Gene Delivery of Alpha-1-Antitrypsin Using Recombinant Adeno-Associated Virus (rAAV)

Methods Mol Biol. 2018:1826:183-196. doi: 10.1007/978-1-4939-8645-3_12.

Abstract

The challenge for alpha-1-antitrypsin (AAT also known as SERPINA1) gene therapy is to achieve long term and high levels of AAT production. Recombinant adeno-associated virus (rAAV) vector has several advantages for AAT gene delivery including no viral genes in the vector, no requirement of integration for long-term transgene expression, low immunogenicity, and wide tropism. AAV-mediated AAT gene therapy has been developed and tested in animal models for AAT deficiency, type 1 diabetes, rheumatoid arthritis, and osteoporosis. AAV-mediated AAT gene therapy has also been tested in clinical studies and has shown promising results. Here we describe the methods of rAAV-AAT vector construction and production as well as AAT gene delivery through (1) liver-directed, (2) muscle-directed, and (3) mesenchymal stem cell (MSC)-mediated routes. We will also describe methods for the evaluation of AAT expression for each delivery approach.

Keywords: AAT deficiency; Adeno-associated virus (AAV); Alpha-1-antitrypsin (AAT); Gene delivery; Serpin.

MeSH terms

  • Animals
  • Arthritis, Rheumatoid* / genetics
  • Arthritis, Rheumatoid* / metabolism
  • Arthritis, Rheumatoid* / pathology
  • Arthritis, Rheumatoid* / therapy
  • Dependovirus*
  • Genetic Therapy / methods*
  • Humans
  • Mice
  • Osteoporosis* / genetics
  • Osteoporosis* / metabolism
  • Osteoporosis* / pathology
  • Osteoporosis* / therapy
  • Transduction, Genetic / methods*
  • alpha 1-Antitrypsin Deficiency* / genetics
  • alpha 1-Antitrypsin Deficiency* / metabolism
  • alpha 1-Antitrypsin Deficiency* / pathology
  • alpha 1-Antitrypsin Deficiency* / therapy
  • alpha 1-Antitrypsin* / biosynthesis
  • alpha 1-Antitrypsin* / genetics

Substances

  • SERPINA1 protein, human
  • alpha 1-Antitrypsin