Global gene expression analysis of human bronchial epithelial cells exposed to cigarette smoke, smoke condensates, or components thereof
View Patent ↗Aspects of the present invention concern the identification of several methods to analyze the genes that are modulated in normal human bronchial epithelial (NHBE) cells after exposure to cigarette smoke condensates (CSC) or cigarette smoke (CS). Embodiments described herein include methods to identify a gene that is modulated in response to exposure to CSC or CS, methods to identify tobacco products that have a reduced potential to contribute to tobacco-related disease, methods to make tobacco products that have a reduced potential to contribute to a tobacco-related disease, methods to identify a subject's predilection to acquire a tobacco related disease, the use of particular genes as biomarkers for tobacco-related disease, and patterns of gene expression or genetic signatures that are unique to each particular tobacco product.
1. A method of selecting a tobacco that produces a reduced expression of Sequestosome 1 in human cells comprising:
(a) providing a first tobacco;
(b) obtaining smoke from said first tobacco;
(c) contacting a first isolated population of human cells with said smoke from said first tobacco;
(d) measuring the level of expression of Sequestosome 1 in said first isolated population of human cells in response to said contact, with said smoke from said first tobacco;
(e) providing a second tobacco;
(f) obtaining smoke from said second tobacco;
(g) contacting a second isolated population of human cells with said smoke from said second tobacco;
(h) measuring the level of expression of Sequestosome 1 in said second isolated population of human cells in response to said contact with said smoke from said second tobacco;
(i) comparing the level of expression of Sequestosome 1 in said first isolated population of human cells measured in step (d) with the level of expression of Sequestosome 1 in said second isolated population of human cells measured in step (h); and
(j) identifying the tobacco that produced a reduced expression of Sequestosome 1 in an isolated population of human cells in response to contact with smoke from said tobacco.
2. The method of claim 1 , wherein said first tobacco and said second tobacco comprise a burley tobacco.
3. The method of claim 1 , wherein said first tobacco and said second tobacco comprise a flue tobacco.
4. The method of claim 1 , wherein said second tobacco comprises a modified tobacco.
5. The method of claim 1 , wherein said first tobacco is provided in a first cigarette and said second tobacco is provided in a second cigarette.
6. The method of claim 5 , wherein said first cigarette and said second cigarette comprise a filter.
7. The method of claim 1 , wherein said first and second isolated populations of human cells are normal human cells of the lung, mouth, or tongue.
8. The method of claim 1 , wherein said first and second isolated populations of human cells are normal human bronchial epithelial (NHBE) cells.
9. The method of claim 1 , further comprising measuring the level of expression of a second gene in said first and second isolated populations of human cells after contacting said first and second isolated populations of human cells with said smoke.
10. The method of claim 9 , wherein said second gene is selected from the group consisting of: Ferritin heavy polypeptide, Heat Shock 70 kD protein 1A, Thioredoxin reductase 1, Heme oxygenase (decycling) 1, and Prostaglandin-endoperoxide synthase 2.
11. The method of claim 1 , further comprising incorporating the tobacco identified in step (j) into a tobacco product.
12. The method of claim 11 , wherein said tobacco product is a cigarette.