Lysosomal Acid Lipase Activity in Nonalcoholic Fatty Liver Disease (NAFLD LAL)

Nonalcoholic Fatty Liver Disease: Crosstalk Between Genetic Predisposition and Epigenetic Lysosomal Acid Lipase Activity Reduction in Blood, Plasma and Platelets

Non-alcoholic fatty liver disease (NAFLD) is a multifactorial disease affecting a quarter of the world population. Pathological accumulation of fat, into the hepatocytes, is the first hit and is due to altered hepatic and extrahepatic lipogenesis, lipolysis and lipophagy of the large lipid droplets.

Lipophagy plays a key role in the onset of NAFLD, in the autolysosomes, small droplets of fat are catabolized by Lysosomal Acid Lipase (LAL) enzyme which hydrolyzes cholesterol esters and triglycerides forming cholesterol and free fatty acids.

Our research group demonstrated that, subjects affected by NAFLD, present a reduced enzymatic activity either compared to patients with chronic liver disease of different etiology, but comparable staging, either compared to healthy control subjects.

Leukocytes are the main site of enzymatic activity in the blood, however, our research group has shown that it can also be detected inside the platelets, demonstrating how the LAL activity can be exchanged between cells.

Furthermore, our group has shown, for the first time, how the intracellular enzymatic activity is reduced, independently of the platelets and leukocytes count and progressively from chronic liver disease up to cirrhosis.

Among factors which contribute to altered lipid metabolism, the genetic predisposition to the accumulation of hepatic fat must be counted. Several variants of genes that code for proteins implicated in the digestion or storage of fats, are involved.

In this study were considered: patatin-like phospholipase domain-containing 3 (PNPLA3), Transmembrane 6 superfamily 2 (TM6SF2) and 17β-Hydroxysteroid dehydrogenase type 13 (HSD17B13).

The rs738409 variant (C> G, p.I148M) of the PNPLA3 gene consists of a protein in which the catalytic site is not entirely accessible to the substrate which, consequently, accumulates in the storage site.

This variant is commonly found in NAFLD subjects and it has been widely reported how the variant carriers progress faster towards severe disease (steatohepatitis) than wild type subjects.

The TM6SF2 gene encodes a membrane transporter involved in the triglycerides movement, the rs58542926 (C> T E167K) variant has been associated with an increased predisposition towards liver fibrosis in NAFLD subjects. This is likely due to the loss of protein function resulting in hepatic retention of triglycerides and cholesterol.

Unlike PNPLA3 and TM6SF2, the rs72613567 (TA> TAA) variant of the HSD17B13 gene has a protective effect against NAFLD progression. It is characterized by a protein loss of function that protects against chronic liver damage and mitigates the progression of the disease although how the protective effect occurs is still under study.

Due to the multifactorial etiology of the disease, to the need of carrying out a targeted surveillance in predisposed genetic subjects and, in order to prevent NALFD progression towards severe pathological forms characterized by an increased mortality, in this study, 316 subjects will be enrolled.

They will be divided as follows: Italian Caucasians, aged> 18 and <70 years, with non-cirrhotic NAFLD and carriers of the PNPLA3 I148M variant, and, 158 Italian Caucasian subjects, aged> 18 and <70 years, with non-cirrhotic NAFLD and carriers of the wild type allele.

The following exclusion criteria will be considered: any type of malignant disease in the past 5 years, any type of inflammatory or autoimmune disease, corticosteroids for systemic use, any type of drug that may affect body weight or body composition, insufficiency kidney (GFR <90 mL / min), heart failure (NYHA classes II-IV), any type of liver disease rather than NAFLD, excessive alcohol intake (> 140 g / week for men and 70 g / week for women), participation in a weight reduction program in the past 3 months, bile salts, cholestyramine in the last 6 months prior to enrollment, previous cholecystectomy, gallbladder disease.

Peripheral blood will be withdrawn in order to measure haematic lipids (total cholesterol and fractions, triglycerides), total blood LAL activity, to perform genetic analysis and finally to evaluate lipase activity into the platelets.

Hepatic elastography will be also executed, in 100 patients, according to the presence/absence of the PNPLA3 variant, in order to weigh up the genetic predisposition on NAFLD development or progression Finally, in subjects who present a lipase activity 30% lower than the normal value (0.88 ± 0.38 (mean ± SD), the methylation of the LIPA promoter will be studied.

Study Overview

• Status: Recruiting
• Conditions: Non-Alcoholic Fatty Liver Disease
• Intervention / Treatment: Diagnostic test: Peripheral blood withdrawn

• Study Type: Observational
• Enrollment (Anticipated): 316

Contacts and Locations

Study Locations

• Italy
  • Rome, Italy, 00185
    • Recruiting
    • Department of Translational and Precision Medicine, Sapienza University of Rome, Umberto I Hospital
    • Contact: Stefano Ginanni Corradini, MD, PhD; Phone Number: +39 0649972086; Email: stefano.corradini@uniroma1.it
    • Principal Investigator: Stefano Ginanni Corradini, MD, PhD
    • Sub-Investigator: Monica Mischitelli, BSD, PhD
    • Sub-Investigator: Flaminia Ferri, MD, PhD
    • Sub-Investigator: Simona Parisse, MD
    • Sub-Investigator: Giulia Tozzi, BSD, PhD
    • Sub-Investigator: Vito Cantisani, MD, PhD
    • Sub-Investigator: Abel Florian

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years to 70 years (Adult, Older Adult)
• Accepts Healthy Volunteers: No
• Genders Eligible for Study: All

• Sampling Method: Probability Sample

Study Population

non-cirrhotic NALFD patients, wild type for the PNPLA3 risk variant and with a LAL value in DBS of 0.88±0.38 (mean±SD) and non-cirrhotic NALFD patients, carriers for PNPLA3 risk variant (heterozygous and mutant homozygous both) with a LAL value in DBS of 0.76±0.38 (mean±SD)

Description

Inclusion Criteria:

• Caucasian Italian subjects
• age > 18 and < 70 years
• non-cirrhotic NAFLD

Exclusion Criteria:

• any malignant disease during the last 5 years
• any inflammatory or autoimmune disease
• corticosteroids for systemic use
• any medication potentially affecting body weight or body composition
• syndromic obesity
• renal failure (GFR<90 ml/min)
• heart failure (NYHA classes II-IV)
• any type liver disease rather than NAFLD
• alcohol intake >140g/ week for men and 70g/ week for women
• participation in a reducing- weight program in the last 3 months
• cholestyramine during the last 6 months before enrollment
• previous cholecystectomy
• gallbladder disease

Study Plan

How is the study designed?

Design Details

• Observational Models: Cohort
• Time Perspectives: Cross-Sectional

Number of groups / cohorts

2

Cohorts and Interventions

Group / Cohort	Intervention / Treatment
non-cirrhotic NAFLD, carriers of the PNPLA3 I148M variant; Italian Caucasians, aged> 18 and <70 years, with non-cirrhotic NAFLD and carriers of the PNPLA3 I148M variant	Diagnostic test: Peripheral blood withdrawn; Peripheral blood will be withdrawn in order to measure haematic lipids (total cholesterol and fractions, triglycerides), total blood LAL activity, to perform genetic analysis and finally to evaluate lipase activity into the platelets. Hepatic elastography will be also executed, in 100 patients, according to the presence/absence of the PNPLA3 variant, in order to weigh up the genetic predisposition on NAFLD development or progression; Other Names: Hepatic elastography
non-cirrhotic NAFLD, carriers of the wild type allele; Italian Caucasian subjects, aged> 18 and <70 years, with non-cirrhotic NAFLD and carriers of the wild type allele	Diagnostic test: Peripheral blood withdrawn; Peripheral blood will be withdrawn in order to measure haematic lipids (total cholesterol and fractions, triglycerides), total blood LAL activity, to perform genetic analysis and finally to evaluate lipase activity into the platelets. Hepatic elastography will be also executed, in 100 patients, according to the presence/absence of the PNPLA3 variant, in order to weigh up the genetic predisposition on NAFLD development or progression; Other Names: Hepatic elastography

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Inhibition enzyme assay to test LAL activity	Activity of LAL enzyme will be measured in 158 subjects affected by non-cirrhotic NAFLD and wild type for the PNPLA3 rs738409 risk variant, in whole blood, plasma and platelets.	enrollment
Inhibition enzyme assay to test LAL activity	Activity of LAL enzyme will be measured in 158 subjects affected by non-cirrhotic NAFLD and carriers for the PNPLA3 rs738409 risk variant, in whole blood, plasma and platelets.	enrollment

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Inhibition enzyme assay to test LAL activity variation	Activity of LAL enzyme will be measured at 18 months respect to enrollment, in 158 subjects affected by non-cirrhotic NAFLD and wild type for the PNPLA3 rs738409 risk variant, in whole blood, plasma and platelets.	18 months
Inhibition enzyme assay to test LAL activity variation	Activity of LAL enzyme will be measured at 18 months respect to enrollment, in 158 subjects affected by non-cirrhotic NAFLD and carriers for the PNPLA3 rs738409 risk variant, in whole blood, plasma and platelets.	18 months

Collaborators and Investigators

• Sponsor: University of Roma La Sapienza
• Collaborators: Alexion Pharmaceuticals

Study record dates

Study Major Dates

• Study Start (Actual): March 1, 2022
• Primary Completion (Anticipated): March 31, 2023
• Study Completion (Anticipated): December 31, 2024

Study Registration Dates

• First Submitted: May 26, 2022
• First Submitted That Met QC Criteria: June 10, 2022
• First Posted (Actual): June 15, 2022

Study Record Updates

• Last Update Posted (Actual): June 29, 2022
• Last Update Submitted That Met QC Criteria: June 22, 2022
• Last Verified: June 1, 2022

More Information

Terms related to this study

• Keywords: Lysosomal acide lipase; PNPLA3; TM6SF2; HSD17B13; Hepatic elastography
• Additional Relevant MeSH Terms: Digestive System Diseases; Metabolic Diseases; Infant, Newborn, Diseases; Genetic Diseases, Inborn; Metabolism, Inborn Errors; Lysosomal Storage Diseases; Lipid Metabolism Disorders; Lipidoses; Lipid Metabolism, Inborn Errors; Cholesterol Ester Storage Disease; Liver Diseases; Fatty Liver; Non-alcoholic Fatty Liver Disease; Wolman Disease
• Other Study ID Numbers: Sapienza Univ&Alexion Pharma

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: No

Drug and device information, study documents

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