Methods for diagnosing and treating asthma
The present invention relates to asthma. Particularly, the present invention relates to clinical screening, diagnosis, prognosis, therapy and prophylaxis, as well as for drug screening and drug development for the treatment of asthma. The present invention relates to a new paradigm in diagnosing, screening, and treating asthma by affecting airway epithelial synchronization.
1. A method for screening for an agent capable of alleviating asthma comprising:
a. inducing an asthmatic airway epithelial cell sample to undergo mitosis;
b. contacting the cell sample with an agent;
c. performing a a cell cycle assay comprising:
application of fluorescent dye or BRDU to the cell sample, and
fluorescence activated cell sorting (FACS) analysis of the cell sample; and
d. comparing the cell cycle synchrony of the cell sample after said performing and contacting to a control asthmatic airway epithelial cell population that has not been in contact with said agent to determine the effect of the agent on the cells, wherein an increase in synchrony is indicative of the agent being useful in alleviating asthma.
2. The method of claim 1 , further comprising the step of determining the release of inflammatory cytokines by the mitotic cells.
3. The method of claim 1 , wherein the agent is selected from the group consisting of proteins, peptides, small molecules, vitamin derivatives, and carbohydrates.
4. The method of claim 1 , wherein said contacting lasts two hours or less.
5. A method for screening for an agent capable of alleviating asthma comprising:
contacting a sample of asthmatic airway epithelial cells with an agent;
performing a cell cycle assay on said sample; and
measuring the percentage of said cells in the same phase of the cell cycle to determine the effect of the agent on the cells, wherein a sample having greater than 60 percent of said cells in the same phase of the cell cycle is indicative of the agent being useful in alleviating asthma.
6. The method of claim 5 , wherein the agent is selected from the group consisting of proteins, peptides, small molecules, vitamin derivatives, and carbohydrates.
7. The method of claim 5 , wherein said contacting lasts two hours or less.
8. A method for screening for an agent capable of alleviating asthma comprising:
contacting a sample of asthmatic airway epithelial cells with an agent;
performing a cell cycle assay on said sample; and
identifying cells in the sample that are paused in the G1-S checkpoint to determine the effect of the agent on the cells, wherein the amount of said cells that are paused in the G1-S checkpoint is indicative of the agent being useful in alleviating asthma.
9. The method of claim 8 , wherein the agent is selected from the group consisting of proteins, peptides, small molecules, vitamin derivatives, and carbohydrates.
10. The method of claim 8 , wherein said contacting lasts two hours or less.