Different ex Vivo and Direct in Vivo DNA Administration Strategies for Growth Hormone Gene Therapy in Dwarf Animals

Abstract

Recombinant growth hormone (GH) was one of the first proteins to be synthesized via DNA recombinant techniques in the late seventies. It was also one of the first proteins to be used in studies of animal models for gene therapy, already in the eighties. This was due to the real therapeutic need for GH, combined with the fact that its detection by well-known immunoassay methods is facile and sensitive. Moreover, evident phenotypic effects can be observed and measured in several animal models (e.g., dwarf mice), some of which have GH deficiencies that closely resemble their human counterparts. GH gene therapy has the potential advantage of circumventing laborious and expensive purification processes, quality control procedures and the repetitive injections that are required in the conventional treatment. The ideal situation would, of course, be to introduce the deficient protein into the circulation via a mechanism that resembles the natural process. These treatments have not yet reached the clinical stage for humans, the major challenge being to achieve a sustainable and regulated in vivo GH secretion. However, several interesting and promising ex vivo and direct in vivo DNA administration strategies for GH gene therapy have been developed and studied using animal models. These studies obviously open the way for the systemic delivery of other therapeutic proteins in addition to GH. The various ex vivo models for GH gene therapy are based on the use of target cells, such as keratinocytes, fibroblasts, endothelial cells, peritoneal mesothelial cells, or skeletal myoblasts. These cells can also be encapsulated to prevent rejection when implanted in allogeneic hosts. The majority of the methodologies are carried out via cell isolation and in vitro cultivation, genetic modification by viral or non-viral vectors containing the GH gene and re-implantation of the secreting cells onto the animal. Primary keratinocytes are one of the most attractive vehicles for gene transfer and gene therapy. They are among the most accessible cells in the body and can be serially propagated in culture; the procedure for their transplantation is already well established, e.g., for burn patients, and the therapy can be reversed by excision of the genetically modified tissue. Cutaneous gene therapy has already been demonstrated to be a powerful