 2019 Nov 11.

Petersen EW, Carey AL, Sacchetti M, Steinberg GR, Macaulay SL, Febbraio MA, Pedersen BK. Acute IL-6 treatment increases fatty acid turnover in elderly humans in vivo and in tissue culture in vitro. Am J Physiol Endocrinol Metab. 2005 Jan;288(1):E155-62. Epub 2004 Sep 21.

van Hall G, Steensberg A, Sacchetti M, Fischer C, Keller C, Schjerling P, Hiscock N, Møller K, Saltin B, Febbraio MA, Pedersen BK. Interleukin-6 stimulates lipolysis and fat oxidation in humans. J Clin Endocrinol Metab. 2003 Jul;88(7):3005-10.

• Responsible Party: Helga Ellingsgaard, Group leader, Rigshospitalet, Denmark
• ClinicalTrials.gov Identifier: NCT04687540
• Other Study ID Numbers: METO
• First Posted: December 29, 2020
• Last Update Posted: April 19, 2021
• Last Verified: April 2021

Individual Participant Data (IPD) Sharing Statement:

• Plan to Share IPD: No

• Studies a U.S. FDA-regulated Drug Product: No
• Studies a U.S. FDA-regulated Device Product: No

Keywords provided by Helga Ellingsgaard, Rigshospitalet, Denmark:

Interleukin-6; Tocilizumab; Postprandial; Postabsorptive; Substrate metabolism; Fat metabolism; Glucose metabolism; Protein metabolism; Isotope dilution technique

Additional relevant MeSH terms:

Obesity; Overnutrition; Nutrition Disorders; Overweight; Body Weight