POST-DOC Nanomedecines to inhibit LasB, a protease involved in antibiotic resistance

POST-DOC

Nanomedecines to inhibit LasB, a protease involved

in antibiotic resistance University of Liège

Project:
Antibiotic resistance is rising to dangerously high levels in all parts of the world. New resistance strains are emerging and spreading globally, threatening our ability to treat common infectious diseases. Pseudomonas aeruginosa is an opportunistic pathogen and its eradication has become increasingly difficult due to its remarkable capacity to develop antibiotics resistance. P. aeruginosa is commonly associated with infections in patients with suppressed immunity, lung inflammatory diseases (e.g., cystic fibrosis, CF) and chronic ulcers. Virulence of P. aeruginosa is largely associated to its ability to secrete into the environment a variety of degradative enzymes. Several proteases can cause extensive tissue damage, interfere with host defences, and promote bacterial propagation and invasion during infection. Of the proteases secreted by P. aeruginosa, LasB is a main virulence factor and its inhibition by small molecules improves the survival rate in different in vivo models. However, none of such small molecule inhibitors has reached the market, due to a lack of potency, stability and/or selectivity. Compared to small molecule inhibitors, VHHs which are the binding domain of heavy-chain only camelid antibodies, have key advantages including high stability, high specificity, no off-target effects, better pharmacokinetics properties and more importantly ability to target cryptic epitope such as enzyme active site via their long fingertip CDR3. The goal of this project is to generate long-acting polymer-VHH conjugates able inhibit LasB proteolytic activities. After the detailed in vitro characterization of the VHH, its inhibitory effects will be tested (i) on the formation of biofilm by P. aeruginosa (ii) in clinical samples from CF patients (iii) and in Galleria mellonella larvae infection model.

This competitive 24-month interdisciplinary project is funded by the Fonds National de la Recherche Scientifique (www.fnrs.be). It will be mainly carried out in the lab of M. Dumoulin at the Centre for Protein Engineering (CIP) which is a multidisciplinary Center of about 50 researchers active in Life Sciences (ULiège, Belgium, hiips://www.cip.uliege.be). The research carried out at CIP covers many areas ranging from molecular to structural biology. Thus, it has a solid experience in the fields of: genetic engineering, bacteriology, protein production and purification, protein characterization, enzymology, protein folding, rapid mixing techniques, fluorescence microscopy and protein crystallography. The lab of M. Dumoulin has been working on VHHs for many years. We select VHHs by phage display and deeply characterize their in vitro properties including affinity, specificity, structure (X-ray crystallography or HDX-MS) and biological activities (e.g., inhibition of enzyme activity, inhibition of fibril formation, etc). We collaborate with various groups to investigate the therapeutic potential of VHHs in vivo. The diseases we are interested in comprise amyloid diseases, infectious diseases and antibiotic resistance, mesothelioma, multiple myeloma and lungs diseases (hiips://www.cip.uliege.be/cms/c_6288623/en/neptuns-nanoantibodies-to-explore-protein-structure-and-functions).

Parts of the project will be performed in collaboration with Prof. Françoise Van Bambeke and Prof Rita Vanbever (UCLouvain) and Prof. Damien Thiry (ULiège). The three groups, which know each other very well, are highly multidisciplinary and opened to collaborations. The 12 month fellowship will start January 8th, the latest and will be renewable once for a total of 24 months.

Your role:
You will select inhibitory VHHs (Nanomedecines) from large immune libraries by phage display using a protocol we have recently developed to specifically select inhibitory VHHs against proteases. You will produce and purify the selected VHHs in large quantities to deeply characterize them in vitro (affinity, specificity, mechanism of inhibition, 3D structure, stability and effect on P. Aeruginosa cultures, etc). Then, the best VHH will be PEGylated to increase its in vivo stability (collaboration with R. Vanberver) and its inhibitory effects will be tested (i) on the formation of biofilm by P. aeruginosa (collaboration with Françoise Van Bambeke), (ii) in clinical samples from CF patients (iii) and in Galleria mellonella larvae infection model (collaboration with Damien Thiry).

Duration: 24months Salary based on candidate’s experience

Start: January 8th 2024

Required knowledge and skills:
– You have a PhD degree in Biochemistry, Molecular and Cellular Biology, Pharmaceutical Biotechnology or Biotechnology.
– You have a strong interest and laboratory experience in molecular biology and construct design, protein engineering, protein expression, purification and characterization, and protein-protein interactions.
– You are highly motivated.
– You have an ability to work independently and as part of a multidisciplinary team.
– You have an excellent organization, communication and team skills


Procedure and contact:
Email to mdumoulin@uliege.be a CV, motivation letter and contact information for three references in A SINGLE file before November 30th 2023.