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      Junction Mapper is a novel computer vision tool to decipher cell–cell contact phenotypes

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          Abstract

          Stable cell–cell contacts underpin tissue architecture and organization. Quantification of junctions of mammalian epithelia requires laborious manual measurements that are a major roadblock for mechanistic studies. We designed Junction Mapper as an open access, semi-automated software that defines the status of adhesiveness via the simultaneous measurement of pre-defined parameters at cell–cell contacts. It identifies contacting interfaces and corners with minimal user input and quantifies length, area and intensity of junction markers. Its ability to measure fragmented junctions is unique. Importantly, junctions that considerably deviate from the contiguous staining and straight contact phenotype seen in epithelia are also successfully quantified (i.e. cardiomyocytes or endothelia). Distinct phenotypes of junction disruption can be clearly differentiated among various oncogenes, depletion of actin regulators or stimulation with other agents. Junction Mapper is thus a powerful, unbiased and highly applicable software for profiling cell–cell adhesion phenotypes and facilitate studies on junction dynamics in health and disease.

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          Most cited references39

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          Ras superfamily GEFs and GAPs: validated and tractable targets for cancer therapy?

          There is now considerable and increasing evidence for a causal role for aberrant activity of the Ras superfamily of small GTPases in human cancers. These GTPases function as GDP-GTP-regulated binary switches that control many fundamental cellular processes. A common mechanism of GTPase deregulation in cancer is the deregulated expression and/or activity of their regulatory proteins, guanine nucleotide exchange factors (GEFs) that promote formation of the active GTP-bound state and GTPase-activating proteins (GAPs) that return the GTPase to its GDP-bound inactive state. In this Review, we assess the association of GEFs and GAPs with cancer and their druggability for cancer therapeutics.
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            The Small GTPases Rho and Rac Are Required for the Establishment of Cadherin-dependent Cell–Cell Contacts

            Cadherins are calcium-dependent cell–cell adhesion molecules that require the interaction of the cytoplasmic tail with the actin cytoskeleton for adhesive activity. Because of the functional relationship between cadherin receptors and actin filament organization, we investigated whether members of the Rho family of small GTPases are necessary for cadherin adhesion. In fibroblasts, the Rho family members Rho and Rac regulate actin polymerization to produce stress fibers and lamellipodia, respectively. In epithelial cells, we demonstrate that Rho and Rac are required for the establishment of cadherin-mediated cell–cell adhesion and the actin reorganization necessary to stabilize the receptors at sites of intercellular junctions. Blocking endogenous Rho or Rac selectively removed cadherin complexes from junctions induced for up to 3 h, while desmosomes were not perturbed. In addition, withdrawal of cadherins from intercellular junctions temporally precedes the removal of CD44 and integrins, other microfilament-associated receptors. Our data showed that the concerted action of Rho and Rac modulate the establishment of cadherin adhesion: a constitutively active form of Rac was not sufficient to stabilize cadherindependent cell–cell contacts when endogenous Rho was inhibited. Upon induction of calcium-dependent intercellular adhesion, there was a rapid accumulation of actin at sites of cell–cell contacts, which was prevented by blocking cadherin function, Rho or Rac activity. However, if cadherin complexes are clustered by specific antibodies attached to beads, actin recruitment to the receptors was perturbed by inhibiting Rac but not Rho. Our results provide new insights into the role of the small GTPases in the cadherin-dependent cell– cell contact formation and the remodelling of actin filaments in epithelial cells.
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              Mechanotransduction in cardiac hypertrophy and failure.

              Cardiac muscle cells have an intrinsic ability to sense and respond to mechanical load through a process known as mechanotransduction. In the heart, this process involves the conversion of mechanical stimuli into biochemical events that induce changes in myocardial structure and function. Mechanotransduction and its downstream effects function initially as adaptive responses that serve as compensatory mechanisms during adaptation to the initial load. However, under prolonged and abnormal loading conditions, the remodeling processes can become maladaptive, leading to altered physiological function and the development of pathological cardiac hypertrophy and heart failure. Although the mechanisms underlying mechanotransduction are far from being fully elucidated, human and mouse genetic studies have highlighted various cytoskeletal and sarcolemmal structures in cardiac myocytes as the likely candidates for load transducers, based on their link to signaling molecules and architectural components important in disease pathogenesis. In this review, we summarize recent developments that have uncovered specific protein complexes linked to mechanotransduction and mechanotransmission within the sarcomere, the intercalated disc, and at the sarcolemma. The protein structures acting as mechanotransducers are the first step in the process that drives physiological and pathological cardiac hypertrophy and remodeling, as well as the transition to heart failure, and may provide better insights into mechanisms driving mechanotransduction-based diseases.
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                Author and article information

                Contributors
                Role: Reviewing Editor
                Role: Senior Editor
                Journal
                eLife
                Elife
                eLife
                eLife
                eLife Sciences Publications, Ltd
                2050-084X
                03 December 2019
                2019
                : 8
                : e45413
                Affiliations
                [1 ]National Heart and Lung Institute, National Institutes of Health LondonUnited Kingdom
                [2 ]deptBioinformatics Data Science Group, Faculty of Medicine Imperial College London LondonUnited Kingdom
                [3 ]deptDepartment Medical Biochemistry, Amsterdam Cardiovascular Sciences Amsterdam UMC, University of Amsterdam AmsterdamNetherlands
                Yale University United States
                Utrecht University Netherlands
                Yale University United States
                University of Cologne Germany
                Author notes
                [‡]

                PAPRSB Institute of Health Sciences, Universiti Brunei, Darussalam, Brunei.

                [§]

                Department of Anatomy, Physiology and Genetics, Oxford University, Oxford, United Kingdom.

                [†]

                These authors contributed equally to this work.

                Author information
                https://orcid.org/0000-0002-3554-6084
                https://orcid.org/0000-0002-8634-6111
                http://orcid.org/0000-0002-3705-7495
                https://orcid.org/0000-0002-1091-475X
                http://orcid.org/0000-0003-1148-9158
                https://orcid.org/0000-0002-5814-7166
                https://orcid.org/0000-0003-0546-7163
                Article
                45413
                10.7554/eLife.45413
                7034980
                31793877
                fcf2b855-63ba-4107-945d-6237f02c4232
                © 2019, Brezovjakova et al

                This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

                History
                : 23 January 2019
                : 02 December 2019
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/501100000265, Medical Research Council;
                Award ID: MR/M026310/1
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/501100000268, Biotechnology and Biological Sciences Research Council;
                Award ID: BB/M022617/1
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/501100000289, Cancer Research UK;
                Award ID: C1282/A11980
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/501100003246, Nederlandse Organisatie voor Wetenschappelijk Onderzoek;
                Award ID: VIDI 016.156.327
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/501100002761, Prime Minister's Office, Brunei Darussalam;
                Award ID: JPLL/A/4:A[2010]/J1(703)
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/100000002, National Institutes of Health;
                Award ID: 1R01HL141855-01
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/501100000274, British Heart Foundation;
                Award ID: RG/17/13/33173
                Award Recipient :
                The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
                Categories
                Tools and Resources
                Cancer Biology
                Cell Biology
                Custom metadata
                Junction Mapper is a powerful new semi-automated software that provides a fingerprint of cell–cell contact morphometry and receptor density alterations.

                Life sciences
                image analysis,computer vision,tissue cohesion,cell-cell contacts,software development,junction regulation,human

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