Abstract:Current approaches to cancer treatment focus on targeting signal transduction pathways. Here, we develop an alternative system for targeting cell mechanics for the discovery of novel therapeutics. We designed a live-cell, high-throughput chemical screen to identify mechanical modulators. We characterized 4-hydroxyacetophenone (4-HAP), which enhances the cortical localization of the mechanoenzyme myosin II, independent of myosin heavy-chain phosphorylation, thus increasing cellular cortical tension. To shift ce… Show more
“…[1] Drugs that modify cytoskeletal or nuclear architecture are often applied in cancer treatment, [8, 36] and recently, 4-hydroxyacetophenone (4-HAP) was found to alter pancreatic cancer mechanical properties by stimulating myosin-II and enhancing its cortical localization, which reduces the invasion and migration of pancreatic cancer cells by stiffening them. [8] Micropipette aspiration was used for the study measuring the cell deformability to find the efficacy of 4-HAP; however, due to its low throughput, only tens of cells were characterized to verify the drug responses. Furthermore, it was suggested that 4-HAP can be applied to other cancers, though with a micropipette aspiration approach, it is limited to test other various cancer cell lines at different stages.…”
Mechanical biomarkers associated with cytoskeletal structures have been reported as powerful label-free cell state identifiers. In order to measure cell mechanical properties, traditional biophysical (e.g., atomic force microscopy, micropipette aspiration, optical stretchers) and microfluidic approaches were mainly employed; however, they critically suffer from low-throughput, low-sensitivity, and/or time-consuming and labor-intensive processes, not allowing techniques to be practically used for cell biology research applications. Here, a novel inertial microfluidic cell stretcher (iMCS) capable of characterizing large populations of single-cell deformability near real-time is presented. The platform inertially controls cell positions in microchannels and deforms cells upon collision at a T-junction with large strain. The cell elongation motions are recorded, and thousands of cell deformability information is visualized near real-time similar to traditional flow cytometry. With a full automation, the entire cell mechanotyping process runs without any human intervention, realizing a user friendly and robust operation. Through iMCS, distinct cell stiffness changes in breast cancer progression and epithelial mesenchymal transition are reported, and the use of the platform for rapid cancer drug discovery is shown as well. The platform returns large populations of single-cell quantitative mechanical properties (e.g., shear modulus) on-the-fly with high statistical significances, enabling actual usages in clinical and biophysical studies.
“…[1] Drugs that modify cytoskeletal or nuclear architecture are often applied in cancer treatment, [8, 36] and recently, 4-hydroxyacetophenone (4-HAP) was found to alter pancreatic cancer mechanical properties by stimulating myosin-II and enhancing its cortical localization, which reduces the invasion and migration of pancreatic cancer cells by stiffening them. [8] Micropipette aspiration was used for the study measuring the cell deformability to find the efficacy of 4-HAP; however, due to its low throughput, only tens of cells were characterized to verify the drug responses. Furthermore, it was suggested that 4-HAP can be applied to other cancers, though with a micropipette aspiration approach, it is limited to test other various cancer cell lines at different stages.…”
Mechanical biomarkers associated with cytoskeletal structures have been reported as powerful label-free cell state identifiers. In order to measure cell mechanical properties, traditional biophysical (e.g., atomic force microscopy, micropipette aspiration, optical stretchers) and microfluidic approaches were mainly employed; however, they critically suffer from low-throughput, low-sensitivity, and/or time-consuming and labor-intensive processes, not allowing techniques to be practically used for cell biology research applications. Here, a novel inertial microfluidic cell stretcher (iMCS) capable of characterizing large populations of single-cell deformability near real-time is presented. The platform inertially controls cell positions in microchannels and deforms cells upon collision at a T-junction with large strain. The cell elongation motions are recorded, and thousands of cell deformability information is visualized near real-time similar to traditional flow cytometry. With a full automation, the entire cell mechanotyping process runs without any human intervention, realizing a user friendly and robust operation. Through iMCS, distinct cell stiffness changes in breast cancer progression and epithelial mesenchymal transition are reported, and the use of the platform for rapid cancer drug discovery is shown as well. The platform returns large populations of single-cell quantitative mechanical properties (e.g., shear modulus) on-the-fly with high statistical significances, enabling actual usages in clinical and biophysical studies.
“…For example, the mechanical response of cells to their environment may be defective in some cancer cells [66]. Restoring this response may reduce the inappropriate movement of the cancer cells [67]. Studying the molecular mechanisms of cell movement in 3D extracellular matrix, rather than on 2D tissue culture plastic, is likely to provide the most efficient way to address these and other important questions (see Outstanding Questions Box) in the future.…”
Primary human fibroblasts are remarkably adaptable, able to migrate in differing types of physiological 3D tissue and on rigid 2D tissue culture surfaces. The crawling behavior of these and other vertebrate cells has been studied intensively, which has helped generate the concept of the cell motility cycle as a comprehensive model of 2D cell migration. However, this model fails to explain how cells force their large nuclei through the confines of a 3D matrix environment and why primary fibroblasts can use more than one mechanism to move in 3D. Recent work shows that the intracellular localization of myosin II activity is governed by cell-matrix interactions to both force the nucleus through the extracellular matrix and dictate the type of protrusions used to migrate in 3D.
“…Furthermore, to treat diseases that include an element of defective mechanotransduction, putative compounds must be tested in the appropriate 3D matrix mechanics to accurately predict responses [116], and active efforts to develop compounds that selectively target key elements of the mechanotransduction pathway will be critical [117]. New mechanistic insights into how cells sense and respond to tissue mechanics will follow from the widespread implementation of in silico modeling [118] to develop complex cell-cell and cell-matrix models [92,97,106,119], and the development of 3D models to systematically tease apart the influence of diverse matrix mechanical properties, biomolecules, and genetic alterations on cell behaviors and fate in a controlled manner [120-123].…”
Section: Concluding Remarks and Future Directionsmentioning
Human tissues are remarkably adaptable and robust, harboring the collective ability to detect and respond to external stresses while maintaining tissue integrity. Following injury, many tissues have the capacity to repair the damage - and restore form and function - by deploying cellular and molecular mechanisms reminiscent of developmental programs. Indeed, it is increasingly clear that cancer and chronic conditions that develop with age arise as a result of cells and tissues re-implementing and deregulating a selection of developmental programs. Therefore, understanding the fundamental molecular mechanisms that drive cell and tissue responses is a necessity when designing therapies to treat human conditions. Extracellular matrix stiffness synergizes with chemical cues to drive single cell and collective cell behavior in culture and acts to establish and maintain tissue homeostasis in the body. This review will highlight recent advances that elucidate the impact of matrix mechanics on cell behavior and fate across these length scales during times of homeostasis and in disease states.
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