This application is a request to continue work to discern mechanisms regulating expression of surfactant protein SP-B gene. SP-B is a hydrophobic polypeptide that enhances the spreading and stability of pulmonary surfactant phospholipids. SP-B synthesis in pulmonary epithelial cells increases prior to birth and is a critical component of the surfactant complex required for adaptation to air breathing. Lack of surfactant at birth is a major cause of morbidity and mortality of premature infants. This application will test the hypothesis that cell-specific, ontogenic and humoral controls of SP-B gene expression are mediated, at least in part, by the cis-active elements of the SP-B gene and their interaction with nuclear transcription proteins. Regions of the SP-B gene conferring cell-specific, ontogenic and the humorally regulated pattern of SP-B expression will be delineated using DNase hypersensitivity, gel retardation analysis, DNA footprinting and by transcriptional assays using chimeric SP-B-reporter gene constructs in transient assays and in transgenic mice. Expression of the SP-B-CAT transgenes will be studied both in vitro and in vivo. Transgenic mice bearing SP-B-CAT constructs will be generated for comparison of CAT expression with that of the endogenous SP-B gene and other cellular markers of epithelial cell maturation including CC10, SP-A and SP-C. Temporal-spatial and humoral regulation of SP-B and SP-B chimeric genes will be assessed. Transgenic mice will be generated expressing human proSP-B mRNA (sense and antisense SP-B mRNAs) using enhancer elements from the human SP-C gene to discern the effects of increased and decreased expression of SP-B on surfactant structure, function and on perinatal adaptation to air breathing. Alternatively, homologous recombination will be used to disrupt the SP-B locus and determine the role of SP-B in lung function in SP-B ablated animals in vivo. Further understanding of the role of SP-B in lung function and the mechanisms regulating SP-B may provide insights critical to developing new therapeutic strategies for prevention of lung disease in premature infants.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37HL038859-14
Application #
6389040
Study Section
Special Emphasis Panel (NSS)
Program Officer
Berberich, Mary Anne
Project Start
1987-07-01
Project End
2003-03-31
Budget Start
2001-04-01
Budget End
2002-03-31
Support Year
14
Fiscal Year
2001
Total Cost
$346,718
Indirect Cost
Name
Cincinnati Children's Hospital Medical Center
Department
Type
DUNS #
071284913
City
Cincinnati
State
OH
Country
United States
Zip Code
45229
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