Conducting airway epithelia are the primary target of inhaled air pollutants that include tobacco smoke. The cellular responses to the inhalation-related injury are quite different among different regions of airways. In trachea and large airway regions, the expression of squamous epithelium (squamous cell metaplasia) is a predominant feature in these lining cells. The mechanisms involved in this change are still unknown. In this proposal, we focus on the regulation of the expression of a squamous cell differentiation marker, small proline-rich protein (spr1), by cigarette smoke in airway epithelial cells. We hypothesize that spr1 gene expression is an early marker of squamous cell differentiation and the regulation of this gene expression by cigarette smoke occurs at the transcriptional level. Specifically, we will compare the effects of environmental tobacco smoke (ETS) and mainstream tobacco smoke (MTS) on the expression of the spr1 gene in primary cultures of monkey/human tracheobronchial epithelial cells that are maintained in a biphasic culture system (Specific Aim 1). We will determine the nature of this regulation at both the transcriptional and post-transcriptional levels (Specific Aim 2). These studies will lead to the characterization of both the cis-elements and trans-factors involved in the regulation (Specific Aim 3). Once these tobacco smoke-responsive elements are identified, we will determine the effects of various known chemicals that are present in tobacco smoke, such as nitrosamine, nicotine, CO, NO, and several carcinogens (Specific Aim 4). These studies will lead to a better understanding of the nature of gene regulation by tobacco smoke. Preliminary studies demonstrated that an increase of spr1 gene expression is closely associated with airway cell squamous differentiation. We recently isolated both the full-length cDNA and the genomic clones of the human spr1 gene. An antibody, which is monospecific to the spr1 protein, has been developed. Using these probes, we demonstrated that vitamin A down-regulates spr1 mRNA level through decreasing the stability of mRNA in cultured cells, while a phorbol ester, phorbol 12-myristate 12-acetate (PMA), up-regulates spr1 gene expression at the transcriptional level. This up-regulation by PMA is associated with the AP-1 binding site found in the 5'-regulatory region of the spr1 gene. A similar approach should be able to determine the nature of cigarette smoke on the regulation of the expression of the squamous cell marker spr1.

Agency
National Institute of Health (NIH)
Institute
National Institute of Environmental Health Sciences (NIEHS)
Type
Research Project (R01)
Project #
1R01ES006230-01
Application #
3254506
Study Section
Lung Biology and Pathology Study Section (LBPA)
Project Start
1993-01-01
Project End
1996-12-31
Budget Start
1993-01-01
Budget End
1993-12-31
Support Year
1
Fiscal Year
1993
Total Cost
Indirect Cost
Name
University of California Davis
Department
Type
Schools of Veterinary Medicine
DUNS #
094878337
City
Davis
State
CA
Country
United States
Zip Code
95618
Chen, Yin; Zhao, Yu Hua; Kalaslavadi, Tejas Baba et al. (2004) Genome-wide search and identification of a novel gel-forming mucin MUC19/Muc19 in glandular tissues. Am J Respir Cell Mol Biol 30:155-65
Di, Yuan-Pu; Harper, Richart; Zhao, Yuhua et al. (2003) Molecular cloning and characterization of spurt, a human novel gene that is retinoic acid-inducible and encodes a secretory protein specific in upper respiratory tracts. J Biol Chem 278:1165-73
Chen, Yin; Thai, Philip; Zhao, Yu-Hua et al. (2003) Stimulation of airway mucin gene expression by interleukin (IL)-17 through IL-6 paracrine/autocrine loop. J Biol Chem 278:17036-43
Kao, Cheng Yuan; Chen, Yin; Zhao, Yu Hua et al. (2003) ORFeome-based search of airway epithelial cell-specific novel human [beta]-defensin genes. Am J Respir Cell Mol Biol 29:71-80
Yoneda, Ken; Chang, Mary Mann-Jong; Chmiel, Ken et al. (2003) Application of high-density DNA microarray to study smoke- and hydrogen peroxide-induced injury and repair in human bronchial epithelial cells. J Am Soc Nephrol 14:S284-9
Chang, Wen-Hsing; Reddy, Sekhar P-M; Di, Yuan-Pu Peter et al. (2002) Regulation of thioredoxin gene expression by vitamin A in human airway epithelial cells. Am J Respir Cell Mol Biol 26:627-35
Soref, C M; Di, Y P; Hayden, L et al. (2001) Characterization of a novel airway epithelial cell-specific short chain alcohol dehydrogenase/reductase gene whose expression is up-regulated by retinoids and is involved in the metabolism of retinol. J Biol Chem 276:24194-202
Chen, Y; Zhao, Y H; Di, Y P et al. (2001) Characterization of human mucin 5B gene expression in airway epithelium and the genomic clone of the amino-terminal and 5'-flanking region. Am J Respir Cell Mol Biol 25:542-53
Chang, M M; Juarez, M; Hyde, D M et al. (2001) Mechanism of dexamethasone-mediated interleukin-8 gene suppression in cultured airway epithelial cells. Am J Physiol Lung Cell Mol Physiol 280:L107-15
Yoneda, K; Peck, K; Chang, M M et al. (2001) Development of high-density DNA microarray membrane for profiling smoke- and hydrogen peroxide-induced genes in a human bronchial epithelial cell line. Am J Respir Crit Care Med 164:S85-9

Showing the most recent 10 out of 33 publications