The Role of Immune Semaphorins in NAFLD

Study Overview
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Brief Summary
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To goal is to identify semaphorins that are associated with NAFLD and to investigate their relationship with variable degrees of steatosis and fibrosis.

Detailed Description
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Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease associated with systemic changes in immune response. Semaphorins were recently recognized as one of the key regulators of immune responses; while some suppress immune cells activation, proliferation, and production of inflammatory cytokines, others stimulate immune responses. We have previously shown that semaphorins are associated with pathogenesis of viral hepatitis and progression of fibrosis. However, their role in NAFLD is unknown. The hypothesis of this project is that semaphorins are regulators of inflammation in patients with NAFLD. This study is designed as a prospective, non-interventional study. The main aims are: (1) to analyze serum concentration of semaphorins in patients with NAFLD; (2) to analyze tissue expression of semaphorins in patients with NAFLD; (3) to analyze semaphorin gene polymorphisms associated with NAFLD. Semaphorins could be a novel diagnostic and prognostic biomarker as well as targets for immune modulation.

Official Title
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The Role of Immune Semaphorins in NAFLD

Conditions
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Non-Alcoholic Fatty Liver Disease, Immune Response, Biomarkers, Liver Fibrosis, Non Alcoholic Steatohepatitis

Intervention / Treatment
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* Diagnostic Test: Evaluation of the degree of fibrosis and steatosis
* Diagnostic Test: Screening for the components of metabolic syndrome
* Diagnostic Test: Measurement of serum semaphorin concentrations
* Diagnostic Test: Identification of semaphorin gene polymorphisms


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Patients diagnosed with NAFLD according to current guidelines (AASLD, EASL) Exclusion Criteria: Immunosuppression Malignancies Autoimmune diseases Pregnancy HIV Chronic viral hepatitis Presence of other chronic liver disease (hemochromatosis, Wilson's disease, toxic hepatitis, deficiency of alpha-1-antitrypsin, liver autoimmune disease) Consumption of alcohol > 20 g/day

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 70 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Patients with Non-alcoholic fatty liver disease<br>120 patients diagnosed with NAFLD | Diagnostic Test: Evaluation of the degree of fibrosis and steatosis<br>* The degree of steatosis will be estimated using the controlled attenuation parameter (CAP), a method for grading steatosis by measuring the degree of ultrasound attenuation by hepatic fat using a process based on simultaneous transient elastography (TE) which measures the degree of fibrosis.<br>Diagnostic Test: Screening for the components of metabolic syndrome<br>* Anthropometric measures including body mass index (BMI), waist circumference (WC), waist hip ratio (WHR), and waist height ratio (WHtR) will be measured in all patients. Results of the routine laboratory tests as part of the standard diagnostic procedure will be collected: bilirubin, AST, ALT, GGT, ALP, albumins, WBC, neutrophil-to-lymphocyte ratio, hemoglobin, platelet count, fasting glucose, triglycerides, cholesterol, HDL and LDL. Non-invasive scores of steatosis/fibrosis will be calculated (APRI, FIB4, NAFLD score). Patients will be screened for the components of metabolic syndrome.<br>Diagnostic Test: Measurement of serum semaphorin concentrations<br>* Semaphorin concentration will be measured in patient sera by ELISA.<br>Diagnostic Test: Identification of semaphorin gene polymorphisms<br>* Semaphorin genes will be sequenced to identify semaphorin gene polymorphisms associated with NAFLD.<br>|
| Controls<br>40 healthy controls | Diagnostic Test: Evaluation of the degree of fibrosis and steatosis<br>* The degree of steatosis will be estimated using the controlled attenuation parameter (CAP), a method for grading steatosis by measuring the degree of ultrasound attenuation by hepatic fat using a process based on simultaneous transient elastography (TE) which measures the degree of fibrosis.<br>Diagnostic Test: Screening for the components of metabolic syndrome<br>* Anthropometric measures including body mass index (BMI), waist circumference (WC), waist hip ratio (WHR), and waist height ratio (WHtR) will be measured in all patients. Results of the routine laboratory tests as part of the standard diagnostic procedure will be collected: bilirubin, AST, ALT, GGT, ALP, albumins, WBC, neutrophil-to-lymphocyte ratio, hemoglobin, platelet count, fasting glucose, triglycerides, cholesterol, HDL and LDL. Non-invasive scores of steatosis/fibrosis will be calculated (APRI, FIB4, NAFLD score). Patients will be screened for the components of metabolic syndrome.<br>Diagnostic Test: Measurement of serum semaphorin concentrations<br>* Semaphorin concentration will be measured in patient sera by ELISA.<br>Diagnostic Test: Identification of semaphorin gene polymorphisms<br>* Semaphorin genes will be sequenced to identify semaphorin gene polymorphisms associated with NAFLD.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Detection of semaphorins in patients with NAFLD | Measurement of semaphorins concentration in serum of patients with NAFLD by enzyme-linked immunosorbent assay (ELISA) | 12 months | 
| Identification of semaphorin gene polymorphism in patients with NAFLD | Sequencing of semaphorin genes in patients with NAFLD using Sanger sequencing | 12 months | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Staging of liver steatosis | The degree of steatosis will be estimated using the controlled attenuation parameter (CAP) in patients with NAFLD | 12 months | 
| Staging of liver fibrosis | The stage of liver fibrosis will be assessed by transient elastography (TE) in patients with NAFLD | 12 months |