Cystic fibrosis is caused by mutation in the cystic fibrosis transmembrane conductance regulator (CFTR). In the lung the site of highest expression of CFTR is in the submucosal gland cells of the airways. These cells express high levels of the polymeric immunoglobulin receptor (pIgR), a receptor that endocytoses IgA at the basolateral surface and transcytoses IgA to the apical surface. We propose to use the pIgR to target vectors for expression CFTR to the submucosal gland cells. We will use monoclonal antibodies directed against the extracellular domain of the pIgR. Intact IgG or monovalant Fab' fragments will be coupled to various vectors. Two vector systems that will be tried are a liganded adenovirus system developed by David Curiel, and a amphiphillic polymer system developed by Frank Szoka. We will use the vectors to deliver and express either a trial reporter gene (beta-galactosidase) or the CFTR, driven by a cytomegalovirus vector. We will start by using Madin-Darby canine kidney cells that have been transfected with the rabbit pIgR. When grown on a permeable support, these, cells form a polarized monolayer that mimics an in vivo epithelial monolayer. If successful we will then use primary cultures of airway submucosal gland cells. Ultimately we will try an intact rodent model.

Project Start
Project End
Budget Start
Budget End
Support Year
4
Fiscal Year
1996
Total Cost
Indirect Cost
Hawgood, Samuel; Ochs, Matthias; Jung, Anja et al. (2002) Sequential targeted deficiency of SP-A and -D leads to progressive alveolar lipoproteinosis and emphysema. Am J Physiol Lung Cell Mol Physiol 283:L1002-10
Lalwani, Anil K; Han, Jay J; Castelein, Caley M et al. (2002) In vitro and in vivo assessment of the ability of adeno-associated virus-brain-derived neurotrophic factor to enhance spiral ganglion cell survival following ototoxic insult. Laryngoscope 112:1325-34
Xu, Z; Jablons, D M; Gruenert, D C (2001) Expression sequence tag-specific full-length cDNA cloning: actin cDNAs. Gene 263:265-72
Gum Jr, J R; Hicks, J W; Gillespie, A M et al. (2001) Mouse intestinal goblet cells expressing SV40 T antigen directed by the MUC2 mucin gene promoter undergo apoptosis upon migration to the villi. Cancer Res 61:3472-9
Colosimo, A; Goncz, K K; Novelli, G et al. (2001) Targeted correction of a defective selectable marker gene in human epithelial cells by small DNA fragments. Mol Ther 3:178-85
Goncz, K K; Colosimo, A; Dallapiccola, B et al. (2001) Expression of DeltaF508 CFTR in normal mouse lung after site-specific modification of CFTR sequences by SFHR. Gene Ther 8:961-5
Colosimo, A; Goncz, K K; Holmes, A R et al. (2000) Transfer and expression of foreign genes in mammalian cells. Biotechniques 29:314-8, 320-2, 324 passim
Camargo, F D; Huey-Louie, D A; Finn, A V et al. (2000) Germline incorporation of a replication-defective adenoviral vector in mice does not alter immune responses to adenoviral vectors. Mol Ther 2:496-504
Laan, M; Cui, Z H; Hoshino, H et al. (1999) Neutrophil recruitment by human IL-17 via C-X-C chemokine release in the airways. J Immunol 162:2347-52
Xu, Y; Hui, S W; Frederik, P et al. (1999) Physicochemical characterization and purification of cationic lipoplexes. Biophys J 77:341-53

Showing the most recent 10 out of 59 publications