Imatinib Alters Agonists-mediated Cytoskeletal Biomechanics in Lung Endothelium

Abstract

The endothelium serves as a size-selective barrier and tightly controls the fluid exchange from the circulation to the surrounding tissues. In this study, a multiplexed microscopy characterization is developed to study the spatio-temporal effects of Abl kinases on endothelial cytoskeletal structure using AFM, SEM, and immunofluorescence. Sphingosine 1-phosphate (S1P) produces significant endothelial barrier enhancement by means of peripheral actin rearrangement. However, Abl kinase inhibition by imatinib reduces rapid redistribution of the important cytoskeletal proteins to the periphery and their association with the cortical actin ring. Herein, it moderates the thickness of the cortical actin ring, and diminishes the increase in elastic modulus at the periphery and cytoplasm. These findings demonstrate that imatinib attenuates multiple cytoskeletal changes associated with S1P-mediated endothelial barrier enhancement and suggest a novel role for Abl kinases in mediating these S1P effects. These observations bridge the gap between molecule dynamics, structure complexity and function connectivity across varied length-scales to improve our understanding on human pulmonary endothelial barrier regulation. Moreover, our study suggests a framework for understanding form-function relationships in other biomechanical subsystems, wherein complex hierarchical organization programmed from the molecular scale to the cellular and tissue levels has an intimate relationship to the overall physiological function.

Polymer-Compatible Low-Temperature Plasma-Enhanced Chemical Vapor Deposition of Graphene on Electroplated Cu for Flexible Hybrid Electronics

“…In fact, the PeakForce error images have been applied to image-detailed structures of fragile cells in biology. 22,23 To investigate the thickness of graphene grown on electroplated Cu substrates, a typical approach is to consider the I(2D)/I(G) ratio from Raman spectroscopy, and a value of I(2D)/I(G) > 1 is commonly considered as a signature for monolayer graphene. 18 On the other hand, it is also well known that the I(2D)/I(G) ratio is sensitive to interlayer coupling, 24 so that it is not always an accurate indicator for the number of graphene layers.…”

“…Comparing the PeakForce error images (Figure ) to the height images (Figure S5), we find that the former can provide a better resolution for the detailed surface morphology. In fact, the PeakForce error images have been applied to image-detailed structures of fragile cells in biology. , …”

Methods for Studying Endometrial Pathology and the Potential of Atomic Force Microscopy in the Research of Endometrium

“…Furthermore, AFM allows cell mapping in terms of its elastic properties. The distribution of Young’s modulus has been determined for both animal and human cells, for example, MMTV-PyMT mouse cells as a model of mammary gland cancer [ 28 ], primary human pulmonary artery endothelial cells [ 29 , 30 , 31 ], or the epithelial-like breast carcinoma cell line [ 32 ]. Elasticity maps selected from the above-mentioned works are shown in Figure 6 .…”

“…Micropipette aspiration Determination of bovine aortic endothelial cells adhesion [24] Optical tweezers Intercellular adhesion of the early embryo epithelial cells determined by cell displacement [25] Assessment of the influence of neighboring cells on the stiffness of breast cancer cells [26] Atomic force microscopy Detecting fibroblast cell inhomogeneities based on point cell pressing [27] Force mapping: Young's modulus maps e.g., of mammary gland cancer [28], primary human pulmonary artery endothelial cells [29][30][31], epithelial-like breast carcinoma cells [32] and elasticity maps e.g., of human aortic endothelial cells [33] Elasticity measurements of human umbilical vein endothelial cells, chondrocytes, fibroblasts, fibrosarcoma and hepatocellular carcinoma cells […”

Cortical Actin Dynamics in Endothelial Permeability

“…Much detail on the mechanobiology of the pulmonary endothelial cytoskeleton has recently been revealed by atomic force microscopy (AFM) (Arce, Whitlock et al 2008, Dudek, Chiang et al 2010, Wang, Bleher et al 2015, Wang, Bleher et al 2017, Wang, Wang et al 2018). Now this technique paired with super-resolution microscopy can correlatively probe the spatiotemporal mechanodynamics of the cytoskeleton in EC with nanometer resolution (Hauser, Wojcik et al 2017).…”