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Biology subjects

Cho, Y.-K.

Publications and source records attributed to Cho, Y.-K..

5 recordsLinked to original sources

Mechanical coupling narrows an epithelial colony before it reaches a bottleneck

A colony migrating into a passage narrower than itself but wider than individuals must adapt its density distribution to pass. Powders, colloids and crowds driven through such an opening jam, since contact interactions provide no pathway for constituents to respond to the constriction before reaching it. Exploring whether a cohesive living tissue can reorganize in advance, we image MDCK epithelial monolayers migrating through microchannels in which a wide channel narrows, and locate response at the taper mouth, a transition zone in which we identify upstream responses. At locations where the local channel width is still unchanged, the monolayer detaches from the side walls and reduces its lateral extent. Particle image velocimetry shows that this is not passive compression; the axial velocity alternates aperiodically in sign, velocity divergence is positive in the transition zone, and tracking the positions of cell nuclei confirms that cells leave the crowded inlet region in both directions. Fluorescence imaging shows a belt of co-enriched E-cadherin, F-actin, and myosin-II at the edges that detach before the inlet, which laser ablation shows to be under elevated tension but dissolves once the tissue passes into the narrow inlet; {beta}-catenin is not co-enriched, indicating that junctions differ in composition in the core and at the edge of colony. This advance proceeds over hours, far more slowly than the viscoelastic reorganization reported for MDCK monolayers, and switches in sign repeatedly, which a linear viscoelastic sheet under sustained constraint cannot do. E-cadherin knockdown abolishes narrowing, flow reversal, density redistribution, and belt assembly.

biophysics↗

Digital Profiling of Tumor Extracellular Vesicle-associated RNAs Directly from Unprocessed Blood Plasma

Tumor-derived extracellular vesicle (tEV)-associated RNAs hold promise as diagnostic biomarkers, but their clinical use is hindered by the rarity of tEVs among non-tumor EVs. Here, we present EV-CLIP, a highly sensitive droplet-based digital method for profiling EV RNA. EV-CLIP utilizes the fusion of EVs with charged liposomes (CLIPs) in a microfluidic chip. Optimized CLIP surface charge enables exceptional sensitivity and selectivity for EV-derived miRNAs and mRNAs. This approach streamlines detection with minimal plasma volume (20 {micro}L) and eliminates the need for prior EV isolation or RNA preparation, preventing loss of EVs or RNA. In testing with 83 patient samples, EV-CLIP detected EGFR L858R and T790M mutations with high AUC values of 1.0000 and 0.9784, respectively. Its success in serial monitoring during chemotherapy highlights its potential for precise quantification of rare EV subpopulations, facilitating the exploration of single EV RNA content and enhancing understanding of diverse EV populations in various disease states. SUMMARYEV-CLIP: A highly sensitive digital method for tumor-derived EV RNA detection using undiluted blood plasma samples, promising for precise tumor management.

bioengineering↗

Machine learning analysis reveals the dynamics of mode transition in dendritic cell migration

Dendritic cells (DCs) patrol the body as immunological sentinels and search for pathogens. Upon stimulation, immature DCs (imDCs) become mature DCs (mDCs), which migrate to the lymph nodes and present antigens to T cells. The migratory behavior is crucial for initiating and controlling immune responses; however, the properties of the highly heterogeneous and dynamic motility phenotype are not fully understood. Here, we established an unsupervised machine learning (ML) strategy to investigate spatiotemporal motility patterns in long-term, two-dimensional cell migration trajectories, and determined the number of motility patterns and how these are related to the maturation status. We identified three distinct migratory modes independent of the cell state: slow-diffusive (SD), slow-persistent (SP), and fast-persistent (FP). We found that maturation-dependent motility changes are emergent properties of the distribution and dynamic transitions of these three modes. Remarkably, imDCs changed their migration modes more frequently, and predominantly followed the SD[->]FP[->]SP[->]SD unicyclic transition, indicating that imDCs rapidly increase their speed during the shift from diffusive to persistent motility; however, persistence progressively declines when switching back to diffusive motility. In contrast, mDCs show no transition directionality. Our ML-promoted motility pattern analysis and history-dependent mode transition investigation may provide new insights into the complex process of biological motility.

biophysics↗

Exhausted mature dendritic cells exhibit a slower and less persistent random motility but retain chemotaxis against CCL19

Dendritic cells (DCs), which are immune sentinels in the peripheral tissues, play a number of roles, including patrolling for pathogens, internalising antigens, transporting antigens to the lymph nodes (LNs), interacting with T cells, and secreting cytokines. The well-coordinated migration of DCs under various immunological or inflammatory conditions is therefore essential to ensure an effective immune response. Upon maturation, DCs migrate faster and more persistently than immature DCs (iDCs), which is believed to facilitate CCR7-dependent chemotaxis. It has been reported that lipopolysaccharide-activated DCs produce IL-12 only transiently, and become resistant to further stimulation through exhaustion. However, little is known about the influence of DC exhaustion on cellular motility. Here, we studied the cellular migration of exhausted DCs in tissue-mimicked confined environments. We found that the speed of exhausted matured DCs (xmDCs) decreased significantly compared to active matured DCs (amDCs) and iDCs. In contrast, the speed fluctuation increased compared to that of amDCs and was similar to that of iDCs. In addition, the diffusivity of the xmDCs was significantly lower than that of the amDCs, which implies that DC exhaustion reduces the space exploration ability. Interestingly, CCR7-dependent chemotaxis against CCL19 in xmDCs was not considerably different from that observed in amDCs. Taken together, we report a unique intrinsic cell migration behavior of xmDCs, which exhibit a slower, less persistent, and less diffusive random motility, which results in the DCs remaining at the site of infection, although a well-preserved CCR7-dependent chemotactic motility is maintained.

cell biology↗

Nanocharacterisation of extracellular vesicles uncovers the correlation between isolation methods, physicochemical composition and their biological function

Extracellular vesicles (EVs) have been lauded as next generation medicines, but very few EV-based therapeutics have progressed to clinical use. Limited clinical translation is largely due to technical barriers that hamper our ability to mass-produce EVs, i.e. to isolate, purify and characterise them effectively. Technical limitations in comprehensive characterisation of EVs leads to unpredicted biological effects of EVs. Here, using a range of optical and non-optical techniques, we showed that the differences in molecular composition of EVs isolated using two isolation methods correlated with the differences in their biological function. Our results demonstrated that the isolation method determines the composition of isolated EVs at single and sub-population levels. Besides the composition, we measured for the first time the dry mass and predicted sedimentation of EVs. These parameters were shown to correlate well with the biological and functional effects of EVs on single cell and cell cultures. We anticipate that our new multiscale characterisation approach, which goes beyond traditional experimental methodology, will support fundamental understanding of EVs as well as elucidate the functional effects of EVs in in vitro and in vivo studies. Our findings and methodology will be pivotal for developing optimal isolation methods and establishing EVs as mainstream therapeutics and diagnostics. This innovative approach is applicable to a wide range of sectors including biopharma and biotechnology as well as to regulatory agencies.

cell biology↗