Abstract

The long term goals of this work are to study functional roles of retinoic acids (RA) and their receptors (RAR) in regulating fetal lung development by controlling lung specific gene expression in respiratory epithelial cells, and to apply the knowledge to clinical treatment. Surfactant protein B (SP-B) maintains alveolar stability by enhancing the rate of spreading of phospholipid at the air-water interface, and therefore is critical to postnatal respiratory adaptation and recovery of respiratory epithelium following oxidant injury. In this application, we propose to determine molecular mechanisms by which RA and RAR stimulate the SP-B gene both in vitro and in vivo.
The specific aims are: 1) Identification of cis-retinoic acid responsive elements (RAREs) on the SP-B promoter. DNA footprint, gel electrophoresis mobility shift assay and site-specific mutagenesis will be performed. The studies will be done in respiratory epithelial cells, including the H441 cell line, the ML3-15 cell line and primary isolates of alveolar Type II epithelial cells; 2) Characterization of the interaction between RAR and the tissue-specific transcription factor, TTF-1. Co-immunoprecipitation, Western blot and the yeast two-hybrid system will be performed to identify the precise interaction domains of both RAR and TTF-1, and their roles in binding and activating cis-acting elements in the SP-B gene; 3) Characterization of cis-RARE to determine the effects of RA/RAR on the temporal and spatial expression of the SP-B gene in transgenic mice. The wild type and RARE site-mutated hSP-B promoters will be linked to the CAT reporter gene and introduced into mice. In situ hybridization of tissue sections and CAT assay from transgenic lungs will be used to assess the in vivo RARE hybridization of tissue sections and CAT assay from transgenic lungs will be used to assess the in vivo RARE functional roles determining SP-B promoter expression levels and specificities in bronchiolar and alveolar Type II epithelial cells.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL061803-03
Application #
6390176
Study Section
Lung Biology and Pathology Study Section (LBPA)
Program Officer
Berberich, Mary Anne
Project Start
1999-08-20
Project End
2003-06-30
Budget Start
2001-07-01
Budget End
2002-06-30
Support Year
3
Fiscal Year
2001
Total Cost
$234,806
Indirect Cost

  Institution

Name
Cincinnati Children's Hospital Medical Center
Department
Type
DUNS #
071284913
City
Cincinnati
State
OH
Country
United States
Zip Code
45229

  Publications

Yan, Cong; Qu, Peng; Du, Hong (2012) Myeloid-specific expression of Stat3C results in conversion of bone marrow mesenchymal stem cells into alveolar type II epithelial cells in the lung. Sci China Life Sci 55:576-90
Wu, Lingyan; Yan, Cong; Czader, Magdalena et al. (2012) Inhibition of PPAR? in myeloid-lineage cells induces systemic inflammation, immunosuppression, and tumorigenesis. Blood 119:115-26
Kirov, Aleksandr; Duarte, Maria; Guay, Justin et al. (2012) Transgenic expression of nonclassically secreted FGF suppresses kidney repair. PLoS One 7:e36485
Wu, Lingyan; Wang, Guixue; Qu, Peng et al. (2011) Overexpression of dominant negative peroxisome proliferator-activated receptor-? (PPAR?) in alveolar type II epithelial cells causes inflammation and T-cell suppression in the lung. Am J Pathol 178:2191-204
Li, Yuan; Qu, Peng; Wu, Lingyan et al. (2011) Api6/AIM/Spýý/CD5L overexpression in alveolar type II epithelial cells induces spontaneous lung adenocarcinoma. Cancer Res 71:5488-99
Qu, Peng; Yan, Cong; Blum, Janice S et al. (2011) Myeloid-specific expression of human lysosomal acid lipase corrects malformation and malfunction of myeloid-derived suppressor cells in lal-/- mice. J Immunol 187:3854-66
Qu, Peng; Yan, Cong; Du, Hong (2011) Matrix metalloproteinase 12 overexpression in myeloid lineage cells plays a key role in modulating myelopoiesis, immune suppression, and lung tumorigenesis. Blood 117:4476-89
Qu, Peng; Shelley, William C; Yoder, Mervin C et al. (2010) Critical roles of lysosomal acid lipase in myelopoiesis. Am J Pathol 176:2394-404
Qu, Peng; Roberts, Jennifer; Li, Yuan et al. (2009) Stat3 downstream genes serve as biomarkers in human lung carcinomas and chronic obstructive pulmonary disease. Lung Cancer 63:341-7
Qu, Peng; Du, Hong; Wilkes, David S et al. (2009) Critical roles of lysosomal acid lipase in T cell development and function. Am J Pathol 174:944-56

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