Theodore Friedmann

Gene transfer into rat airway epithelial cells using retroviral vectors

Primary cultures of epithelial cells from adult rat tracheas were maintained in vitro on collagen matrices and were exposed to a murine retrovirus vector expressing theE. coli β-galactosidase gene. Infection was carried out on cells grown as monolayers under medium and on cells grown on raised platforms. Cells maintained at an air-medium interface were highly susceptible to infection with the vector, showing an efficiency of infection of 20–25%, compared with an efficiency of <1% for cells grown under medium. Infected β-galactosidase-expressing cells were seeded into denuded tracheas and were capable of partially repopulating the denuded tracheas grafted subcutaneously into host rats. The susceptibility of these cells to retroviral infection suggests an approach to the treatment of some pulmonary genetic disorders such as cystic fibrosis.

An approach to the genetic correction of defects and disorders of the central nervous system

Publisher Summary This chapter discusses an approach to the genetic correction of defects and disorders of the central nervous system. The level of understanding of pathogenesis has led not only to improved detection and diagnosis but also to a conceptually new approach to treatment of some kinds of genetic diseases, which is the concept of gene therapy. For the first time, it has become possible to imagine treating disease by restoring precisely the function that is defective, the mutant gene, to achieve a definitive correction of a disease phenotype. An animal model for the degeneration of cholinergic neurons that occurs in Alzheimers disease is available in the rat. The genetic correction of some or many central nervous system disorders may require the establishment or reestablishment of faithful intercellular synaptic connections. Model systems to study these possibilities have not yet been developed and exploited because of the paucity of replicating nontransformed cell culture systems and the refractoriness of nonreplicating neuronal cells to viral infection.