Nonviral Delivery in Gene Therapy

NONVIRAL DELIVERY IN GENE THERAPY

However sophisticated a viral delivery system may be, nonviral vectors are inherently safer. Nonetheless, they have been relatively neglected because viruses were more efficient. However, several unfortunate incidents have occurred with viral vectors, especially retroviruses, including the occasional appearance of leukemia-like disease. This has resulted in renewed interest in nonviral delivery systems.

About 75% of gene therapy trials have used viral vectors. A variety of alternative approaches have also been investigated, though few have been effective or widely used so far. These include:

(a) Use of naked nucleic acid (DNA or less often RNA). Many animal cells can be transformed directly with purified DNA. The therapeutic gene may be inserted into a plasmid and the plasmid DNA used directly. Some 10% to 20% of gene therapy trials have used unprotected nucleic acid.

(b)Particle bombardment. DNA is fired through the cell walls and membranes on metal particles. This method was originally developed to get DNA into plants and is therefore discussed in Chapter 14. However, it has also been used to make transgenic animals and is occasionally used for humans.

(c)Receptor-mediated uptake. DNA is attached to a protein that is recognized by a cell surface receptor. When the protein enters the cell, the DNA is taken in with it.

(d)Polymer-complexed DNA. Binding to a positively charged polymer, such as polyethyleneimine, protects the negatively charged DNA. Such complexes are often taken up by cells in culture and may in principle be used for ex vivo gene therapy.

(e) Encapsulated cells. Whole cells engineered to express and secrete a needed protein may be encapsulated in a porous polymeric coat and injected locally. Foreign cells excreting nerve growth factor have been injected into the brains of aging rats. The rats showed some improvement in cognitive ability, suggesting that this approach may be of value in treating conditions such as Alzheimer’s disease.

(f) Liposomes are spherical vesicles composed of phospholipid. They have been used in around 10% of gene therapy trials (see later discussion).