Description of the thesis topic

BEFORE APPLYING, PLEASE CHECK THAT YOU MEET THE MOBILITY REQUIREMENTS LISTED IN THE 'CONSTRAINTS AND RISKS' SECTION.

Titre: Elucidation of the molecular basis of BMP1 inhibition by its endogenous inhibitor and development of an anti-fibrotic molecule

Introduction
BMP1 (Bone Morphogenetic Protein 1) is an extracellular metalloprotease playing critical roles in extracellular matrix assembly and growth factor activation (1). It is involved in several physiological processes such as development, growth and skin wound healing. The dysregulation of its activity can however lead to pathological conditions, including wound healing disorders (e.g. chronic wounds and scarring). The lab of C. Moali is interested in understanding the mechanisms of regulation of BMP1 activity, at the molecular, cellular and tissue levels, with the goal to develop innovative therapeutic approaches to treat wound healing disorders.

Background
Previous work has shown that BMP1 regulation relies on two main proteins, PCPE1 and PCPE2, with both overlapping and divergent functions. One major difference between the PCPE proteins is that only PCPE2 can inhibit BMP1 proteolytic activity (2, 3). This finding was unexpected because PCPE1 and PCPE2 are very similar in terms of sequences and structures.

Objectives
The main objectives of the PhD project will be (i) to elucidate the molecular basis of BMP1 inhibition by PCPE2 and (ii) to use this knowledge to design an anti-fibrotic molecule capable of blocking collagen proteolytic maturation by BMP1.

Methodology
We will use systematic sequence swapping between PCPE1 and PCPE2 to identify the important residues for PCPE2 inhibitory function (molecular biology, activity assays, surface plasmon resonance). In parallel, we will try to solve the structure of the BMP1/PCPE2 complex by cryo-electron microscopy with the help of previously developed Nanobodies® to stabilize flexible regions. Finally, we will engineer an optimized inhibitor of BMP1 with high potency, stability and specificity and test its therapeutic potential in a wound-on-a-chip model of skin scarring.

Expected Results
This project will lead to the elucidation of a new mechanism of protease inhibition and to the development of a specific and potent inhibitor of BMP1 with anti-fibrotic properties.

Expected skills
- Master degree in biochemistry and structural biology
- Practical experience in at least one of these techniques: molecular biology (PCR, cloning), mammalian cell culture, protein purification, methods to analyze protein-protein interactions (ELISA, SPR, BLI, etc), crystallography, cryo-electron microscopy
- Basic knowledge in extracellular matrix, skin biology and proteolysis
- Excellent command of English (written and spoken)
- Writing skills
- Ability to work in a team and in a multicultural context
- Highly autonomous, organized, rigorous and responsive

Work Context

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 LBTI (Lyon, France), a laboratory with 120 staff members specializing in cutting-edge research in tissue biology and in the development of new therapeutic strategies in the field of tissue repair. In addition, the doctoral candidate will experience a 3-week placement at the University of Manchester (UK) in the lab of Clair Baldock for cryo-electron microscopy training and a 3-month placement at Mimetas (Leiden, The Netherlands) in the lab of Dorota Durek to test an engineered protease inhibitor in an in vitro model of skin scarring.

More information on the scientific project is available here:https://sdrive.cnrs.fr/s/wHipMCHC77pgrsc

The position is located in a sector under the protection of scientific and technical potential (PPST), and therefore requires, in accordance with the regulations, that your arrival is authorized by the competent authority of the MESR.

Constraints and risks

Applicants should not have resided or carried out their main activity (work, studies, etc.) in the country of the host institution for more than 12 months in the 36 months prior to their recruitment date.

The selected candidate will work in BSL2 laboratories and with laboratory chemicals. Individual protection will be provided by the host lab and during secondments.

Additional Information

In addition to the CV and cover letter, the application must include (in a single PDF file attached to the CV):

• A summary of the applicant's Master thesis (max 1 page);

• A transcript along with a scanned copy of your Master's Degree certificate. If the degree has not been awarded by the application closing date, the candidate should submit a declaration signed by their supervisor or University official stating that the degree will be obtained by the time of PhD enrolment.

• Two letters of recommendation from relevant and appropriate referees or the names and contact details of two Referees (as former Supervisors/PIs).