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Saudi Journal of Kidney Diseases and Transplantation
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Table of Contents   
ORIGINAL ARTICLE  
Year : 2014  |  Volume : 25  |  Issue : 3  |  Page : 577-581
Single nucleotide polymorphisms at erythropoietin, superoxide dismutase 1, splicing factor, arginine/serin-rich 15 and plasmacytoma variant translocation genes association with diabetic nephropathy


1 Department of Medical Laboratory Sciences, Faculty of Allied Health Sciences, Health Science Center, Kuwait
2 Department of Pathology, Faculty of Medicine, Health Science Center, Kuwait
3 Department of Internal Medicine, Farwaniyah Hospital, Kuwait
4 Nephrology Department, Mubarak Al-Kabeer Hospital, Kuwait

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Date of Web Publication9-May-2014
 

   Abstract 

A number of genes have been identified in diabetic nephropathy. Association between diabetes-associated nephropathy and polymorphisms in the erythropoietin (EPO) gene, variants in the superoxide dismutase 1 (SOD1) gene and plasmacytoma variant translocation 1 (PVT1) gene have been identified. The EPO, SOD1:SFRS15 and PVT1 genes were genotyped using the single nucleotide polymorphism (SNP) technique in 38 diabetic nephropathy patients (Group 1) compared with 64 diabetic type 2 subjects without nephropathy (Group 2) at the Mubarak Alkabeer Hospital, Kuwait. The frequency of the risk allele T of the EPO (rs1617640) gene was high in both groups (0.96 in Group 1 and 0.92 in Group 2). Similarly, SNPs of the PVT1 (rs2720709) gene showed a higher frequency of the risk allele G in both groups (0.70 in the Group 1 and 0.68 in Group 2). Although the frequency of the risk allele A was higher than the frequency of the non-risk allele C of the SOD1:SFRS15 gene in both groups, the lowest probability value was observed in those gene SNPs (P = 0.05). We observed that the A allele of the SOD1:SFRS15 gene (rs17880135) was more frequently present in Group 1 (0.75) compared with Group 2 (0.62). Susceptibility to diabetes-associated nephropathy is partially mediated by genetic predisposition, and screening tests may open the gate for new therapeutic approaches.

How to cite this article:
Alwohhaib M, Alwaheeb S, Alyatama N, Dashti AA, Abdelghani A, Hussain N. Single nucleotide polymorphisms at erythropoietin, superoxide dismutase 1, splicing factor, arginine/serin-rich 15 and plasmacytoma variant translocation genes association with diabetic nephropathy. Saudi J Kidney Dis Transpl 2014;25:577-81

How to cite this URL:
Alwohhaib M, Alwaheeb S, Alyatama N, Dashti AA, Abdelghani A, Hussain N. Single nucleotide polymorphisms at erythropoietin, superoxide dismutase 1, splicing factor, arginine/serin-rich 15 and plasmacytoma variant translocation genes association with diabetic nephropathy. Saudi J Kidney Dis Transpl [serial online] 2014 [cited 2019 Oct 20];25:577-81. Available from: http://www.sjkdt.org/text.asp?2014/25/3/577/132190

   Introduction Top


Diabetic nephropathy is a serious complication of diabetes mellitus, affecting 20-40% of patients with type 1 or type 2 diabetes, with a greater chance of occurrence within the first 15 years of the onset of the disease. [1],[2],[3] Furthermore, a strong familial component for the development of diabetic nephropathy was shown from studying siblings. [3] Certain ethnic groups seem to be at greater risk of developing nephropathy. [4],[5],[6],[7],[8] Associated groups of genes have been proposed as candidates that play a role in the genetic susceptibility to diabetic nephropathy. Products of a wide range of genes might mediate a sequence of pathological changes in the renal tissues that will cause end-stage renal disease. [9] Association between diabetes-associated nephropathy and polymorphisms in the erythropoietin (EPO) gene [10] and variants of the superoxide dismutase-1 (SOD1) have been identified. [11] Plasmacytoma variant translocation-1 (PVT1) gene was found to be associated with nephropathy related to type 1 and 2 diabetes. [12],[13]

EPO gene is a potent angiogenic factor observed in the diabetic human and mouse eye. [10] The T allele of single nucleotide polymerphism (SNP) rs1617640 in the promoter of the EPO gene was found to be significantly associated with end-stage renal disease in three European-American cohorts. [10]

The association between rs17880135 in the 3' region of SOD1 and the incidence of both severe nephropathy and persistent microalbuminuria was identified in patients with type 1 diabetes. [11] Strong evidence for a similar association was found between the rs2720709 of PVT1 gene and end-stage renal disease in type 2 diabetes subjects using SNPs. [13] Allelic discrimination using 5'Nuclease assay with fluorogenic probes provides a rapid and sensitive method for detecting known mutants or polymorphisms. [14],[15]

The aim of our study was to determine the profile of the EPO, SOD1 and PVT1 genes in diabetic nephropathy patients in our population. We used SNPs to determine the allele frequency of biallelic polymorphisms in the DNA samples of the diabetic nephropathy patients.


   Subjects and Methods Top


The EPO, SOD1:SFRS15 and PVT1 genes were genotyped using the SNP technique in 38 diabetic nephropathy patients who were on dialysis because of diabetic nephropathy, and we compared them with 64 diabetic subjects without evidence of diabetic nephropathy.

Genotyping of SNPs was performed on extracted DNA from the two groups of patients. After polymerase chain reaction (PCR) amplification of the samples, real-time PCR was performed before applying the allelic discrimination using a TaqMan® GTXpress™ Master Mix protocol (Applied Biosystems).

Genotyping frequencies for the risk and non-risk alleles of the three genes investigated were obtained by direct counting.


   Statistical Analysis Top


Analysis was performed using the chi-squared test calculated on 2 × 2 contingency tables using the Vassar Stats online calculator. Odds ratio (OR) with 95% confidence intervals (95% CIs) was calculated using the same program. Pearson's probability values were calculated using the same program. P-values less than 0.05 were considered as statistically significant.


   Results Top


The SNPs of the EPO, SOD1:SFRS15 and PVT1 genes were genotyped. No significant association was found in the EPO and PVT1 frequencies and any of the diabetic groups tested [Table 1]. The frequency of the risk allele T of the EPO gene was high in both groups (0.96 in the nephropathy group and 0.92 in the group without nephropathy). Similarly, SNPs of the PVT1 gene showed a higher frequency of the risk allele G in both diabetic groups (0.70 in the diabetic nephropathy group and 0.68 in the group without nephropathy). Although the frequency of the risk allele A was higher than the frequency of the non-risk allele C of the SOD1:SFRS15 gene in both groups, the lowest probability value was observed in that gene SNP (P = 0.05). We observed that the A allele of the SOD1:SFRS15 gene (rs17880135) was more frequently present in diabetic nephropathy individuals. The association of the SOD1: SFRS15 SNPs was observed more commonly in the diabetic nephropathy group (frequency of risk allele = 0.75) compared with the group without nephropathy (frequency of risk allele = 0.62).
Table 1: Association of the EPO, SOD1;SFRS15 and PVT1 gene SNPs with diabetes nephropathy (76 patients) and diabetes without nephropathy (128 patients).

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   Discussion Top


The results of our study showed the frequencies of the risk alleles of the EPO, SOD1 and PVT1 genes in diabetic nephropathy patients compared with diabetic patients without nephropathy.

EPO is a glycoprotein that plays a major role in the stimulation of bone marrow stem cells and erythropoiesis. It has also shown proliferation, migration and angiogenesis in vascular endothelial cells exposed to hypoxia. [16],[17] Expression of EPO receptors in vascular endothelial cells has been demonstrated. [10] SNPs in EPO affects its expression. Our study confirms the findings of Tong et al, in that SNPs at rs167640 in the promoter of EPO were found, where the T allele is significantly associated with end-stage renal disease. [10]

Alkateb et al found an association between rs17880135 in the SOD1 gene and the incidence of both severe nephropathy and persistent microalbuminuria. [11] Our results show a higher frequency of the A allele than the C allele in the diabetic subjects studied with and without nephropathy. However, the frequency of the A allele was more commonly associated with the diabetic nephropathy group (P = 0.05). SOD1 (Copper-zinc superoxide dismutase) is an anti-oxidant enzyme present in the cytosol, nuclei, peroxisomes and inner mitochondrial spaces. Studies show that overexpression of SOD1 reduces renal cell injury. [18],[19] Consistent with that, diabetic renal disease develops faster when SOD1 is not expressed adequately. [20] Our finding confirms the association of rs17800135 in SOD1 with diabetic nephropathy. Accordingly, adjusting the inadequate expression of SOD1 in diabetic patients prior to the development of the disease may delay or prevent the nephropathic effect of the mutant gene.

One of the susceptibility genes linked to the development of diabetic nephropathy is the PVT1 gene. PVT1 is located on 8q24, [12],[21] and it is well known for its participation in recurrent translocations between this region and chromosomes 2 and 22. [22],[23] PVT1 is expressed at high levels in the kidney; however, its role is not yet known. Some studies have focused on the investigation of PVT1 gene or its gene product. PVT1 has an impact on disregulated cell growth, particularly mesangial cell expansion, which is a hallmark of diabetic kidney disease. [24],[25] PVT1 may mediate the development and progression of diabetic nephropathy through mechanisms involving extracellular matrix proteins accumulation in the glomerular mesangium. [24],[25],[26],[27] PVT1 is co-amplified with the transcription factor MYC and plays a role in cell cycle progression, apoptosis and cellular transformation. [22],[28] Hanson et al found a strong association of rs2720709 SNPs of the PVT1 with diabetes. [13] Their results associated the G allele frequency with the diabetes type 2 subjects and the A allele with the end-stage renal disease subjects. In our study, the frequency of the G allele was high in both study groups. The high frequency of the G allele suggests that this alteration may be related to the development of diabetes type 2 with or without nephropathy. However, variants in PVT1 gene rs2720709 were found by Millis et al to be associated with end-stage renal disease related to type 1 diabetes due to the high frequency of the G allele. [14] Type 1 and type 2 diabetes share common complications resulting from either disease that often follows a similar course of progression. [26],[29]

The limitations of our study include the small number of investigated patients from one center. Nevertheless, we consider this study to be an initial step in studying the genetics of diabetes and its complications.

We conclude that our study found that the gene SOD1:SFRS15 was the only one from the investigated genes that showed a higher association of the risk allele A with diabetes nephropathy. Further confirmatory large studies should be carried out to clarify the roles of the different genes.


   Acknowledgments Top


The authors thank the Kuwait University for funding this study. Special thanks are also due to Dr. Fahad Al-Mulla for his advice during the project and facilitating the conduct of the SNPs in the Research Core Facility Laboratory in the health science center of Kuwait University.

 
   References Top

1.Kroleweski AS, Warram JH, Rand LI, Kahn CR. Epidemiologic approach to the etiology of type I diabetes mellitus and its complications. N Engl J Med 1987;317:1390-8.  Back to cited text no. 1
    
2.Nelson RG, Newman JM, Knowler WC, et al. incidence of end-stage renal disease in type 2 (non-insulin dependent) diabetes mellitus in Pima Indians. Diabetologia 1988;31:730-6.  Back to cited text no. 2
    
3.Ritz E, Orth S. Nephropathy in patients with type 2 diabetes mellitus. N Engl J Med 1999; 341:1127-33.  Back to cited text no. 3
    
4.Petitt DJ, Saad MF, Bernnett PH, Nelson RG, Knwoler WC. Familial predisposition to renal disease in two generations of Pima Indians with type 2 (non-insulin dependent) diabetes mellitus. Diabetologia 1990;33:438-43.  Back to cited text no. 4
    
5.Seaquist ER, Goetz FC, Rich S, Barbosa J. Familial clustering of diabetic kidney disease: Evidence for genetic susceptibility to diabetic nephropathy. N Engl J Med 1989;320:1161-5.  Back to cited text no. 5
    
6.Canani LH, Gerchman F, Gross JL. Familial clustering of diabetic nephropathy in Brazilian type 2 diabetic patients. Diabetes 1999;48:909-13.  Back to cited text no. 6
    
7.Mooyaart AL, van Valkengoed IG, Shaw PK, et al. Lower frequency of the 5/5 homozygous CNDP1 genotype in South Asian Surinamese. Diabetes Res Clin Pract 2009;85:272-9.  Back to cited text no. 7
    
8.Quinn M, Angelico MC, Warram JH, Krolewski AS. Familial factors determine the development of diabetic nephropathy in patients with IDDM. Diabetologia 1996;39:940-5.  Back to cited text no. 8
    
9.Ewens KG, George RA, Sharma K, Ziyadeh FN, Spielman RS. Assessment of 115 candidate genes for diabetic nephropathy by transmission/disequilibrium test. Diabetes 2005;54: 3305-18.  Back to cited text no. 9
    
10.Tong Z, Yang Z, Patel S, et al. Promoter polymorphism of the erythropoietin gene in severe diabetic eye and kidney complications. Proc Natl Acad Sci USA 2008;105:6998-7003.  Back to cited text no. 10
    
11.Al-kateb H, Boright AP, Mirea L, et al. Multiple superoxide dismutase 1/splicing factor serine alanine 15 variants are associated with the development and progression of diabetic nephropathy. Diabetes 2008;57:218-28.  Back to cited text no. 11
    
12.Millis MP, Bowen D, Kingsley C, Watanabe RM, Wolford JK. Variants in the plasma-cytoma variant translocation gene (PVT1) are associated with end stage renal disease attributed to type 1 diabetes. Diabetes 2007;56: 3027-32.  Back to cited text no. 12
    
13.Hanson RL, Bogardus C, Duggan D, et al. A search for variants associated with young-onset type 2 diabetes in American Indians in a 100 K genotyping array. Diabetes 2007;56:3045-52.  Back to cited text no. 13
    
14.Livak KJ, Marmaro J, Todd JA. Towards fully automated genome-wide polymorphism screening [letter]. Nat Genet 1995;9:341-2.  Back to cited text no. 14
    
15.Livak KJ. Allelic discrimination using fluorogenic probles and the 5'nuclease assay. Genet Anal 1999;14:143-9.  Back to cited text no. 15
    
16.Yamaji R, Okada T, Moriya M, et al. Brain capillary endothelial cells express two forms of erythropoietin receptor mRNA. Eur J Biochem 1996;239:494-500.  Back to cited text no. 16
    
17.Anagnostou A, Lee ES, Kessimian N, Levinson R, Steiner M. Erythropoietin has a mitogenic and positive chemotactic effect on endothelial cells. Proc Natl Acad Sci USA 1990;87:5978-82.  Back to cited text no. 17
    
18.DeRubertis FR, Craven PA, Melhem MF, Salah EM. Attenuation of renal injury in db/db mice over expression dismutase: Evidence for reduced superoxidenitric oxide interaction. Diabetes 2004;53:762-8.  Back to cited text no. 18
    
19.Craven PA, Melhem MF, Philips SL, DeRubertis FR. Overexpression of Cu2+/Zn2+ superoxide dismutase protects against early diabetic glomerular injury in transgenic mice. Diabetes 2001;50:2114-25.  Back to cited text no. 19
    
20.Derubertis FR, Craven PA, Melhem MF. Acceleration of diabetic renal injury in the superoxide dismutase knockout mouse: Effects of tempol. Metabolism 2007;56:1256-64.  Back to cited text no. 20
    
21.Imperatore G, Knowler WC, Pettitt DJ, Kobes S, Bennett PH, Hanson RL. Segregation analysis of diabetic nephropathy in Pima Indians. Diabetes 2000;49:1049-56.  Back to cited text no. 21
    
22.Graham M, Adams JM. Chromosome 8 break-point far 3' of the c-myc oncogene in a Burkitt's lymphoma 2;8 variant translocation is equivalent to the murine pvt-1 locus. Embo J 1986;5:2845-51.  Back to cited text no. 22
    
23.Mengle-Gaw L, Rabitts TH. A human chromosome 8 region with abnormalities in B cell, HTLV-I+ T cell and c-myc amplified tumours. Embo J 1987;6:1959-65.  Back to cited text no. 23
    
24.Alvares ML, DiStefano JK. Functional characterization of plasmacytoma variant translocation 1 gene (PVT) in diabetic nephropathy. PloS One 2011;6:e18671.  Back to cited text no. 24
    
25.Mason RM, Wahab NA. Extracellular matrix metabolism in diabetic nephropathy. J Am Soc Nephrol 2003;14:1358-73.  Back to cited text no. 25
    
26.Tsilbary EC. Macrovascular basement membranes in diabetes mellitus. J Pathol 2003;200: 537-46.  Back to cited text no. 26
    
27.Haneda M, Koya D, Isono M, Kikkawa R. Overview of glucose signalling in mesangial cells in diabetic nephropathy. J Am Soc Nephrol 2003;14:1374-82.  Back to cited text no. 27
    
28.Shtivelman E, Bishop JM. The PVT gene frequency amplifies with MYC in tumor cells. Mol Cell Biol 1989;9:1148-54.  Back to cited text no. 28
    
29.Caramori ML, Mauer M. Diabetes and nephropathy. Curr Opin Nephrol Hypertens 2003;12: 273-82.  Back to cited text no. 29
    

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Correspondence Address:
Dr. Maisaa Alwohhaib
Department of Medical Laboratory Sciences, Faculty of Allied Health Sciences, Health Science Center
Kuwait
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DOI: 10.4103/1319-2442.132190

PMID: 24821155

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    Abstract
   Introduction
   Subjects and Methods
   Statistical Analysis
   Results
   Discussion
   Acknowledgments
    References
    Article Tables
 

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