Biology subjects

van Batenburg-Sherwood, J.

Publications and source records attributed to van Batenburg-Sherwood, J..

6 recordsLinked to original sources

bioRxiv2026

Engineered microvessel networks -- Longitudinal transcriptomic profiling validates chip-grown tissues as a robust model for studying vasculature

Vascularised Organ-on-Chip (OoC) models are promising New Approach Methodologies (NAMs), but their translation requires validation for defined Contexts of Use (CoU). Here, we combine microfluidic microvessel networks (MVNs) with longitudinal bulk and single-cell RNA sequencing to determine how cell provenance influences engineered vascular phenotype. Stromal cell identity produced distinct network architectures and transcriptional trajectories, demonstrating that support cells, commonly treated as interchangeable, represent an important CoU variable. Primary human pancreatic microvascular endothelial cells failed to form stable perfusable networks, but supplementation with 10% Human Umbilical Vein Endothelial Cells (HUVECs) or Blood Outgrowth Endothelial Cells (BOECs) rescued self-assembly. Single-cell transcriptomics identified specialised endothelial populations associated with vascular morphogenesis, including tip, stalk, phalanx, endothelial to mesenchymal transition (EndMT) and pericyte-like endothelial cells. Failed networks lacked coherent endothelial specialisation, whereas rescued networks restored stabilising cell states and revealed FLRT2--UNC5B signalling within pericyte-like HUVECs, recapitulating a pathway implicated in angiogenic development in vivo. These findings establish cell provenance as a critical determinant of microvascular NAM performance and demonstrate how transcriptomic validation can define and strengthen the biological relevance of microvessel-on-chip models for specific CoUs.

bioengineering↗

bioRxiv2026

Fluid shear magnitude, mechanical history and actin-sensitive coupling shape Piezo1 signalling

Mechanical forces shape cell behaviour, yet how Piezo1-associated responses encode force magnitude, recent mechanical history and cytoskeletal context under repeated fluid shear remains unclear. We combined defined fluid shear stress (FSS) with GenEPi, a genetically encoded reporter of Piezo1-proximal Ca2+ entry, in engineered XLGenEPi-HEK293T monolayers to resolve mechanically evoked responses at single-cell and population scales. GenEPi responses increased with FSS magnitude but showed diminishing response gain at higher shear. Repeated submaximal FSS produced history-dependent attenuation and reduced temporal coordination, with higher FSS accelerating the early phase of attenuation. Low-dose Latrunculin A, without significantly reducing bulk F-actin abundance, converted sustained FSS-evoked signalling into a brief, decay-dominated response while largely preserving peak responses to Yoda1. These findings show that Piezo1-associated signalling is shaped over minutes by force magnitude, recent mechanical history and actin-sensitive mechanical coupling, providing a quantitative framework for understanding mechanotransduction under sustained and repeated mechanical stimulation.

biophysics↗

bioRxiv2026

Organ on chip model of respiratory vascular interactions under COPD relevant oxidative stress

Oxidative stress-induced airway injury contributes to chronic obstructive pulmonary disease (COPD). Cardiovascular complications increase COPD morbidity and mortality, but mechanistic links between airway injury and vascular dysfunction remain unclear, largely due to limitations of in vitro models that fail to replicate the multicellular lung environment. We developed REVAS, a modular organ-on-chip platform to study human respiratory-vascular cell-cell interactions at baseline and under oxidative stress conditions. REVAS consists of two respiratory chips hosting airway epithelium and microvascular endothelium, and a vascular chip hosting pulmonary artery endothelial cells co-cultured with vascular support cells, including smooth muscle cells, pericytes and fibroblasts. We studied effects of vascular support and respiratory cells on vascular endothelial phenotype at baseline and under H2O2-induced epithelial oxidative stress using functional assays, proteomic and transcriptomic analyses. Multicellular environment enhanced vascular endothelial barrier function and promoted respiratory and vascular cell differentiation at baseline. Mural cells altered endothelial cell-matrix interactions, metabolism and cytoskeletal remodelling, while respiratory cells promoted endothelial aerobic respiration and quiescent phenotype. Epithelial oxidative stress triggered inflammatory gene expression across all respiratory and vascular cells alongside apoptotic, reparative and pro-angiogenic signalling in endothelial and mural cells, accompanied by increased release of COPD-relevant cytokines and chemokines, including IL-6, TNF-/{beta}, IL-8, CCL5, CXCL9, PDGF, TGF-{beta}. Comparative analyses with COPD endothelial datasets confirmed that REVAS recapitulates key features of disease-associated endothelial dysfunction. These findings demonstrate that airway epithelial injury drives downstream vascular responses linked to inflammation and vascular remodelling, establishing REVAS as a human-relevant platform for mechanistic and therapeutic evaluation of cell-cell interactions in COPD and related lung diseases. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=164 HEIGHT=200 SRC="FIGDIR/small/730087v1_ufig1.gif" ALT="Figure 1"> View larger version (59K): org.highwire.dtl.DTLVardef@e29fd8org.highwire.dtl.DTLVardef@6c3d22org.highwire.dtl.DTLVardef@21a53forg.highwire.dtl.DTLVardef@e7f432_HPS_FORMAT_FIGEXP M_FIG C_FIG REVAS: a microfluidic platform developed to model multicellular interactions between airway epithelium and pulmonary vasculature under basal and oxidative stress. COPD: Chronic Obstructive Pulmonary Disease; EMT: endothelial-to-mesenchymal transition; HsEpCs: human small airway epithelial cells; HPMVECs: human pulmonary microvascular endothelial cells; HPAECs: human pulmonary artery endothelial cells; HPASMCs: human pulmonary artery smooth mucle cells; HPFs: human pulmonary fibcroblasts; HPCs: human pericytes.

bioengineering↗

bioRxiv2025

Corticosteroids elevate intraocular pressure through suppression of TREK-1 signaling

Clinicians are often forced into the dilemma of whether to battle ocular inflammation or preserve vision imperiled by elevated intraocular pressure (IOP). Anti-inflammatory treatments utilizing glucocorticosteroid regimens may induce glaucoma by chronically elevating IOP via increased trabecular meshwork (TM) resistance to the flow of aqueous humor, but it is not known whether pressure transduction itself is impacted by steroids and how changes in TM mechanosignaling affect conventional outflow resistance and IOP. To address this, we investigated the role of TREK-1 (TWIK-related potassium channel-1), a mechanosensitive K+ channel, in regulation of outflow facility, transmembrane signaling and dexamethasone (DEX)-induced ocular hypertension (OHT). The expression of tandem-pore potassium channels in mouse TM cells was dominated by Trek-1 (Kcnk2) mRNA, with residual expression of Traak, Tresk2 and Twik3 and vanishingly low levels of Task1 and Trek2. DEX suppressed Trek1 transcription by [~]80% but did not affect expression of Trpv4 and Piezo1 genes. Chronic DEX administration depolarized the membrane potential of TM cells and elevated IOP in mice whereas the selective TREK-1 agonist ML-402 lowered IOP in rodent OHT models. ML-402 doubled the outflow facility in perfused mouse eyes at all applied pressures and hyperpolarized DEX-treated TM cells. These in vitro, ex vivo and in vivo results implicate TREK-1 channels in homeostatic regulation of TM mechanosignaling, conventional outflow regulation and IOP homeostasis. Suppression of TREK-1 signaling by corticosteroids underlies OHT and could contribute to steroid glaucoma but this can be obviated by pharmacological stimulation of the channel with cornea-permeant ML-402 eye drops.

physiology↗

bioRxiv2025

A Microfluidic Blood Vessel-On-Chip Model of Thrombosis

BackgroundThrombus formation is regulated by the interplay between endothelial cells (EC), platelets and coagulation factors. However, most in vitro assays used to study thrombosis and develop anti-thrombotic therapies do not include the endothelium. ObjectiveTo develop a thrombus-on-chip model that includes endothelium and whole blood and allows for manipulation of extracellular matrix (ECM), shear stress and the addition of multiple cell types. MethodsA cylindrical vessel was created in a collagen matrix using the needle-based fabrication technique in a microfluidic device. Human umbilical vein EC (HUVEC) or endothelial colony-forming cells (ECFC) were cultured in the channel, with continuous monodirectional media turnover. Immunostaining and permeability measurement validated confluence and integrity of the monolayer. To investigate thrombosis, whole blood from healthy donors was perfused through TNF--activated or untreated EC-lined vessels. Image analysis and time-lapse microscopy were used to quantify labelled platelet adhesion and fibrin deposition. ResultsTNF- treatment resulted in increased platelet adhesion and fibrin deposition compared to control vessels. TNF-induced endothelial activation was confirmed by upregulation of adhesion molecules ICAM-1, E-selectin and tissue factor (TF). Thrombus formation in TNF- treated vessels was inhibited by an anti-TF antibody. In ECFC vessels, platelet adhesion and fibrin deposition were comparable to HUVEC, supporting feasibility of patient-based studies. ConclusionsWe developed a perfused thrombus-on-chip model that combines key elements of thrombus formation including endothelium. The model is amenable to independent control of microenvironmental stimuli, crosstalk with tissue-specific cells, and the inclusion of patients own cells and blood for precision medicine studies. ESSENTIALS- The endothelium is a key contributor to thrombosis - Standard in vitro methods to study thrombosis do not include the endothelium - We have developed a thrombus-on-a-chip model of thromboinflammation to measure live platelet adhesion and fibrin deposition on activated endothelium - Using endothelial colony forming cells (ECFC), we show that the method is suitable for patient studies with autologous endothelium and blood

bioengineering↗

bioRxiv2023

Aging and intraocular pressure homeostasis in mice

Age and elevated intraocular pressure (IOP) are the two primary risk factors for glaucoma, an optic neuropathy that is the leading cause of irreversible blindness. In most people, IOP is tightly regulated over a lifetime by the conventional outflow tissues. However, the mechanistic contributions of age to conventional outflow dysregulation, elevated IOP and glaucoma are unknown. To address this gap in knowledge, we studied how age affects the morphology, biomechanical properties and function of conventional outflow tissues in C57BL/6 mice, which have an outflow system similar to humans. As reported in humans, we observed that IOP in mice was maintained within a tight range over their lifespan. Remarkably, despite a constellation of age-related changes to the conventional outflow tissues that would be expected to hinder aqueous drainage and impair homeostatic function (decreased cellularity, increased pigment accumulation, increased cellular senescence and increased stiffness), outflow facility, a measure of conventional outflow tissue fluid conductivity, was stable with age. We conclude that the murine conventional outflow system has significant functional reserve in healthy eyes. However, these age-related changes, when combined with other underlying factors, such as genetic susceptibility, are expected to increase risk for ocular hypertension and glaucoma.

physiology↗