Biology subjects

van Es, R. M.

Publications and source records attributed to van Es, R. M..

3 recordsLinked to original sources

bioRxiv2025

Bioengineering Developmentally Inspired Matrix Vesicles as Designer Nanotherapeutics for Bone Regeneration

Extracellular vesicles (EVs) are emerging as promising acellular nanotherapeutics for musculoskeletal repair. Matrix vesicles, a matrix-bound subset of EVs, are essential mediators of endochondral ossification in bone development and fracture repair. This study aims to design bioengineered matrix vesicles from hypertrophic cartilage microtissues to drive endochondral ossification for bone repair. Human mesenchymal stromal cell (hBMSC) microtissues were differentiated with/without BMP2 in chondrogenic or hypertrophic medium. Isolated matrix vesicles were characterized for physiochemical properties and biological functionality. BMP2 and hypertrophic conditioning significantly increased vesicle yield (1.5-fold), alkaline phosphatase activity (3.24-fold), calcium binding capacity (8.82-fold), and growth factor content (BMP2, VEGF). These vesicles promoted proliferation, migration, and mineralization of hBMSCs and enhanced angiogenesis in human endothelial colony forming cells (hECFCs), with BMP2 and hypertrophically conditioned vesicles showing the most pronounced effects. Proteomics analysis confirmed the enrichment of proteins involved in extracellular matrix remodelling, mineral deposition and vascularization within these hypertrophically engineered vesicles. These findings demonstrate that hypertrophic induction of cartilaginous microtissues substantially improves the yield and therapeutic potential of matrix vesicles. Taken together, this research unveils a powerful strategy to bioengineer developmentally inspired vesicles that not only recapitulate key cues of endochondral ossification but offers a tailorable, multifunctional nanotherapeutic platform for improved bone regeneration strategies. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=112 SRC="FIGDIR/small/684111v1_ufig1.gif" ALT="Figure 1"> View larger version (37K): org.highwire.dtl.DTLVardef@17327a9org.highwire.dtl.DTLVardef@1310583org.highwire.dtl.DTLVardef@160f095org.highwire.dtl.DTLVardef@152eff0_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioengineering↗

bioRxiv2025

Phosphoproteomics Maps Calcineurin-NFAT-DSCR1.4 Signaling as Druggable Axis in Gαq-R183Q-Driven Capillary Malformations

Vascular malformations are congenital lesions caused by somatic and germline mutations that disrupt developmental signaling pathways. Capillary malformations (CMs) typically present as port-wine stains in the skin and can also affect ocular and cerebral tissues in Sturge Weber Syndrome (SWS), leading to aesthetic, ophthalmic, and neurological complications. CMs are caused by a somatic mutation in the GNAQ gene in endothelial cells, leading to a p.R183Q substitution in the Gq protein. The underlying mechanisms of Gq-R183Q-driven CMs formation remain unclear. To address this, we generated CRISPR/Cas9-engineered human dermal microvascular endothelial cells lacking endogenous Gq, whilst expressing the Gq-R183Q mutant instead. The Gq-R183Q mutation strongly impaired endothelial cell migration and angiogenic sprouting capacity compared to wild-type controls. Next, using SILAC-based quantitative proteomics, we investigated the Gq-R183Q-induced changes in the endothelial phosphoproteome. These analyses revealed prominent activation of the calcineurin-NFAT signaling pathway in Gq-R183Q-expressing endothelial cells, leading to dephosphorylation of NFAT1 and NFAT2 and the selective expression of their transcriptional target DSCR1.4. Immunofluorescence of patient-derived skin biopsies confirmed deregulation of NFAT1/2 and the expression of DSRC1 in endothelial cells, validating their potential importance in CMs. We further demonstrate that pharmacological inhibition of calcineurin with tacrolimus (FK506) could partially restore NFAT signaling in Gq-R183Q endothelial cells. Intriguingly, the genetic depletion of the NFAT target DSCR1 in Gq-R183Q cells fully restored calcineurin/NFAT signaling to normal levels, enabling proper endothelial migration and sprouting. In summary, we uncovered a calcineurin-NFAT-DSCR1.4 signal transduction axis that is driven by Gq-R183Q and established its importance for endothelial angiogenic properties. These findings highlight the calcineurin/NFAT signaling axis as a promising therapeutic target to restore endothelial function in CMs.

cell biology↗

bioRxiv2025

E-cadherin mechanotransduction activates EGFR-ERK signaling in epithelial cells by inducing ADAM-mediated ligand shedding

The behavior of cells is governed by signals originating from their local environment, including mechanical forces that cells experience. Forces are transduced by mechanosensitive proteins, which can impinge on signaling cascades that are also activated by growth factor receptors upon ligand binding. We investigated the crosstalk between these mechanical and biochemical signals in the regulation of intracellular signaling networks in epithelial monolayers. Phosphoproteomic and transcriptomic analyses on epithelial monolayers subjected to mechanical strain revealed ERK signaling as a predominant strain-activated hub, initiated at the level of the upstream epidermal growth factor receptor (EGFR). Strain-induced EGFR-ERK signaling depends on mechanosensitive E-cadherin adhesions. Proximity labeling identified the metalloproteinase ADAM17, an enzyme that mediates shedding of soluble EGFR ligands, to be closely associated with E-cadherin. We developed a novel probe for monitoring ADAM-mediated shedding, which demonstrated that mechanical strain induced ADAM activation. Mechanically-induced ADAM activation was essential for mechanosensitive signaling from E-cadherin adhesions towards EGFR-ERK. Collectively, our data demonstrate that mechanical strain transduced by E-cadherin adhesion triggers the shedding of EGFR ligands that stimulate downstream downstream ERK activity. Our findings illustrate how mechanical signals and biochemical ligands can operate within a single, linear signaling cascade. Significance statementCells integrate different types of information that they receive from their local environment to regulate their behavior. This includes biochemical signals, such as growth factors binding to their dedicated receptor. Similarly, cells respond to mechanical forces that they are subjected to. Although biochemical and mechanical signals can elicit similar signaling responses in cells, the interplay between these types of signals is not well understood. Here we unveil that mechanical strain of epithelia modulates the activity of the EGFR-ERK signaling pathway by controlling the availability of growth factors that bind and activate EGFR. This finding demonstrates that biochemical and mechanical signals do not act in a segregated fashion, but rather can function in a linear cascade, shedding light on fundamental principles governing cellular regulation.

cell biology↗