Samanta, B. | bioRxiv Science

bioRxiv2025

C2-α-dicarbonyls selectively impair morphodynamic interactions between cells and extracellular matrix

Ageing and diabetes, risk factors for cardiovascular diseases and cancer, are characterized by abnormally high tissue levels of several dicarbonyl species. Although all dicarbonyls disrupt biomolecular structure and function by forming advanced glycation end products, understanding the relative toxicity of each can help design targeted control strategies. Although known for cells, the comparative effects of distinct dicarbonyls on extracellular matrices (ECM) have yet to be benchmarked. Here, we investigate how glycation by three well known dicarbonyls: the C2--dicarbonyls methylglyoxal (MGO) and glyoxal (GO), and the C6--dicarbonyl 3-deoxyglucosone (3-DG), affects two ECMs, Collagen I and laminin-rich basement membrane (lrBM), and alters their morphodynamic interactions with two human cells: untransformed endothelial TeloHAEC and aggressive triple negative breast cancer MDA-MB-231. On dicarbonyl-treated lrBM, endothelial and cancer cell adhesion, collective cord morphogenesis, cancer cell motility, shape polarity and its entropy are perturbed at low MGO levels, high GO levels, and are unchanged for 3-DG. Alcian Blue staining of lrBM shows dissolution of matrix at low MGO and high GO levels with no effect on 3-DG exposure. On collagen I substrata, adhesion for both cells is disrupted at low MGO levels, at high GO levels but is unchanged for 3-DG. Interestingly at high concentrations of MGO and GO, the latter decreases motility and increases elongation entropy more than the former; 3-DG has no effect. Picrosirius staining suggests that at high levels GO-driven collagen hyperpolymerization is similar to MGO with no effect of 3-DG. Our observations therefore suggest qualitative and quantitative distinctions in the effect of C2- and C-6--dicarbonyls on ECM homoeostasis and ECM-driven cellular morphodynamics.

cell biology↗

bioRxiv2024

Dicarbonyl stress enhances tumor intravasation

Metastasis of cancer is a multi-step process that involves the migration of transformed cells from their native organ into a vascular channel, followed by their dissemination to prospective sites of colonization. The entry of tumor cells into blood or lymph, known as intravasation involves their breaching the stromal and endothelial extracellular matrix (ECM) and the endothelial barriers. How the kinetics of these cell-ECM interactions are confounded by chronic inflammatory stresses seen in comorbid risk factors of cancer such as diabetes and aging remain ill-investigated. Here, we construct and deploy a histopathology-motivated, imaging-tractable, microfluidic multi-organ-on-chip platform, that seamlessly integrates two tissue environments: that of a breast tumor and a vascular channel, to study the problem. The former comprises invasive triple-negative MDA-MB-231 breast cancer cells embedded within a three-dimensional fibrillar Collagen I milieu. The latter consists of a monolayer of TeloHAEC, immortalized human aortic endothelial cells arranged on laminin-rich basement membrane ECM, both of which concentrically line a hollow channel, wherein unidirectional fluid flows are implemented. The chip showcases the complexity of intravasation, wherein tumor cells and endothelia cooperate to form anastomotic structures. The formation of such structures is regulated by fluid flow in the vascular channel and is associated with cancer cell migration and entry into the vascular channel. Disseminated cancer cells are observed to enter, get adhered within, and flow through the vascular channel. Exposure to methylglyoxal (MG), a mediator of dicarbonyl stress associated with diabetic circulatory milieu, leads to greater cancer cell intravasation and flow through the vascular channel. This could be driven not just by MG-induced endothelial senescence and shedding, but also by the effect of MG on the chip ECM: we demonstrate it can degrade basement membrane and pathologically crosslink Collagen I, diminishing their cell adhesiveness. Our results thus show how dicarbonyl stress may attenuate homoeostatic barriers to cancer intravasation, exacerbating metastasis.

bioengineering↗