Differential gene expression analysis in patients with primary hyperhidrosis

Abstract

Hyperhidrosis, a condition of excessive sweat generation, is believed to be genetic, but it is yet not known well whether primary hyperhidrosis is regulated by a single gene or multiple genes. The prospective genes that regulate primary hyperhidrosis are known to be ITPR2, TMEM16A, FOXA1 and AQP5. Of these four genes, ITPR2 gene plays a significant role in primary hyperhidrosis; thus in the current research sequence, analysis of this gene was carried out. Following inclusion and exclusion criteria, whole blood samples were taken from four patients inflicted with primary hyperhidrosis and two from healthy persons as control. From the blood samples, white blood cells were separated by reacting the samples with ammonium chloride (NH₄Cl) solution to lyse red blood cells. Then total RNA was extracted using the trizol reagent. For synthesizing cDNA, one script reverse transcriptase cDNA synthesis kit was employed. The cDNA samples were quantified using a Qubit Flourometer and their quality was appraised on agarose gel. Real time PCR was performed to carry out the quantitative analysis of these four genes. Of four genes, the expression of three genes, i.e., TMEM16A, ITPR2 and FOXA1, was found to be significantly high in the primary hyperhidrosis patients, whereas low expression of the gene AQP5 was noted compared to that of the control samples. Microarray data analysis on anhidrosis was done using GEO2R database as well as the anhidrosis datasets available on Gene Exression Omnibus (GEO). From this analysis, the low expression of FOXA1 in anhidrosis was confirmed. Based on the highest expression of ITPR2 in hyperhidrosis, it was selected for PCR amplification as well as sequencing. The sequence analysis of ITPR2 showed 100% identity without any mutation.These key genes will further help us to devise new treatment modalities.