Defining Single Molecular Forces Required for Notch Activation Using Nano Yoyo

Chowdhury, Farhan; Li, Isaac T. S.; Ngo, Thuy; Leslie, Benjamin J.; Kim, Byoung Choul; Sokoloski, Joshua E.; Weiland, Elizabeth; Wang, Xuefeng; Chemla, Yann R.; Lohman, Timothy M.; Ha, Taekjip

doi:10.1021/acs.nanolett.6b01403

Cited by 78 publications

(76 citation statements)

References 50 publications

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“…1a) [11,12,15]. Glass surfaces were passivated with biotinylated bovine serum albumin (BSA) prior to coating with neutravidin [12,16,17]. TGTs of varying T tol or cyclic-RGDfK peptide (ruptures at significantly higher tension forces, >100 pN [12,18]) were then immobilized to the passivated surface through neutravidin-biotin linker (Fig.…”

Section: Resultsmentioning

confidence: 99%

Cdc42-dependent modulation of rigidity sensing and cell spreading in tumor repopulating cells

Chowdhury

Doğanay

Leslie

et al. 2018

Biochemical and Biophysical Research Communications

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Recently, a robust mechanical method has been established to isolate a small subpopulation of highly tumorigenic tumor repopulating cells (TRCs) from parental melanoma cells. In order to characterize the molecular and mechanical properties of TRCs, we utilized the tension gauge tether (TGT) single-molecule platform and investigated force requirements during early cell spreading events. TRCs required the peak single molecular tension of around 40 pN through integrins for initial adhesion like the parental control cells, but unlike the control cells, they did not spread and formed very few mature focal adhesions (FAs). Single molecule resolution RNA quantification of three Rho GTPases showed that downregulation of Cdc42, but not Rac1, is responsible for the unusual biophysical features of TRCs and that a threshold level of Cdc42 transcripts per unit cell area is required to initiate cell spreading. Cdc42 overexpression rescued TRC spreading through FA formation and restored the sensitivity to tension cues such that TRCs, like parental control cells, increase cell spreading with increasing single-molecular tension cues. Our single molecule studies identified an unusual biophysical feature of suppressed spreading of TRCs that may enable us to distinguish TRC population from a pool of heterogeneous tumor cell population.

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Section: Resultsmentioning

confidence: 99%

Cdc42-dependent modulation of rigidity sensing and cell spreading in tumor repopulating cells

Chowdhury

Doğanay

Leslie

et al. 2018

Biochemical and Biophysical Research Communications

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“…For cells that do not contain granules, the cell surface in principle can be tagged sparsely by functionalized gold nanoparticles, which strongly scatter light and are visible under darkfield microscopy. Quantitative understanding of the adhesion behavior of rolling cells is important not only for fundamental cell biology, but also for practical applications including cancer diagnostics, screening and treatment [9][10][11] . The approaches presented here represent a first step toward a comprehensive label-free cell surface adhesion mapping and characterization technique.…”

Section: Discussionmentioning

confidence: 99%

“…Rolling adhesion behavior is also exhibited by circulating tumor cells (CTCs) which is believed to enhance cancer metastasis [5][6][7][8] . Therefore, quantitative understanding of rolling adhesion is necessary to enable practical applications such as cancer screening and treatment [9][10][11] . At the molecular level, this adhesion is mediated by catch-bond-like interactions 12,13 between P- 14 and E-selectins 15 expressed on endothelial cells lining blood vessels and P-selectin glycoprotein ligand-1 (PSGL-1) found at microvilli tips of leukocytes 16 .…”

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Mapping Cell Surface Adhesion by Rotation Tracking and Adhesion Footprinting

Chemla

2017

Biophysical Journal

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Rolling adhesion, in which cells passively roll along surfaces under shear flow, is a critical process involved in inflammatory responses and cancer metastasis. Surface adhesion properties regulated by adhesion receptors and membrane tethers are critical in understanding cell rolling behavior. Locally, adhesion molecules are distributed at the tips of membrane tethers. However, how functional adhesion properties are globally distributed on the individual cell's surface is unknown. Here, we developed a label-free technique to determine the spatial distribution of adhesive properties on rolling cell surfaces. Using dark-field imaging and particle tracking, we extract the rotational motion of individual rolling cells. The rotational information allows us to construct an adhesion map along the contact circumference of a single cell. To complement this approach, we also developed a fluorescent adhesion footprint assay to record the molecular adhesion events from cell rolling. Applying the combination of the two methods on human promyelocytic leukemia cells, our results surprisingly reveal that adhesion is non-uniformly distributed in patches on the cell surfaces. Our label-free adhesion mapping methods are applicable to the variety of cell types that undergo rolling adhesion and provide a quantitative picture of cell surface adhesion at the functional and molecular level.Rolling adhesion is a common process by which cells attach themselves to surfaces under shear flow, such as in the circulatory system. Leukocytes in the blood utilize this mechanism to locate inflammation sites throughout the body. During an inflammation response, endothelial cells lining the blood vessels surrounding an infection site express adhesion proteins called selectins that are specific to leukocyte surface receptors. As the first step of the leukocyte adhesion cascade, leukocytes captured via selectin-specific interactions passively roll on the blood vessel wall under blood flow toward the inflammation site in a process known as rolling adhesion [1][2][3] . Malfunction of any adhesion molecules involved in this process leads to severe immune disorders such as the leukocyte adhesion deficiencies (LAD) 4 . Rolling adhesion behavior is also exhibited by circulating tumor cells (CTCs) which is believed to enhance cancer metastasis [5][6][7][8] . Therefore, quantitative understanding of rolling adhesion is necessary to enable practical applications such as cancer screening and treatment [9][10][11] . At the molecular level, this adhesion is mediated by catch-bond-like interactions 12,13 between P-14 and E-selectins 15 expressed on endothelial cells lining blood vessels and P-selectin glycoprotein ligand-1 (PSGL-1) found at microvilli tips of leukocytes 16 . Despite our understanding of the individual components, how the molecular details of adhesion bonds scale to cell-surface adhesion and rolling behavior remains poorly understood 2,17,18 . Here, we developed a label-free method that maps the functional adhesion sites and strengths on a cell ...

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“…Two separate models for the requirement of endocytosis have been suggested: the first model suggests that recycling is important for maturation of the ligands to become active, whereas according to the second model, transendocytosis of the extracellular domain of the Notch receptor (NECD) is needed to produce a strain on the receptor. This strain is termed the pulling force (41,42,44,45) and is thought to reveal the cleavage site and initiate proteolytic processing of the receptor with subsequent release of the NICD.…”

mentioning

confidence: 99%

Selective regulation of Notch ligands during angiogenesis is mediated by vimentin

Antfolk

Sjöqvist

Cheng

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Notch signaling is a key regulator of angiogenesis, in which sprouting is regulated by an equilibrium between inhibitory Dll4-Notch signaling and promoting Jagged-Notch signaling. Whereas Fringe proteins modify Notch receptors and strengthen their activation by Dll4 ligands, other mechanisms balancing Jagged and Dll4 signaling are yet to be described. The intermediate filament protein vimentin, which has been previously shown to affect vascular integrity and regenerative signaling, is here shown to regulate ligand-specific Notch signaling. Vimentin interacts with Jagged, impedes basal recycling endocytosis of ligands, but is required for efficient receptor ligand transendocytosis and Notch activation upon receptor binding. Analyses of Notch signal activation by using chimeric ligands with swapped intracellular domains (ICDs), demonstrated that the Jagged ICD binds to vimentin and contributes to signaling strength. Vimentin also suppresses expression of Fringe proteins, whereas depletion of vimentin enhances Fringe levels to promote Dll4 signaling. In line with these data, the vasculature in vimentin knockout (VimKO) embryos and placental tissue is underdeveloped with reduced branching. Disrupted angiogenesis in aortic rings from VimKO mice and in endothelial 3D sprouting assays can be rescued by reactivating Notch signaling by recombinant Jagged ligands. Taken together, we reveal a function of vimentin and demonstrate that vimentin regulates Notch ligand signaling activities during angiogenesis.

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Defining Single Molecular Forces Required for Notch Activation Using Nano Yoyo

Cited by 78 publications

References 50 publications

Cdc42-dependent modulation of rigidity sensing and cell spreading in tumor repopulating cells

Cdc42-dependent modulation of rigidity sensing and cell spreading in tumor repopulating cells

Mapping Cell Surface Adhesion by Rotation Tracking and Adhesion Footprinting

Selective regulation of Notch ligands during angiogenesis is mediated by vimentin

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