Dietary Gluten-Induced Gut Dysbiosis is Accompanied by Selective Upregulation of microRNAs with Intestinal Tight Junction and Bacteria-Binding Motifs in Rhesus Macaque Model of Celiac Disease

The study describes miRNA expression in intact jejunum following feeding of gluten containing monkey chow to gluten sensitive (GS) rhesus macaques. Gluten feeding induced several inflammation associated miRNAs validated and predicted to directly target intestinal tight junction proteins. Upregulation of these microRNAs was accompanied significantly reduced mRNA expression of claudin-1, claudin-3 and occludin and severe gut dysbiosis. These findings suggest that miRNA mediated downregulation of intestinal epithelial tight junction proteins could increase intestinal permeability and facilitate translocation of dysbiotic bacteria into the systemic circulation. Four age and weight matched gluten sensitive (GS) Indian rhesus macaques were fed gluten containing monkey chow. Five rhesus macaques served as normal healthy controls. Jejunal segments were collected at necropsy from both groups. All animals were necropsied at 60 days post SIV infection. ~100 ng of total RNA was first reverse transcribed and preamplified according to the manufacturer’s recommendation. microRNA expression profiling was performed using TaqMan ®OpenArray® Human microRNA panels. Data analysis was performed using ExpressionSuite® software. Data was normalized to two endogenous controls (RNU44 and RNU48). Delta CT values were calculated by subtracting individual miRNA CT values from an average of both endogenous controls. Comparisons were made between gluten sensitive and normal healthy control macaques. Four age and weight matched gluten sensitive Indian rhesus macaques were fed gluten containing monkey chow. Five rhesus macaques served as normal healthy controls. Jejunal segments were collected at necropsy from both groups. All animals were necropsied at 60 days post SIV infection. ~100 ng of total RNA was first reverse transcribed and preamplified according to the manufacturer’s recommendation. microRNA expression profiling was performed using TaqMan ®OpenArray® Human microRNA panels. Data analysis was performed using ExpressionSuite® software. Data was normalized to two endogenous controls (RNU44 and RNU48). Delta CT values were calculated by subtracting individual miRNA CT values from an average of both endogenous controls. Comparisons were made between gluten sensitive and normal healthy control macaques. Four age and weight matched gluten sensitive Indian rhesus macaques were fed gluten containing monkey chow. Five rhesus macaques served as normal healthy controls. Jejunal segments were collected at necropsy from both groups. All animals were necropsied at 60 days post SIV infection. ~100 ng of total RNA was first reverse transcribed and preamplified according to the manufacturer’s recommendation. microRNA expression profiling was performed using TaqMan ®OpenArray® Human microRNA panels. Data analysis was performed using ExpressionSuite® software. Data was normalized to two endogenous controls (RNU44 and RNU48). Delta CT values were calculated by subtracting individual miRNA CT values from an average of both endogenous controls. Comparisons were made between gluten sensitive and normal healthy control macaques.