Altered cellular localisation and expression, together with unconventional protein trafficking, of prion protein, PrP^C, in type 1 diabetes

Normal cellular prion protein (PrP^C) is a conserved mammalian glycoprotein found on the outer plasma membrane leaflet through a glycophosphatidylinositol anchor. Although PrP^C is expressed by a wide range of tissues throughout the body, the complete repertoire of its functions has not been fully determined. The misfolded pathogenic isoform PrP^Sc (the scrapie form of PrP) is a causative agent of neurodegenerative prion diseases. The aim of this study is to evaluate PrP^C localisation, expression and trafficking in pancreases from organ donors with and without type 1 diabetes and to infer PrP^C function through studies on interacting protein partners.

In order to evaluate localisation and trafficking of PrP^C in the human pancreas, 12 non-diabetic, 12 type 1 diabetic and 12 autoantibody-positive organ donor tissue samples were analysed using immunofluorescence analysis. Furthermore, total RNA was isolated from 29 non-diabetic, 29 type 1 diabetic and 24 autoantibody-positive donors to estimate PrP^C expression in the human pancreas. Additionally, we performed PrP^C-specific immunoblot analysis on total pancreatic protein from non-diabetic and type 1 diabetic organ donors to test whether changes in PrP^C mRNA levels leads to a concomitant increase in PrP^C protein levels in human pancreases.

In non-diabetic and type 1 diabetic pancreases (the latter displaying both insulin-positive [INS(+)] and -negative [INS(−)] islets), we found PrP^C in islets co-registering with beta cells in all INS(+) islets and, strikingly, unexpected activation of PrP^C in alpha cells within diabetic INS(−) islets. We found PrP^C localised to the plasma membrane and endoplasmic reticulum (ER) but not the Golgi, defining two cellular pools and an unconventional protein trafficking mechanism bypassing the Golgi. We demonstrate PrP^C co-registration with established protein partners, neural cell adhesion molecule 1 (NCAM1) and stress-inducible phosphoprotein 1 (STI1; encoded by STIP1) on the plasma membrane and ER, respectively, linking PrP^C function with cyto-protection, signalling, differentiation and morphogenesis. We demonstrate that both PRNP (encoding PrP^C) and STIP1 gene expression are dramatically altered in type 1 diabetic and autoantibody-positive pancreases.

As the first study to address PrP^C expression in non-diabetic and type 1 diabetic human pancreas, we provide new insights for PrP^C in the pathogenesis of type 1 diabetes. We evaluated the cell-type specific expression of PrP^C in the human pancreas and discovered possible connections with potential interacting proteins that we speculate might address mechanisms relevant to the role of PrP^C in the human pancreas.