Maria Rosalia Pasca

Dr. Maria Rosalia Pasca EDUCATION • 21/03/1997: Degree in Natural Sciences (110/110), University of Bari. • 04/02/2002: PhD in Genetics and Molecular Evolution, University of Bari. PROFESSIONAL EXPERIENCES • 1998-2002: PhD student in the Laboratory of Genetics of Microorganisms, University of Bari (supervisor: Prof. M.S. Ciampi). • 12/1998-01/1999: Stages in the Laboratory of Molecular Genetics, University of Camerino (supervisor: Prof. C. Gualerzi). • 1999, 2001: Stages in the Laboratory of Molecular Genetics, University of Ancona (supervisor: Prof. A. La Teana). • 01-12/2000: Stage in the Laboratory of Molecular Genetics of CNRS de Gif-sur-Yvette, Paris, France (supervisor: Dr. L. Bossi). • 04/2002-09/2006: Post-Doc in the Laboratory of Molecular Microbiology, University of Pavia (supervisor: Prof. G. Riccardi). DIDACTICS Official teachings • 2007-2008: In the SILSIS Speciality School (University of Pavia), Professor of the following courses: Didactics of Microbiology (20 h) and Didactics of Microbiology (15 h) and Food Microbiology Laboratories (15 h) for 13A, 57A, 60A classes, University of Pavia. • From 2008-2009: Professor of the course Pathogen Identifications (3 CFU), Master degree in Experimental and Applied Biology (Bioanalytic course), University of Pavia. • 2009-2010: Professor of the course “Microbiological techniques” (6 CFU), Degree in Biological Sciences (Industrial course), University of Pavia. • 2009-2010: Professor of the course “Microbiological techniques” (20 h), Speciality School in Microbiology and Virology, University of Pavia. • From 2010-11: Professor of the course “Laboratory of cellular Methodologies (1 CFU), Degree in Biological Sciences (Biomolecular course), University of Pavia. • From 2010-11: Professor of the course “Environmental Microbiology” (3 CFU), Master degree in Experimental and Applied Biology (Environmental biology and Biodiversity course), University of Pavia. • From 2008: member staff of PhD School in Biomolecular and Genetics sciences, University of Pavia (Coordinator: Prof. Antonio Torroni). LECTURE Pasca M. R. (28/05/2009) “New drugs and a new cellular target against tuberculosis”. I.N.M.I. L. Spallanzani I.R.C.C.S., Roma. REFEREE'S ACTIVITY She has been referee for “Journal of enzyme inhibition and medicinal chemistry” and “Microbial drug resistance”. ACADEMIC POSITION From 10/2006: Researcher in Microbiology (BIO/19) (confirmed from 10/2009), Department of Genetics and Microbiology, University of Pavia. MAIN FIELDS OF RESEARCH 1. Identification of targets for new antitubercular drugs (EC-VI and –VII framework). Tuberculosis (TB) remains the leading cause of mortality due to a bacterial pathogen, Mycobacterium tuberculosis (Mtb). Moreover, Mtb strains resistant to several drugs (MDR-TB and XDR-TB) are becoming a threat to public health worldwide. Consequently, there is an urgent necessity of new anti-TB drugs. We have identified the target of the benzothiazinones (BTZ), very effective drugs against Mtb, that is Rv3790, an enzyme involved in the biosynthesis of arabinogalactan, a cell wall component (PCT/EP2008/001088) (Makarov et al., 2009; Manina et al., 2010b). We have characterized 240 Mtb clinical isolates and verified that the BTZ are effective also against MDR and XDR strains (Pasca et al., 2010). We have also identified a Mycobacterium smegmatis nitroreductase, NfnB, able to convert the effective molecule BTZ-NO2 into its less active derivative BTZ-NH2 (PCT/EP2008/009231) (Manina et al., 2010a). The study of mechanism of action of new anti-TB drugs is in progress. Collaborations: Cole ST (EPFL, Lausanne, Switzerland), Makarov V (Bakh Institute of Biochemistry, Russian Academy of Science, Moscow, Russia), Mikusova K (Department of Biochemistry, Comenius University, Bratislava, Slovakia). 2. The role of RND efflux pumps in the intrinsic antibiotic resistance of Burkholderia cenocepacia (Fibrosi Cistica 2004, 2006, 2009). Burkholderia cenocepacia is a Gram-negative opportunistic pathogen responsible for serious infections in cystic fibrosis patients. Eradication of these infections is complicated by its intrinsic drug-resistance. The resistance mechanisms in Gram-negative bacteria comprise efflux systems that extrude several drugs out of the cells. We studied the RND drug efflux transporters in B. cenocepacia J2315, a MDR clinical isolate. Using an in silico analysis, we discovered 14 genes encoding putative RND efflux pumps in the B. cenocepacia J2315 genome (Guglierame et al., 2006). In order to investigate the contribution of these transporters in drug resistance, three rnd operons were inactivated in J2315 strain. The inactivation of orf4 operon conferred more sensitivity to some antibiotics and we demonstrated that this operon codified for an efflux pump (Buroni et al., 2009). We compared the transcription profile of mutant strains with that of the wild-type by microarray analysis. We found that one of the efflux pumps we investigated contributes to the antibiotic resistance of B. cenocepacia. Interestingly, motility and chemotaxis-related genes appeared to be up-regulated in two mutants. Moreover, biofilm production was enhanced in all mutants. Together our results indicate that efflux pumps play a wider role than just in drug resistance in B. cenocepacia (Bazzini S, Udine C, Sass A, Pasca MR, Longo F, Emiliani G, Fondi M, Perrin E, Decorosi F, Viti C, Giovannetti L, De Rossi E, Leoni L, Fani R, Riccardi G, Mahenthiralingam E, Buroni S. Deciphering the role of RND efflux transporters in Burkholderia cenocepacia. Submitted to PloS One). Collaborations: Mahenthiralingam E (Cardiff University, Cardiff, UK), Fani R (Department of Evolutionary Biology, University of Florence, Firenze, Italy), Leoni L (Department of Biology, University Roma Tre, Rome, Italy). PERSONAL BIBLIOGRAPHY SHE IS AUTHOR OF 18 PEER-REVIEWED ARTICLES, 25 INTERNATIONAL AND NATIONAL COMMUNICATIONS TO CONGRESSES AND 2 INTERNATIONAL PATENT APPLICATIONS. PUBLICATIONS 1. Federici F, Vitali B, Gotti R, Pasca MR, Gobbi S, Peck AB, Brigidi P (2004) Characterization and heterologous expression of the oxalyl-CoA decarboxylase gene from Bifidobacterium lactis. Applied Environmental Microbiology 70: 5066-5073 (IF = 3.818) 2. Pasca MR, Guglierame P, Arcesi F, Bellinzoni M, De Rossi E, Riccardi G (2004) Rv2786c-2687c-2688c, an ABC fluoroquinolone efflux pump in Mycobacterium tuberculosis. Antimicrobial Agents and Chemiotherapy 48: 3175-3178 (IF = 4.379) 3. Bellinzoni M, Buroni S, Pasca MR, Guglierame P, Arcesi F, De Rossi E, Riccardi G (2005) Glutamine amidotransferase activity of NAD+ synthetase from Mycobacterium tuberculosis depends on an amino-terminal nitrilase domain. Research in Microbiology 156: 173-177 (IF = 2.426) 4. Pasca MR, Guglierame P, De Rossi E, Zara F, Riccardi G (2005) The mmpL7 gene of Mycobacterium tuberculosis is responsible for isoniazid efflux in Mycobacterium smegmatis. Antimicrobial Agents and Chemiotherapy 49: 4775-4777 (IF = 4.379). 5. Guglierame P, Pasca MR, De Rossi E, Buroni S, Arrigo P, Manina G, Riccardi G (2006) Efflux pump genes of the resistance-nodulation-division family in Burkholderia cenocepacia genome. BMC Microbiology 6:66 (IF = 3.329). 6. Buroni S, Manina G, Guglierame P, Pasca MR, Riccardi G, De Rossi E (2006) LfrR is a repressor that regulates expression of the efflux pump LfrA in Mycobacterium smegmatis. Antimicrobial Agents and Chemoterapy 50: 4044-4052 (IF = 4.819). 7. Maciag A, Dainese E, Rodriguez MG, Milano A, Provvedi R, Pasca MR, Smith I, Palù G, Riccardi G, Manganelli R (2007) Global analysis of Mycobacterium tuberculosis FurB regulon. Journal of Bacteriology 189: 730-740 (IF = 4.035). 8. Riccardi G, Milano A, Pasca MR, Nies DH (2008) Genomic analysis of zinc homeostasis in Mycobacterium tuberculosis. FEMS Microbiology Letters 287:1-7 (IF = 2.354). 9. Milano A, Pasca MR, Provvedi R, Lucarelli AP, Manina G, Ribeiro AL, Manganelli R, Riccardi G (2009) Azole resistance in Mycobacterium tuberculosis is mediated by the MmpL5-MmpS5 efflux system. Tuberculosis (Edinb) 89:84-90 (Primo nome) (IF = 3.425). 10. * Makarov V, Manina G, Mikusova K, Möllmann U, Ryabova O, Saint-Joanis B, Dhar N, Pasca MR, Buroni S, Lucarelli AP, Milano A, De Rossi E, Belanova M, Bobovska A, Dianiskova P, Kordulakova J, Sala C, Fullam E, Schneider P, McKinney JD, Brodin P, Christophe T, Waddell S, Butcher P, Albrethsen J, Rosenkrands I, Brosch R, Nandi V, Bharath S, Gaonkar S, Shandil RK, Balasubramanian V, Balganesh T, Tyagi S, Grosset J, Riccardi G, Cole ST (2009) Benzothiazinones kill Mycobacterium tuberculosis by blocking arabinan synthesis. Science 324:801-804 (IF = 30.268). 11. Riccardi G, Pasca MR, Buroni S (2009) Mycobacterium tuberculosis: drug resistance and future perspectives. Future Microbiology 4:597-614 (IF = 2.859). 12. Dalla Valle C, Pasca MR, De Vitis D, Capra Marzani F, Emmi V, Marone P (2009) Control of MRSA infection and colonisation in an intensive care unit by GeneOhm MRSA assay and culture methods. BMC Infectious Diseases 9:137 (IF = 2.55). 13. Buroni S, Pasca MR, Flannagan RS, Bazzini S, Milano A, Bertani I, Venturi V, Valvano MA, Riccardi G (2009) Assessment of three Resistance-Nodulation-Cell Division drug efflux transporters of Burkholderia cenocepacia in intrinsic antibiotic resistance. BMC Microbiology 9:200 (IF = 3.329). 14. Pasca MR, Degiacomi G, Ribeiro AL, Zara F, De Mori P, Mirrione M, Brerra R, Pagani L, Pucillo L, Troupioti P, Makarov V, Cole ST, Riccardi G (2010) Clinical isolates of Mycobacterium tuberculosis in four european hospitals are uniformly susceptible to benzothiazinones. Antimicrobial Agents and Chemoterapy 54:1616-1618 (IF = 4.819). 15. Perrin E, Fondi M, Papaleo MC, Maida I, Buroni S, Pasca MR, Riccardi G, Fani R (2010) Exploring the HME and HAE1 efflux systems in the genus Burkholderia. BMC Evolutionary Biology 10:164 (IF=4.29). 16. Manina G, Bellinzoni M, Pasca MR, Neres J, Milano A, de Jesus Lopes Ribeiro AL, Buroni S, Škovierová H, Dianišková P, Mikušová K, Marák J, Makarov V, Giganti D, Haouz A, Lucarelli AP, Degiacomi G, Piazza A, Chiarelli LR, De Rossi E, Salina E, Cole ST, Alzari PM, Riccardi G (2010a) Biological and structural characterization of the Mycobacterium smegmatis nitroreductase NfnB, and role in benzothiazinone resistance. Molecular Microbiology 77:1172-1185 (Primo nome) (IF=5.602). 17. Manina G, Pasca MR, Buroni S, De Rossi E, Riccardi G (2010b) Decaprenylphosphoryl-β-D-ribose 2’-epimerase from Mycobacterium tuberculosis is a magic drug target. Current medicinal chemistry 17:3099-3108 (IF=4.90). 18. Lucarelli AP, Buroni S, Pasca MR, Rizzi M, Valentini G, Riccardi G, Chiarelli LR (2010) Mycobacterium tuberculosis phosphoribosyl pyrophosphate synthetase: biochemical features of a crucial enzyme for mycobacterial cell wall biosynthesis. PloS One 5:e15494 (IF=4.383). 19. Bazzini S, Udine C, Sass A, Pasca MR, Longo F, Emiliani G, Fondi M, Perrin E, Decorosi F, Viti C, Giovannetti L, De Rossi E, Leoni L, Fani R, Riccardi G, Mahenthiralingam E, Buroni S (2011) Deciphering the role of RND efflux transporters in Burkholderia cenocepacia. PloS One (IF=4.383). In press. 20. Pasca MR, Dalla Valle C, de Jesus Lopes Ribeiro AL, Buroni S, Papaleo MC, Bazzini S, Udine C, Incandela ML, Daffara S, Fani R, Riccardi G, Marone P. Evaluation of fluoroquinolone resistance mechanisms in Pseudomonas aeruginosa MDR clinical isolates. Submitted to Microbial Drug Resistance (IF=1.989). *THIS ARTICLE HAS BEEN CITED AS ONE OF THE KEY PAPER PUBLISHED IN 2009 (SEE: NATURE MEDICINES 15: 1349). PATENTS 1. Riccardi G, Manina G, Pasca MR (2008) “An effective new drug target for the treatment of tuberculosis” (PCT/EP2008/001088) (licensed to Sentinel Diagnostics). 2. Riccardi G, Manina G, Pasca MR (2008) “Nitroreductase NfnB from Mycobacterium smegmatis” (PCT/EP2008/009231).