Final Activity Report Summary - CHRONINFLAM (Identification of genes associated with chronic inflammation.)
The objectives of this project were to identify a gene that determined whether an inflammation, such as rheumatoid arthritis, would be chronic or self-limiting, to understand the biological role of the gene and to use the acquired knowledge to develop new, more effective drugs for treatment of chronic inflammations.
To achieve this goal, a rat model was used in which a small, isolated genetic region was shown to heal rats from chronic, arthritic inflammation. When the project started this genetic region was pretty large, coding for several hundreds of genes. During the year supported by the Marie Curie reintegration grant, a smaller genetic region within the original region was isolated and shown to exert the effect. This new region encoded only 30 genes approximately. The region was further divided into smaller parts, which were tested for effects on disease by the time of the project completion. From these data it was anticipated to be possible to determine which part encoded three to four genes that were protective. This number of genes would be possible to analyse in detail in order to decide which of them were responsible for the protection. The latter would be possible within a few months beyond the project timeframe.
While crossing rats identified the gene, new powerful bio-techniques, like microarray, were used to make qualified guesses of the identity of the protective gene. These guesses could be used to speed up the process of gene identification by pointing out genetic fragments worthwhile to test for protection of disease. Through microarray technique it was possible to scan differences in the way protected and non-protected rats used their genome in a large-scale, unbiased way.
During the year of project support by the Marie Curie reintegration grant a number of such differentially used genes suggested by microarray were confirmed by an independent method, namely real-time polymerase chain reaction (PCR). This resulted in a list of possible candidate genes, all encoded from the smaller protective genetic fragment. The list contained a gene important for the development of blood cells, a gene important for new formation of blood vessels and a gene regulating the production of acute phase proteins, i.e. proteins produced at a different level during inflammation compared to levels in health. The candidate genes were further analysed by deoxyribonucleic acid (DNA) sequencing. The gene regulating new formation of blood vessels had a number of mutations, i.e. mistakes, in the DNA code of the non-protected rat compared to the protected rat. To further examine which of these genes could be the protective one, various new methodologies were set up to analyse the genes protein products. Methods for analysis of both their properties and functions were set up. Preliminary data pointed out either the new vessel-controlling gene or the gene regulating the production of acute phase proteins as most likely. However, more data had to be produced to obtain final conclusions.
In summary, during the year supported by the Marie Curie reintegration grant, the project reached several important scientific milestones on its way towards identification of chronic inflammation controlling genes.
To achieve this goal, a rat model was used in which a small, isolated genetic region was shown to heal rats from chronic, arthritic inflammation. When the project started this genetic region was pretty large, coding for several hundreds of genes. During the year supported by the Marie Curie reintegration grant, a smaller genetic region within the original region was isolated and shown to exert the effect. This new region encoded only 30 genes approximately. The region was further divided into smaller parts, which were tested for effects on disease by the time of the project completion. From these data it was anticipated to be possible to determine which part encoded three to four genes that were protective. This number of genes would be possible to analyse in detail in order to decide which of them were responsible for the protection. The latter would be possible within a few months beyond the project timeframe.
While crossing rats identified the gene, new powerful bio-techniques, like microarray, were used to make qualified guesses of the identity of the protective gene. These guesses could be used to speed up the process of gene identification by pointing out genetic fragments worthwhile to test for protection of disease. Through microarray technique it was possible to scan differences in the way protected and non-protected rats used their genome in a large-scale, unbiased way.
During the year of project support by the Marie Curie reintegration grant a number of such differentially used genes suggested by microarray were confirmed by an independent method, namely real-time polymerase chain reaction (PCR). This resulted in a list of possible candidate genes, all encoded from the smaller protective genetic fragment. The list contained a gene important for the development of blood cells, a gene important for new formation of blood vessels and a gene regulating the production of acute phase proteins, i.e. proteins produced at a different level during inflammation compared to levels in health. The candidate genes were further analysed by deoxyribonucleic acid (DNA) sequencing. The gene regulating new formation of blood vessels had a number of mutations, i.e. mistakes, in the DNA code of the non-protected rat compared to the protected rat. To further examine which of these genes could be the protective one, various new methodologies were set up to analyse the genes protein products. Methods for analysis of both their properties and functions were set up. Preliminary data pointed out either the new vessel-controlling gene or the gene regulating the production of acute phase proteins as most likely. However, more data had to be produced to obtain final conclusions.
In summary, during the year supported by the Marie Curie reintegration grant, the project reached several important scientific milestones on its way towards identification of chronic inflammation controlling genes.