Search bioRxiv⌕ Search

Biology subjects

Nicolai, A.

Publications and source records attributed to Nicolai, A..

2 recordsLinked to original sources

Examining the Atypical Structure Adopted by the 37-51 Segment of α-synuclein in the Early Stage of Aggregation

The aggregation of -synuclein into insoluble filaments is a hallmark of Parkinsons disease. Upon membrane binding, -synuclein adopts a helical conformation, whereas during neurodegeneration it misfolds and aggregates into {beta}-sheet-rich structures. However, the factors that trigger the misfolding and aggregation remain poorly understood. In this study, coarse-grained molecular dynamics simulations are used to characterize the structural ensembles of -synuclein monomers and dimers at atomic resolution during the early stage of aggregation, covering an effective timescale of 29.7 ms. The simulations reveal that a short sequence spanning residues 37-51 predominantly adopts three distinct conformations: {beta}-hairpin, braid, and random coil. Here, focus is placed on a braid conformation, which, to the best of our knowledge, has not been reported previously in either experimental or computational studies. Although we could not definitively rule out the possibility that the coarse-grained force field overestimates the stability of the braid, two all-atom simulations demonstrated that this structure is sterically feasible and metastable. Furthermore, in monomers, the braid appears to hinder the formation of long-range intramolecular interactions, leaving the N-terminus, NAC (non-amyloid-{beta} component) region, and C-terminus solvent-exposed and potentially facilitating fibril nucleation. In dimers, however, the braid appears to impede intermolecular contacts, especially homotypic intermolecular interactions, thereby slowing aggregation. Upon unfolding of the braid structure, intermolecular contacts, including homotypic interactions, increase noticeably.

biophysics↗

Piezo1 activation increases the release of therapeutic extracellular vesicles after mechanical stimulation in bioreactors

Enhancing protocols and methods for producing therapeutic extracellular vesicles (EVs) in bioreactors is crucial to achieve scalable production while ensuring both quality and quantity. Studies have shown that mechanical stress can promote EV release, although the underlying mechanisms remain largely unclear. Here we investigated which mechanisms are responsible for the increase of EV production under shear stress. EVs were produced from adipose tissue-derived stromal cells (ASCs), also described as mesenchymal stromal cells (MSCs) that can support the regeneration of injured tissues via several paracrine factors. The cultures were treated with GsMTx4, an inhibitor which blocks the Piezo1 ion channels, or with YODA1, its agonist, to assess if mechanical or shear forces pathways are involved in enhancing the EV release. EVs were quantified and characterized after high shear (HS) stimulation compared with no shear stress stimulation (3D) and standard cultures methods (2D). These experiments showed that, after mechanical stimulation by shear stress, EV production increased in bioreactors and this effect was blocked by the inhibition of Piezo1 ion channels with GsMTx4 (88%) with no impact on cell viability. Consistently, the agonist YODA1 increased the EV production (149%). The implications of these findings are significant, especially for regenerative medicine and cellular therapies, where the efficient production of high-quality EVs is crucial. By understanding turbulence-induced shear stress and the natural mechanotransductive pathways within cells, it may be possible to optimize the production of therapeutic EVs, paving the way for more effective treatments in the future.

cell biology↗