- Email: [email protected]
Altered Immune Modulation is Key to Alcoholic Liver Disease Severity and NAFLD Development: A Tribal Dominated Northeast India Based Study
JOURNAL OF CLINICAL AND EXPERIMENTAL HEPATOLOGY
CyclinD1 and hTERT in CaGB and CL cases compared to controls; while downregulation of p53 was observed in the same cases. Conclusion: Deregulation of b-catenin, CyclinD1 and p53 is pivotal for the development of gall bladder disease and carcinogenesis. Deregulation of Wnt/b-catenin and telomerase signalling promotes gall bladder disease development and progression to carcinogenesis. Reactivation of hTERT is an early event in gall bladder disease development and may be used as a selective biomarker to access disease predisposition. b-catenin may be a used as a therapeutic target to control disease development and progression. Corresponding author: Sujoy Bose. Email: [email protected]
ALTERED IMMUNE MODULATION IS KEY TO ALCOHOLIC LIVER DISEASE SEVERITY AND NAFLD DEVELOPMENT: A TRIBAL DOMINATED NORTHEAST INDIA BASED STUDY Tarun Basumatary*, Moumita Bose*, Manash Kalitay, Anjan Kumar Saikiaz, Jagat Singh Terronx, Ajit Kakatix, Shyam Sundar Swargiary*, Ramie H Begumy, Sujoy Bose* *Department of Biotechnology, Gauhati University, Guwahati, Assam, India, yDepartment of Life Science and Bioinformatics, Assam University, Diphu campus, Diphu Assam, India, z Medicinal and Gastroenterological unit, Gauhati University, Guwahati, Assam, India, and xCivil Hospital, Diphu, Assam, India Aim: To delineate the role of differential expression/modulation of important cytokines in the development of ALD and its progression to advance liver disease, and non-alcoholic fatty liver disease (NAFLD). Methods: The study included tribal patients who were clinically proved of having alcoholic liver disease (ALD) of different severity grade, NAFLD cases, along with comparative cohorts of alcohol consumers without liver disease, age and sex-matched healthy-controls from different parts of Northeast India. Differential cytokine profile was studied by Magpix magnetic bead based
Table 1 Altered Th1/Th2 Expression in Different Stratified Groups(in pg/ml). Diagnosis
Case nos.
IFN g
IL-10
IL12p70
IL-2
IL-4
IL-6
Control
180
1.341 0.862
2.334 1.48
3.744 5.39
0.742 0.258
44.53 35.38
544.624 441.94
Chronic ALD
150
3.164 2.89
10.743 8.96
3.125 3.04
1.134 2.245
4.175 ± 3.51
380.121 236.918
ALD-cirr
60
8.770 ± 8.558
2.267 ± 1.26
19.322 ± 22.79
1.092 1.29
2.837 ± 0.88
35.796 ± 39.78
Alc-no dis
148
3.587 2.031
9.635 8.877
1.877 1.904
1.015 0.988
41.257 66.80
957.318 835.327
NAFLD
60
2.688 3.686
2.51 ± 1.26
79.775 ± 86.78
55.18 ± 16.19
18.94 ± 15.01
19.08 ± 17.65
Journal of Clinical and Experimental Hepatology | March 2014 | Vol. 4 | No. S2 | S30–S38
S37
AFLD and NAFLD
Background: Activation and reactivation of proto-oncogene b-catenin and telomerase respectively are critical in the development of cancers of various cellular aetiologies. Limited data is available on the role of these key signal transducers in the development of gall bladder diseases and progression to gall bladder carcinoma (CaGB). Aim: To study the modulation of b-catenin and associated key signal transducing molecules and their role in the development and progression of gall bladder diseases and carcinoma. Methods: Surgically ressected tissues of clinically and histopathologically proven cases of gallbladder disease patients [Cholelithiasis (CL, n = 25), Cholecystitis (CS, n = 20) and CaGB (n = 20) along with autopsy based controls (n = 10)] were collected from Central Hospital, NF Railway, Guwahati; and RIMS, Manipur. b-catenin, CyclinD1, p53 and human telomerase reverse transcriptase (hTERT) mRNA expres-sion by Real Time-PCR, using b-actin as internal control. Mutation analysis of b-catenin exon3 region was performed by PCR-sequencing based analysis. Protein expression and co-localization studies were performed by immunohistochemistry/IF/confocal microscopy and/or western blotting. Statistical analysis was performed by SPSS v13.0 software. Results: b-catenin mRNA expression was up-regulated chronologically in the gallbladder diseases (control
ABSTRACTS
multiplex ELISA and confirmed by FACS analysis (Th1/ Th2 kit, BD). Results: There is a distinct role of higher Th1-modulation in ALD and NAFLD development. Importantly, altered expression of important Th2 cytokines like IL-6 and antiinflammatory cytokine IL-10 plays a major role in development of cirrhosis and NAFLD; whereas gradient downregulation in IL-4 expression correlated with disease development and severity (Table1). Higher expression of TNF-a
22ND ANNUAL CONFERENCE–2014
at mRNA and protein levels correlated with disease severity. Conclusion: Th1 biased modulation is critical for ALD development and severity, and NAFLD. Hence new strategies to target cytokine metabolism as a form of therapy for ALD and NAFLD may be suitable approach to attenuate liver injury. Corresponding author: Sujoy Bose. Email: [email protected]
AFLD and NAFLD
S38
© 2014, INASL
CyclinD1 and hTERT in CaGB and CL cases compared to controls; while downregulation of p53 was observed in the same cases. Conclusion: Deregulation of b-catenin, CyclinD1 and p53 is pivotal for the development of gall bladder disease and carcinogenesis. Deregulation of Wnt/b-catenin and telomerase signalling promotes gall bladder disease development and progression to carcinogenesis. Reactivation of hTERT is an early event in gall bladder disease development and may be used as a selective biomarker to access disease predisposition. b-catenin may be a used as a therapeutic target to control disease development and progression. Corresponding author: Sujoy Bose. Email: [email protected]
ALTERED IMMUNE MODULATION IS KEY TO ALCOHOLIC LIVER DISEASE SEVERITY AND NAFLD DEVELOPMENT: A TRIBAL DOMINATED NORTHEAST INDIA BASED STUDY Tarun Basumatary*, Moumita Bose*, Manash Kalitay, Anjan Kumar Saikiaz, Jagat Singh Terronx, Ajit Kakatix, Shyam Sundar Swargiary*, Ramie H Begumy, Sujoy Bose* *Department of Biotechnology, Gauhati University, Guwahati, Assam, India, yDepartment of Life Science and Bioinformatics, Assam University, Diphu campus, Diphu Assam, India, z Medicinal and Gastroenterological unit, Gauhati University, Guwahati, Assam, India, and xCivil Hospital, Diphu, Assam, India Aim: To delineate the role of differential expression/modulation of important cytokines in the development of ALD and its progression to advance liver disease, and non-alcoholic fatty liver disease (NAFLD). Methods: The study included tribal patients who were clinically proved of having alcoholic liver disease (ALD) of different severity grade, NAFLD cases, along with comparative cohorts of alcohol consumers without liver disease, age and sex-matched healthy-controls from different parts of Northeast India. Differential cytokine profile was studied by Magpix magnetic bead based
Table 1 Altered Th1/Th2 Expression in Different Stratified Groups(in pg/ml). Diagnosis
Case nos.
IFN g
IL-10
IL12p70
IL-2
IL-4
IL-6
Control
180
1.341 0.862
2.334 1.48
3.744 5.39
0.742 0.258
44.53 35.38
544.624 441.94
Chronic ALD
150
3.164 2.89
10.743 8.96
3.125 3.04
1.134 2.245
4.175 ± 3.51
380.121 236.918
ALD-cirr
60
8.770 ± 8.558
2.267 ± 1.26
19.322 ± 22.79
1.092 1.29
2.837 ± 0.88
35.796 ± 39.78
Alc-no dis
148
3.587 2.031
9.635 8.877
1.877 1.904
1.015 0.988
41.257 66.80
957.318 835.327
NAFLD
60
2.688 3.686
2.51 ± 1.26
79.775 ± 86.78
55.18 ± 16.19
18.94 ± 15.01
19.08 ± 17.65
Journal of Clinical and Experimental Hepatology | March 2014 | Vol. 4 | No. S2 | S30–S38
S37
AFLD and NAFLD
Background: Activation and reactivation of proto-oncogene b-catenin and telomerase respectively are critical in the development of cancers of various cellular aetiologies. Limited data is available on the role of these key signal transducers in the development of gall bladder diseases and progression to gall bladder carcinoma (CaGB). Aim: To study the modulation of b-catenin and associated key signal transducing molecules and their role in the development and progression of gall bladder diseases and carcinoma. Methods: Surgically ressected tissues of clinically and histopathologically proven cases of gallbladder disease patients [Cholelithiasis (CL, n = 25), Cholecystitis (CS, n = 20) and CaGB (n = 20) along with autopsy based controls (n = 10)] were collected from Central Hospital, NF Railway, Guwahati; and RIMS, Manipur. b-catenin, CyclinD1, p53 and human telomerase reverse transcriptase (hTERT) mRNA expres-sion by Real Time-PCR, using b-actin as internal control. Mutation analysis of b-catenin exon3 region was performed by PCR-sequencing based analysis. Protein expression and co-localization studies were performed by immunohistochemistry/IF/confocal microscopy and/or western blotting. Statistical analysis was performed by SPSS v13.0 software. Results: b-catenin mRNA expression was up-regulated chronologically in the gallbladder diseases (control
ABSTRACTS
multiplex ELISA and confirmed by FACS analysis (Th1/ Th2 kit, BD). Results: There is a distinct role of higher Th1-modulation in ALD and NAFLD development. Importantly, altered expression of important Th2 cytokines like IL-6 and antiinflammatory cytokine IL-10 plays a major role in development of cirrhosis and NAFLD; whereas gradient downregulation in IL-4 expression correlated with disease development and severity (Table1). Higher expression of TNF-a
22ND ANNUAL CONFERENCE–2014
at mRNA and protein levels correlated with disease severity. Conclusion: Th1 biased modulation is critical for ALD development and severity, and NAFLD. Hence new strategies to target cytokine metabolism as a form of therapy for ALD and NAFLD may be suitable approach to attenuate liver injury. Corresponding author: Sujoy Bose. Email: [email protected]
AFLD and NAFLD
S38
© 2014, INASL