Biology subjects

Manneville, C.

Publications and source records attributed to Manneville, C..

3 recordsLinked to original sources

bioRxiv2026

GPNMB enables lysosomal adaptation required for myelin clearance and nerve regeneration after peripheral nerve injury

Recovery from peripheral nerve injury involves coordinated axonal and immune repair programs that promote tissue regeneration. However, the mechanisms regulating immune-mediated repair remain incompletely understood. Here, we demonstrate that glycoprotein non-metastatic melanoma protein B (GPNMB) is a key regulator of repair processes following peripheral nerve injury. In a mouse sciatic nerve crush model, loss of Gpnmb impaired functional recovery after injury. MRI and histological analyses revealed persistent myelin debris accumulation, defective debris clearance, and impaired macrophage lysosomal function within injured nerves. Mechanistic studies showed that GPNMB-deficient human iPSC-derived macrophages exhibited impaired lysosomal adaptation, reduced lysosomal fitness, and defective processing of myelin-associated cargo, resulting in inefficient degradation of injury-associated material. These defects were accompanied by impaired acquisition of repair-associated cellular programs. Together, our findings identify GPNMB as a critical regulator of lysosome-dependent repair programs during nerve regeneration and demonstrate that efficient GPNMB-dependent myelin processing is required for functional recovery following peripheral nerve injury.

neuroscience↗

bioRxiv2024

Development of a p62 biodegrader for autophagy targeted degradation

Harnessing autophagy for targeted degradation is a promising extension to proteasome-based targeted protein degradation because of the capacity and versatility of lysosomes to degrade large and complex cargo, thus broadening the scope of therapeutic targets. While small-molecule degraders recruiting the autophagy machinery to targets are starting to emerge, it remains unclear which component of the autophagy lysosomal pathway is most efficacious to induce selective target degradation. Here, we describe two orthogonal induced-proximity strategies to identify and prioritize autophagy effectors that are sufficient to degrade organelles as well as soluble targets. We show that induced proximity of different effectors such as autophagy cargo receptors, ATG8-like proteins or the kinases ULK1 and TBK1 are sufficient to trigger mitophagy. In contrast, self-oligomerizing autophagy cargo receptors outperform ATG8- like effectors and autophagy-related kinases in clearing a soluble cytosolic protein. We further explore the importance of avidity for targeted degradation via autophagy and reveal that the PB1 domain of p62 fused to a LIR peptide is a minimal degron to induce the degradation of mitochondria as well as cytosolic proteins. By developing a novel and highly selective intrabody against the autophagy cargo receptor p62 into a heterobifunctional degrader, we demonstrate that recruitment of endogenous p62 is sufficient to clear mitochondria. This biodegrader, however, is unable to induce degradation of soluble cytosolic proteins due to its inhibitory effect on p62 self-oligomerization. Our study highlights the importance of avidity and suggests that autophagy cargo receptors are attractive entry points for the development of heterobifunctional degraders for complex targets such as organelles or protein aggregates.

synthetic biology↗

bioRxiv2019

A simple and efficient CRISPR technique for protein tagging

Genetic knock-in using homology directed repair is an inefficient process, requiring selection of few modified cells and hindering its application to primary cells. Here we describe Homology independent gene Tagging (HiTag), a method to tag a protein of interest by CRISPR in up to 66% transfected cells with one single electroporation. The technique has proven effective in various cell types, can be used to knock in a fluorescent protein for live cell imaging, to modify the cellular location of a target protein and to monitor levels of a protein of interest by a luciferase assay in primary cells.

cell biology↗