Clinical Trial Details
— Status: Not yet recruiting
Administrative data
| NCT number |
NCT02507882 |
| Other study ID # |
ELH - 2015 - IL28 |
| Secondary ID |
|
| Status |
Not yet recruiting |
| Phase |
N/A
|
| First received |
July 22, 2015 |
| Last updated |
July 22, 2015 |
| Start date |
January 2016 |
Study information
| Verified date |
July 2015 |
| Source |
Egyptian Liver Hospital |
| Contact |
Waleed Samir Abdelrazek, PhD |
| Phone |
+20507942901 |
| Email |
waleed_samir1[@]yahoo.com |
| Is FDA regulated |
No |
| Health authority |
Egypt: Egyptian Liver Hospital |
| Study type |
Interventional
|
Clinical Trial Summary
Hepatocellular carcinoma (HCC) is the third leading cause of cancer deaths worldwide as well
as in Egypt. Despite improvements in HCC therapy, the prognosis for HCC patients remains
poor. Today molecular, genomic and epigenomic aberrations in tumors are being deeply
investigated. Many biomarkers were associated to HCC onset, and they could be useful for
clinicians, but all show some limitations and no one is so early to predict the HCC onset.
It is estimated that 51.5% of HCC cases can be attributed to HCV infection. Moreover, there
is a large occult reservoir of HCV caused chronic liver disease in approximately 9 % in the
Egyptian with estimated 6 million HCV chronic infections and estimated 150 000 new
infections per year. Among them, we have to mention the polymorphism of IL28B gene
rs12979860 C/T. and rs 4803217. The IL-28B gene encodes interferon-lambda 3 (IFN-λ3), which
belongs to the type III IFN family. IFN-λ interacts with a transmembrane receptor inducing a
potent antiviral response. In experimental model of HCV type III IFN was able to inhibit
viral replication. IL-28B polymorphisms are linked to the efficiency of the inflammatory
process during HCV infection, and to the mechanisms that HCV adopts to escape by innate and
adaptive immunity.
During the last years, a number of studies have assessed the association between the IL-28B
polymorphisms and risk of HCC and liver cirrhosis (LC) occurrence in various populations;
however, results obtained are still inconclusive.
Interestingly, some polymorphisms located at the 3' untranslated region (UTR) of IL28B, e.g.
rs 4803217, seem to interfere with the binding of miRNA, to date recognized as important
post-transcriptional regulators. In the last years miRNAs acquired a growing relevance as
potential biomarkers for several diseases including cancer, and many researches are focusing
on understanding their role in cancer.
Thus the objectives of the current proposal are to determine through investigating a cohort
of 405 patients, whether IL28B rs12979860 and rs4803217 polymorphisms are associated to the
risk of HCC in chronic hepatitis C (CHC) patients and, above all, to identify their role as
predictor marker of HCC in CHC, when associated to miRNAs modulation. Data obtained by our
work could be helpful in HCC diagnosis, thus leading to the improvement of the patients
prognosis. The proposed activities are going to be implemented through a partnership us as
Egyptian Liver Research Institute and Hospital (ELRIAH)- Dakhlya- Egypt and Non- Egyptian
Partners.
Description:
Introduction
Hepatocellular carcinoma (HCC) is the third leading cause of cancer deaths worldwide.
Despite improvements in HCC therapy, the prognosis for HCC patients remains poor due to a
high incidence of recurrence. An improved understanding of the pathogenesis of HCC
development would facilitate the development of more effective outcomes for the diagnosis
and treatment of HCC at earlier stages.
Currently, molecular alterations in tumors are being scrutinized at a genome-wide scale,
covering different dimensions, such as gene expression, epigenetic changes, chromosomal
aberrations, and more recently, next generation sequencing. A large number of molecular
markers are associated with the development of HCC. that could be useful in the clinic;
however, racial differences have been reported, and these need to be examined more
thoroughly.
It is estimated that 51.5% of HCC cases can be attributed to HCV infection. Among them, we
have to mention the polymorphisms of IL28B gene, e.g. rs12979860 C/T and rs4803217. The
IL-28B gene encodes interferon-lamda 3 (IFN-λ3), which belongs to the type III IFN family
including IFN-λ1, IFN-λ2, and IFN-λ3. IFN-λ interacting with a trans membrane receptor
induces potent antiviral responses mediated by the activation of the JAK-STAT and MAPK
pathways. IL-28B polymorphisms are linked to the efficiency of the inflammatory process
during HCV infection and to the mechanisms that HCV adopts to escape by innate and adap¬tive
immunity. Interestingly, since type III IFN has been found to inhibit both HBV and HCV
replication in experimental models, During the last years, a number of studies have assessed
the association between the IL-28B polymorphisms and risk of HCC and liver cirrhosis (LC)
development in different populations; however, the results are inconsistent and
inconclusive. For these reasons, a deep comprehension of the impact of IL28B polymorphisms
in HCC occurrence is mandatory.
Interestingly, some polymorphisms located at the 3' untranslated region (UTR) of IL28B, e.g.
rs 4803217, seem to interfere with the binding of miRNA, to date recognized as important
post-transcriptional regulators.
miRNA, in fact, are small, interfering, non-coding RNA that are 21-30 nucleotides in length,
that can promote the modulation of more than 200 mRNAs and are widely associated with human
cancers. In the last years miRNA acquired a growing relevance as potential biomarkers for
several diseases including cancer.
In particular, the discovery of extracellular miRNA, which are stable in circulation, drives
a lot of researchers on their evaluation aiming to the identification of new useful and less
invasive diagnostic markers also for HCC.
Objectives
The overall goal is to investigate the Impact of IL-28B rs12979860 and rs4803217 gene
polymorphisms associated with miRNAs deregulation on HCV-related hepatocellular carcinoma
Specific Objectives:
1. To determine the association of IL 28B polymorphism(s) with the risk of HCC in chronic
hepatitis C patients.
2. To screen a wide panel of miRNAs to uncover the deregulated ones during chronic
hepatitis C progression toward HCC
3. To identify an association among IL 28B polymorphism(s) and the major deregulated
miRNAs as predictor marker of HCC in chronic hepatitis C patients
Research Approach and Methodology
Patients and methods:
Patients:
This study will include 405 Subjects divided into 3 groups:
Group 1. This group will include patients with chronic hepatitis C (no. =135) Group2. This
group will include patients with chronic hepatitis C (no. =135) with cirrhosis (F4).
Group3. This group will include patients with HCV related HCC (no. =135) This is confirmed
by presence of focal lesion detected by Imaging (computed tomography (CT) and ultrasound),
and elevated serum AFP.
• The basis of the sample size calculation is the following:
- Alpha Feto Protein: ses 41-60% , sp 80-94%
- Prevalence of HCC among HCV infected patients: 5-10%
- Calculation is based on specificity of alpha feto protein at average 87%
A total of 405 cases with average 135 cases /group is required, based on confidence 90 and
margin of error 5 %, prevalence of HCC among HCV infected patients 7 %
Methods:
1. Serum α fetoprotein levels were routinely tested in all patients with decompensated
cirrhosis. AFP levels were generally elevated in cases that were proved to be HCC,
however, the levels ranged from 15 to 650 ng/ml. The diagnosis of HCC was confirmed by
a 4 phase multidetector computed tomography (CT) scan or dynamic contrast enhanced
magnetic resonance imaging (MRI).
2. HCV RNA quantification. Plasma will be obtained and HCV RNA determined by RT-PCR of
plasma using Cobas AmpliPrep/COBAS TaqMan HCV Test (Roche Diagnostics, Branchburg,
(NJ).
3. IL28B Polymorphism:
SNP rs12979860 and 4803217 will be determined in whole blood by allelic discrimination
using specific probes by real time PCR.
4. miRNA quantification
RNAs will be extracted from serum using miRNeasy Mini Kit (Quiagen) according to the
manufacturer's instruction.
The RNA purity will be assessed by the RNA concentration and quantified by NanoDrop
ND-1000 (Nanodrop, United States).
cDNA will be obtained by miScript II Reverse Transcription Kits (Quiagen) A
Preamplification will be performed using miScript PreAMP PCR Kits (Quiagen) Real Time
PCRarray will be done using miScript miRNA PCR Arrays, with SYBR Green PCR Master Mix
(Quiagen).
5. Statistical Analysis:
Statistical analyses will be performed using SPSS Statistics software. P-values, 0.05 were
considered significant.
IL28B SNPs comparisons will be done by stratifying patients according to rs12979860CC and
rs12979860CT/TT genotypes and 4803217 Analysis miRNA PCR Array will be done by specific Data
Analysis Software specifically supplied by Quiagen.
