DOCSLIB.ORG

Three-Dimensional Biomimetic Vascular Model Reveals a Rhoa, Rac1, and N-Cadherin Balance in Mural Cell–Endothelial Cell-Regulated Barrier Function

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Three-Dimensional Biomimetic Vascular Model Reveals a Rhoa, Rac1, and N-Cadherin Balance in Mural Cell–Endothelial Cell-Regulated Barrier Function Three-dimensional biomimetic vascular model reveals a RhoA, Rac1, and N-cadherin balance in mural cell–endothelial cell-regulated barrier function Stella Alimpertia,b,c,1, Teodelinda Mirabellaa,b,c,1, Varnica Bajaja,b, William Polachecka,b,c, Dana M. Pironed, Jeremy Duffielde,f,g, Jeroen Eyckmansa,b,c, Richard K. Assoianh,i, and Christopher S. Chena,b,c,2 aDepartment of Biomedical Engineering, Boston University, Boston, MA 02215; bBiological Design Center, Boston University, Boston, MA 02215; cThe Wyss Institute for Biologically Inspired Engineering at Harvard, Harvard University, Boston, MA 02115; dDepartment of Science, Mount St. Mary’s University, Emmitsburg, MD 21727; eDivision of Nephrology, Department of Medicine, University of Washington, Seattle, WA 98195; fDepartment of Pathology, University of Washington, Seattle, WA 98195; gInstitute of Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA 98195; hDepartment of Systems Pharmacology and Translational Therapeutics, University of Pennsylvania, Philadelphia, PA 19104; and iProgram in Translational Biomechanics, Institute of Translational Medicine and Therapeutics, University of Pennsylvania, Philadelphia, PA 19104 Edited by Shu Chien, University of California, San Diego, La Jolla, CA, and approved July 7, 2017 (received for review November 4, 2016) The integrity of the endothelial barrier between circulating blood blood vessel contractility, vascular integrity, and, overall, organ and tissue is important for blood vessel function and, ultimately, for homeostasis (10–14). For example, for proper organ function in organ homeostasis. Here, we developed a vessel-on-a-chip with kidney and brain, where barrier properties in the microvasculature perfused endothelialized channels lined with human bone marrow need to be finely tuned, an appropriate mural cell–EC ratio is stromal cells, which adopt a mural cell-like phenotype that recapit- important (15–17). Whereas mural cells are thought to be im- ulates barrier function of the vasculature. In this model, barrier portant in vivo, much of our understanding of barrier function function is compromised upon exposure to inflammatory factors regulation is focused solely on the endothelium (18, 19). This may α such as LPS, thrombin, and TNF , as has been observed in vivo. In- be, in part, because barrier function is widely studied in endo- terestingly, we observed a rapid physical withdrawal of mural cells thelial monolayers (20, 21) and is also due to the lack of an in vitro from the endothelium that was accompanied by an inhibition of system that incorporates both ECs and mural cells, which can be endogenous Rac1 activity and increase in RhoA activity in the mural exposed to fluid flow (22). cells themselves upon inflammation. Using a system to chemically Here, we have developed a biomimetic 3D platform that reca- induce activity in exogenously expressed Rac1 or RhoA within mi- pitulates perivascular-mediated barrier function in an engineered nutes of stimulation, we demonstrated RhoA activation induced loss microvessel with a perfusable lumen and then investigated whether of mural cell coverage on the endothelium and reduced endothelial inflammatory cues would impact the endothelial–mural cell in- barrier function, and this effect was abrogated when Rac1 was si- multaneously activated. We further showed that N-cadherin expres- teraction. We showed that both primary human bone marrow stro- sion in mural cells plays a key role in barrier function, as CRISPR- mal cells (hBMSCs) and pericytes reside perivascularly and adopt mediated knockout of N-cadherininthemuralcellsledtolossof mural functions within the engineered vasculature. Interestingly, we barrier function, and overexpression of N-cadherin in CHO cells pro- found that vascular inflammation induces a Ras homolog family moted barrier function. In summary, this bicellular model demon- member A (RhoA)-dependent detachment of mural cells and re- strates the continuous and rapid modulation of adhesive duces N-cadherin–mediated coverage of vessels, leading to vascular interactions between endothelial and mural cells and its impact permeability. Furthermore, we were able to substantiate these on vascular barrier function and highlights an in vitro platform to study the biology of perivascular–endothelial interactions. Significance 3D culture | mural cells | vascular inflammation | RhoGTPases | N-cadherin Organ homeostasis requires integrity of blood vessels; alter- ations or disruption of the vascular barrier between blood and arrier properties of blood vessels play a major role in organ tissue contribute to numerous diseases. Endothelial cells and Bfunction and homeostasis (1). Upon tissue injury, activation of mural cells are two key cell types, which play significant roles for inflammatory cascades causes a loss of barrier function, which the maintenance of barrier function. Here, we present a 3D results in fluid leakage into the interstitial tissue, ultimately lead- bicellular vascular model to mimic this barrier function and study ing to edema. When blood and platelets are exposed to the un- the role of mural cells in vascular inflammation. Importantly, by derlying extracellular matrix, clotting and further inflammation using this 3D model we identified RhoA, Rac1, and N-cadherin as – can also be triggered. Thus, in acute settings, such as in ischemic important regulators in mural endothelial cell-mediated vascular or hemorrhagic stroke, massive changes in barrier function can barrier function. Given the recognized fundamental importance compromise vital organs (2, 3). Even mild but prolonged loss of of this barrier in numerous disease settings, this in vitro micro- barrier function can ultimately result in compromised perfusion, physiological system presented herein could provide a tool for studying vascular barrier function in 3D microenvironments. chronic inflammation, and tissue damage (4–7). Although endothelial cells (ECs) are the primary effectors of Author contributions: S.A., T.M., R.K.A., and C.S.C. designed research; S.A., T.M., and V.B. barrier control, it has long been appreciated that there is a sup- performed research; W.P., D.M.P., and J.D. contributed new reagents/analytic tools; S.A. porting mesenchymal population known as mural cells that are and T.M. analyzed data; and S.A., T.M., J.E., and C.S.C. wrote the paper. important for modulating endothelial function. Vascular mural The authors declare no conflict of interest. cells are a heterogeneous cell population consisting of vascular This article is a PNAS Direct Submission. smooth muscle cells and pericytes, which were defined originally Freely available online through the PNAS open access option. based on their anatomical location, covering the abluminal surface 1S.A. and T.M. contributed equally to this work. and sharing the basement membrane with the endothelium of 2To whom correspondence should be addressed. Email: [email protected]. capillaries, arterioles, and venules (8, 9). The functional role of This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10. mural cells is to modulate vessel function by regulating blood flow, 1073/pnas.1618333114/-/DCSupplemental. 8758–8763 | PNAS | August 15, 2017 | vol. 114 | no. 33 www.pnas.org/cgi/doi/10.1073/pnas.1618333114 Downloaded by guest on September 24, 2021 findings in an in vivo model of LPS-driven skin inflammation. Fig. S3) and quantified permeability by measuring the extravasa- Hence, our 3D bicellular system enables us to deconvolute the tion of fluorescently labeled 10 kDa or 70 kDa dextran from the contribution of different blood vessel cell types on vascular barrier vessel lumen (34) (Fig. 1C). We found that the presence of function and to detect molecular targets for potential therapeutics to hBMSCs in ratios of 1:1 and 1:5 relative to HUVECs significantly treat blood vessel diseases. reduced leakage of fluorescent dextran into the interstitial space compared with control microvessels containing ECs alone. Further Results decreasing the ratio of hBMSCs-HUVECs (from 1:1 to 1:100) Bicellular Perfusable 3D Platform Recapitulates Pericyte Role in resulted in dye extravasation, indicating increased vascular per- Regulating Vascular Barrier Function. To study the role of peri- meability (Fig. 1C). To further validate that hBMSCs are indeed vascular cells in regulating barrier function, we used a microfluidic an appropriate cell type to study barrier function, we compared the device in which human bone marrow stromal cells (hBMSCs), a vascular permeability of endothelialized channels covered with cell population that exhibit mural cell characteristics (23–28), lined human kidney pericytes (PCs), hBMSCs, and human lung fibro- a perfusable endothelialized channel. The device was assembled blasts (hFs). Whereas hFs reduced permeability compared with based largely on previously described models of endothelialized HUVECs alone, the effect was much smaller than the near com- channels (29), in this case by casting a single, hollowed cylindrical plete reduction in permeability seen with either hBMSCs or PCs channel (160 μm diameter) into type-I collagen within a poly- (SI Appendix,Fig.S4). In contrast to hFs, hBMSCs do not exhibit dimethylsiloxane (PDMS) mold containing a bulk chamber hosting migratory behavior, and they cover the endothelium by sharing the the vessel and reservoir chambers for introducing media into the basement membrane with ECs (SI Appendix,Fig.S5A and B), thus system. hBMSCs were seeded 4
Load more

Recommended publications
  • Human ADAM12 Quantikine ELISA
    Quantikine® ELISA Human ADAM12 Immunoassay Catalog Number DAD120 For the quantitative determination of A Disintegrin And Metalloproteinase domain- containing protein 12 (ADAM12) concentrations in cell culture supernates, serum, plasma, and urine. This package insert must be read in its entirety before using this product. For research use only. Not for use in diagnostic procedures. TABLE OF CONTENTS SECTION PAGE INTRODUCTION .....................................................................................................................................................................1 PRINCIPLE OF THE ASSAY ...................................................................................................................................................2 LIMITATIONS OF THE PROCEDURE .................................................................................................................................2 TECHNICAL HINTS .................................................................................................................................................................2 MATERIALS PROVIDED & STORAGE CONDITIONS ...................................................................................................3 OTHER SUPPLIES REQUIRED .............................................................................................................................................3 PRECAUTIONS .........................................................................................................................................................................4
    [Show full text]
  • Human N-Cadherin / CD325 / CDH2 Protein (His & Fc Tag)
    Human N-Cadherin / CD325 / CDH2 Protein (His & Fc Tag) Catalog Number: 11039-H03H General Information SDS-PAGE: Gene Name Synonym: CD325; CDHN; CDw325; NCAD Protein Construction: A DNA sequence encoding the human CDH2 (NP_001783.2) (Met 1-Ala 724) was fused with the C-terminal polyhistidine-tagged Fc region of human IgG1 at the C-terminus. Source: Human Expression Host: HEK293 Cells QC Testing Purity: > 70 % as determined by SDS-PAGE Endotoxin: Protein Description < 1.0 EU per μg of the protein as determined by the LAL method Cadherins are calcium dependent cell adhesion proteins, and they preferentially interact with themselves in a homophilic manner in Stability: connecting cells. Cadherin 2 (CDH2), also known as N-Cadherin (neuronal) (NCAD), is a single-pass tranmembrane protein and a cadherin containing ℃ Samples are stable for up to twelve months from date of receipt at -70 5 cadherin domains. N-Cadherin displays a ubiquitous expression pattern but with different expression levels between endocrine cell types. CDH2 Asp 160 Predicted N terminal: (NCAD) has been shown to play an essential role in normal neuronal Molecular Mass: development, which is implicated in an array of processes including neuronal differentiation and migration, and axon growth and fasciculation. The secreted recombinant human CDH2 is a disulfide-linked homodimeric In addition, N-Cadherin expression was upregulated in human HSC during protein. The reduced monomer comprises 813 amino acids and has a activation in culture, and function or expression blocking of N-Cadherin predicted molecular mass of 89.9 kDa. As a result of glycosylation, it promoted apoptosis. During apoptosis, N-Cadherin was cleaved into 20- migrates as an approximately 114 and 119 kDa band in SDS-PAGE under 100 kDa fragments.
    [Show full text]
  • The N-Cadherin Interactome in Primary Cardiomyocytes As Defined Using Quantitative Proximity Proteomics Yang Li1,*, Chelsea D
    © 2019. Published by The Company of Biologists Ltd | Journal of Cell Science (2019) 132, jcs221606. doi:10.1242/jcs.221606 TOOLS AND RESOURCES The N-cadherin interactome in primary cardiomyocytes as defined using quantitative proximity proteomics Yang Li1,*, Chelsea D. Merkel1,*, Xuemei Zeng2, Jonathon A. Heier1, Pamela S. Cantrell2, Mai Sun2, Donna B. Stolz1, Simon C. Watkins1, Nathan A. Yates1,2,3 and Adam V. Kwiatkowski1,‡ ABSTRACT requires multiple adhesion, cytoskeletal and signaling proteins, The junctional complexes that couple cardiomyocytes must transmit and mutations in these proteins can cause cardiomyopathies (Ehler, the mechanical forces of contraction while maintaining adhesive 2018). However, the molecular composition of ICD junctional homeostasis. The adherens junction (AJ) connects the actomyosin complexes remains poorly defined. – networks of neighboring cardiomyocytes and is required for proper The core of the AJ is the cadherin catenin complex (Halbleib and heart function. Yet little is known about the molecular composition of the Nelson, 2006; Ratheesh and Yap, 2012). Classical cadherins are cardiomyocyte AJ or how it is organized to function under mechanical single-pass transmembrane proteins with an extracellular domain that load. Here, we define the architecture, dynamics and proteome of mediates calcium-dependent homotypic interactions. The adhesive the cardiomyocyte AJ. Mouse neonatal cardiomyocytes assemble properties of classical cadherins are driven by the recruitment of stable AJs along intercellular contacts with organizational and cytosolic catenin proteins to the cadherin tail, with p120-catenin β structural hallmarks similar to mature contacts. We combine (CTNND1) binding to the juxta-membrane domain and -catenin β quantitative mass spectrometry with proximity labeling to identify the (CTNNB1) binding to the distal part of the tail.
    [Show full text]
  • Supplementary Table 1: Adhesion Genes Data Set
    Supplementary Table 1: Adhesion genes data set PROBE Entrez Gene ID Celera Gene ID Gene_Symbol Gene_Name 160832 1 hCG201364.3 A1BG alpha-1-B glycoprotein 223658 1 hCG201364.3 A1BG alpha-1-B glycoprotein 212988 102 hCG40040.3 ADAM10 ADAM metallopeptidase domain 10 133411 4185 hCG28232.2 ADAM11 ADAM metallopeptidase domain 11 110695 8038 hCG40937.4 ADAM12 ADAM metallopeptidase domain 12 (meltrin alpha) 195222 8038 hCG40937.4 ADAM12 ADAM metallopeptidase domain 12 (meltrin alpha) 165344 8751 hCG20021.3 ADAM15 ADAM metallopeptidase domain 15 (metargidin) 189065 6868 null ADAM17 ADAM metallopeptidase domain 17 (tumor necrosis factor, alpha, converting enzyme) 108119 8728 hCG15398.4 ADAM19 ADAM metallopeptidase domain 19 (meltrin beta) 117763 8748 hCG20675.3 ADAM20 ADAM metallopeptidase domain 20 126448 8747 hCG1785634.2 ADAM21 ADAM metallopeptidase domain 21 208981 8747 hCG1785634.2|hCG2042897 ADAM21 ADAM metallopeptidase domain 21 180903 53616 hCG17212.4 ADAM22 ADAM metallopeptidase domain 22 177272 8745 hCG1811623.1 ADAM23 ADAM metallopeptidase domain 23 102384 10863 hCG1818505.1 ADAM28 ADAM metallopeptidase domain 28 119968 11086 hCG1786734.2 ADAM29 ADAM metallopeptidase domain 29 205542 11085 hCG1997196.1 ADAM30 ADAM metallopeptidase domain 30 148417 80332 hCG39255.4 ADAM33 ADAM metallopeptidase domain 33 140492 8756 hCG1789002.2 ADAM7 ADAM metallopeptidase domain 7 122603 101 hCG1816947.1 ADAM8 ADAM metallopeptidase domain 8 183965 8754 hCG1996391 ADAM9 ADAM metallopeptidase domain 9 (meltrin gamma) 129974 27299 hCG15447.3 ADAMDEC1 ADAM-like,
    [Show full text]
  • Selective Inhibition of ADAM12 Catalytic Activity Through
    Selective inhibition of ADAM12 catalytic activity through engineering of tissue inhibitor of metalloproteinases (TIMP)-2 Marie Kveiborg, Jonas Jacobsen, Meng-Huee Lee, Hideaki Nagase, Ulla M Wewer, Gillian Murphy To cite this version: Marie Kveiborg, Jonas Jacobsen, Meng-Huee Lee, Hideaki Nagase, Ulla M Wewer, et al.. Selective inhibition of ADAM12 catalytic activity through engineering of tissue inhibitor of metalloproteinases (TIMP)-2. Biochemical Journal, Portland Press, 2010, 430 (1), pp.79-86. 10.1042/BJ20100649. hal-00506526 HAL Id: hal-00506526 https://hal.archives-ouvertes.fr/hal-00506526 Submitted on 28 Jul 2010 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Biochemical Journal Immediate Publication. Published on 10 Jun 2010 as manuscript BJ20100649 Selective inhibition of ADAM12 catalytic activity through engineering of tissue inhibitor of metalloproteinases (TIMP)-2 Marie Kveiborg*,§, Jonas Jacobsen*,§, Meng-Huee Lee†, Hideaki Nagase‡, Ulla M. Wewer*,¶, and Gillian Murphy†,¶ *Department of Biomedical Sciences and Biotech Research and Innovation Centre (BRIC), University of Copenhagen, Ole Maaløes Vej 5, 2200 Copenhagen, Denmark †Department of Oncology, Cambridge University, Cancer Research Institute, Li Ka Shing Centre, Cambridge CB2 ORE, UK ‡Kennedy Institute of Rheumatology Division, Faculty of Medicine, Imperial College London, 65 Aspenlea Road, London W6 8LH, UK §,¶These authors contributed equally to the study.
    [Show full text]
  • And MUC1 in Mammary Epithelial Cells
    Linköping University Post Print Loss of ICAM-1 signaling induces psoriasin (S100A7) and MUC1 in mammary epithelial cells Stina Petersson, E. Shubbar, M. Yhr, A. Kovacs and Charlotta Enerbäck N.B.: When citing this work, cite the original article. The original publication is available at www.springerlink.com: Stina Petersson, E. Shubbar, M. Yhr, A. Kovacs and Charlotta Enerbäck, Loss of ICAM-1 signaling induces psoriasin (S100A7) and MUC1 in mammary epithelial cells, 2011, Breast Cancer Research and Treatment, (125), 1, 13-25. http://dx.doi.org/10.1007/s10549-010-0820-4 Copyright: Springer Science Business Media http://www.springerlink.com/ Postprint available at: Linköping University Electronic Press http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-63954 1 Loss of ICAM-1 signaling induces psoriasin (S100A7) and MUC1 in mammary epithelial cells Petersson S1, Shubbar E1, Yhr M1, Kovacs A2 and Enerbäck C3 Departments of 1Clinical Genetics and 2Pathology, Sahlgrenska University Hospital, SE-413 45 Gothenburg, Sweden; 3Department of Clinical and Experimental Medicine, Division of Cell Biology and Dermatology, Linköping University, SE-581 85 Linköping, Sweden E-mail: [email protected] E-mail: maria.yhr@ clingen.gu.se E-mail: [email protected] E-mail: [email protected] Correspondence: Stina Petersson, Department of Clinical Genetics, Sahlgrenska University Hospital, SE-413 45 Gothenburg, Sweden E-mail: [email protected] 2 Abstract Psoriasin (S100A7), a member of the S100 gene family, is highly expressed in high-grade comedo ductal carcinoma in situ (DCIS), with a higher risk of local recurrence. Psoriasin is therefore a potential biomarker for DCIS with a poor prognosis.
    [Show full text]
  • Development and Validation of a Protein-Based Risk Score for Cardiovascular Outcomes Among Patients with Stable Coronary Heart Disease
    Supplementary Online Content Ganz P, Heidecker B, Hveem K, et al. Development and validation of a protein-based risk score for cardiovascular outcomes among patients with stable coronary heart disease. JAMA. doi: 10.1001/jama.2016.5951 eTable 1. List of 1130 Proteins Measured by Somalogic’s Modified Aptamer-Based Proteomic Assay eTable 2. Coefficients for Weibull Recalibration Model Applied to 9-Protein Model eFigure 1. Median Protein Levels in Derivation and Validation Cohort eTable 3. Coefficients for the Recalibration Model Applied to Refit Framingham eFigure 2. Calibration Plots for the Refit Framingham Model eTable 4. List of 200 Proteins Associated With the Risk of MI, Stroke, Heart Failure, and Death eFigure 3. Hazard Ratios of Lasso Selected Proteins for Primary End Point of MI, Stroke, Heart Failure, and Death eFigure 4. 9-Protein Prognostic Model Hazard Ratios Adjusted for Framingham Variables eFigure 5. 9-Protein Risk Scores by Event Type This supplementary material has been provided by the authors to give readers additional information about their work. Downloaded From: https://jamanetwork.com/ on 10/02/2021 Supplemental Material Table of Contents 1 Study Design and Data Processing ......................................................................................................... 3 2 Table of 1130 Proteins Measured .......................................................................................................... 4 3 Variable Selection and Statistical Modeling ........................................................................................
    [Show full text]
  • Cell Adhesion Molecules in Normal Skin and Melanoma
    biomolecules Review Cell Adhesion Molecules in Normal Skin and Melanoma Cian D’Arcy and Christina Kiel * Systems Biology Ireland & UCD Charles Institute of Dermatology, School of Medicine, University College Dublin, D04 V1W8 Dublin, Ireland; [email protected] * Correspondence: [email protected]; Tel.: +353-1-716-6344 Abstract: Cell adhesion molecules (CAMs) of the cadherin, integrin, immunoglobulin, and selectin protein families are indispensable for the formation and maintenance of multicellular tissues, espe- cially epithelia. In the epidermis, they are involved in cell–cell contacts and in cellular interactions with the extracellular matrix (ECM), thereby contributing to the structural integrity and barrier for- mation of the skin. Bulk and single cell RNA sequencing data show that >170 CAMs are expressed in the healthy human skin, with high expression levels in melanocytes, keratinocytes, endothelial, and smooth muscle cells. Alterations in expression levels of CAMs are involved in melanoma propagation, interaction with the microenvironment, and metastasis. Recent mechanistic analyses together with protein and gene expression data provide a better picture of the role of CAMs in the context of skin physiology and melanoma. Here, we review progress in the field and discuss molecular mechanisms in light of gene expression profiles, including recent single cell RNA expression information. We highlight key adhesion molecules in melanoma, which can guide the identification of pathways and Citation: D’Arcy, C.; Kiel, C. Cell strategies for novel anti-melanoma therapies. Adhesion Molecules in Normal Skin and Melanoma. Biomolecules 2021, 11, Keywords: cadherins; GTEx consortium; Human Protein Atlas; integrins; melanocytes; single cell 1213. https://doi.org/10.3390/ RNA sequencing; selectins; tumour microenvironment biom11081213 Academic Editor: Sang-Han Lee 1.
    [Show full text]
  • CDH2 and CDH11 Act As Regulators of Stem Cell Fate Decisions Stella Alimperti A, Stelios T
    Stem Cell Research (2015) 14, 270–282 Available online at www.sciencedirect.com ScienceDirect www.elsevier.com/locate/scr REVIEW CDH2 and CDH11 act as regulators of stem cell fate decisions Stella Alimperti a, Stelios T. Andreadis a,b,⁎ a Bioengineering Laboratory, Department of Chemical and Biological Engineering, University at Buffalo, State University of New York, Amherst, NY 14260-4200, USA b Center of Excellence in Bioinformatics and Life Sciences, Buffalo, NY 14203, USA Received 18 September 2014; received in revised form 24 January 2015; accepted 10 February 2015 Abstract Accumulating evidence suggests that the mechanical and biochemical signals originating from cell–cell adhesion are critical for stem cell lineage specification. In this review, we focus on the role of cadherin mediated signaling in development and stem cell differentiation, with emphasis on two well-known cadherins, cadherin-2 (CDH2) (N-cadherin) and cadherin-11 (CDH11) (OB-cadherin). We summarize the existing knowledge regarding the role of CDH2 and CDH11 during development and differentiation in vivo and in vitro. We also discuss engineering strategies to control stem cell fate decisions by fine-tuning the extent of cell–cell adhesion through surface chemistry and microtopology. These studies may be greatly facilitated by novel strategies that enable monitoring of stem cell specification in real time. We expect that better understanding of how intercellular adhesion signaling affects lineage specification may impact biomaterial and scaffold design to control stem cell fate decisions in three-dimensional context with potential implications for tissue engineering and regenerative medicine. © 2015 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
    [Show full text]
  • In Prostate Cancer Tissues of Men with Type 2 Diabetes
    biomedicines Article Increased Expressions of Matrix Metalloproteinases (MMPs) in Prostate Cancer Tissues of Men with Type 2 Diabetes Andras Franko 1,2,3 , Lucia Berti 2,3 , Jörg Hennenlotter 4 , Steffen Rausch 4, Marcus O. Scharpf 5, Martin Hrabe˘ de Angelis 3,6, Arnulf Stenzl 4 , Andreas Peter 2,3,7, Andreas L. Birkenfeld 1,2,3, Stefan Z. Lutz 1,8, Hans-Ulrich Häring 1,2,3 and Martin Heni 1,2,3,7,* 1 Department of Internal Medicine IV, Division of Diabetology, Endocrinology and Nephrology, University Hospital Tübingen, 72076 Tübingen, Germany; [email protected] (A.F.); [email protected] (A.L.B.); [email protected] (H.-U.H.) 2 Institute for Diabetes Research and Metabolic Diseases of the Helmholtz Centre Munich at the University of Tübingen, 72076 Tübingen, Germany; [email protected] 3 German Center for Diabetes Research (DZD), 85764 Neuherberg, Germany 4 Department of Urology, University Hospital Tübingen, 72076 Tübingen, Germany; [email protected] (J.H.); steff[email protected] (S.R.); [email protected] (A.S.) 5 Institute of Pathology, University Hospital Tübingen, 72076 Tübingen, Germany; [email protected] 6 Institute of Experimental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health, 85764 Neuherberg, Germany; [email protected] 7 Institute for Clinical Chemistry and Pathobiochemistry, Department for Diagnostic Laboratory Medicine, University Hospital Tübingen, 72076 Tübingen, Germany; [email protected] 8 Clinic for Geriatric and Orthopedic Rehabilitation Bad Sebastiansweiler, 72116 Mössingen, Germany; [email protected] * Correspondence: [email protected]; Tel.: +49-7071-29-82714 Received: 3 October 2020; Accepted: 12 November 2020; Published: 16 November 2020 Abstract: Type 2 diabetes (T2D) is associated with worse prognosis of prostate cancer (PCa).
    [Show full text]
  • N Cadherin (CDH2) Mouse Monoclonal Antibody [Clone ID: OTI3B10] Product Data
    OriGene Technologies, Inc. 9620 Medical Center Drive, Ste 200 Rockville, MD 20850, US Phone: +1-888-267-4436 [email protected] EU: [email protected] CN: [email protected] Product datasheet for TA503775 N Cadherin (CDH2) Mouse Monoclonal Antibody [Clone ID: OTI3B10] Product data: Product Type: Primary Antibodies Clone Name: OTI3B10 Applications: FC, IHC, WB Recommend Dilution: WB 1:2000, IHC 1:150, FLOW 1:100 Reactivity: Human Host: Mouse Isotype: IgG2a Clonality: Monoclonal Immunogen: Full length human recombinant protein of human CDH2(NP_001783) produced in HEK293T cell. Formulation: PBS (PH 7.3) containing 1% BSA, 50% glycerol and 0.02% sodium azide. Concentration: 1 mg/ml Purification: Purified from mouse ascites fluids or tissue culture supernatant by affinity chromatography (protein A/G) Predicted Protein Size: 97.2 kDa Gene Name: cadherin 2 Database Link: NP_001783 Entrez Gene 1000 Human Background: This gene is a classical cadherin from the cadherin superfamily. The encoded protein is a calcium dependent cell-cell adhesion glycoprotein comprised of five extracellular cadherin repeats, a transmembrane region and a highly conserved cytoplasmic tail. The protein functions during gastrulation and is required for establishment of left-right asymmetry. At certain central nervous system synapses, presynaptic to postsynaptic adhesion is mediated at least in part by this gene product. [provided by RefSeq] Synonyms: CD325; CDHN; CDw325; NCAD Protein Families: Druggable Genome, ES Cell Differentiation/IPS, Transmembrane Protein Pathways: Arrhythmogenic right ventricular cardiomyopathy (ARVC), Cell adhesion molecules (CAMs) This product is to be used for laboratory only. Not for diagnostic or therapeutic use. View online » ©2020 OriGene Technologies, Inc., 9620 Medical Center Drive, Ste 200, Rockville, MD 20850, US 1 / 3 N Cadherin (CDH2) Mouse Monoclonal Antibody [Clone ID: OTI3B10] – TA503775 Product images: HEK293T cells were transfected with the pCMV6- ENTRY control (Left lane) or pCMV6-ENTRY CDH2 ([RC207170], Right lane) cDNA for 48 hrs and lysed.
    [Show full text]
  • Novel Alternatively Spliced ADAM8 Isoforms Contribute to the Aggressive Bone Metastatic Phenotype of Lung Cancer
    Oncogene (2010) 29, 3758–3769 & 2010 Macmillan Publishers Limited All rights reserved 0950-9232/10 www.nature.com/onc ORIGINAL ARTICLE Novel alternatively spliced ADAM8 isoforms contribute to the aggressive bone metastatic phenotype of lung cancer I Herna´ndez1, JL Moreno2, C Zandueta1, L Montuenga3,4 and F Lecanda1 1Adhesion and Metastasis Laboratory, Division of Oncology, University of Navarra, Pamplona, Spain; 2Department of Orthopaedics, University of Maryland School of Medicine, Baltimore, MD, USA; 3Department of Histology and Pathology, School of Medicine, University of Navarra, Pamplona, Spain and 4Biomarkers Laboratory, Center for Applied Biomedical Research (CIMA), University of Navarra, Pamplona, Spain ADAMs (a disintegrin and metalloprotease) are trans- survival rates for lung cancer are o15% in all developed membrane proteins involved in a variety of physiological countries. It is estimated that 30–40% of lung cancer processes and tumorigenesis. Recently, ADAM8 has been patients with advanced NSCLC suffer from bone associated with poor prognosis of lung cancer. However, metastasis (Coleman, 1997). Patients with bone metas- its contribution to tumorigenesis in the context of lung tasis experience pain, metabolic syndromes and spinal cancer metastasis remains unknown. Native ADAM8 cord compression associated with pathological fractures expression levels were lower in lung cancer cell lines. as a consequence of osteolytic lesions. In contrast, we identified and characterized two novel ADAMs (a disintegrin and metalloprotease) form a spliced isoforms encoding truncated proteins, D18a and large family of cell-surface proteins, which are char- D140, which were present in several tumor cell lines and not acterized by disintegrin and metalloproteinase domains, in normal cells. Overexpression of D18a protein resulted in that possess adhesive properties and proteolytic activ- enhanced invasive activity in vitro.ADAM8anditsD140 ities, respectively (Lu et al., 2007).
    [Show full text]