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Silvani, G.

Publications and source records attributed to Silvani, G..

3 recordsLinked to original sources

Engineered 3D hydrogel model reveals divergence of adhesion-migration balance in Glioblastoma under simulated microgravity

Glioblastoma is an aggressive brain cancer whose cells can switch between different modes of invasion in response to their surroundings, making the disease difficult to predict and treat. How physical forces influence this adaptability remains poorly understood. Here, we used simulated microgravity together with engineered hydrogels that independently control adhesion, degradability, and mechanical properties to test how gravity affects glioblastoma invasion. Microgravity strongly reduced invasion and shifted cells from elongated, protrusive behavior to a more cohesive state. Proteomic analysis showed reduced invasive signaling together with increased cell-matrix and cell-cell adhesion, consistent with a redistribution of contractile forces toward the cell edge. Under normal gravity, blocking CD44, integrin {beta}1, or N-cadherin reduced matrix-dependent invasion. In contrast, under microgravity, inhibiting these same adhesion pathways restored invasion, indicating that microgravity traps cells in an overly adhesive, cohesive state that limits movement rather than motility itself. These findings show that gravity is an important regulator of cancer cell plasticity and reveal a mechanically induced vulnerability in glioblastoma invasion. More broadly, combining defined biomaterials with gravitational modulation provides a new way to study how physical forces shape tumor behavior.

cancer biology↗

Capillary constrictions prime cancer cell tumorigenicity through PIEZO1

Metastasis is a hallmark of cancer and is responsible for the majority of cancer-related deaths. Evidence suggests that even a single cancer cell can spread and seed a secondary tumour. However, not all circulating tumour cells have this ability, which implies that dissemination and distal growth require adaptative mechanisms during circulation. Here we report that constriction during microcapillary transit will trigger reprogramming of melanoma cells to a tumorigenic cancer stem cell-like state. Using a microfluidic device mimicking physiological flow rates and gradual capillary narrowing, we showed that compression through narrow channels lead to cell and nuclear deformation, rapid changes in chromatin state and increased calcium handling through the mechanosensor PIEZO1. Within minutes of microcapillary transit, cells show increased regulation of transcripts associated with metabolic reprogramming and metastatic processes, which culminates in the adoption of cancer stem cell-like properties. Squeezed cells displayed elevated melanoma stem cell markers, increased propensity for trans-endothelium invasion, and characteristics of enhanced tumorigenicity in vitro and in vivo. Pharmacological disruption of channel activity inhibited the stem cell-like state, while the selective PIEZO1 activator Yoda1 primed this state irrespective of constriction. Finally, deletion of PIEZO1 led to complete abrogation of the constriction-induced stem cell-like state. Together, this work demonstrates that compressive forces during circulation can reprogram circulating cancer cells to tumorigenic stem cell-like states that are primed for extravasation and metastatic colonization.

cancer biology↗

An acoustic platform for single-cell, high-throughput measurements of the viscoelastic properties of cells

Cellular processes including adhesion, migration and differentiation are governed by the distinct mechanical properties of each cell. Importantly, the mechanical properties of individual cells can vary depending on local physical and biochemical cues in a time-dependent manner resulting in significant inter-cell heterogeneity. While several different methods have been developed to interrogate the mechanical properties of single cells, throughput to capture this heterogeneity remains an issue. While new high-throughput techniques are slowly emerging, they are primarily aimed at characterizing cells in suspension, whereas high-throughput measurements of adherent cells have proven to be more challenging. Here, we demonstrate single-cell, high-throughput characterization of adherent cells using acoustic force spectroscopy. We demonstrate that cells undergo marked changes in viscoelasticity as a function of temperature, the measurements of which are facilitated by a closed microfluidic culturing environment that can rapidly change temperature between 21 {degrees}C and 37 {degrees}C. In addition, we show quantitative differences in cells exposed to different pharmacological treatments specifically targeting the membrane-cytoskeleton interface. Further, we utilize the high-throughput format of the AFS to rapidly probe, in excess of 1000 cells, three different cell-lines expressing different levels of a mechanosensitive protein, Piezo1, demonstrating the ability to differentiate between cells based on protein expression levels.

bioengineering↗