Direct Versus Indirect Effect of Amino Acids on Hepatokines (Diaakine)

Direct Versus Indirect Effect of Amino Acids on Hepatokines in Healthy Subjects and Patients With Non-alcoholic Fatty Liver Disease

Liver hormones are key metabolic regulators and increased in metabolic diseases, including fatty liver disease. The underlying mechanisms driving the elevated levels are currently unknown and presents a major challenge in understanding the interplay between liver hormones and fatty liver disease. The project aims to investigate what stimulates the liver to secrete its hormones and why the secretion is increased in patients with fatty liver disease. The investigator (Associate Prof. Nicolai J Wewer Albrechtsen) will investigate the direct and indirect effects of an amino acid amino infusion on the secretion of hepatokines in individuals with and without metabolic dysfunction-associated steatotic liver disease (MASLD).

Study Overview

• Status: Not yet recruiting
• Conditions: Obesity; Non-Alcoholic Fatty Liver Disease
• Intervention / Treatment: Other: Evaluating the acute effect of an amino acid infusion with and without a concomitant infusion of the somatostatin analogue octreotide to eliminate endogenous production of glucagon

Detailed Description

The liver secretes signaling molecules, (termed hepatokines) to the blood circulation which are powerful metabolic regulators and biomarkers of liver disease. Some of the more studied hepatokines include follistatin, fibroblast growth factor 21 (FGF21) and growth differentiation factor 15 (GDF15) and they have been sown to improve glucose tolerance, reduce liver fat content and regulate appetite.

In dysregulated metabolic conditions, including obesity, MASLD and type 2 diabetes, the circulating levels of hepatokines are increased. It could be speculated that the body increases hepatokine levels as a feedback mechanism to combat dysregulated metabolism. However, the underlying mechanisms driving the elevated levels in metabolic disease are currently unknown. The secretion of follistatin, FGF21 and GDF15 from the liver has been suggested to be stimulated by glucagon and amino acids. In dysregulated metabolic diseases, circulating levels of glucagon and amino acids are often increased and are highly dependent on hepatic steatosis. Increased levels of hepatokines observed in dysregulated metabolic individuals could therefore be attributed to an increase in circulating glucagon, amino acids, or a combination of both.

The study aims to explore the direct and indirect effect of amino acids on the regulation of hepatokines in individuals with and without MASLD. The study evaluates the acute effect of an amino acid infusion with and without a concomitant infusion of the somatostatin analogue octreotide to eliminate endogenous production of glucagon, thus isolating the direct effect of amino acids. ,

The investigators hypothesizes that an amino acid infusion will increase the secretion of hepatokines independent of glucagon.

• Study Type: Interventional
• Enrollment (Estimated): 30
• Phase: Not Applicable

Contacts and Locations

• Study Contact: Name: Nicolai J Wewer Albrechtsen; Phone Number: +4521700880; Email: nicolai.albrechtsen@regionh.dk
• Study Contact Backup: Name: Michael M Richter; Phone Number: +4522154336; Email: michael.martin.richter.02@regionh.dk

Study Locations

• Denmark
  • Copenhagen, Denmark
    • Bispebjerg University Hospital

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Adult; Older Adult
• Accepts Healthy Volunteers: Yes

Description

Group 1: (Lean controls)

Inclusion Criteria:

• Male or female between 25-65 years of age at time of screening
• Body mass index of 18.6-25 kg/m2

Exclusion Criteria:

• Contraindications for MRI-scan
• Severe liver disease (estimated by FIB4 score > 3.25)
• Type 2 diabetes according to ADA criteria
• Significant history of alcoholism or drug/chemical abuse as per investigators judgement
• Amino acid related diseases
• Kidney disease
• Cardiac problems
• Cancer within the past 1 year
• Anemia
• Pregnancy or breast feeding
• Smoking
• Any medicine, acute illness (within the last two weeks) or other circumstances that in the opinion of the investigator might endanger the participants' safety or compliance with the protocol

Group 2 (individuals with hepatic steatosis):

Inclusion Criteria:

• Male or female between 25-65 years of age at time of screening
• Body mass index of 25-40 kg/m2
• Hepatic non-alcoholic steatosis verified by liver biopsy, fibroscan or ultrasound

Exclusion Criteria:

• Contraindications for MRI-scan
• Severe liver disease (estimated by FIB4 score > 3.25)
• Type 2 diabetes according to ADA criteria
• Significant history of alcoholism or drug/chemical abuse as per investigators judgement
• Amino acid related diseases
• Kidney disease
• Cardiac problems
• Cancer within the past 1 year
• Anemia
• Pregnancy or breast feeding
• Smoking
• Any medicine, acute illness (within the last two weeks) or other circumstances that in the opinion of the investigator might endanger the participants' safety or compliance with the protocol

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Basic Science
• Allocation: N/A
• Interventional Model: Single Group Assignment
• Masking: None (Open Label)

Number of Arms

1

Arms and Interventions

Participant Group / Arm

Intervention / Treatment

Experimental: Evaluating the direct and indirect effect of amino acids on the regulation of hepatokines; Participants will be subjected to four experimental days

Other: Evaluating the acute effect of an amino acid infusion with and without a concomitant infusion of the somatostatin analogue octreotide to eliminate endogenous production of glucagon; The experimental days consist of four study days: Assessment of liver fat and visceral fat by magnetic resonance imaging (MRI; 6-point Dixon) (study day A); Somatostatin infusion (4 hours) plus amino acid infusion (45 minutes) (study day B); Saline infusion (4 hours) plus amino acid infusion (45 minutes) (study day C); Somatostatin infusion (4 hours) plus saline infusion (45 minutes) (study day D) The subjects will participate in the experimental days (study day B to D) in randomized order on three different days. For study day B to D, at timepoint t = -75, subjects will receive either; a 240-minute intravenous infusion of a somatostatin analogue (at 200 ng/kg/min (infusion rate will not exceed 1000 µg/hour) or saline. After 75 minutes (timepoint t = 0), the subjects will receive a 45-minute intravenous infusion of amino acids or saline at 3.885 ml/kg/hour. In total, blood will be sampled 11 times over a period of 6 hours and 15 minutes.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
The direct (amino acid + somatostatin) versus the indirect (amino acid + placebo) effect of amino acids on circulating levels of follistatin.	Defined as the difference in incremental AUC0-300 min (iAUC) of follistatin between study day B and study day C in healthy individuals.	From blood sample at minute 0 until blood sample at minute 300.

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
The direct (amino acid + somatostatin) versus the indirect (amino acid + placebo) effect of amino acids on circulating levels of FGF21 and GDF15	Defined as the difference in iAUC0-300 min between study day B and study day C in healthy individuals.	From blood sample at minute 0 until blood sample at minute 300.
The direct versus the indirect effect of amino acids on circulating levels of hepatokines (follistatin, FGF21 and GDF15).	Defined as the difference in iAUC0-300 min of hepatokines between study day B and study day C in individuals with MASLD.	From blood sample at minute 0 until blood sample at minute 300
Differences between healthy and MASLD in the increase in hepatokines during study day B (direct effect of amino acids)	Defined as the differences in iAUC0-300 min of hepatokines between healthy and MASLD	From blood sample at minute 0 until blood sample at minute 300
Differences between healthy and MASLD in the increase in hepatokines during study day C (indirect effect of amino acids).	Defined as the differences in iAUC0-300 min of hepatokines between healthy and MASLD	From blood sample at minute 0 until blood sample at minute 300
The inhibitory effect of somatostatin versus the stimulatory effect of amino acids on glucagon, insulin and C-peptide levels during study day B in healthy and MASLD.	Defined as the difference between iAUC -75-0 min and iAUC0-45 min in both groups	From blood sample at minute -75 until blood sample at minute 45
Differences in glucagon, insulin and C-peptide concentrations between study day B and C in healthy and MASLD	Defined as the difference between iAUC0-300 min during study day B and C in healthy and MASLD	From blood sample at minute 0 until blood sample at minute 300

Collaborators and Investigators

• Sponsor: University of Copenhagen
• Collaborators: Bispebjerg Hospital

Study record dates

Study Major Dates

• Study Start (Estimated): February 1, 2024
• Primary Completion (Estimated): November 1, 2026
• Study Completion (Estimated): December 1, 2026

Study Registration Dates

• First Submitted: January 25, 2024
• First Submitted That Met QC Criteria: January 25, 2024
• First Posted (Estimated): February 2, 2024

Study Record Updates

• Last Update Posted (Estimated): February 2, 2024
• Last Update Submitted That Met QC Criteria: January 25, 2024
• Last Verified: January 1, 2024

More Information

Terms related to this study

• Additional Relevant MeSH Terms: Digestive System Diseases; Liver Diseases; Fatty Liver; Non-alcoholic Fatty Liver Disease; Physiological Effects of Drugs; Antineoplastic Agents; Gastrointestinal Agents; Hormones; Hormones, Hormone Substitutes, and Hormone Antagonists; Antineoplastic Agents, Hormonal; Glucagon; Octreotide; Somatostatin
• Other Study ID Numbers: Diaakine

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO
• IPD Plan Description: We have not decided to share IPD as this also would require approval from the ethical and data approval committee.

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No