Improved Baculovirus Vectors for Transduction and Gene Expression in Human Pancreatic Islet Cells

Viruses. 2018 Oct 20;10(10):574. doi: 10.3390/v10100574.

Abstract

Pancreatic islet transplantation is a promising treatment for type 1 diabetes mellitus offering improved glycaemic control by restoring insulin production. Improved human pancreatic islet isolation has led to higher islet transplantation success. However, as many as 50% of islets are lost after transplantation due to immune responses and cellular injury, gene therapy presents a novel strategy to protect pancreatic islets for improved survival post-transplantation. To date, most of the vectors used in clinical trials and gene therapy studies have been derived from mammalian viruses such as adeno-associated or retrovirus. However, baculovirus BacMam vectors provide an attractive and safe alternative. Here, a novel BacMam was constructed containing a frameshift mutation within fp25, which results in virus stocks with higher infectious titres. This improved in vitro transduction when compared to control BacMams. Additionally, incorporating a truncated vesicular stomatitis virus G protein increased transduction efficacy and production of EGFP and BCL2 in human kidney (HK-2) and pancreatic islet β cells (EndoC βH3). Lastly, we have shown that our optimized BacMam vector can deliver and express egfp in intact pancreatic islet cells from human cadaveric donors. These results confirm that BacMam vectors are a viable choice for providing delivery of transgenes to pancreatic islet cells.

Keywords: BacMam; baculovirus; gene therapy; high-titre virus; human pancreatic islet cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Baculoviridae / genetics*
  • Baculoviridae / physiology
  • Diabetes Mellitus, Type 1 / therapy*
  • Gene Transfer Techniques
  • Genetic Therapy / instrumentation*
  • Genetic Therapy / methods
  • Genetic Vectors / genetics
  • Genetic Vectors / physiology
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Insulin-Secreting Cells / virology*
  • Islets of Langerhans / virology
  • Transduction, Genetic*

Substances

  • enhanced green fluorescent protein
  • Green Fluorescent Proteins