1985 — 1989 |
Whitsett, Jeffrey A |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Regulation of Surfactant Release in Type Ii Lung Cell @ University of Cincinnati
Pulmonary immaturity and the accompanying lack of surfactant results in hyaline membrane disease, a major factor in morbidity and mortality associated with premature birth. This disorder is related to lack of synthesis and secretion of lamellar bodies from Type II cells. Proposed research will seek to characterize molecular and humoral mechanisms controlling synthesis and release of surfactant from rat Type II cells in culture. The proposal is based on the hypothesis that catecholamine dependent surfactant release is mediated by activation of c-AMP protein kinase and subsequent protein phosphorylation, enhancing synthesis, processing and secretion of lamellar bodies. We will test whether the dramatic ontogenic appearance of surfactant release prior to birth is related to synthesis of developmentally and humorally regulated cellular components which mediate surfactant synthesis and secretion. Two major areas of work are proposed. The first seeks to associate activation of c-AMP and Ca2+ dependent protein kinases, specific protein phosphorylation (in 32P-phosphate labelled Type II cells) and secretogogue dependent apoprotein and phospholipid secretion from Type II cells. Developmental aspects of the c-AMP surfactant release system will be assessed in fetal Type II epithelial cells. The second area of proposed work will use polyclonal and monoclonal antibodies prepared for surfactant associated apoprotein to characterize synthesis, intracellular processing and secretion of apoprotein (A). Factors controlling the developmental appearance of apoprotein will be determined during late gestation. Molecular characteristics of apoprotein, rates of synthesis and processing (proteolysis, glycosylation and association with phospholipid) will be assessed using pulse-chase techniques after labelling with (35S) methionine, (3H) leucine, (14C) carbohydrates and (3H)choline. Ontogenic regulation of synthesis and processing will be determined in association with phospholipid synthesis and appearance of lamellar bodies. Purification and translation of apoprotein (A) mRNA will be assessed to determine whether developmental appearance of apoprotein (A) production is controlled at transcriptional, translational or post-translational levels. cDNA probes will be prepared to determine gene and protein sequence and to assess regulatory aspects of apo A synthesis.
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0.958 |
1985 — 1986 |
Whitsett, Jeffrey A |
K04Activity Code Description: Undocumented code - click on the grant title for more information. |
C-Amp Induced Surfactant Release/Type Ii Penumocyte @ University of Cincinnati |
0.958 |
1990 — 1999 |
Whitsett, Jeffrey A |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Regulation of Surfactant Secretion in Type Ii Lung Cell @ University of Cincinnati
Pulmonary immaturity and the accompanying lack of surfactant results in hyaline membrane disease, a major factor in morbidity and mortality associated with premature birth. This disorder is related to lack of synthesis and secretion of lamellar bodies from Type II cells. Proposed research will seek to characterize molecular and humoral mechanisms controlling synthesis and release of surfactant from rat Type II cells in culture. The proposal is based on the hypothesis that catecholamine dependent surfactant release is mediated by activation of c-AMP protein kinase and subsequent protein phosphorylation, enhancing synthesis, processing and secretion of lamellar bodies. We will test whether the dramatic ontogenic appearance of surfactant release prior to birth is related to synthesis of developmentally and humorally regulated cellular components which mediate surfactant synthesis and secretion. Two major areas of work are proposed. The first seeks to associate activation of c-AMP and Ca2+ dependent protein kinases, specific protein phosphorylation (in 32P-phosphate labelled Type II cells) and secretogogue dependent apoprotein and phospholipid secretion from Type II cells. Developmental aspects of the c-AMP surfactant release system will be assessed in fetal Type II epithelial cells. The second area of proposed work will use polyclonal and monoclonal antibodies prepared for surfactant associated apoprotein to characterize synthesis, intracellular processing and secretion of apoprotein (A). Factors controlling the developmental appearance of apoprotein will be determined during late gestation. Molecular characteristics of apoprotein, rates of synthesis and processing (proteolysis, glycosylation and association with phospholipid) will be assessed using pulse-chase techniques after labelling with (35S) methionine, (3H) leucine, (14C) carbohydrates and (3H)choline. Ontogenic regulation of synthesis and processing will be determined in association with phospholipid synthesis and appearance of lamellar bodies. Purification and translation of apoprotein (A) mRNA will be assessed to determine whether developmental appearance of apoprotein (A) production is controlled at transcriptional, translational or post-translational levels. cDNA probes will be prepared to determine gene and protein sequence and to assess regulatory aspects of apo A synthesis.
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0.958 |
1996 — 2000 |
Whitsett, Jeffrey A |
P01Activity Code Description: For the support of a broadly based, multidisciplinary, often long-term research program which has a specific major objective or a basic theme. A program project generally involves the organized efforts of relatively large groups, members of which are conducting research projects designed to elucidate the various aspects or components of this objective. Each research project is usually under the leadership of an established investigator. The grant can provide support for certain basic resources used by these groups in the program, including clinical components, the sharing of which facilitates the total research effort. A program project is directed toward a range of problems having a central research focus, in contrast to the usually narrower thrust of the traditional research project. Each project supported through this mechanism should contribute or be directly related to the common theme of the total research effort. These scientifically meritorious projects should demonstrate an essential element of unity and interdependence, i.e., a system of research activities and projects directed toward a well-defined research program goal. |
Molecular Mechanisms Determining Lung Development and Disease @ University of Cincinnati
The long-term goals of this component seek to 1) elucidate the hierarchy of transcriptional controls that determine the commitment of progenitor cells that produce the differentiated cells of the developing and mature respiratory epithelium and 2) utilize transcriptional control elements governing lung epithelial cell gene expression to determine the pathogenesis of acute and chronic lung disorders such as respiratory distress syndrome, pulmonary fibrosis, bronchopulmonary dysplasia and other disorders of surfactant homeostasis. Our analysis of the transcriptional controls of surfactant proteins A, B, C and CC10 provided critical insight into the role of thyroid transcription factor 1 (TTF-1), a 38 kilosdalton homeodomain, containing protein of the NK-2 family, and the HNF-3 family of nuclear transcription proteins in lung-specific gene expression. We hypothesize that these proteins bind to and activate promoter elements that control the temporal, spatial and humoral influences on lung epithelial cell gene expression. The present application will now determine the role of TTF-1 and HNF-3alpha in lung development, differentiation and gene expression using gene ablation and gene addition experiments in vitro and in vivo. The mechanisms controlling TTF-1 gene expression in the developing lung epithelium will be discerned. Downstream targets activated by TTF-1 will be identified by differential display to determine their roles in gene expression and surfactant homeostasis in developing and mature lung.
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0.958 |