Lack of association between CD45 C77G polymorphism and multiple sclerosis in Kashmir

Multiple sclerosis (MS) is a severe disabling and demyelinating disease of the nervous system. Its etiology involves profound genetic component. The latest contender known to have been correlated with MS is protein tyrosine phosphatase receptor (PTPRC or CD45); however, to date its role remains contentious. The aim of the current study was to examine the association of functionally significant exon 4 C77G polymorphism of CD45 with MS in Kashmiri population from Indian subcontinent. The preliminary findings of our study revealed absence of C77G in majority of the cases as well as controls. These findings strongly suggest that the alterations in CD45 are sporadically associated with the genesis of MS. In conclusion, results from our study are in accordance with some of the international studies; however, more studies with large datasets from Kashmir as well as other ethnic populations are warranted to validate the above preliminary findings and demonstrate the role of CD45 C77G polymorphism in MS pathogenesis. | Volume – 1 | Issue – 6 | Sep Oct 2017 6470 | www.ijtsrd.com | Volume Journal of Trend in Scientific and Development (IJTSRD) International Open Access Journal , Amrina Shafi , Mudasir A Mir 3a -e-Kashmir University of Agricultural (Tel.: +91-979-708-8530; E-mail: inshazahoor11@gmail.com -type C

Multiple sclerosis (MS) is a severe disabling and disease of the nervous system. Its etiology involves profound genetic component. The latest contender known to have been correlated with MS is protein tyrosine phosphatase receptor-type C (PTPRC or CD45); however, to date its role remains aim of the current study was to examine the association of functionally significant exon 4 C77G polymorphism of CD45 with MS in Kashmiri population from Indian subcontinent. The preliminary findings of our study revealed absence of cases as well as controls. These findings strongly suggest that the alterations in CD45 are sporadically associated with the genesis of MS. In conclusion, results from our study are in accordance with some of the international studies; es with large datasets from Kashmir as well as other ethnic populations are warranted to validate the above preliminary findings and demonstrate the role of CD45 C77G

Keywords:
MS; genetics; CD45; C77G; polymorphism; Kashmir INTRODUCTION Multiple sclerosis (MS) is a chronic autoimmune inflammatory disease which primarily affects central nervous system (CNS) of the body. Owing to its inexplicable etiology, it has been found to develop due to complex interactions between environmental and genetic factors; however, genetic components seem to play a major rule in its pathogenesis [1 The variations in different candidate genes of MS play an important role in disease susceptibility [4]. By and large MS susceptibility has been linked to the Major Histocompatibility Complex (MHC) [5,6], although there are other genes as well which have been found to be associated with its development [7,8]. Incidentally, few previous studies have reported a polymorphism in exon 4 of protein tyrosine phosphatase receptor-type C (PTPRC or CD45), which results in replacement of C to G at nucleotide 77 [9][10][11]; albeit it is not conclusive and it has been complicated due to controversial data from some studies [12][13][14][15] Multiple sclerosis (MS) is a chronic autoimmune inflammatory disease which primarily affects central nervous system (CNS) of the body. Owing to its etiology, it has been found to develop due to complex interactions between environmental and genetic factors; however, genetic components seem to play a major rule in its pathogenesis [1][2][3]. The variations in different candidate genes of MS play ant role in disease susceptibility [4]. By and large MS susceptibility has been linked to the Major Histocompatibility Complex (MHC) [5,6], although there are other genes as well which have been found to be associated with its development [7,8].
ally, few previous studies have reported a polymorphism in exon 4 of protein tyrosine type C (PTPRC or CD45), which results in replacement of C to G at nucleotide 11]; albeit it is not conclusive and it has been o controversial data from some CD45 [OMIM 151460; Entrez Gene International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470 Page: 1334 5788] is located on chromosome 1 at position q31.3-q32.1 and belongs to the family of protein tyrosine phosphatase (PTP). It plays a critical role in regulating protein-tyrosine kinases involved in signal transduction thereby activating T and B cells [16,17]. Keeping in view its essentiality for the functioning of immune system, we investigated the association between CD45 and MS in an Indian population, covering the patients from Kashmir region.

Sequencing
For examining C77G polymorphism in CD45, the amplified products were sequenced commercially through the services of SciGenom Labs, Pvt., Ltd., Kerala, India (http://www.scigenom.com). Nucleotide sequences of the amplicons were viewed and analyzed for alterations by comparing with NCBI reference sequence using ClustalX Version 2 [19,20]

III. Results and Discussion
The present study comprised of 20 RRMS patients as well as contro patients from its northern region Kashmir. All the subjects were of the Kashmiri ethnicity. Agarose gel electrophoresis revealed successful extraction of DNA from blood samples of both cases as well as controls (Fig. 1). The extracted DNA was used as a template for amplification of CD45 gene and PCR products on analysis showed specific bands on the agarose gel as per expected size of 199 bp (Fig. 2). sequence alignment and chromatograms of nucleoti cases as well as controls (Fig. 3).  The present study comprised of 20 RRMS patients as well as controls from the Indian data set, covering the patients from its northern region Kashmir. All the subjects were of the Kashmiri ethnicity. Agarose gel electrophoresis revealed successful extraction of DNA from blood samples of both cases as well as controls g. 1). The extracted DNA was used as a template for amplification of CD45 gene and PCR products on analysis showed specific bands on the agarose gel as per expected size of 199 bp (Fig. 2). sequence alignment and chromatograms of nucleotide sequences of samples, C77G was found to be absent in ls from the Indian data set, covering the patients from its northern region Kashmir. All the subjects were of the Kashmiri ethnicity. Agarose gel electrophoresis revealed successful extraction of DNA from blood samples of both cases as well as controls g. 1). The extracted DNA was used as a template for amplification of CD45 gene and PCR products on analysis showed specific bands on the agarose gel as per expected size of 199 bp (Fig. 2). After analysis of de sequences of samples, C77G was found to be absent in The development of MS is a complex and multifactorial process mediated by various genetic and environmental factors involving numerous related pathways [1,3,8,21]. It has been seen that genetic alterations contribute to the genesis of MS [8]. To a large extent, progress has been attained in recognizing the candidate genes and gene defects that underlie inherited predisposition to MS. Therefore, screening of MS candidate genes for genetic variations can further clarify the relationship between genetics of MS and its development. Even though there is paucity of reports on the association between MS and CD45, some previous studies have reported C77G polymorphism in exon 4 of CD45 among MS patients from different ethnic backgrounds [9][10][11]. Keeping in view the association between C77G polymorphism of CD45 with MS, we screened this variant in ethnic cohort of Kashmiri patients as our population being small, isolated, and containing conserved gene pool makes it a right candidate for gene analysis study. Sequence analysis indicated that no alteration exists in CD45 in MS patients from Kashmir valley of India. This study strongly suggests that alterations present in MS patients vary from population to population and it can be inferred that alterations found in one population may not be necessarily present in MS patients from other geographic area as both environmental as well as genetic factors influence MS development. Thus, it can be suggested that alterations in CD45 are infrequently associated with the genesis of MS and its association with MS susceptibility strongly depends on environmental factors.

IV. Conclusion
To the best of our knowledge, this is the first study from India in which we examined the presence of C77G polymorphism in CD45 gene in Kashmiri population. Our preliminary findings clearly revealed absence of C77G polymorphism in MS patients as well as controls. However, keeping in view primary limitation of our study due to very small sample size, it becomes mandatory to validate our findings by taking into account large datasets from Kashmir region of India.