Article Text

Genetically engineered pigs manifesting pancreatic agenesis with severe diabetes
  1. Masaki Nagaya1,2,
  2. Koki Hasegawa3,
  3. Masahito Watanabe1,3,
  4. Kazuaki Nakano1,3,
  5. Kazutoshi Okamoto3,
  6. Takeshi Yamada3,
  7. Ayuko Uchikura1,3,
  8. Kenji Osafune4,
  9. Harumasa Yokota5,
  10. Taiji Nagaoka5,
  11. Hitomi Matsunari1,3,
  12. Kazuhiro Umeyama1,3,
  13. Eiji Kobayashi6,
  14. Hiromitsu Nakauchi7,8,
  15. Hiroshi Nagashima1,3
  1. 1Meiji University International Institute for Bio-Resource Research, Meiji University – Ikuta Campus, Kawasaki, Japan
  2. 2Department of Immunology, St. Marianna University School of Medicine, Kawasaki, Japan
  3. 3Laboratory of Medical Bioengineering, Department of Life Sciences, School of Agriculture, Meiji University, Kawasaki, Japan
  4. 4Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan
  5. 5Division of Ophthalmology, Department of Visual Sciences, Nihon University School of Medicine, Tokyo, Japan
  6. 6Department of Organ Fabrication, Keio University, School of Medicine, Tokyo, Japan
  7. 7Division of Stem Cell Therapy, Institute of Medical Science, Institute of Medical Science, The University of Tokyo, Tokyo, Japan
  8. 8Institute for Stem Cell Biology and Regenerative Medicine, Department of Genetics, Stanford University School of Medicine, Stanford, California, USA
  1. Correspondence to Professor Masaki Nagaya; m2nagaya{at}gmail.com; Professor Hiroshi Nagashima; hnagas{at}meiji.ac.jp

Abstract

Introduction Pancreatic duodenum homeobox 1 (Pdx1) expression is crucial for pancreatic organogenesis and is a key regulator of insulin gene expression. Hairy and enhancer of split 1 (Hes1) controls tissue morphogenesis by maintaining undifferentiated cells. Hes1 encodes a basic helix loop helix (bHLH) transcriptional repressor and functionally antagonizes positive bHLH genes, such as the endocrine determination gene neurogenin-3. Here, we generated a new pig model for diabetes by genetic engineering Pdx1 and Hes1 genes.

Research design and methods A transgenic (Tg) chimera pig with germ cells carrying a construct expressing Hes1 under the control of the Pdx1 promoter was used to mate with wild-type gilts to obtain Tg piglets.

Results The Tg pigs showed perinatal death; however, this phenotype could be rescued by insulin treatment. The duodenal and splenic lobes of the Tg pigs were slender and did not fully develop, whereas the connective lobe was absent. β cells were not detected, even in the adult pancreas, although other endocrine cells were detected, and exocrine cells functioned normally. The pigs showed no irregularities in any organs, except diabetes-associated pathological alterations, such as retinopathy and renal damage.

Conclusion Pdx1-Hes1 Tg pigs were an attractive model for the analysis of pancreatic development and testing of novel treatment strategies for diabetes.

  • diabetes mellitus
  • experimental
  • insulin
  • animals
  • genetically modified
  • animal experimentation
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Footnotes

  • MN and KH contributed equally.

  • Contributors MN and HirosN conception and design, financial support, collection and assembly of data, data analysis and interpretation, manuscript writing and final approval of manuscript. KH, MW, KN, KaO, TY, AU, HM and KU collection and assembly of data, data analysis and interpretation. HY and TN collection and assembly of data, data analysis and interpretation. KeO, EK and HiromN data analysis and interpretation.

  • Funding This research work was supported by Grants-in-Aid from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (17K10527 and 20K08944 to MN), the Japan Agency for Medical Research and Development (Leading Advanced Projects for Medical Innovation, Generation of Functional Organs using Developmental Niche (to HirosN), Japan Science and Technology Agency (Exploratory Research for Advanced Technology, NAKAUCHI Stem Cell and Organ Regeneration (to HirosN), and the Meiji University International Institute for Bio-Resource Research (to MN and HirosN).

  • Disclaimer The funding organizations had no roles in the study’s design, data collection and analysis, the decision to publish, or the manuscript’s preparation.

  • Competing interests None declared.

  • Patient consent for publication Not required.

  • Ethics approval All animal experiments in this study were approved by Meiji University’s Institutional Animal Care and Use Committee (IACUC07-0005, 14–0008, 19–0003).

  • Provenance and peer review Not commissioned; externally peer reviewed.

  • Data availability statement All data relevant to the study are included in the article or uploaded as supplemental information.

  • Supplemental material This content has been supplied by the author(s). It has not been vetted by BMJ Publishing Group Limited (BMJ) and may not have been peer-reviewed. Any opinions or recommendations discussed are solely those of the author(s) and are not endorsed by BMJ. BMJ disclaims all liability and responsibility arising from any reliance placed on the content. Where the content includes any translated material, BMJ does not warrant the accuracy and reliability of the translations (including but not limited to local regulations, clinical guidelines, terminology, drug names and drug dosages), and is not responsible for any error and/or omissions arising from translation and adaptation or otherwise.

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