Fatty Acids, Genes and Microbiota in Fatty Liver

Nonalcoholic Steatohepatitis Clinical Trial

Official title:

Non-alcoholic Steatohepatitis Versus Simple Hepatic Steatosis: Is There a Difference in the Nutritional Factors Influencing Lipid Perioxidation and Inflammation?

Clinical Trial Details — Status: Recruiting

Administrative data

• NCT number: NCT02148471
• Other study ID #: 03-0505-A
• Status: Recruiting
• Phase: N/A
• First received: May 23, 2014
• Last updated: November 25, 2014
• Start date: October 2003
• Est. completion date: June 2016

Study information

• Verified date: November 2014
• Source: University Health Network, Toronto
• Contact: Johane Allard, MD,FRCPC
• Phone: 416-340-5159
• Email: johane.allard[@]uhn.on.ca
• Is FDA regulated: No
• Health authority: Canada: Health Canada
• Study type: Observational

Clinical Trial Summary

The first aim of this study is to assess oxidative stress and nutritional status in patients with elevated liver enzymes who were found to have either simple steatosis (SS) or nonalcoholic steatohepatitis (NASH) or normal histological findings on liver biopsy by measuring liver lipid peroxides and tumor necrosis factor (TNF)-α, liver pathology and immunohistochemistry, liver function tests, liver and red blood cell membrane fatty composition, insulin resistance (IR) parameters, plasma lipid peroxides, plasma antioxidant vitamins and antioxidant power, lipid profile, subject demographics, medical history and medication use. The second aim is to detect differences in hepatic gene expression (messenger RNA, mRNA) and epigenetic regulation (micro RNA, miRNA) between patients with SS or NASH and healthy controls, in addition to determine in patients with non-alcoholic fatty liver disease (NAFLD = SS+NASH combined) whether there is an association between hepatic n-3 PUFA content and gene expression. The third aim is to determine the intestinal microbiome (microbial composition and metagenome) in patients with SS or NASH and healthy controls.

Description:

NASH is associated with obesity, diabetes and hyperlipidemia. Fat accumulation in the liver is likely due to variable degrees of disordered fatty-acid metabolism and insulin resistance (IR). Liver steatosis, especially polyunsaturated fatty acids (PUFA) in the liver, increases lipid peroxidation and is associated with a reduction in the antioxidant defense system. This oxidative stress can lead to increased production of pro-inflammatory cytokines (TNF-α, transforming growth factor-beta) contributing to the development of steatohepatitis and fibrosis.TNF-α - may further contribute to IR. In addition, changes in fatty acid composition within the liver may influence lipid metabolism and inflammation. In particular, n-3 PUFA have an effect on the insulin sensitivity, transcription of antioxidant genes, inflammatory response and production of reactive oxygen species. Differences might be seen on the gene expression level (mRNA) and also in epigenetic regulation (miRNA).

Microbiota composition might influence energy metabolism, and inflammatory tone and IR through increased endotoxemia and therefore could also play a role in the development of NAFLD.

Recruitment information / eligibility

• Status: Recruiting
• Enrollment: 165
• Est. completion date: June 2016
• Est. primary completion date: September 2015
• Accepts healthy volunteers: Accepts Healthy Volunteers
• Gender: Both
• Age group: 18 Years to 70 Years

Eligibility

Inclusion criteria:

• Male and female patients, age >18 y

• A liver biopsy with a diagnosis of SS or NASH OR No signs of steatosis, fibrosis or any other kind of liver disease on histology (minimal findings) OR For healthy control subjects, those with normal liver enzymes and normal liver imaging on ultrasound

• alcohol consumption (<20g of ethanol per day);

• absence of any other possible cause for liver dysfunction.

Exclusion criteria:

• any other liver disease apart from NAFLD

• anticipated need for liver transplantation in one year or complications of liver disease;

• any reasons contraindicating a liver biopsy (patients) or liver donation (healthy donors)

• chronic gastrointestinal diseases, previous gastrointestinal surgery modifying the anatomy, patients with diabetes requiring insulin.

• medications known to precipitate steatohepatitis (corticosteroids, high dose estrogens, methotrexate, amiodarone, spironolactone, sulfasalazine, perhexiline maleate, diethylamino- ethoxyhexestrol (DH), tamoxifen, diethylstilbestrol, naproxen or oxacillin) or regular intake of non-steroidal anti-inflammatory drugs (except for low dose aspirin), use of ursodeoxycholic acid or any experimental drug in the 6 months prior to entry.

• regular intake of prebiotics, probiotics, antibiotics, or laxatives; in the 3 months prior to study entry

• Pregnant or lactating

Study Design

Observational Model: Cohort, Time Perspective: Cross-Sectional

Related Conditions & MeSH terms

• Fatty Liver
• Liver Diseases
• Nonalcoholic Fatty Liver Disease
• Nonalcoholic Steatohepatitis
• Steatosis

Locations

Country	Name	City	State
Canada	Toronto General Hospital	Toronto	Ontario

Sponsors (4)

Lead Sponsor	Collaborator
Johane Allard	American College of Gastroenterology, Canadian Institutes of Health Research (CIHR), Canadian Liver Foundation

Country where clinical trial is conducted

Canada, 

References & Publications (8)

Allard JP, Aghdassi E, Mohammed S, Raman M, Avand G, Arendt BM, Jalali P, Kandasamy T, Prayitno N, Sherman M, Guindi M, Ma DW, Heathcote JE. Nutritional assessment and hepatic fatty acid composition in non-alcoholic fatty liver disease (NAFLD): a cross-se — View Citation

Arendt BM, Ma DW, Simons B, Noureldin SA, Therapondos G, Guindi M, Sherman M, Allard JP. Nonalcoholic fatty liver disease is associated with lower hepatic and erythrocyte ratios of phosphatidylcholine to phosphatidylethanolamine. Appl Physiol Nutr Metab. — View Citation

Da Silva HE, Arendt BM, Noureldin SA, Therapondos G, Guindi M, Allard JP. A cross-sectional study assessing dietary intake and physical activity in Canadian patients with nonalcoholic fatty liver disease vs healthy controls. J Acad Nutr Diet. 2014 Aug;114 — View Citation

Monteiro J, Leslie M, Moghadasian MH, Arendt BM, Allard JP, Ma DW. The role of n - 6 and n - 3 polyunsaturated fatty acids in the manifestation of the metabolic syndrome in cardiovascular disease and non-alcoholic fatty liver disease. Food Funct. 2014 Mar;5(3):426-35. doi: 10.1039/c3fo60551e. Review. — View Citation

Mouzaki M, Allard J. Non-alcoholic steatohepatitis: the therapeutic challenge of a global epidemic. Ann Gastroenterol. 2012;25(3):207-217. Review. — View Citation

Mouzaki M, Allard JP. The role of nutrients in the development, progression, and treatment of nonalcoholic fatty liver disease. J Clin Gastroenterol. 2012 Jul;46(6):457-67. doi: 10.1097/MCG.0b013e31824cf51e. Review. — View Citation

Mouzaki M, Comelli EM, Arendt BM, Bonengel J, Fung SK, Fischer SE, McGilvray ID, Allard JP. Intestinal microbiota in patients with nonalcoholic fatty liver disease. Hepatology. 2013 Jul;58(1):120-7. doi: 10.1002/hep.26319. Epub 2013 May 14. — View Citation

Schwenger KJ, Allard JP. Clinical approaches to non-alcoholic fatty liver disease. World J Gastroenterol. 2014 Feb 21;20(7):1712-23. doi: 10.3748/wjg.v20.i7.1712. Review. — View Citation

Outcome

Type	Measure	Description	Time frame	Safety issue
Other	Hepatic phospholipid composition	Gas chromatography	Baseline	No
Other	Red blood cell fatty acid and phospholipid composition	Gas chromatography	Baseline	No
Other	Plasma fatty acid composition	Gas chromatography	Baseline	No
Other	Plasma lipid peroxides	Test kit	Baseline	No
Other	Plasma antioxidant vitamins	Vitamin C colorimetric, alpha- and gamma-tocopherol and beta-carotene by high-performance liquid chromatography	Baseline	No
Other	Serum antioxidant power	Test kit	Baseline	No
Other	TNF-alpha in the liver	Enzyme linked immunosorbent assay	Baseline	No
Other	Immunohistochemistry	Staining for malondialdehyde, alpha-smooth muscle actin, transforming growth factor beta	Baseline	No
Other	Free choline in serum	liquid chromatography/electrospray ionization-isotope dilution mass spectrometry (LC/ESI-IDMS)	Baseline	No
Other	Bacterial DNA in plasma	Quantitative polymerase chain reaction for bacterial 16S rDNA	Baseline	No
Other	Insulin resistance	Fasting glucose and insulin to calculate insulin resistance (HOMA-IR), C-peptide, hemoglobin A1c, all by standard laboratory methods	Baseline	No
Other	Plasma ethanol	standard laboratory measurement	Baseline	No
Other	Anthropometry	Weight, height, skinfolds, bioelectrical impedance analysis	Baseline	No
Other	Food intake	7-day food records	Baseline	No
Other	Physical activity	7 day activity logs	Baseline	No
Other	Factors influencing intestinal microbiota	Environmental questionnaire	Baseline	No
Other	Liver function tests	Alanine transaminase, aspartate transaminase, alkaline phosphatase, standard laboratory tests	Baseline	No
Primary	Hepatic fatty acid composition in total lipids in liver biopsy	Gas chromatography	Baseline	No
Primary	Hepatic gene expression	mRNA by microarray	Baseline	No
Primary	Intestinal microbiota composition	Illumina 16S technology	Baseline	No
Secondary	Lipid peroxides in the liver	Test kit	Baseline	No
Secondary	Hepatic liver antioxidant power	Test kit	Baseline	No
Secondary	Hepatic microRNA expression in the liver	NanoString	Baseline	No
Secondary	Intestinal microbiota - specific organisms and groups	Quantitative real-time polymerase chain reaction	Baseline	No
Secondary	Intestinal microbiome on a genetic level	Illumina sequencing technology	Baseline	No
Secondary	Short-chain fatty acids in stool	Gas chromatography	Baseline	No
Secondary	Plasma endotoxin	Limulus assay	Baseline	No