Sains Malaysiana 52(3)(2023): 851-862

http://doi.org/10.17576/jsm-2023-5203-13

 

Investigation of Gene Variation among Non-Alcoholic Fatty Liver Disease Patients in Gaza Strip: A Preliminary Study

(Kajian Variasi Gen dalam Kalangan Pesakit Hati Berlemak Bukan Alkohol di Semenanjung Gaza: Suatu Kajian Awal)

 

ABEER AL-QATATI 1, ALI AL-BELTAJI2 & MAZEN ALZAHARNA2,*

 

1Clinical Laboratory Sciences Department, Faculty of Science, The University of Jordan, Amman, Jordan

2Medical Laboratory Sciences Dept., Faculty of Health Sciences, Islamic University of Gaza, P.O. Box 108, Gaza city, Palestine

 

Diserahkan: 18 Oktober 2022/Diterima: 7 Februari 2023

 

Abstract

The prevalent hepatic presentation of the metabolic syndrome is a non-alcoholic fatty liver disease (NAFLD), one of the most common types of chronic liver illnesses. Patients with NAFLD may develop liver damage depending on their genetic heritage. In this preliminary study, our main aim was to detect the genetic association of p85α (Met326Ile), PNPLA3 (C>G), IL28B275 (A>G), and IL28B860 (C>T) single nucleotide polymorphisms (SNPs) with steatosis and NASH in patients from Gaza Strip. We performed an SNP analysis by RFLP-PCR in 33 cases of steatosis and 28 cases of non-alcoholic steatohepatitis (NASH), in addition to 29 age- and sex-matched controls. We found that only the mutant T allele of IL28B860 was significantly associated with an increased risk of steatosis (P = 0.04). The other studied alleles and genotypes were not significantly associated with increased or decreased risk of steatosis, NASH, or combined steatosis or NASH groups. Among all of the studied variables (age, sex, diabetes, and BMI), only BMI was significantly associated with an increased risk of steatosis as well as NASH. A linkage disequilibrium analysis showed that the association between the two SNPs of IL28B860 and IL28B275 was significant. Having the TG haplotype increased the risk of steatosis by 2.97 fold and the risk of combined steatosis or NASH by 2.44 fold. This haplotype increased the risk of NASH, but the effect was not significant.

 

Keywords: NAFLD; NASH; preliminary study; single nucleotide polymorphism; steatosis

 

Abstrak

Persembahan hati yang lazim bagi sindrom metabolik ialah penyakit hati berlemak bukan alkohol (NAFLD), salah satu jenis penyakit hati kronik yang paling biasa. Pesakit dengan NAFLD mungkin mengalami kerosakan hati bergantung kepada warisan genetik mereka. Dalam kajian awal ini, matlamat utama kami adalah untuk mengesan perkaitan genetik p85α (Met326Ile), PNPLA3 (C> G), IL28B275 (A> G) dan IL28B860 (C> T) polimorfisme nukleotida tunggal (SNP) dengan steatosis dan NASH pada pesakit dari Semenanjung Gaza. Kami menjalankan analisis SNP dengan RFLP-PCR dalam 33 kes steatosis dan 28 kes steatohepatitis bukan alkohol (NASH), sebagai tambahan kepada 29 kawalan padanan umur dan jantina. Kami mendapati bahawa hanya alel T mutan IL28B860 dikaitkan dengan peningkatan risiko steatosis (P = 0.04). Kajian lain Alel dan genotip tidak dikaitkan dengan peningkatan atau penurunan risiko steatosis, NASH, atau gabungan steatosis atau kumpulan NASH. Antara semua pemboleh ubah yang dikaji (umur, jantina, diabetes dan BMI), hanya BMI dikaitkan dengan peningkatan risiko steatosis serta NASH. Analisis ketidakseimbangan kaitan menunjukkan bahawa perkaitan antara dua SNP IL28B860 dan IL28B275 adalah signifikan. Mempunyai haplotip TG meningkatkan risiko steatosis sebanyak 2.97 kali ganda dan risiko gabungan steatosis atau NASH sebanyak 2.44 kali ganda. Haplotip ini meningkatkan risiko NASH, tetapi kesannya tidak ketara.

 

Kata kunci: Kajian awal; NAFLD; NASH; polimorfisme nukleotida tunggal; steatosis

 

RUJUKAN

Abe, H., Ochi, H., Maekawa, T., Hayes, C.N., Tsuge, M., Miki, D., Mitsui, F., Hiraga, N., Imamura, M. & Takahashi, S. 2010. Common variation of IL28 affects gamma-GTP levels and inflammation of the liver in chronically infected hepatitis C virus patients. Journal of Hepatology 53(3): 439-443.

Agundez, J.A., Garcia-Martin, E., Maestro, M.L., Cuenca, F., Martinez, C., Ortega, L., Carballo, M., Vidaurreta, M., Agreda, M. & Diaz-Zelaya, G. 2012. Relation of IL28B gene polymorphism with biochemical and histological features in hepatitis C virus-induced liver disease. PLoS ONE 7(5): e37998.

Anstee, Q.M. & Day, C.P. 2015. The genetics of nonalcoholic fatty liver disease: Spotlight on PNPLA3 and TM6SF2. Seminars in Liver Disease 35(3): 270-290.

Baier, L.J., Wiedrich, C., Hanson, R.L. & Bogardus, C. 1998. Variant in the regulatory subunit of phosphatidylinositol 3-kinase (p85alpha): Preliminary evidence indicates a potential role of this variant in the acute insulin response and type 2 diabetes in Pima women. Diabetes 47(6): 973-975.

Chen, S., Yan, W., Huang, J., Ge, D., Yao, Z. & Gu, D. 2005. Association analysis of the variant in the regulatory subunit of phosphoinositide 3‐kinase (p85α) with Type 2 diabetes mellitus and hypertension in the Chinese Han population. Diabetic Medicine 22(6): 737-743.

Clark, P.J., Thompson, A.J., Zhu, M., Vock, D.M., Zhu, Q., Ge, D., Patel, K., Harrison, S.A., Urban, T.J. & Naggie, S. 2012. Interleukin 28B polymorphisms are the only common genetic variants associated with low‐density lipoprotein cholesterol (LDL‐C) in genotype‐1 chronic hepatitis C and determine the association between LDL‐C and treatment response. Journal of Viral Hepatitis 19(5): 332-340.

Clark, P.J., Thompson, A.J., Zhu, Q., Vock, D.M., Zhu, M., Patel, K., Harrison, S.A., Naggie, S., Ge, D., Tillmann, H.L., Urban, T.J., Shianna, K., Fellay, J., Goodman, Z., Noviello, S., Pedicone, L.D., Afdhal, N., Sulkowski, M., Albrecht, J.K., Goldstein, D.B., McHutchison, J.G. & Muir, A.J. 2012. The association of genetic variants with hepatic steatosis in patients with genotype 1 chronic hepatitis C infection. Digestive Diseases and Sciences 57(8): 2213-2221.

Dutta, A.K. 2011. A new PCR-RFLP method for diagnosing PNPLA3 RS738409 polymorphism.

Engwa, G.A., Nwalo, F.N., Chiezey, V.O., Unachukwu, M.N., Ojo, O.O. & Ubi, B.E. 2018. Assessment of the Pro12Ala polymorphism in the PPAR-γ2 gene among type 2 diabetes patients in a Nigerian population. Journal of Clinical Medicine 7(4): 69.

Ficarella, R., Laviola, L. & Giorgino, F. 2015. Lipodystrophic diabetes mellitus: A lesson for other forms of diabetes? Current Diabetes Reports 15(3): 1-10.

Fruman, D.A., Meyers, R.E. & Cantley, L.C. 1998. Phosphoinositide kinases. Annual Review of Biochemistry 67: 481.

Ge, D., Fellay, J., Thompson, A.J., Simon, J.S., Shianna, K.V., Urban, T.J., Heinzen, E.L., Qiu, P., Bertelsen, A.H. & Muir, A.J. 2009. Genetic variation in IL28B predicts hepatitis C treatment-induced viral clearance. Nature Biotechnology 461(7262): 399-401.

Hansen, L., Zethelius, B., Berglund, L., Reneland, R., Hansen, T., Berne, C., Lithell, H., Hemmings, B.A. & Pedersen, O. 2001. In vitro and in vivo studies of a naturally occurring variant of the human p85α regulatory subunit of the phosphoinositide 3-kinase: Inhibition of protein kinase B and relationships with type 2 diabetes, insulin secretion, glucose disappearance constant, and insulin sensitivity. Diabetes and Metabolism 50(3): 690-693.

Hansen, T., Andersen, C.B., Echwald, S.M., Urhammer, S.A., Clausen, J.O., Vestergaard, H., Owens, D., Hansen, L. & Pedersen, O. 1997. Identification of a common amino acid polymorphism in the p85α regulatory subunit of phosphatidylinositol 3-kinase: Effects on glucose disappearance constant, glucose effectiveness, and the insulin sensitivity index. Diabetes 46(3): 494-501.

Honda, M., Sakai, A., Yamashita, T., Nakamoto, Y., Mizukoshi, E., Sakai, Y., Yamashita, T., Nakamura, M., Shirasaki, T. & Horimoto, K. 2010. Hepatic ISG expression is associated with genetic variation in interleukin 28B and the outcome of IFN therapy for chronic hepatitis C. Gastroenterology 139(2): 499-509.

Kawanishi, M., Tamori, Y., Masugi, J., Mori, H., Ito, C., Hansen, T., Andersen, C.B., Pedersen, O. & Kasuga, M. 1997. Prevalence of a polymorphism of the phosphatidylinositol 3-Kinase p85α regulatory subunit (Codon 326Met→ ile) in Japanese NIDDM patients. Diabetes Care 20(6): 1043.

Lankarani, K.B., Ghaffarpasand, F., Mahmoodi, M., Lotfi, M., Zamiri, N., Heydari, S.T., Fallahzadeh, M.K., Maharlouei, N., Babaeinejad, M. & Mehravar, S. 2013. Non alcoholic fatty liver disease in southern Iran: A population based study. Hepatitis Monthly 13(5): e9248.

Li, J.H., Lao, X.Q., Tillmann, H.L., Rowell, J., Patel, K., Thompson, A., Suchindran, S., Muir, A.J., Guyton, J.R. & Gardner, S.D., McHutchison, J.G. & McCarthy, J.J. 2010. Interferon‐lambda genotype and low serum low‐density lipoprotein cholesterol levels in patients with chronic hepatitis C infection.  Hepatology 51(6): 1904-1911.

Li, L., Liu, D‐W., Yan, H‐Y., Wang, Z‐Y., Zhao, S‐H. & Wang, B. 2016. Obesity is an independent risk factor for non‐alcoholic fatty liver disease: Evidence from a meta‐analysis of 21 cohort studies. Obesity Reviews 17(6): 510-519.

Ludwig, J., Viggiano, T.R., Mcgill, D.B. & Oh, B.J. 1980. Nonalcoholic steatohepatitis: Mayo clinic experiences with a hitherto unnamed disease. Mayo Clinic Proceedings 55(7): 434-438.

Matsuda, S., Kobayashi, M. & Kitagishi, Y. 2013. Roles for PI3K/AKT/PTEN pathway in cell signaling of nonalcoholic fatty liver disease. International Scholarly Research Notices 2013: Article ID. 472432.

Petta, S., Grimaudo, S., Cammà, C., Cabibi, D., Marco, V.D., Licata, G., Pipitone, R.M. & Craxì, A. 2012. IL28B and PNPLA3 polymorphisms affect histological liver damage in patients with non-alcoholic fatty liver disease. Journal of Hepatology 56(6): 1356-1362.

Pirazzi, C., Adiels, M., Burza, M.A., Mancina, R.M., Levin, M., Ståhlman, M., Taskinen, M-R., Orho-Melander, M., Perman, J. & Pujia, A. 2012. Patatin-like phospholipase domain-containing 3 (PNPLA3) I148M (rs738409) affects hepatic VLDL secretion in humans and in vitro. Journal of Hepatology 57(6): 1276-1282.

Rüstemoğlu, A., Yalcin, D., Günal, Ö., Çelik, B., Barut, Ş. & Ateş, Ö. 2016. Interleukin 28B rs12979860 CT, rs12980275 GA, rs8099917 GT and TT genotypes are the predictors of rapid viral response in hepatitis C virus-infected patients. Viral Hepat. J. 22(3): 97-102.

Schreuder, T.C.M.A., Verwer, B.J., van Nieuwkerk, C.M.J. & Mulder, C.J.J. 2008. Nonalcoholic fatty liver disease: An overview of current insights in pathogenesis, diagnosis and treatment. World Journal of Gastroenterology: WJG 14(16): 2474-2486.

Schwimmer, J.B., Celedon, M.A., Lavine, J.E., Salem, R., Campbell, N., Schork, N.J., Shiehmorteza, M., Yokoo, T., Chavez, A. & Middleton, M.S. 2009. Heritability of nonalcoholic fatty liver disease. Gastroenterology 136(5): 1585-1592.

Shibata, M., Kihara, Y., Taguchi, M., Tashiro, M. & Otsuki, M. 2007. Nonalcoholic fatty liver disease is a risk factor for type 2 diabetes in middle-aged Japanese men. Diabetes Care 30(11): 2940-2944.

Sookoian, S. & Pirola, C.J. 2011. Meta‐analysis of the influence of I148M variant of patatin‐like phospholipase domain containing 3 gene (PNPLA3) on the susceptibility and histological severity of nonalcoholic fatty liver disease. Hepatology 53(6): 1883-1894.

Tai, C-M., Huang, C-K., Tu, H-P., Hwang, J-C., Chang, C-Y. & Yu, M-L. 2015. PNPLA3 genotype increases susceptibility of nonalcoholic steatohepatitis among obese patients with nonalcoholic fatty liver disease. Surgery for Obesity and Related Diseases 11(4): 888-894.

Tanase, D.M., Gosav, E.M., Costea, C.F., Ciocoiu, M., Lacatusu, C.M., Maranduca, M.A., Ouatu, A. & Floria, M. 2020. The intricate relationship between type 2 diabetes mellitus (T2DM), insulin resistance (IR), and nonalcoholic fatty liver disease (NAFLD). Journal of Diabetes Research 2020: 3920196.

Thompson, A.J., Clark, P.J., Zhu, M., Zhu, Q., Ge, D., Sulkowski, M.S., Muir, A.J., Tillmann, H.L., Patel, K. & Naggie, S. 2010. Genome wide-association study identifies IL28B polymorphism to be associated with baseline ALT and hepatic necro-inflammatory activity in chronic hepatitis C patients enrolled in the IDEAL study. Hepatology 52(4): 1220A-1221A.

Tilg, H. & Moschen, A. 2010. Update on nonalcoholic fatty liver disease: Genes involved in nonalcoholic fatty liver disease and associated inflammation. Current Opinion in Clinical Nutrition Metabolic Care 13(4): 391-396.

Tillmann, H.L., Patel, K., Muir, A.J., Guy, C.D., Li, J.H., Lao, X.Q., Thompson, A., Clark, P.J., Gardner, S.D. & McHutchison, J.G. 2011. Beneficial IL28B genotype associated with lower frequency of hepatic steatosis in patients with chronic hepatitis C. Journal of Hepatology 55(6): 1195-1200.

Xia, M-F., Lin, H-D., Chen, L-Y., Wu, L., Ma, H., Li, Q., Aleteng, Q., Hu, Y., He, W-Y. & Gao, J. 2019. The PNPLA3 rs738409 C> G variant interacts with changes in body weight over time to aggravate liver steatosis, but reduces the risk of incident type 2 diabetes. Diabetologia 62(4): 644-654.

Xu, R., Tao, A., Zhang, S., Deng, Y. & Chen, G. 2015. Association between patatin-like phospholipase domain containing 3 gene (PNPLA3) polymorphisms and nonalcoholic fatty liver disease: A HuGE review and meta-analysis. Scientific Reports 5(1): 1-11.

Younes, R. & Bugianesi, E. 2019. NASH in lean individuals. Seminars in Liver Disease 39(1): 86-95.

Zhang, L., You, W., Zhang, H., Peng, R., Zhu, Q., Yao, A., Li, X., Zhou, Y., Wang, X. & Pu, L. 2015. PNPLA3 polymorphisms (rs738409) and non‐alcoholic fatty liver disease risk and related phenotypes: A meta‐analysis. Journal of Gastroenterology and Hepatology 30(5): 821-829.

 

*Pengarang untuk surat-menyurat; email: mzaharna@iugaza.edu.ps

   

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