Université Lyon 1, The Molecular Microbiology and Structural Biochemistry Lab. (MMSB)
Mono- and bi-specific nanobodies targeting the CUB domains of PCPE-1 reduce the proteolytic processing of fibrillar procollagens
Mol. Biol. 436(16):168667.doi: 10.1016/j.jmb.2024.168667
Lagoutte P, Bourhis JM, Mariano N, Gueguen-Chaignon V, Vandroux D, Moali C, Vadon-Le Goff S.
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Priscillia Lagoutte (31 years old) obtained a Bachelor of Science in Biochemistry and Molecular biology at the "Université de Bourgogne Dijon" and a Master of Science in Biotechnology and Biochemical Engineering at the "Université Claude Bernard Lyon 1". She continued with a PhD in Biochemistry and Molecular biology at the Technologic Institute of Microbiology Bioaster under the supervision of Dr. Bettina Werle (PhD defense, 2018). During her PhD, she developed a method for in vitro generation of binders (Affibody, vHH) and generated binders able to distinguish between β-lactamases involved in antibiotic resistance. In 2018, she began a post-doc in Lyon at the Tissue Biology and Therapeutic Engineering Laboratory (LBTI) in Dr Catherine Moali’s team. Her project was to elucidate and characterize nanobodies (vHH) against the procollagen C-proteinase enhancer (PCPE-1), a protein involved in the maturation of fibrillar collagens. Her work, patented and published in JMB, showed the ability to reduce collagen maturation by targeting PCPE-1 with vHHs and highlighted the potential of vHHs selected against PCPE-1 as molecule to detect or treat fibrosis. She also worked on another project focused on solving the structure and understanding the C-terminal maturation complex of fibrillar collagen through cryo-EM and protein engineering approaches, in collaboration with Oxford Particle Imaging Centre (OPIC, STRUBI).
Since 2022, Priscillia Lagoutte has been an Assistant Professor at the "Université Claude Bernard Lyon 1", working on type IV secretion system of Helicobacter pylori in the Molecular Microbiology and Structural Biochemistry Laboratory (MMSB) in Dr Laurent Terradot’s team.
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Résumé de l'article
The excessive deposition of fibrillar collagens is a hallmark of fibrosis. Collagen fibril formation requires proteolytic maturations by Procollagen N- and C-proteinases (PNPs and PCPs) to remove the N- and C-propeptides which maintain procollagens in the soluble form. Procollagen C-Proteinase Enhancer-1 (PCPE-1, a glycoprotein composed of two CUB domains and one NTR domain) is a regulatory protein that activates the C-terminal processing of procollagens by the main PCPs. It is often up-regulated in fibrotic diseases and represents a promising target for the development of novel anti-fibrotic strategies.
Here, our objective was to develop the first antagonists of PCPE-1, based on the nanobody scaffold. Using both an in vivoselection through the immunization of a llama and an in vitro selection with a synthetic library, we generated 18 nanobodies directed against the CUB domains of PCPE-1, which carry its enhancing activity. Among them, I5 from the immune library and H4 from the synthetic library have a high affinity for PCPE-1 and inhibit its interaction with procollagens. The crystal structure of the complex formed by PCPE-1, H4 and I5 showed that they have distinct epitopes and enabled the design of a biparatopic fusion, the diabody diab-D1. Diab-D1 has a sub-nanomolar affinity for PCPE-1 and is a potent antagonist of its activity, preventing the stimulation of procollagen cleavage in vitro. Moreover, Diab-D1 is also effective in reducing the proteolytic maturation of procollagen I in cultures of human dermal fibroblasts and hence holds great promise as a tool to modulate collagen deposition in fibrotic conditions.