Abstract

Recent findings indicating a Cl ion transport defect in human airway submucosal gland cells suggest that these cells represent an important target for gene therapy. The selective accumulation of CF mRNA in these cells further suggests the presence of gland cell-specific regulatory elements in the CFTR gene. The availability of human gland cell cultures should facilitate the identification of these elements, providing information needed for the design of optimal gene delivery vectors. In the proposed studies, human gland cell cultures will be used to: (l) analyze endogenous CFTR gene transcription using Northern blots, primer extension, and S1 nuclease protection assays, (2) survey potential regulatory regions of the CFTR gene by chromatin nuclease hypersensitivity assays, and (3) identify CFTR promoter elements that specify gland cell-specific expression by means of transfections with CFTR promoter-reporter gene constructs. Information obtained from these studies should permit construction of a minimal length CFTR-based promoter able to direct high level selective expression in gland cells.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Center Core Grants (P30)
Project #
5P30DK047766-05
Application #
6105617
Study Section
Project Start
1997-09-01
Project End
1999-08-31
Budget Start
1996-10-01
Budget End
1997-09-30
Support Year
5
Fiscal Year
1997
Total Cost
Indirect Cost

  Institution

Name
University of California San Francisco
Department
Type
DUNS #
073133571
City
San Francisco
State
CA
Country
United States
Zip Code
94143

  Publications

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
Hirota, S; de Ilarduya, C T; Barron, L G et al. (1999) Simple mixing device to reproducibly prepare cationic lipid-DNA complexes (lipoplexes). Biotechniques 27:286-90
Holmes, A R; Dohrman, A F; Ellison, A R et al. (1999) Intracellular compartmentalization of DNA fragments in cultured airway epithelial cells mediated by cationic lipids. Pharm Res 16:1020-5

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