Laminin-associated integrins mediate Diffuse Intrinsic Pontine Glioma infiltration and therapy response within a neural assembloid model.

Diffuse Intrinsic Pontine Glioma (DIPG) is a highly aggressive and fatal pediatric brain cancer. One pre-requisite for tumor cells to infiltrate is adhesion to extracellular matrix (ECM) components. However, it remains largely unknown which ECM proteins are critical in enabling DIPG adhesion and migration and which integrin receptors mediate these processes. Here, we identify laminin as a key ECM protein that supports robust DIPG cell adhesion and migration. To study DIPG infiltration, we developed a DIPG-neural assembloid model, which is composed of a DIPG spheroid fused to a human induced pluripotent stem cell-derived neural organoid. Using this assembloid model, we demonstrate that knockdown of laminin-associated integrins significantly impedes DIPG infiltration. Moreover, laminin-associated integrin knockdown improves DIPG response to radiation and HDAC inhibitor treatment within the DIPG-neural assembloids. These findings reveal the critical role of laminin-associated integrins in mediating DIPG progression and drug response. The results also provide evidence that disrupting integrin receptors may offer a novel therapeutic strategy to enhance DIPG treatment outcomes. Finally, these results establish DIPG-neural assembloid models as a powerful tool to study DIPG disease progression and enable drug discovery.

Dynamically Crosslinked Poly(ethylene-glycol) Hydrogels Reveal a Critical Role of Viscoelasticity in Modulating Glioblastoma Fates and Drug Responses in 3D.

Glioblastoma multiforme (GBM) is the most prevalent and aggressive brain tumor in adults. Hydrogels have been employed as 3D in vitro culture models to elucidate how matrix cues such as stiffness and degradation drive GBM progression and drug responses. Recently, viscoelasticity has been identified as an important niche cue in regulating stem cell differentiation and morphogenesis in 3D. Brain is a viscoelastic tissue, yet how viscoelasticity modulates GBM fate and drug response remains largely unknown. Using dynamic hydrazone crosslinking chemistry, a poly(ethylene-glycol)-based hydrogel system with brain-mimicking stiffness and tunable stress relaxation is reported to interrogate the role of viscoelasticity on GBM fates in 3D. The hydrogel design allows tuning stress relaxation without changing stiffness, biochemical ligand density, or diffusion. The results reveal that increasing stress relaxation promotes invasive GBM behavior, such as cell spreading, migration, and GBM stem-like cell marker expression. Furthermore, increasing stress relaxation enhances GBM proliferation and drug sensitivity. Stress-relaxation induced changes on GBM fates and drug response are found to be mediated through the cytoskeleton and transient receptor potential vanilloid-type 4. These results highlight the importance of incorporating viscoelasticity into 3D in vitro GBM models and provide novel insights into how viscoelasticity modulates GBM cell fates.

Interfacial energy driven distinctive pattern formation during the drying of blood droplets.

HYPOTHESIS: Dried blood droplet morphology may potentially serve as an alternative biomarker for several patho-physiological conditions. The deviant properties of the red blood cells and the abnormal composition of diseased samples are hypothesized to manifest through unique cell-cell and cell-substrate interactions leading to different morphological patterns. Identifying distinctive morphological trait from a large sample size and proposing confirmatory explanations are necessary to establish the signatory pattern as a potential biomarker to differentiate healthy and diseased samples. EXPERIMENTS: Comprehensive experimental investigation was undertaken to identify the signatory dried blood droplet patterns. The corresponding image based analysis was in turn used to differentiate the blood samples with a specific haematological disorder "Thalassaemia" from healthy ones. Relevant theoretical analysis explored the role of cell-surface and cell-cell interactions pertinent to the formation of the distinct dried patterns. FINDINGS: The differences observed in the dried blood patterns, specifically the radial crack lengths, were found to eventuate from the differences in the overall interaction energies of the system. A first-generation theoretical analysis, with the mean field approximation, also confirmed similar outcome and justified the role of the different physico-chemical properties of red blood cells in diseased samples resulting in shorter radial cracks.

Analysis of the Distinct Pattern Formation of Globular Proteins in the Presence of Micro- and Nanoparticles.

Pattern formation during evaporation of biofluids has numerous biomedical applications, e.g., in disease identification. The drying of a bidisperse colloidal droplet involves formation of coffee ring patterns owing to the deposition of constituent particles. In the present study, we examine the distinctly different pattern formations during the drying of a colloidal solution depending on the nature of the constituent proteins. The pattern formations of two oppositely charged proteins, namely HSA and lysozyme, have been studied in the presence of fluorescence polystyrene beads of two different sizes (providing better image contrast for further analysis). The variation of pattern formation has been studied by varying the concentrations of the proteins as well as the particles. Furthermore, using image analysis, the patterns are segmented into different regions for quantification. To explain the variations in the patterns, we delve into the interplay of the interactions, especially the capillary and the DLVO forces (between the particles and the substrate). The developed methodology based on the coffee ring effect may be used to identify individual proteins.