PhD position funded by Horizon Europe at the Univ. of Liège, Centre of Protein Engineering - MMP-9 Inhibitory Nano-antibodies for the Treatment of Impaired Wound Healing

This PhD position is part of the European MSCA REMOD-HEALING project whiwh aims at Targeting Extracellular Matrix Remodelling in Cutaneous Wounds to Promote Better Healing.

REMOD-HEALING is a Doctoral Network funded by the Horizon Europe Programme of the European Union. The REMOD-HEALING Consortium has funding for 12 full-time (100%) doctoral scholarships (locations across Europe and Costa Rica) within the context of the recently approved Marie Sklodowska-Curie Action Doctoral Network (MSCA-DN).

REMOD-HEALING is a research network of leading European and Latin American scientists from academia and industry, who want to tackle the burden of pathological wounds by exploiting therapeutic targets and biomarkers associated with extracellular matrix remodelling in cutaneous wounds. Through its research and training activities, the REMOD-HEALING project will contribute to scientific advancement and innovation in wound healing and regenerative medicine through a coordinated, interdisciplinary effort, ultimately leading to societal and economic benefits (more information coming soon on REMOD-HEALING website: remod-healing.eu).

Participating in REMOD-HEALING offers doctoral candidates many unique opportunities, including:

• A 36 month project as Marie Sklodowska Curie trainee in one of the participating institutions with the objective of receiving a doctoral degree (PhD).

• State-of-the-art, exciting research in an international consortium with highly integrated research projects.

• Expert training in basic and applied research, along with a thorough understanding of the process involved in transitioning from basic to pre-clinical research.

• Research training periods in another consortium member's lab lasting from a few weeks up to three months, performed in a different EU country than the country where most of the project will take place.

• Training in both academic and industrial research environments.

• Salary according to EU guidelines for Marie Sklodowska Curie trainees, including mobility payments and family allowances where applicable.

Several summer/winter schools are also planned to strengthen doctoral students' knowledge in scientific fields, as well as in innovation and transferable skills (scientific communication, intellectual property, writing grant applications, etc.).

The thesis project described here will mainly take place at NEPTUNS at the Centre for Protein Engineering (CIP) at the University of Liège (Belgium).

Title of the project: MMP-9 Inhibitory Nano-antibodies for the Treatment of Impaired Wound Healing

Introduction

The skin regulates body homeostasis and serves as a primary defence against physical, chemical and biological assaults. Any breach in this primary defence must be rapidly repaired to avoid haemorrhage, infection or water loss. This is achieved through wound healing, the physiological process leading to the coordinated restoration of the skin barrier integrity after injury. However, a large number of factors (wound size and location, exogenous pathogenic agents, aging, co-morbidities etc) can affect wound healing and prevent its normal resolution, leading to wounds that never heal (i.e. chronic wounds) or that heal in an aberrant manner (e.g. with scarring or dermonecrosis). These defective wound healing conditions represent a major health problem, affecting up to 6% of the adult population in Europe. Elevated levels of the metalloproteinase MMP9, due to excessive activation of neutrophils, contribute to chronic wounds. MMP9 constitutes therefore a therapeutic target.

Nano-antibodies (also referred to as Nanobodies® or VHHs) are single-domain antibody fragments derived from heavy-chain only antibodies produced by camelids. Despite their small size, the affinity of Nano-antibodies for their target is comparable to that of classical antibodies. Because of their small size, Nano-antibodies display a series of remarkable properties including high stability and solubility, easy production in E. coli, easy modification by genetic engineering, high tissue penetration and an ability to target, via a long CDR3 (Complementarity Determining Region 3), cryptic epitopes generally inaccessible to conventional antibodies, such as enzymes active site. The objective of this project is to generate Nano-antibodies specifically inhibiting MMP9 and assess their efficiency in a mouse model of delayed healing.

Objectives

The goals of this project are (i) to generate anti-MMP9 inhibitory Nano-antibodies; (ii) to deeply characterize their in vitro properties  (affinity, specificity, inhibition constant and mechanism of inhibition, stability, binding epitopes, structure of the VHH-MMP9 complexes),(iii) to demonstrate their therapeutic potential in pathological wounds from human or mouse origins and in a mouse model of delayed healing and (iv) develop an ELISA assay to quantify MMP9 in biological samples.

Expected Results

Selection of a highly specific anti-MMP9 inhibitory Nano-antibody demonstrating therapeutic efficiency.

The thesis project described here will mainly take place at NEPTUNS at the Centre for Protein Engineering (CIP) at the University of Liège (Belgium).

CIP is a highly interdisciplinary Centre providing complementary approaches for the analysis of structure and function relationships of proteins. Ten closely interacting research teams constitute the framework of the Centre, offering cutting-edge expertise from isolated proteins to integrated cellular networks. 

The NEPTUNS (Nano-antibodies to Explore Protein Structure and Functions) lab has an expertise in the generation,selection and characterisation of Nano-antibodies against a wide range of antigens; it has in particular developed original protocols to select Nano-antibodies inhibiting enzymes including proteases (Cawez et al., 2023, doi: 10.1128/aac.01499-22; Redeghieri et al., 2024, doi: 10.1002/pro.5227).

The CIP has developed 3 technological platforms: Protein Factory to express and purify recombinant proteins (www.proteinfactory.uliege.be), Robotein® for the high throughput production and characterization of proteins (www.robotein.uliege.be) and AlpaNano for the generation and selection of Nano-antibodies  (www.alpanano.uliege.be). The equipment and expertise of the three platforms will be used for the realisation of the project.  

In addition, the doctoral candidate will experience a 3 month training at the University of Freiburg (Germany) in the lab of Dr. Alexander Nyström to test the therapeutic efficiency of the anti-MMP9 Nano-antibodies in a mouse model of delayed healing and a 3 weeks training  at Nordic Bioscience( Herlev, Denmark) in the team of Dr. Signe Holm Nielsen to establish a Nano-antibodies based ELISA assay to detect active MMP9.

The candidate will obtain the PhD degree in Life Sciences from the University of Liège (Doctoral school of Biochimie, Biologie moléculaire et cellulaire, Bioinformatique et modélisation). The daily activities of the PhD candidate consist mainly in research but it is also expected that the candidate participates to the REMOD-HEALING training activities. The candidate has the possibility to develop collaborations with other members of the REMOD-HEALING consortium and to visit their laboratory in order to broaden his fields of expertise.

Candidates must have a Master in Biochemistry, Pharmaceutical biotechnology or Biotechnology. 

They have a strong interest in molecular biology, protein engineering, protein expression, protein purification and protein characterization, and protein-protein interactions. Knowledge in phage display and experience with mice experiments are beneficial but not mandatory.

Candidates should be highly motivated and creative; they should have an ability to work independently and as part of a multidisciplinary team. Moreover, they should have an excellent organization, communication and team skill.

The candidate must have an excellent proficiency of English