Plasma-derived exosome characterization reveals a distinct microRNA signature in long duration Type 1 diabetes

Type 1 diabetes mellitus (T1DM) results from an autoimmune attack against the insulin-producing ß cells which leads to chronic hyperglycemia. Exosomes are lipid vesicles derived from cellular multivesicular bodies that are enriched in specific miRNAs, potentially providing a disease-specific diagnostic signature. To assess the value of exosome miRNAs as biomarkers for T1DM, miRNA expression in plasma-derived exosomes was measured. Nanoparticle tracking analysis and transmission electron microscopy confirmed the presence of plasma-derived exosomes (EXOs) isolated by differential centrifugation. Total RNA extracted from plasma-derived EXOs of 12 T1DM and 12 control subjects was hybridized onto Nanostring human v2 miRNA microarray array and expression data were analyzed on nSolver analysis software. We found 7 different miRNAs (1 up-regulated and 6 down-regulated), that were differentially expressed in T1DM. The selected candidate miRNAs were validated by qRT-PCR analysis of cohorts of 24 T1DM and 24 control subjects. Most of the deregulated miRNAs are involved in progression of T1DM. These findings highlight the potential of EXOs miRNA profiling in the diagnosis as well as new insights into the molecular mechanisms involved in T1DM.

[1]  L. Eliasson,et al.  Role of non‐coding RNAs in pancreatic beta‐cell development and physiology , 2014, Acta physiologica.

[2]  J. D. de Fijter,et al.  Circulating MicroRNAs Associate With Diabetic Nephropathy and Systemic Microvascular Damage and Normalize After Simultaneous Pancreas–Kidney Transplantation , 2015, American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons.

[3]  S. Rome,et al.  miRNA-375 a Sensor of Glucotoxicity Is Altered in the Serum of Children with Newly Diagnosed Type 1 Diabetes , 2016, Journal of diabetes research.

[4]  I. Knezevic,et al.  A Novel Cardiomyocyte-enriched MicroRNA, miR-378, Targets Insulin-like Growth Factor 1 Receptor , 2012, The Journal of Biological Chemistry.

[5]  P. Hougaard,et al.  Circulating Levels of MicroRNA from Children with Newly Diagnosed Type 1 Diabetes and Healthy Controls: Evidence That miR-25 Associates to Residual Beta-Cell Function and Glycaemic Control during Disease Progression , 2012, Experimental diabetes research.

[6]  Chisayo Kozuka,et al.  MicroRNA-378 Regulates Adiponectin Expression in Adipose Tissue: A New Plausible Mechanism , 2014, PloS one.

[7]  Piero Marchetti,et al.  Phasic insulin release and metabolic regulation in type 2 diabetes. , 2002, Diabetes.

[8]  Krisztina V. Vukman,et al.  Low-density lipoprotein mimics blood plasma-derived exosomes and microvesicles during isolation and detection , 2016, Scientific Reports.

[9]  She Chen,et al.  Characterization and proteomic analysis of ovarian cancer-derived exosomes. , 2013, Journal of proteomics.

[10]  E. Sijbrands,et al.  Study on inflammation-related genes and microRNAs, with special emphasis on the vascular repair factor HGF and miR-574-3p, in monocytes and serum of patients with T2D , 2016, Diabetology & Metabolic Syndrome.

[11]  Yi Li,et al.  Identification of Endogenous Controls for Analyzing Serum Exosomal miRNA in Patients with Hepatitis B or Hepatocellular Carcinoma , 2015, Disease markers.

[12]  Robert J Freishtat,et al.  Adipocyte-derived Exosomal miRNAs: A Novel Mechanism for Obesity-Related Disease , 2014, Pediatric Research.

[13]  I. Gérin,et al.  Roles for miRNA-378/378* in adipocyte gene expression and lipogenesis. , 2010, American journal of physiology. Endocrinology and metabolism.

[14]  Lesley Cheng,et al.  Exosomes provide a protective and enriched source of miRNA for biomarker profiling compared to intracellular and cell-free blood , 2014, Journal of extracellular vesicles.

[15]  A. Cooke,et al.  Type 1 diabetes: translating mechanistic observations into effective clinical outcomes , 2013, Nature Reviews Immunology.

[16]  Laurence Zitvogel,et al.  Exosomes: composition, biogenesis and function , 2002, Nature Reviews Immunology.

[17]  Aniruddha Chatterjee,et al.  A cross comparison of technologies for the detection of microRNAs in clinical FFPE samples of hepatoblastoma patients , 2015, Scientific Reports.

[18]  D. Schatz,et al.  Autoimmune markers in diabetes. , 2011, Clinical chemistry.

[19]  P. Poulsen,et al.  Electronic Supplementary Material -methods , 2022 .

[20]  J. Lötvall,et al.  Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells , 2007, Nature Cell Biology.

[21]  Sugunavathi Sepramaniam,et al.  miR-25 and miR-92a regulate insulin I biosynthesis in rats , 2013, RNA biology.

[22]  R. Planas,et al.  Gene expression profiles for the human pancreas and purified islets in Type 1 diabetes: new findings at clinical onset and in long‐standing diabetes , 2010, Clinical and experimental immunology.

[23]  P. Buchwald,et al.  Experimental evaluation and computational modeling of the effects of encapsulation on the time-profile of glucose-stimulated insulin release of pancreatic islets , 2015, Biomedical engineering online.

[24]  T. V. van Berkel,et al.  Nonalcoholic fatty liver disease is associated with an altered hepatocyte microRNA profile in LDL receptor knockout mice. , 2012, The Journal of nutritional biochemistry.

[25]  G. Camussi,et al.  Isolation, Characterization and Potential Role in Beta Cell-Endothelium Cross-Talk of Extracellular Vesicles Released from Human Pancreatic Islets , 2014, PloS one.

[26]  Fang Wei,et al.  Detection of Tumor Cell-Specific mRNA and Protein in Exosome-Like Microvesicles from Blood and Saliva , 2014, PloS one.

[27]  G. Raposo,et al.  As we wait: coping with an imperfect nomenclature for extracellular vesicles , 2013, Journal of extracellular vesicles.

[28]  P. Hougaard,et al.  Erratum to “Circulating Levels of MicroRNA from Children with Newly Diagnosed Type 1 Diabetes and Healthy Controls: Evidence That miR-25 Associates to Residual Beta-Cell Function and Glycaemic Control during Disease Progression” , 2012, Experimental Diabetes Research.

[29]  T. Jiang,et al.  Exosomal levels of miRNA-21 from cerebrospinal fluids associated with poor prognosis and tumor recurrence of glioma patients , 2015, Oncotarget.

[30]  Hamid Cheshmi.  Glioblastoma microvesicles transport RNA and proteins that promote tumour growth and provide diagnostic biomarkers , 2011 .

[31]  Jeffrey P. Krischer,et al.  Pancreatic Islet Autoantibodies as Predictors of Type 1 Diabetes in the Diabetes Prevention Trial–Type 1 , 2009, Diabetes Care.

[32]  Emily K. Sims,et al.  Minireview: Emerging Roles for Extracellular Vesicles in Diabetes and Related Metabolic Disorders. , 2015, Molecular endocrinology.

[33]  S. Rome,et al.  Horizontal transfer of exosomal microRNAs transduce apoptotic signals between pancreatic beta-cells , 2015, Cell Communication and Signaling.

[34]  N. Rajewsky,et al.  A pancreatic islet-specific microRNA regulates insulin secretion , 2004, Nature.