Combating Antibiotics Resistant Pneumococci by Novel Strategies Based on in vivo and in vitro Host-Pathogen Interactions
This collaborative project is aimed at understanding pneumococcus-host interactions for developing novel combat strategies.
The objective is to provide the basic knowledge needed for future development of novel prevention, diagnostic and treatment tools against pneumococcal infections. Emphasis will be placed on studies into fundamental molecular aspects of the interactions between S. pneumoniae and the human host, in particular the cell biology of bacteria. Experimental approaches will include the use of cell cultures and animal models of pneumococcal diseases, the study of virulence mechanisms, as well as the molecular epidemiology of drug resistance. CAREPNEUMO brings together 12 research organizations and one SME to achieve the objectives of the call.
is a collaborative research project within the Seventh Framework Programme funded by European Commission
from 01.03. 2009-28.02.2012.
Contract No. 22311
The need to meet the challenge of Streptococcus pneumoniae
Streptococcus pneumoniae is a major cause of life-threatening infections such as pneumonia, meningitis and septicemia. The disease burden is high both in developed and developing countries, and the high-risk groups include children, elderly persons and immuno compromised patients. Approximately one million children under 5 years of age die from pneumococcal infections every year. In spite of the availability of a large number of antibiotics mortality and morbidity due to S. pneumoniae infections remain very high. The worldwide emergence of multi drug resistant S. pneumoniae strains is a big hindrance in treating pneumococcal invasive diseases.
Another problem in combating pneumococcal infections is the genetic diversity of circulating strains. Based on the capsular polysaccharide there are more than 90 structurally and chemically different serotypes. It is known that different serotypes are prevalent in different geographical regions of the world. Although a 7-valent conjugate vaccine is currently available and has proved to be successful in preventing invasive disease caused by the vaccine serotype strains, the worldwide coverage is limited. Moreover, the use of this vaccine has resulted in a replacement by non-vaccine serotypes, which may make the current vaccine ineffective in the near future. The development of novel combat strategies is urgently needed and is a big challenge for the international pneumococcal community. This is also the global aim of the present proposal.
There are three prerequisites to achieve this goal. It is important firstly to understand the distribution of serotypes and the occurrence of antibiotic resistance, secondly to identify and characterize new intervention targets by studying host-pathogen interactions, and thirdly to validate these targets in in vitro and in vivo infection models in order to identify potential candidates for prevention and therapy.
This proposal would apply a multi-disciplinary approach that includes epidemiology, host-pathogen interactions, infection models and intervention strategies to combat antibiotic resistance problems among S. pneumoniae.
The objectives of this proposal were defined in order to obtain new knowledge on the molecular epidemiology of pneumococcal diseases and antibiotic resistance in different parts of the world, apply state-of-the-art technology to study host-pathogen interactions and apply innovative technologies for improving existing intervention strategies. Moreover, it would contribute towards the implementation of measures for prevention, control and treatment of pneumococcal diseases, especially those caused by multi-drug resistant strains. To achieve the purpose of this proposal, we have assembled a team with expertise required to fulfil the following objectives:
The major tasks of this objective will be to undertake monitoring of prevalent S. pneumoniae serotypes and their resistance profiles in various countries. This will also include the epidemiology of the serotype replacement in areas where pneumococcal vaccine is in use. Since the prevalence of serotypes as well as the magnitude and quality of drug resistance is different in different parts of the world, this objective will address the epidemiology of S. pneumoniae in six countries, which include an Asian and a South American country. These studies will determine the frequency of emergence of antibiotical resistance, particularly in those serotypes that have potential to spread rapidly in the community. The second part of this objective will be to determine the changes in pneumococcal disease after introduction of the 7-valent pneumococcal conjugate vaccine in different regions.
A prerequisite for developing new control strategies is the detailed understanding of the interaction between pneumococcus and the host cells. The analysis of host-pathogen interaction will include the prevalent virulent strains, which will be continuously identified in the work packages dealing with the epidemiological studies. The molecular mechanisms of host-pathogen interactions will be elucidated by applying genomics and cell biological approaches. Identification and characterization of the factors involved will be an important part of objective 2. The biological functions of these factors will be determined by using pathogenicity tests involving cell cultures as well as animal models of pneumococcal diseases.
Development of improved vaccine and intervention strategies
This objective deals with the development of new therapeutic and vaccine tools against pneumococcal infections based on the knowledge generated in objectives 1 and 2. For designing new therapeutics, the molecular basis of pathogen recognition through specific components of pneumococcal cell wall by host proteins will be identified by using a structural biological approach. In the area of vaccine development, this objective will deal with a novel polysaccharide glycolipid conjugate vaccine as well as the development of a protein-based universal vaccine.
- Silva-Martín N, Retamosa MG, Maestro B, Bartual SG, Rodes MJ, García P, Sanz JM, Hermoso JA. (2014) Crystal structures of CbpF complexed with atropine and ipratropium reveal clues for the design of novel antimicrobials against Streptococcus pneumoniae.Biochim Biophys Acta. 1840(1):129-135. doi: 10.1016/j.bbagen.2013.09.006.
- Ribes S, Riegelmann J, Redlich S, Maestro B, de Waal B, Meijer EW, Sanz JM, Nau R. (2013) Multivalent Choline Dendrimers Increase Phagocytosis of Streptococcus pneumoniae R6 by Microglial Cells.Chemotherapy. 59(2):138-42. doi: 10.1159/000353439
- Chan WT, Moreno-Cordoba I, Yeo CC, Espinosa M (2012) Toxin-antitoxin genes of the Gram-positive pathogen Streptococcus pneumoniae: So few and yet so many.Microbiol Mol Biol Rev 76: 773-791
2. Moreno-Cordoba I, Diago-Navarro E, Barendregt A, Heck AJ, Alfonso C, Diaz-Orejas R, Nieto C, Espinosa M (2012) The toxin-antitoxin proteins relBE2Spn of Streptococcus pneumoniae: characterization and association to their DNA target.Proteins 80: 1834-1846
- Asmat TM, Klingbeil K, Jensch I, Burchhardt G, Hammerschmidt S (2012) Heterologous expression of pneumococcal virulence factor PspC on the surface of Lactococcus lactis confers adhesive properties.Microbiology 158: 771-780
- Djukic M, Munz M, Sorgel F, Holzgrabe U, Eiffert H, Nau R (2012) Overton's rule helps to estimate the penetration of anti-infectives into patients' cerebrospinal fluid.Antimicrobial Agents and Chemotherapy 56: 979-988
- Gamez G, Hammerschmidt S (2012) Combat pneumococcal infections: adhesins as candidates for protein-based vaccine development.Current Drug Targets 13: 323-337
- Haertel T, Eylert E, Schulz C, Petruschka L, Gierok P, Grubmuller S, Lalk M, Eisenreich W, Hammerschmidt S (2012) Characterization of sentral carbon metabolism of Streptococcus pneumoniae by isotopologue profiling. The Journal of Biological Chemistry 287: 4260-4274
- Horacio AN, Diamantino-Miranda J, Aguiar SI, Ramirez M, Melo-Cristino J (2012) Serotype changes in adult invasive pneumococcal infections in Portugal did not reduce the high fraction of potentially vaccine preventable infections.Vaccine 30: 218-224
- Lioy VS, Machon C, Tabone M, Gonzalez-Pastor JE, Daugelavicius R, Ayora S, Alonso JC (2012) The zeta toxin induces a set of protective responses and dormancy.PloS one 7: e30282
- Lorenzo-Diaz F, Solano-Collado V, Lurz R, Bravo A, Espinosa M (2012) Autoregulation of the synthesis of the MobM relaxase encoded by the promiscuous plasmid pMV158.Journal of Bacteriology 194: 1789-1799
- Luettge M, Fulde M, Talay SR, Nerlich A, Rohde M, Preissner KT, Hammerschmidt S, Steinert M, Mitchell TJ, Chhatwal GS, Bergmann S (2012) Streptococcus pneumoniae induces exocytosis of Weibel-Palade bodies in pulmonary endothelial cells.Cellular Microbiology 14: 210-225
- Redlich S, Ribes S, Schutze S, Czesnik D, Nau R (2012) Palmitoylethanolamide stimulates phagocytosis of Escherichia coli K1 and Streptococcus pneumoniae R6 by microglial cells.Journal of Neuroimmunology 244: 32-34
- Ruiz-Maso JA, Lopez-Aguilar C, Nieto C, Sanz M, Buron P, Espinosa M, del Solar G (2012) Construction of a plasmid vector based on the pMV158 replicon for cloning and inducible gene expression in Streptococcus pneumoniae.Plasmid 67: 53-59
- Artola-Recolons C, Llarrull LI, Lastochkin E, Mobashery S, Hermoso JA (2011) Crystallization and preliminary X-ray diffraction analysis of the lytic transglycosylase MltE from Escherichia coli.Acta Crystallographica Section F, Structural Biology and Crystallization Communications 67: 161-163
- Asmat TM, Agarwal V, Rath S, Hildebrandt JP, Hammerschmidt S (2011) Streptococcus pneumoniae infection of host epithelial cells via polymeric immunoglobulin receptor transiently induces calcium release from intracellular stores.The Journal of Biological Chemistry 286: 17861-17869
- Chan WT, Nieto C, Harikrishna JA, Khoo SK, Othman RY, Espinosa M, Yeo CC (2011) Genetic regulation of the yefM-yoeB toxin-antitoxin locus of Streptococcus pneumoniae.Journal of Bacteriology 193: 4612-4625
- Garcia MT, Blazquez MA, Ferrandiz MJ, Sanz MJ, Silva-Martin N, Hermoso JA, de la Campa AG (2011) New alkaloid antibiotics that target the DNA topoisomerase I of Streptococcuspneumoniae.The Journal of Biological Chemistry 286: 6402-6413
- Haertel T, Klein M, Koedel U, Rohde M, Petruschka L, Hammerschmidt S (2011) Impact of glutamine transporters on pneumococcal fitness under infection-related conditions.Infection and Immunity 79: 44-58
- Hernandez-Rocamora VM, Reulen SW, de Waal B, Meijer EW, Sanz JM, Merkx M (2011) Choline dendrimers as generic scaffolds for the non-covalent synthesis of multivalent protein assemblies. Chem Commun (Camb) 47: 5997-5999
- Kreikemeyer B, Gamez G, Margarit I, Giard JC, Hammerschmidt S, Hartke A, Podbielski A (2011) Genomic organization, structure, regulation and pathogenic role of pilus constituents in major pathogenic Streptococci and Enterococci. International Journal of Medical Microbiology (IJMM) 301: 240-251
- Lorenzo-Diaz F, Dostal L, Coll M, Schildbach JF, Menendez M, Espinosa M (2011) The MobM relaxase domain of plasmid pMV158: thermal stability and activity upon Mn2+ and specific DNA binding. Nucleic Acids Research 39: 4315-4329
- Maestro B, Novakova L, Hesek D, Lee M, Leyva E, Mobashery S, Sanz JM, Branny P (2011) Recognition of peptidoglycan and beta-lactam antibiotics by the extracellular domain of the Ser/Thr protein kinase StkP from Streptococcus pneumoniae.FEBS Letters 585: 357-363
- Maestro B, Santiveri CM, Jimenez MA, Sanz JM (2011) Structural autonomy of a beta-hairpin peptide derived from the pneumococcal choline-binding protein LytA.Protein Engineering, Design & Selection (PEDS) 24: 113-122
- Skoczynska A, Sadowy E, Bojarska K, Strzelecki J, Kuch A, Golebiewska A, Wasko I, Forys M, van der Linden M, Hryniewicz W (2011) The current status of invasive pneumococcal disease in Poland.Vaccine 29: 2199-2205
- Agarwal V, Asmat TM, Dierdorf NI, Hauck CR, Hammerschmidt S (2010) Polymeric immunoglobulin receptor-mediated invasion of Streptococcus pneumoniae into host cells requires a coordinate signaling of SRC family of protein-tyrosine kinases, ERK, and c-Jun N-terminal kinase. The Journal of Biological Chemistry 285: 35615-35623
- Aguiar SI, Brito MJ, Goncalo-Marques J, Melo-Cristino J, Ramirez M (2010) Serotypes 1, 7F and 19A became the leading causes of pediatric invasive pneumococcal infections in Portugal after 7 years of heptavalent conjugate vaccine use.Vaccine 28: 5167-5173
- Eldholm V, Johnsborg O, Straume D, Ohnstad HS, Berg KH, Hermoso JA, Havarstein LS (2010) Pneumococcal CbpD is a murein hydrolase that requires a dual cell envelope binding specificity to kill target cells during fratricide.Molecular Microbiology 76: 905-917
- Jensch I, Gamez G, Rothe M, Ebert S, Fulde M, Somplatzki D, Bergmann S, Petruschka L, Rohde M, Nau R, Hammerschmidt S (2010) PavB is a surface-exposed adhesin of Streptococcus pneumoniae contributing to nasopharyngeal colonization and airways infections.Molecular Microbiology 77: 22-43
- Kaur SJ, Nerlich A, Bergmann S, Rohde M, Fulde M, Zahner D, Hanski E, Zinkernagel A, Nizet V, Chhatwal GS, Talay SR (2010) The CXC chemokine-degrading protease SpyCep of Streptococcus pyogenes promotes its uptake into endothelial cells.The Journal of Biological Chemistry 285: 27798-27805
- Kuch A, Sadowy E, Skoczynska A, Hryniewicz W (2010) First report of Streptococcus pneumoniae serotype 6D isolates from invasive infections.Vaccine 28: 6406-6407
- Lioy VS, Pratto F, de la Hoz AB, Ayora S, Alonso JC (2010) Plasmid pSM19035, a model to study stable maintenance in Firmicutes.Plasmid 64: 1-17
- Lioy VS, Rey O, Balsa D, Pellicer T, Alonso JC (2010) A toxin-antitoxin module as a target for antimicrobial development.Plasmid 63: 31-39
- Nau R, Sorgel F, Eiffert H (2010) Penetration of drugs through the blood-cerebrospinal fluid/blood-brain barrier for treatment of central nervous system infections.Clinical Microbiology Reviews 23: 858-883
- Nieto C, Sadowy E, de la Campa AG, Hryniewicz W, Espinosa M (2010) The relBE2Spn toxin-antitoxin system of Streptococcus pneumoniae: role in antibiotic tolerance and functional conservation in clinical isolates.PloS One 5: e11289
- Perez-Dorado I, Gonzalez A, Morales M, Sanles R, Striker W, Vollmer W, Mobashery S, Garcia JL, Martinez-Ripoll M, Garcia P, Hermoso JA (2010) Insights into pneumococcal fratricide from the crystal structures of the modular killing factor LytC.Nature Structural & Molecular Biology 17: 576-581
- Perez-Dorado I, Sanles R, Gonzalez A, Garcia P, Garcia JL, Martinez-Ripoll M, Hermoso JA (2010) Crystallization of the pneumococcal autolysin LytC: in-house phasing using novel lanthanide complexes.Acta crytallographica Section F, Structural Biology and Crystallization Communications 66: 448-451
- Ribes S, Adam N, Ebert S, Regen T, Bunkowski S, Hanisch UK, Nau R (2010) The viral TLR3 agonist poly(I:C) stimulates phagocytosis and intracellular killing of Escherichia coli by microglial cells.Neuroscience Letters 482: 17-20
- Ribes S, Ebert S, Regen T, Agarwal A, Tauber SC, Czesnik D, Spreer A, Bunkowski S, Eiffert H, Hanisch UK, Hammerschmidt S, Nau R (2010) Toll-like receptor stimulation enhances phagocytosis and intracellular killing of nonencapsulated and encapsulated Streptococcus pneumoniae by murine microglia.Infection and Immunity 78: 865-871
- Ribes S, Ebert S, Regen T, Czesnik D, Scheffel J, Zeug A, Bunkowski S, Eiffert H, Hanisch UK, Hammerschmidt S, Nau R (2010) Fibronectin stimulates Escherichia coli phagocytosis by microglial cells.Glia 58: 367-376
- Ruiz-Cruz S, Solano-Collado V, Espinosa M, Bravo A (2010) Novel plasmid-based genetic tools for the study of promoters and terminators in Streptococcus pneumoniae and Enterococcusfaecalis.Journal of Microbiological Methods 83: 156-163
- Sadowy E, Kuch A, Gniadkowski M, Hryniewicz W (2010) Expansion and evolution of the Streptococcus pneumoniae Spain9V-ST156 clonal complex in Poland.Antimicrobial Agents and Chemotherapy 54: 1720-1727
- Silva-Martin N, Molina R, Angulo I, Mancheno JM, Garcia P, Hermoso JA (2010) Crystallization and preliminary crystallographic analysis of the catalytic module of endolysin from Cp-7, a phage infecting Streptococcus pneumoniae. Acta Crystallographica Section F, Structural Biology and Crystallization Communications 66: 670-673
- Zahlten J, Steinicke R, Opitz B, Eitel J, N'Guessan P D, Vinzing M, Witzenrath M, Schmeck B, Hammerschmidt S, Suttorp N, Hippenstiel S (2010) TLR2- and nucleotide-binding oligomerization domain 2-dependent Kruppel-like factor 2 expression downregulates NF-kappa B-related gene expression.Journal of Immunology 185: 597-604
- Carrolo M, Pinto FR, Melo-Cristino J, Ramirez M (2009) Pherotypes are driving genetic differentiation within Streptococcus pneumoniae.BMC Microbiology 9: 191
- Lorenzo-Diaz F, Espinosa M (2009) Lagging-strand DNA replication origins are required for conjugal transfer of the promiscuous plasmid pMV158.Journal of Bacteriology 191: 720-727
- Lorenzo-Diaz F, Espinosa M (2009) Large-scale filter mating assay for intra- and inter-specific conjugal transfer of the promiscuous plasmid pMV158 in Gram-positive bacteria.Plasmid 61: 65-70