DOCSLIB.ORG

Fibrosis and Immune Cell Infiltration Are Separate Events Regulated By

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Fibrosis and Immune Cell Infiltration Are Separate Events Regulated By BASIC RESEARCH www.jasn.org Fibrosis and Immune Cell Infiltration Are Separate Events Regulated by Cell-Specific Receptor Notch3 Expression Sabine Brandt,1,2 Tobias M. Ballhause ,1 Anja Bernhardt,1,2 Annika Becker,1 Delia Salaru,1 Hien Minh Le-Deffge,1 Alexander Fehr,1,2 Yan Fu,2,3 Lars Philipsen ,2,3 Sonja Djudjaj,4 Andreas J. Müller,2,3,5 Rafael Kramann ,6,7 Mahmoud Ibrahim,6 Robert Geffers,8 Chris Siebel,9 Berend Isermann,2,10 Florian H. Heidel,11,12,13 Jonathan A. Lindquist ,1,2 and Peter R. Mertens1,2 Due to the number of contributing authors, the affiliations are listed at the end of this article. ABSTRACT Background Kidney injuries that result in chronic inflammation initiate crosstalk between stressed resident cells and infiltrating immune cells. In animal models, whole-body receptor Notch3 deficiency protects from leukocyte infiltration and organ fibrosis. However, the relative contribution of Notch3 expression in tissue versus infiltrating immune cells is unknown. Methods Chimeric mice deficient for Notch3 in hematopoietic cells and/or resident tissue cells were generated, and kidney fibrosis and inflammation after unilateral ureteral obstruction (UUO) were analyzed. Adoptive transfer of labeled bone marrow–derived cells validated the results in a murine Leishmania ear infection model. In vitro adhesion assays, integrin activation, and extracellular matrix production were analyzed. Results Fibrosis follows UUO, but inflammatory cell infiltration mostly depends upon Notch3 expression in hematopoietic cells, which coincides with an enhanced proinflammatory milieu (e.g., CCL2 and CCL5 1 upregulation). Notch3 expression on CD45 leukocytes plays a prominent role in efficient cell transmi- gration. Functionally, leukocyte adhesion and integrin activation are abrogated in the absence of receptor Notch3. Chimeric animal models also reveal that tubulointerstitial fibrosis develops, even in the absence of prominent leukocyte infiltrates after ureteral obstruction. Deleting Notch3 receptors on resident cells blunts kidney fibrosis, ablates NF-kB signaling, and lessens matrix deposition. Conclusions Cell-specific receptor Notch3 signaling independently orchestrates leukocyte infiltration and organ fibrosis. Interference with Notch3 signaling may present a novel therapeutic approach in inflamma- tory as well as fibrotic diseases. JASN 31: ccc–ccc, 2020. doi: https://doi.org/10.1681/ASN.2019121289 Chronic inflammation of solid organs often results in destruction of organ architecture and perpetu- ated fibrosis, with functional deficits and ultimately Received December 13, 2019. Accepted July 12, 2020. organ failure.1,2 The list of injuries causing inflam- Published online ahead of print. Publication date available at mation includes acute and chronic infections, toxic www.jasn.org. agents, autoimmune diseases, and metabolic alter- Correspondence: Dr. Peter R. Mertens, Clinic of Nephrology and 3–6 ations (e.g., hyperglycemia and uremia). Key Hypertension, Diabetes and Endocrinology, Otto-von-Guericke events after primary insults are stress responses of University Magdeburg, Leipziger Strasse 44, 39120 Magdeburg, resident organ cells and leukocyte recruitment. The Germany. Email: [email protected] composition of the infiltrate depends on the timing Copyright © 2020 by the American Society of Nephrology JASN 31: ccc–ccc, 2020 ISSN : 1046-6673/3111-ccc 1 BASIC RESEARCH www.jasn.org and severity of the immune response, which encompasses de- Significance Statement fense mechanisms, such as complement activation, phagocy- tosis, and antigen presentation.7 Approximately 15% of adults In patients with CKD, receptor Notch3 is strongly upregulated. within the United States (37 million people) have CKD.8 De- Conversely, in experimental kidney disease models, Notch3 de- fi spite the enormity of this problem, therapeutic options are ciency protects from organ damage. To determine whether Notch3 on immune cells or tissue-resident cells participates in the scarce and mostly ineffective. Therefore, an improved under- inflammatory response, animals with bone marrow chimerism were standing of the cellular and molecular mechanisms leading to generated. These animal strains do not exhibit phenotypic differ- inflammation and renal fibrosis is essential to develop strate- ences in the absence of disease. However, after unilateral ureteral gies to combat these processes, because fibrosis-related organ obstruction, distinct alterations in the immune response and organ fi diseases account for almost 50% of all deaths.9–12 Currently, brosis become apparent. Notch3 receptors expressed by immune fi cells are of relevance for transmigration into tissue; the receptors established models of renal brosis propose that kidney injury expressed by resident kidney cells orchestrate organ fibrosis. These leads to immune cell infiltration, fibroblast activation, extra- events seem to be separable and distinct. cellular matrix (ECM) deposition, microvascular rarefaction, and tubular atrophy.9 Inflammation is considered the driving force in fibrotic diseases, with an intimate crosstalk between were housed and bred in the Central Animal Facility of the resident and infiltrating cells; therefore, targeting immune cell Otto-von-Guericke University Magdeburg Medical Faculty. fi infiltration is deemed to be a therapeutic option.13–15 How- The mice were kept under speci c pathogen-free conditions ever, it is not known whether immune cell recruitment is the in individual, ventilated cages (Techniplast, Buguggiate, Italy). culprit of organ fibrosis, or a bystander that modulates repar- All experiments and procedures were conducted in accor- ative processes.1 dance with the German National Guidelines for the Use of In CKD, the Notch receptor family plays a key role in both Experimental Animals (Animal Protection Act) and were ap- inflammatory responses and organ fibrosis.16–19 Animals proved by the state of Sachsen-Anhalt (Aktenzeichen UniMD deficient in receptors Notch1 and -3 demonstrate similar 42502-2-1135 and UniMD 42502-2-1253). All mouse strains fi protective effects on organ fibrosis. Notch3-deficient mice were con rmed by PCR genotyping that was performed as 18 fl exhibit significantly reduced kidney fibrosis (.50%), fewer previously described. Brie y, the mouse tail was lysed with m m myofibroblasts (approximately 50%), and fewer infiltrating 200 l direct PCR tail lysis buffer and 50 g of proteinase immune cells (approximately 30% of total leukocytes) com- K. PCR was performed using primer pairs for WT (forward, pared with wild-type (WT) mice after ureteral obstruc- CCATGAGGATGCTATCTGTGAC; reverse, CACATTGGC tion.18 The inflammatory chemokines CCL2 and CCL5 are ACAAGAATGAGCC) and Notch3 knockout (forward, TCG reduced by .50% in Notch3-deficient mice.18 Similar find- CCTTCTTGACGAGTTCT; reverse, GCGATGCAATTTCCT ings are reported in Notch3-deficient animals undergoing CATTT). The PCR cycler performed 35 cycles at 94°C, 60°C, nephrotoxic serum nephritis and ischemia reperfusion. and 72°C (each for 60 seconds). Products were separated on Thus, genetic ablation of receptor Notch3 protects mice 1% agarose/ethidium bromide gels. from kidney damage.18–20 However, it is not clear whether this protection is mediated Leishmania major Infection by Notch3 deficiency on immune cells or by a lack of Notch3 Leishmania major inoculation was performed as described.23 expression on tissue-resident cells. Here, we use bone marrow Briefly, dsRed-expressing parasites were grown at 26°C, for a (BM) chimeras to clarify cell type–specific roles of receptor maximum of five passages, in M119 medium supplemented Notch3 and discover that Notch3 receptors are not only rele- with 10% heat-inactivated FBS, 0.1 mM adenine, 1 mg/ml biotin, vant for leukocyte transmigration, but are also key molecules 5mg/mlhemin,and2mg/ml biopterin (all from Sigma-Aldrich, 5 for the fibrogenic niche. Unexpectedly, organ fibrosis is not Darmstadt, Germany). For infection, 10 stationary-phase pro- reduced in the absence of prominent leukocyte infiltration and mastigotes were resuspended in 10 ml PBS and injected into the vice versa. ear dermis. Experimental Kidney Disease METHODS Unilateral ureter obstruction (UUO) was performed with sex- matched, 12- to 16-week-old mice. The right ureters of WT Animals and knockout mice were ligated for 5 and 14 days. Both kid- Studies were performed using Notch3-knockout animals and neys were collected for preparation of cortical RNA, protein their WT littermates in a C57BL/6J (CD45.2) background lysates, immunohistochemistry, and flow cytometry (FACS). (kindly provided by Dr. Anne Joutel, Institut National de la Blood sampling by heart puncture was subjected to complete Santé et de la Recherche Médicale).21,22 The commercially blood cell counting (ADVIA 120; Bayer Diagnostics Munich, available CD45.1 mice are C57BL/6J background. The health Germany) and plasma specimens were stored at 280°C until of the mice corresponded to the Federation of European Lab- further analysis. Contralateral kidneys were harvested for oratory Animal Science Association guidelines. The animals comparative analyses. 2 JASN JASN 31: ccc–ccc,2020 www.jasn.org BASIC RESEARCH Table 1. List of primary antibodies Primary Antibodies Catalog No. Dilution Company Dye Immunhistochemistry and immunofluorescence Gli1 (IF) AF3455 1:200 R&D Systems Notch3 (IHC) ab23426 1:200 Abcam F4/80 (IHC) MON3099 1:100 Monosan Western blotting Notch3 sc-5539 1:500 Santa Cruz TNC ab108930 1:500 Abcam Tubulin T5168 1:1000 Sigma Phospho–NF-kB–p65 (S536) 3033 1:1000 Cell Signaling Phospho-IkBa
Load more

Recommended publications
  • Association of Notch and Hedgehog Pathway Activation with Prognosis
    ANTICANCER RESEARCH 39 : 2129-2138 (2019) doi:10.21873/anticanres.13326 Association of Notch and Hedgehog Pathway Activation With Prognosis in Early-stage Colorectal Cancer GRIGORIOS RALLIS 1, TRIANTAFYLLIA KOLETSA 2, ZENIA SARIDAKI 3, KYRIAKI MANOUSOU 4, GEORGIA-ANGELIKI KOLIOU 4, IOANNIS KOSTOPOULOS 2, VASSILIKI KOTOULA 2,5 , THOMAS MAKATSORIS 6, HELEN P. KOUREA 7, GEORGIA RAPTOU 2, SOFIA CHRISAFI 3, EPAMINONTAS SAMANTAS 8, KLEO PAPAPARASKEVA 9, ELISSAVET PAZARLI 10 , PAVLOS PAPAKOSTAS 11 , GEORGIA KAFIRI 12 , DAVIDE MAURI 13 , ALEXANDRA PAPOUDOU-BAI 14 , CHRISTOS CHRISTODOULOU 15 , KALLIOPI PETRAKI 16 , NIKOLAOS DOMBROS 17 , DIMITRIOS PECTASIDES 18 and GEORGE FOUNTZILAS 5,17 1Department of Medical Oncology, School of Health Sciences, Faculty of Medicine, Papageorgiou Hospital, Aristotle University of Thessaloniki, Thessaloniki, Greece; 2Department of Pathology, School of Health Sciences, Faculty of Medicine, Aristotle University of Thessaloniki, Thessaloniki, Greece; 3Asklepios Oncology Department, Heraklion, Greece; 4Section of Biostatistics, Hellenic Cooperative Oncology Group, Data Office, Athens, Greece; 5Laboratory of Molecular Oncology, Hellenic Foundation for Cancer Research/Aristotle University of Thessaloniki, Thessaloniki, Greece; 6Division of Oncology, Department of Medicine, University Hospital, University of Patras Medical School, Patras, Greece; 7Department of Pathology, University Hospital of Patras, Patras, Greece; 8Third Department of Medical Oncology, Agii Anargiri Cancer Hospital, Athens, Greece; 9Department of Pathology,
    [Show full text]
  • Coronary Arterial Development Is Regulated by a Dll4-Jag1-Ephrinb2 Signaling Cascade
    RESEARCH ARTICLE Coronary arterial development is regulated by a Dll4-Jag1-EphrinB2 signaling cascade Stanislao Igor Travisano1,2, Vera Lucia Oliveira1,2, Bele´ n Prados1,2, Joaquim Grego-Bessa1,2, Rebeca Pin˜ eiro-Sabarı´s1,2, Vanesa Bou1,2, Manuel J Go´ mez3, Fa´ tima Sa´ nchez-Cabo3, Donal MacGrogan1,2*, Jose´ Luis de la Pompa1,2* 1Intercellular Signalling in Cardiovascular Development and Disease Laboratory, Centro Nacional de Investigaciones Cardiovasculares Carlos III (CNIC), Madrid, Spain; 2CIBER de Enfermedades Cardiovasculares, Madrid, Spain; 3Bioinformatics Unit, Centro Nacional de Investigaciones Cardiovasculares, Madrid, Spain Abstract Coronaries are essential for myocardial growth and heart function. Notch is crucial for mouse embryonic angiogenesis, but its role in coronary development remains uncertain. We show Jag1, Dll4 and activated Notch1 receptor expression in sinus venosus (SV) endocardium. Endocardial Jag1 removal blocks SV capillary sprouting, while Dll4 inactivation stimulates excessive capillary growth, suggesting that ligand antagonism regulates coronary primary plexus formation. Later endothelial ligand removal, or forced expression of Dll4 or the glycosyltransferase Mfng, blocks coronary plexus remodeling, arterial differentiation, and perivascular cell maturation. Endocardial deletion of Efnb2 phenocopies the coronary arterial defects of Notch mutants. Angiogenic rescue experiments in ventricular explants, or in primary human endothelial cells, indicate that EphrinB2 is a critical effector of antagonistic Dll4 and Jag1 functions in arterial morphogenesis. Thus, coronary arterial precursors are specified in the SV prior to primary coronary plexus formation and subsequent arterial differentiation depends on a Dll4-Jag1-EphrinB2 signaling *For correspondence: [email protected] (DMG); cascade. [email protected] (JLP) Competing interests: The authors declare that no Introduction competing interests exist.
    [Show full text]
  • Updates on the Role of Molecular Alterations and NOTCH Signalling in the Development of Neuroendocrine Neoplasms
    Journal of Clinical Medicine Review Updates on the Role of Molecular Alterations and NOTCH Signalling in the Development of Neuroendocrine Neoplasms 1,2, 1, 3, 4 Claudia von Arx y , Monica Capozzi y, Elena López-Jiménez y, Alessandro Ottaiano , Fabiana Tatangelo 5 , Annabella Di Mauro 5, Guglielmo Nasti 4, Maria Lina Tornesello 6,* and Salvatore Tafuto 1,* On behalf of ENETs (European NeuroEndocrine Tumor Society) Center of Excellence of Naples, Italy 1 Department of Abdominal Oncology, Istituto Nazionale Tumori, IRCCS Fondazione “G. Pascale”, 80131 Naples, Italy 2 Department of Surgery and Cancer, Imperial College London, London W12 0HS, UK 3 Cancer Cell Metabolism Group. Centre for Haematology, Immunology and Inflammation Department, Imperial College London, London W12 0HS, UK 4 SSD Innovative Therapies for Abdominal Metastases—Department of Abdominal Oncology, Istituto Nazionale Tumori, IRCCS—Fondazione “G. Pascale”, 80131 Naples, Italy 5 Department of Pathology, Istituto Nazionale Tumori, IRCCS—Fondazione “G. Pascale”, 80131 Naples, Italy 6 Unit of Molecular Biology and Viral Oncology, Department of Research, Istituto Nazionale Tumori IRCCS Fondazione Pascale, 80131 Naples, Italy * Correspondence: [email protected] (M.L.T.); [email protected] (S.T.) These authors contributed to this paper equally. y Received: 10 July 2019; Accepted: 20 August 2019; Published: 22 August 2019 Abstract: Neuroendocrine neoplasms (NENs) comprise a heterogeneous group of rare malignancies, mainly originating from hormone-secreting cells, which are widespread in human tissues. The identification of mutations in ATRX/DAXX genes in sporadic NENs, as well as the high burden of mutations scattered throughout the multiple endocrine neoplasia type 1 (MEN-1) gene in both sporadic and inherited syndromes, provided new insights into the molecular biology of tumour development.
    [Show full text]
  • NOTCH3 Gene Notch 3
    NOTCH3 gene notch 3 Normal Function The NOTCH3 gene provides instructions for making a protein with one end (the intracellular end) that remains inside the cell, a middle (transmembrane) section that spans the cell membrane, and another end (the extracellular end) that projects from the outer surface of the cell. The NOTCH3 protein is called a receptor protein because certain other proteins, called ligands, attach (bind) to the extracellular end of NOTCH3, fitting like a key into a lock. This binding causes detachment of the intracellular end of the NOTCH3 protein, called the NOTCH3 intracellular domain, or NICD. The NICD enters the cell nucleus and helps control the activity (transcription) of other genes. The NOTCH3 protein plays a key role in the function and survival of vascular smooth muscle cells, which are muscle cells that surround blood vessels. This protein is thought to be essential for the maintenance of blood vessels, including those that supply blood to the brain. Health Conditions Related to Genetic Changes Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy More than 270 mutations in the NOTCH3 gene have been found to cause cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy, commonly known as CADASIL. Almost all of these mutations change a single protein building block (amino acid) in the NOTCH3 protein. The amino acid involved in most mutations is cysteine. The addition or deletion of a cysteine molecule in a certain area of the NOTCH3 protein, known as the EGF-like domain, presumably affects NOTCH3 function in vascular smooth muscle cells. Disruption of NOTCH3 functioning can lead to the self-destruction (apoptosis) of these cells.
    [Show full text]
  • Repressor Activity in Notch 3 3927
    Development 126, 3925-3935 (1999) 3925 Printed in Great Britain © The Company of Biologists Limited 1999 DEV9635 The Notch 3 intracellular domain represses Notch 1-mediated activation through Hairy/Enhancer of split (HES) promoters Paul Beatus, Johan Lundkvist, Camilla Öberg and Urban Lendahl* Department of Cell and Molecular Biology, Medical Nobel Institute, Karolinska Institute, S-171 77 Stockholm, Sweden *Author for correspondence (e-mail: [email protected]) Accepted 9 June; published on WWW 5 August 1999 SUMMARY The Notch signaling pathway is important for cellular levels. First, Notch 3 IC competes with Notch 1 IC for differentiation. The current view is that the Notch receptor access to RBP-Jk and does not activate transcription when is cleaved intracellularly upon ligand activation. The positioned close to a promoter. Second, Notch 3 IC appears intracellular Notch domain then translocates to the to compete with Notch 1 IC for a common coactivator nucleus, binds to Suppressor of Hairless (RBP-Jk in present in limiting amounts. In conclusion, this is the first mammals), and acts as a transactivator of Enhancer of Split example of a Notch IC that functions as a repressor in (HES in mammals) gene expression. In this report we show Enhancer of Split/HES upregulation, and shows that that the Notch 3 intracellular domain (IC), in contrast to mammalian Notch receptors have acquired distinct all other analysed Notch ICs, is a poor activator, and in fact functions during evolution. acts as a repressor by blocking the ability of the Notch 1 IC to activate expression through the HES-1 and HES-5 promoters.
    [Show full text]
  • Off the Beaten Pathway: the Complex Cross Talk Between Notch and NF-Kb Clodia Osipo1,2, Todd E Golde3, Barbara a Osborne4 and Lucio a Miele1,2,5
    Laboratory Investigation (2008) 88, 11–17 & 2008 USCAP, Inc All rights reserved 0023-6837/08 $30.00 PATHOBIOLOGY IN FOCUS Off the beaten pathway: the complex cross talk between Notch and NF-kB Clodia Osipo1,2, Todd E Golde3, Barbara A Osborne4 and Lucio A Miele1,2,5 The canonical Notch pathway that has been well characterized over the past 25 years is relatively simple compared to the plethora of recently published data suggesting non-canonical signaling mechanisms and cross talk with other pathways. The manner in which other pathways cross talk with Notch signaling appears to be extraordinarily complex and, not surprisingly, context-dependent. While the physiological relevance of many of these interactions remains to be estab- lished, there is little doubt that Notch signaling is integrated with numerous other pathways in ways that appear increasingly complex. Among the most intricate cross talks described for Notch is its interaction with the NF-kB pathway, another major cell fate regulatory network involved in development, immunity, and cancer. Numerous reports over the last 11 years have described multiple cross talk mechanisms between Notch and NF-kB in diverse experimental models. This article will provide a brief overview of the published evidence for Notch–NF-kB cross talk, focusing on vertebrate systems. Laboratory Investigation (2008) 88, 11–17; doi:10.1038/labinvest.3700700; published online 3 December 2007 KEYWORDS: Notch signaling; NF-kB cross talk; non-canonical signaling; IKK kinases CANONICAL NOTCH SIGNALING endocytosed into the ligand-expressing cell.10 This unmasks Canonical Notch signaling has been recently reviewed by the HD and triggers an extracellular cleavage in it by ADAM several authors,1–6 and the reader is referred to these reviews (a disintegrin and metalloproteinase) 10 or 17,1,2 followed by for detailed information and additional references.
    [Show full text]
  • Notch 2 (M-20): Sc-7423
    SAN TA C RUZ BI OTEC HNOL OG Y, INC . Notch 2 (M-20): sc-7423 BACKGROUND SELECT PRODUCT CITATIONS The LIN-12/Notch family of transmembrane receptors is believed to play a 1. Nijjar, S.S., et al. 2002. Altered Notch ligand expression in human liver central role in development by regulating cell fate decisions. To date, four disease: further evidence for a role of the Notch signaling pathway in notch homologs have been identified in mammals and have been designated hepatic neovascularization and biliary ductular defects. Am. J. Pathol. 160: Notch 1, Notch 2, Notch 3 and Notch 4. The notch genes are expressed in a 1695-1703. variety of tissues in both the embryonic and adult organism, suggesting that 2. Tsunematsu, R., et al. 2004. Mouse Fbw7/SEL-10/Cdc4 is required for the genes are involved in multiple signaling pathways. The notch proteins Notch degradation during vascular development. J. Biol. Chem. 279: have been found to be overexpressed or rearranged in human tumors. Ligands 9417-9423. for notch include Jagged1, Jagged2 and Delta. Jagged can activate notch and prevent myoblast differentiation by inhibiting the expression of muscle 3. Parr, C., et al. 2004. The possible correlation of Notch receptors, Notch 1 regulatory and structural genes. Jagged2 is thought to be involved in the and Notch 2, with clinical outcome and tumour clinicopathological development of various tissues whose development is dependent upon epithe - para- meters in human breast cancers. Int. J. Mol. Med. 14: 779-786. lial-mesenchymal interactions. Normal Delta expression is restricted to the 4.
    [Show full text]
  • Corporate Medical Policy Template
    Corporate Medical Policy Genetic Testing of CADASIL Syndrome AHS – M2069 “Notification” File Name: genetic_testing_of_cadasil_syndrome Origination: 01/01/2019 Last CAP Review: N/A Next CAP Review: 01/01/2020 Last Review: 01/01/2019 Policy Effective April 1, 2019 Description of Procedure or Service Definitions Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is the most common form of hereditary cerebral angiopathy. It is caused by mutations in the NOTCH3 gene located on chromosome 19p13. CADASIL resulting in a clinical syndrome of migraines (frequently with aura), progressive strokes, and cognitive decline in adults leading to severe functional impairment by the seventh decade of life(Zhu & Nahas, 2016). ***Note: This Medical Policy is complex and technical. For questions concerning the technical language and/or specific clinical indications for its use, please consult your physician. Policy BCBSNC will provide coverage for genetic testing of CADASIL syndrome when it is determined to be medically necessary because the medical criteria and guidelines shown below are met. Benefits Application This medical policy relates only to the services or supplies described herein. Please refer to the Member's Benefit Booklet for availability of benefits. Member's benefits may vary according to benefit design; therefore member benefit language should be reviewed before applying the terms of this medical policy. When Genetic Testing of CADASIL Syndrome is covered 1. Genetic testing to confirm the diagnosis of CADASIL syndrome is considered medically necessary under the following conditions: A. Clinical signs, symptoms, and imaging results are consistent with CADASIL, indicating that the pre-test probability of CADASIL is at least in the moderate to high range (See policy guidelines for further details) B.
    [Show full text]
  • Notch3 Intracellular Domain Accumulates in Hepg2 Cell Line
    ANTICANCER RESEARCH 26: 2123-2128 (2006) Notch3 Intracellular Domain Accumulates in HepG2 Cell Line CATIA GIOVANNINI1,2, MICHELA LACCHINI2, LAURA GRAMANTIERI1,2, PASQUALE CHIECO2 and LUIGI BOLONDI1,2 1Department of Internal Medicine and Gastroenterology, University of Bologna and S. Orsola-Malpighi University Hospital, 40138 Bologna; 2Center for Applied Biomedical Research (CRBA), S. Orsola-Malpighi University Hospital, 40138 Bologna, Italy Abstract. Background: By mediating local cell-cell epithelial growth factor repeats and a lin-12 Notch repeat interactions, the Notch signaling pathway seems to control a as well as a transmembrane subunit (NTM 97-120 kDa) variety of processes from cell fate decisions during containing a short extracellular fragment and an development, to stem cell renewal and to differentiation in intracellular domain (NICD 65-110 kDa) (1). These non- many adult tissues. Hence, perturbed Notch signaling may covalently associated subunits are presented as a be involved both in the development and the spread of heterodimeric functional receptor at the cell surface. Even cancer. The expression and the functional role of some though ligand-independent activation of the Notch major components of the Notch signaling pathway in signaling pathway has been described (2), Notch receptors human hepatocellular carcinoma (HCC) are poorly are mainly activated by transmembrane ligands expressed characterized. Materials and Methods: Notch3, HES1, on the surface of neighboring cells. Five ligands of Notch Jagged1 and Delta1 were analyzed both at the RNA and receptors have been described in vertebrates: Delta-like 1, protein levels in the HepG2 liver cell line derived from 3 and 4 and Jagged 1 and 2 (3, 4).
    [Show full text]
  • Association of a Notch 3 Gene Polymorphism with Migraine Susceptibility
    Association of a Notch 3 gene polymorphism with migraine susceptibility Author Menon, Saras, Cox, Hannah, Kuwahata, Melissa, Quinlan, Sharon, MacMillan, J, Haupt, Larisa, Lea, Rodney, Griffiths, Lyn Published 2010 Journal Title Cephalalgia DOI https://doi.org/10.1177/0333102410381143 Copyright Statement © 2010 International Headache Society. This is the author-manuscript version of the paper. Reproduced in accordance with the copyright policy of the publisher. Please refer to the journal's website for access to the definitive, published version. Downloaded from http://hdl.handle.net/10072/36796 Griffith Research Online https://research-repository.griffith.edu.au Association of a Notch 3 gene polymorphism with migraine susceptibility S. Menon1, H.C. Cox 1, M. Kuwahata1 , S. Quinlan1 , J. C. MacMillan 2 , L. M. Haupt1 , R.A. Lea 1,3, and L. R. Griffiths 1 1Genomics Research Centre, Griffith Institute of Health and Medical Research, Griffith University Gold Coast, PMB 50, Gold Coast Mail Centre, Queensland, Australia, 4215 2Department of Medicine, University of Queensland Graduate School of Medicine, and Queensland Institute of Medical Research, Herston, Brisbane, Australia 3Institute of Environmental Science and Research, 34 Kenepuru Drive, Porirua Wellington, New Zealand Communicating Author: Professor Lyn Griffiths Genomics Research Centre Griffith Institute of Health and Medical Research Griffith University, Gold Coast Queensland, Australia, 4215 Email: [email protected] Telephone: +61-7-55528664 Fax: +61-7-55948908 Abstract Cerebral Autosomal Dominant Arteriopathy with Sub cortical infarcts and leucoencephalopathy (CADASIL) shares common symptoms with migraine. Most CADASIL causative mutations occur in exons 3 and 4 of the Notch 3 gene. This study investigated the role of C381T (rs 3815188) and G684A (rs 1043994) single nucleotide polymorphisms (SNP) in exons 3 and 4 respectively of the Notch 3 gene in migraine.
    [Show full text]
  • Cadasil – Genetic and Ultrastructural Diagnosis Case Report
    Dement Neuropsychol 2015 December;9(4):428-432 Case Report Cadasil – genetic and ultrastructural diagnosis Case report Julio Cesar Vasconcelos da Silva1, Leila Chimelli2, Felipe Kenji Sudo3, Eliasz Engelhardt4 ABSTRACT. Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy (CADASIL) is a hereditary disorder which affects the cerebral vasculature due to mutations in the NOTCH 3 gene. The diagnosis may be established through genetic testing for detection of these mutations and/or by skin biopsy. We report a case of the disorder in a female patient, who presented recurrent transient ischemic attacks that evolved to progressive subcortical dementia. Neuroimaging disclosed extensive leukoaraiosis and lacunar infarcts. The genetic analysis for NOTCH 3 was confirmatory. The ultrastructural examination of the skin biopsy sample, initially negative, confirmed the presence of characteristic changes (presence of granular osmiophilic material inclusions [GOM]), after the analysis of new sections of the same specimen. The present findings indicate that negative findings on ultrastructural examinations of biopsy should not exclude the diagnosis of the disease and that further analyses of the sample may be necessary to detect the presence of GOM. Key words: CADASIL, skin biopsy, granular osmiophilic material, NOTCH 3. CADASIL: DIAGNÓSTICO GENÉTICO E ULTRAESTRUTURAL. RELATO DE CASO RESUMO. Arteriopatia Cerebral Autossômica Dominante com Infartos Subcorticais e Leucoencefalopatia (CADASIL) é uma desordem hereditária que acomete a vasculatura cerebral devido a mutações no gene do NOTCH 3. O diagnóstico pode ser estabelecido através de testes genéticos para a detecção das mutações e/ou de biópsia de pele. É relatado um caso da desordem em paciente feminino, que apresentou ataques isquêmicos transitórios recorrentes, evoluindo para um quadro progressivo de demência subcortical.
    [Show full text]
  • Prognostic Significance of Notch Ligands in Patients with Non‑Small Cell Lung Cancer
    506 ONCOLOGY LETTERS 13: 506-510, 2017 Prognostic significance of Notch ligands in patients with non‑small cell lung cancer 1 2 JOANNA PANCEWICZ-WOJTKIEWICZ , ANDRZEJ ELJASZEWICZ , 3 1 3 OKSANA KOWALCZUK , WIESLAWA NIKLINSKA , RADOSLAW CHARKIEWICZ , 4 1 2 MIROSLAW KOZŁOWSKI , AGNIESZKA MIASKO and MARCIN MONIUSZKO Departments of 1Histology and Embryology, 2Regenerative Medicine and Immune Regulation, 3Clinical Molecular Biology and 4Thoracic Surgery, Medical University of Bialystok, 15-269 Bialystok, Poland Received June 14, 2016; Accepted September 29, 2016 DOI: 10.3892/ol.2016.5420 Abstract. The Notch signaling pathway is deregulated in cancer patients are diagnosed with non-small-cell lung cancer numerous solid types of cancer including non-small cell (NSCLC) (3). Currently, lung cancer therapy is mainly based lung cancer (NSCLC). However, the profile of Notch ligand on Tumor-Node-Metastasis (TNM) disease staging and expression remains unclear. Therefore, the present study tumor histological classification. However, despite progress aimed to determine the profile of Notch ligands in NSCLC in surgical techniques, chemotherapy and radiotherapy, the patients and to investigate whether quantitative assessment 5-year survival rate of patients with lung cancer remains low of Notch ligand expression may have prognostic significance (~16%) (4,5). Therefore, there is a continuous need to identify in NSCLC patients. The study was performed in 61 pairs of specific and sensitive biomarkers that may improve cancer tumor and matched unaffected lung tissue specimens obtained patient management. Such markers should allow prediction from patients with various stages of NSCLC, which were and prognostication of patient survival, disease free survival analyzed by reverse transcription-polymerase chain reac- or treatment response (6).
    [Show full text]