Ignite Creation Date:
2024-05-06 @ 7:16 AM
Last Modification Date:
2024-10-26 @ 11:47 AM
Study NCT ID:
NCT02507882
Status:
UNKNOWN
Last Update Posted:
2015-07-24
First Post:
2015-07-22
Brief Title:
Impact of IL-28B rs12979860 and rs4803217 Gene Polymorphisms Associated With miRNAs Deregulation on HCV-related Hepatocellular Carcinoma
Sponsor:
DrWaleed Samir
Organization:
Egyptian Liver Hospital
Study Overview
Official Title:
Impact of IL-28B rs12979860 and rs4803217 Gene Polymorphisms Associated With miRNAs Deregulation on HCV-related Hepatocellular Carcinoma
Status:
UNKNOWN
Status Verified Date:
2015-07
Last Known Status:
NOT_YET_RECRUITING
Delayed Posting:
No
If Stopped, Why?:
Not Stopped
Has Expanded Access:
False
If Expanded Access, NCT#:
N/A
Has Expanded Access, NCT# Status:
N/A
Acronym:
None
Brief 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 515 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 CT 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 eg 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
It is estimated that 515 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 CT 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 eg 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
Detailed 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 515 of HCC cases can be attributed to HCV infection Among them we have to mention the polymorphisms of IL28B gene eg rs12979860 CT 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 adaptive 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 eg 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 polymorphisms 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 polymorphisms 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 ngml 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 AmpliPrepCOBAS 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 manufacturers 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 005 were considered significant
IL28B SNPs comparisons will be done by stratifying patients according to rs12979860CC and rs12979860CTTT genotypes and 4803217 Analysis miRNA PCR Array will be done by specific Data Analysis Software specifically supplied by Quiagen
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 515 of HCC cases can be attributed to HCV infection Among them we have to mention the polymorphisms of IL28B gene eg rs12979860 CT 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 adaptive 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 eg 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 polymorphisms 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 polymorphisms 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 ngml 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 AmpliPrepCOBAS 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 manufacturers 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 005 were considered significant
IL28B SNPs comparisons will be done by stratifying patients according to rs12979860CC and rs12979860CTTT genotypes and 4803217 Analysis miRNA PCR Array will be done by specific Data Analysis Software specifically supplied by Quiagen
Study Oversight
Has Oversight DMC:
None
Is a FDA Regulated Drug?:
None
Is a FDA Regulated Device?:
None
Is an Unapproved Device?:
None
Is a PPSD?:
None
Is a US Export?:
None
Is an FDA AA801 Violation?:
None