Universitą degli studi di Pavia
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Incandela research activity
Identification of targets for new anti-tuberculosis drugs
Tuberculosis (TB), an infectious disease caused by Mycobacterium tuberculosis, is still a leading cause of death in developing countries, and a re-emerging disease in developed countries. The increasing spread of M. tuberculosis multidrug resistant strains (MDR, XDR and TDR) represents a serious threat to public health, particularly in immunocompromised patients. The management of this disease is challenging and requires the development of a pipeline of new drugs with novel mechanisms of action and, moreover, the identification of new drug targets. Within the project “New Medicines for Tuberculosis” (NM4TB), the laboratory headed by Prof. Giovanna Riccardi, at the University of Pavia, identified in 2009 the therapeutic target of two new drug classes against TB: the Benzothiazinones (BTZ) and the Dinitrobenzamides derivatives (DNB). Their anti-tuberculosis activity has been demonstrated in vitro and in vivo. The cellular target of BTZs and DNBs is the Rv3790 (DprE1) protein of M. tuberculosis which is involved in the biosynthesis of arabinogalactan, an essential component of the mycobacterial cell-wall. Additionally, it was also identified a nitroreductase from M. smegmatis (NfnB) responsible for the transformation of the BTZ in a less active derivative. I have dealt with the characterization of several mutants of M. smegmatis that over-express the nitroreductase NfnB. Also, I'm characterizing some mutants of M. smegmatis resistant to BTZ which present a different mechanism of resistance to BTZ.
Development of a new diagnostic kit for screening of infections caused by pathogenic mycobacteria
The agents that cause tuberculosis in humans and animals, include among others Mycobacterium tuberculosis, Mycobacterium africanum, Mycobacterium bovis and Mycobacterium canettii (belonging to the Mycobacterium tuberculosis complex or MTBC). In addition, infections caused by group of nontuberculous mycobacteria (NTM), are due to different clinical problems. So it is an urgent need to identify new diagnostic tools. For this reason, I am working to create a rapid assay for molecular screening and identification of strains belonging to the genus Mycobacterium, resistant to BTZ and DNB. This assay involves the use of a single tube of Real-Time PCR, a single pair of primers and three probes for the identification of sensitivity, resistance and genus. The assay exploits a new technology developed by Sentinel CH SpA (STAT-NAT, STabilized Amplification Technology) that allows the room temperature storage of the complete and ready-to-use PCR mix for at least one year. It is based on the analysis of the Rv3790 gene of M. tuberculosis and the ortholog genes of other 19 mycobacteria species (MTBC and NTM).
Tuberculosis (TB), an infectious disease caused by Mycobacterium tuberculosis, is still a leading cause of death in developing countries, and a re-emerging disease in developed countries. The increasing spread of M. tuberculosis multidrug resistant strains (MDR, XDR and TDR) represents a serious threat to public health, particularly in immunocompromised patients. The management of this disease is challenging and requires the development of a pipeline of new drugs with novel mechanisms of action and, moreover, the identification of new drug targets. Within the project “New Medicines for Tuberculosis” (NM4TB), the laboratory headed by Prof. Giovanna Riccardi, at the University of Pavia, identified in 2009 the therapeutic target of two new drug classes against TB: the Benzothiazinones (BTZ) and the Dinitrobenzamides derivatives (DNB). Their anti-tuberculosis activity has been demonstrated in vitro and in vivo. The cellular target of BTZs and DNBs is the Rv3790 (DprE1) protein of M. tuberculosis which is involved in the biosynthesis of arabinogalactan, an essential component of the mycobacterial cell-wall. Additionally, it was also identified a nitroreductase from M. smegmatis (NfnB) responsible for the transformation of the BTZ in a less active derivative. I have dealt with the characterization of several mutants of M. smegmatis that over-express the nitroreductase NfnB. Also, I'm characterizing some mutants of M. smegmatis resistant to BTZ which present a different mechanism of resistance to BTZ.
Development of a new diagnostic kit for screening of infections caused by pathogenic mycobacteria
The agents that cause tuberculosis in humans and animals, include among others Mycobacterium tuberculosis, Mycobacterium africanum, Mycobacterium bovis and Mycobacterium canettii (belonging to the Mycobacterium tuberculosis complex or MTBC). In addition, infections caused by group of nontuberculous mycobacteria (NTM), are due to different clinical problems. So it is an urgent need to identify new diagnostic tools. For this reason, I am working to create a rapid assay for molecular screening and identification of strains belonging to the genus Mycobacterium, resistant to BTZ and DNB. This assay involves the use of a single tube of Real-Time PCR, a single pair of primers and three probes for the identification of sensitivity, resistance and genus. The assay exploits a new technology developed by Sentinel CH SpA (STAT-NAT, STabilized Amplification Technology) that allows the room temperature storage of the complete and ready-to-use PCR mix for at least one year. It is based on the analysis of the Rv3790 gene of M. tuberculosis and the ortholog genes of other 19 mycobacteria species (MTBC and NTM).
