TN-SEQ

Identification of genetic determinants involved in Streptococcus pneumoniae pathogenesis

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 Obiettivo del progetto (Objective)

'Respiratory tract infections are a leading cause of global mortality and morbidity. It has been estimated by the WHO that annually 4-5 million people die of pneumonia. Streptococcus pneumoniae, aptly named because it is the most important bacterial cause of pneumonia, is likely primarily responsible for these deaths. Infection by S. pneumoniae is a complex process dependent on a number of essential pathways, of which members could form ideal candidates for drug design. To identify microbial genes required for pathogenesis, we plan to use an insertion knockout strategy that allows rapid identification of disrupted genes. The host lab successfully performed micro array based approaches and proof-of-concept next generation sequencing methodologies to determine which genes are essential under a number of relevant in vitro and in vivo conditions. To identify shared essential pathways we will use statistical analysis, pathway analysis, network reconstruction and functional category enrichment methods on existing data and data generated within this project to determine the most ideal candidates for drug design. These key genes do not necessarily have to be detected initially in all screenings because of the noise inherent to the techniques used or limited survival of the mutant library. Genes coding for products with the appropriate characteristics will be subsequently tested in an animal model. Vaccines and novel drugs generated from this approach will provide alternatives to continue treatment and prevention of life-threatening chronic and acute bacterial infectious diseases.'

Introduzione (Teaser)

Pneumonia annually kills millions of people around the world. This respiratory tract infection is usually treated with antibiotics or vaccines but emerging multi-drug-resistant strains pose an even greater threat.

Descrizione progetto (Article)

Caused primarily by Streptococcus pneumoniae, the incidence of non-vaccine drug-resistant strains are on the rise. This increases the need to find effective broad-range vaccines to lower infection and improve patient survival, particularly in the case of infants.

Scientists of the EU-funded project 'Identification of genetic determinants involved in Streptococcus pneumoniae pathogenesis' (TN-SEQ) worked on identifying genes associated with the growth and pathogenesis of this bacterium. They used an insertion knockout strategy for this purpose.

Using micro arrays and next generation sequencing methodologies such as transposon sequencing, researchers pinpointed genes critical for S. pneumoniae survival and pathogenesis. Associated pathways were identified using statistical analysis, pathway analysis, network reconstruction and functional category enrichment methods. Moreover, they successfully determined factors affecting S. pneumoniae persistence and virulence in the host. This helped in shortlisting promising drug and vaccine candidates for further investigation and validation after generating knockouts.

Project outcomes have helped determine promising antibiotic drug targets through proof of concept studies. These warrant further investigation and successful outcomes in future clinical trials would mitigate the severity of this life-threatening pathogen.