Co-localization of acinar markers and insulin in pancreatic cells of subjects with type 2 diabetes

To search for clues suggesting that beta cells may generate by transdifferentiation in humans, we assessed the presence of cells double positive for exocrine (amylase, carboxypeptidase A) and endocrine (insulin) markers in the pancreas of non-diabetic individuals (ND) and patients with type 2 diabetes (T2D). Samples from twelve ND and twelve matched T2D multiorgan donors were studied by electron microscopy, including amylase and insulin immunogold labeling; carboxypeptidase A immunofluorescence light microscopy assessment was also performed. In the pancreas from four T2D donors, cells containing both zymogen-like and insulin-like granules were observed, scattered in the exocrine compartment. Nature of granules was confirmed by immunogold labeling for amylase and insulin. Double positive cells ranged from 0.82 to 1.74 per mm2, corresponding to 0.26±0.045% of the counted exocrine cells. Intriguingly, cells of the innate immune systems (mast cells and/or macrophages) were adjacent to 33.3±13.6% of these hybrid cells. No cells showing co-localization of amylase and insulin were found in ND samples by electron microscopy. Similarly, cells containing both carboxypeptidase A and insulin were more frequently observed in the diabetic pancreata. These results demonstrate more abundant presence of cells containing both acinar markers and insulin in the pancreas of T2D subjects, which suggests possible conversion from one cellular type to the other and specific association with the diseased condition.

[1]  Dirk Trauner,et al.  Beta Cell Hubs Dictate Pancreatic Islet Responses to Glucose , 2016, Cell metabolism.

[2]  Olle Korsgren,et al.  Identification of proliferative and mature β-cells in the islets of Langerhans , 2016, Nature.

[3]  L. Bouwens,et al.  Acinar phenotype is preserved in human exocrine pancreas cells cultured at low temperature: implications for lineage-tracing of β-cell neogenesis , 2016, Bioscience reports.

[4]  P. Marchetti.  Islet inflammation in type 2 diabetes , 2016, Diabetologia.

[5]  K. Prasadan,et al.  Intraislet Pancreatic Ducts Can Give Rise to Insulin-Positive Cells. , 2016, Endocrinology.

[6]  C. Ricordi,et al.  BMP-7 Induces Adult Human Pancreatic Exocrine-to-Endocrine Conversion , 2015, Diabetes.

[7]  U. Boggi,et al.  Mast cells infiltrate pancreatic islets in human type 1 diabetes , 2015, Diabetologia.

[8]  Michael J. Parsons,et al.  Centroacinar Cells Are Progenitors That Contribute to Endocrine Pancreas Regeneration , 2015, Diabetes.

[9]  David L. Morris.  Minireview: Emerging Concepts in Islet Macrophage Biology in Type 2 Diabetes. , 2015, Molecular endocrinology.

[10]  H. Itoh,et al.  Effects of Glucocorticoid Treatment on β- and α-Cell Mass in Japanese Adults With and Without Diabetes , 2015, Diabetes.

[11]  L. Bouwens,et al.  Reprogramming of human pancreatic exocrine cells to β-like cells , 2014, Cell Death and Differentiation.

[12]  N. Rosenthal,et al.  Preparing the ground for tissue regeneration: from mechanism to therapy , 2014, Nature Medicine.

[13]  C. Ling,et al.  β-Cell Failure in Type 2 Diabetes: Postulated Mechanisms and Prospects for Prevention and Treatment , 2014, Diabetes Care.

[14]  J. Holst,et al.  Insulin Resistance Alters Islet Morphology in Nondiabetic Humans , 2014, Diabetes.

[15]  U. Boggi,et al.  Are we overestimating the loss of beta cells in type 2 diabetes? , 2014, Diabetologia.

[16]  Y. Dor,et al.  Transient cytokine treatment induces acinar cell reprogramming and regenerates functional beta cell mass in diabetic mice , 2013, Nature Biotechnology.

[17]  V. Periwal,et al.  Regional Differences in Islet Distribution in the Human Pancreas - Preferential Beta-Cell Loss in the Head Region in Patients with Type 2 Diabetes , 2013, PloS one.

[18]  C. Mummery,et al.  Conversion of Mature Human β-Cells Into Glucagon-Producing α-Cells , 2013, Diabetes.

[19]  M. Atkinson,et al.  Reponse to Comments on: Butler et al. Marked Expansion of Exocrine and Endocrine Pancreas With Incretin Therapy in Humans With Increased Exocrine Pancreas Dysplasia and the Potential for Glucagon-Producing Neuroendocrine Tumors. Diabetes 2013;62:2595–2604 , 2013, Diabetes.

[20]  M. Bugliani,et al.  Palmitate Activates Autophagy in INS-1E β-Cells and in Isolated Rat and Human Pancreatic Islets , 2012, PloS one.

[21]  P. Herrera,et al.  β-Cell regeneration: the pancreatic intrinsic faculty , 2011, Trends in Endocrinology & Metabolism.

[22]  Dennis C. Sgroi,et al.  Gene Expression Profiles of Beta-Cell Enriched Tissue Obtained by Laser Capture Microdissection from Subjects with Type 2 Diabetes , 2010, PloS one.

[23]  P. Halban,et al.  Current status of islet cell replacement and regeneration therapy. , 2010, The Journal of clinical endocrinology and metabolism.

[24]  L. Rosenberg,et al.  {beta}-Cell mass dynamics and islet cell plasticity in human type 2 diabetes. , 2010, Endocrinology.

[25]  P. Herrera,et al.  Conversion of Adult Pancreatic α-cells to β-cells After Extreme β-cell Loss , 2010, Nature.

[26]  R. Gianani.  Beta cell regeneration in human pancreas , 2010, Seminars in Immunopathology.

[27]  C. Sempoux,et al.  Pancreatic β‐cell mass in European subjects with type 2 diabetes , 2008, Diabetes, obesity & metabolism.

[28]  Douglas A. Melton,et al.  In vivo reprogramming of adult pancreatic exocrine cells to β-cells , 2008, Nature.

[29]  H. Ellingsgaard,et al.  Islet Inflammation in Type 2 Diabetes , 2008, Diabetes Care.

[30]  M. Polak,et al.  A transient microenvironment loaded mainly with macrophages in the early developing human pancreas. , 2006, The Journal of endocrinology.

[31]  A. Dvorak.  Ultrastructural Studies of Human Basophils and Mast Cells , 2005, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[32]  U. Boggi,et al.  Functional and molecular defects of pancreatic islets in human type 2 diabetes. , 2005, Diabetes.

[33]  L. Orci,et al.  Ultrastructural studies of the hyperplastic islets of langerhans of spiny mice (acomys cahirinus) before and during the development of hyperglycemia , 1967, Diabetologia.

[34]  Douglas A. Melton,et al.  Adult pancreatic β-cells are formed by self-duplication rather than stem-cell differentiation , 2004, Nature.

[35]  J. Miyazaki,et al.  Analysis of insulin-producing cells during in vitro differentiation from feeder-free embryonic stem cells. , 2003, Diabetes.

[36]  S. Bonner-Weir,et al.  In vitro cultivation of human islets from expanded ductal tissue. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[37]  L. Cossel.  Electron microscopic demonstration of intermediate cells in the healthy adult human pancreas , 1986, Virchows Archiv. B, Cell pathology including molecular pathology.

[38]  H. Verlohren,et al.  Ultrastructural, immunohistological, and clinical findings in the pancreas in insulin-dependent diabetes mellitus (IDDM) of long duration. , 1983, Zentralblatt fur allgemeine Pathologie u. pathologische Anatomie.

[39]  T. Seemayer,et al.  Dynamic time course studies of the spontaneously diabetic BB Wistar rat. III. Light-microscopic and ultrastructural observations of pancreatic islets of Langerhans. , 1982, The American journal of pathology.

[40]  M. Bendayan,et al.  Double immunocytochemical labeling applying the protein A-gold technique. , 1982, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[41]  J. Reddy,et al.  Immunohistochemical localization of pancreatic exocrine enzymes in normal and neoplastic pancreatic acinar epithelium of rat. , 1981, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[42]  Sturrock Rr.  A semithin light microscopic, transmission electron microscopic and scanning electron microscopic study of macrophages in the lateral ventricle of mice from embryonic to adult life. , 1979 .

[43]  M. Bendayan,et al.  Immunohistochemical localization of exocrine enzymes in normal rat pancreas. , 1979, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[44]  R. Sturrock.  A semithin light microscopic, transmission electron microscopic and scanning electron microscopic study of macrophages in the lateral ventricle of mice from embryonic to adult life. , 1979, Journal of anatomy.