DOCSLIB.ORG

Suv420h2 Ko Mice Show Enlarged Spleen

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Suv420h2 Ko Mice Show Enlarged Spleen Aus dem Adolf-Butenandt-Institut der Ludwig-Maximilians-Universität München Lehrstuhl:Molekularbiologie Direktor: Prof. Dr. Peter B. Becker Arbeitsgruppe: Prof. Dr. Gunnar Schotta The role of repressive chromatin functions during haematopoiesis Dissertation zum Erwerb des Doktorgrades der Naturwissenschaften (Dr. rer. nat.) an der Medizinischen Fakultät der Ludwig-Maximilians-Universität München vorgelegt von Alessandra Pasquarella aus Potenza 2014 II III Gedruckt mit Genehmigung der Medizinischen Fakultät der Ludwig-Maximilians-Universität München Betreuer: Prof. Dr. Gunnar Schotta Zweitgutachter: Prof. Dr. Rainer Haas Dekan: Prof. Dr. med. Dr. h. c. M. Reiser, FACR, FRCR Tag der mündlichen Prüfung: 16.04.2015 IV V Eidesstattliche Versicherung Ich erkläre hiermit an Eides statt, dass ich die vorliegende Dissertation mit dem Thema “The role of repressive chromatin functions during haematopoiesis” selbständig verfasst, mich außer der angegebenen keiner weiteren Hilfsmittel bedient und alle Erkenntnisse, die aus dem Schrifttum ganz oder annähernd übernommen sind, als solche kenntlich gemacht und nach ihrer Herkunft unter Bezeichnung der Fundstelle einzeln nachgewiesen habe. Ich erkläre des Weiteren, dass die hier vorgelegte Dissertation nicht in gleicher oder in ähnlicher Form bei einer anderen Stelle zur Erlangung eines akademischen Grades eingereicht wurde. ____________________ _____________________________ Ort, Datum Unterschrift Alessandra Pasquarella VI My doctoral work focused on the description and analysis of the role of repressive histone modifications during haematopoiesis. This thesis includes the three main projects which I followed during my PhD. Although two of them still require further investigation to be suitable for publication, the results described in the section “SETDB1-mediated silencing of MuLVs is essential for B cell development” have already been assembled in a manuscript which is ready for submission. VII Table of contents I. ABSTRACT .................................................................................................... 1 II. ZUSAMMENFASSUNG ............................................................................ 3 1 INTRODUCTION .......................................................................................... 6 1.1 Epigenetic modifications: several ways to regulate gene expression ....................... 6 1.1.1 DNA methylation ...................................................................................................... 6 1.1.2 Nucleosome positioning and histone modifications ................................................. 8 1.1.3 Histone modifications during development ............................................................ 10 1.2 Haematopoiesis: a good tool to study chromatin functions ................................... 14 1.2.1 Central haematopoiesis ........................................................................................... 15 1.2.2 Peripheral haematopoiesis ....................................................................................... 24 1.3 The histone methyltransferase SETDB1 ............................................................... 31 1.4 The histone methyltransferases SUV420H ............................................................ 33 2 RESULTS (I) .................................................................................................35 2.1 Spontaneous germinal center activation in Suv420h2 knockout (ko) mice............ 35 2.1.1 Suv420h2 ko mice show enlarged spleen ................................................................ 35 2.1.2 Suv420h2 ko B cells spontaneously initiated the germinal center reaction ............ 35 2.1.3 Suv420h2 ko B cells response to external stimuli is comparable to wild type cells 40 2.1.4 Follicular helper T cells are overrepresented in Suv420h2 ko spleen ..................... 41 2.1.5 Release of apoptotic checkpoints exacerbates germinal center activation in Suv420h2 ko mice .................................................................................................................. 43 2.1.6 Suv420h2 ko; VavBcl2 mice have increased proportion of follicular helper T cells 45 2.2 Discussion (I) ........................................................................................................ 47 3 RESULTS (II) ...............................................................................................51 3.1 SETDB1-mediated silencing of MuLVs is essential for B cell development .......... 51 VIII 3.1.1 Loss of Setdb1 impairs B cell development............................................................ 51 3.1.2 Intrinsic role of Setdb1 during B cell development ................................................ 56 3.1.3 Setdb1Mb1 B cells ectopically transcribe murine retrotransposable elements.......... 58 3.1.4 SETDB1 represses retrotransposons in pro B cells ................................................ 62 3.1.5 MuLVs derepression results in activation of neighbouring genes .......................... 64 3.1.6 Setdb1 loss in pro B cells induces apoptosis........................................................... 65 3.1.7 Ovexpression of antiapoptotic Bcl2 partially rescues B cell defects in Setdb1Mb1 mice 66 3.2 Discussion (II) ....................................................................................................... 69 4 RESULTS (III) ............................................................................................. 74 4.1 The role of the histone methyltransferase SETDB1 during haematopoiesis .......... 74 4.1.1 Setdb1Vav mice are underdeveloped and show impaired hematopoietic organs ...... 74 4.1.2 Lack of Setdb1 during hematopoietic development results in complete loss of lymphocytes and expansion of myeloid-erythroid lineage in the periphery ......................... 76 4.1.3 Setdb1Vav bone marrow completely abolishes B cell development in favour of myeloid-erythroid expansion ................................................................................................. 76 4.1.4 Common lymphoid progenitors (CLPs) are not altered in Setdb1Vav mice while granulocytes macrophage progenitors (GMPs) are overrepresented..................................... 78 4.1.5 HSCs compartment collapses 4 weeks after birth in Setdb1Vav mice...................... 79 4.2 Discussion (III) ...................................................................................................... 81 5 APPENDIX .................................................................................................. 86 5.1 Appendix (Results I) ............................................................................................. 86 5.2 Appendix (Results II) ............................................................................................ 88 6 MATERIAL AND METHODS ................................................................ 112 6.1 Materials ............................................................................................................. 112 6.1.1 Mice ...................................................................................................................... 112 6.1.2 Cell lines ............................................................................................................... 112 6.1.3 Antibodies and dyes .............................................................................................. 112 6.1.4 Technical devices and material ............................................................................. 113 6.1.5 Kits ........................................................................................................................ 114 IX 6.1.6 Tissue culture media, cytokines and immunostimulants ...................................... 115 6.1.7 Reagents and buffers ............................................................................................. 115 6.1.8 Oligonucleotides ................................................................................................... 117 6.1.9 Software and databases ......................................................................................... 118 6.2 Methods .............................................................................................................. 119 6.2.1 Mice and cell lines ................................................................................................ 119 6.2.2 Flow cytometry and cell sorting ............................................................................ 119 6.2.3 Red blood cell lysis ............................................................................................... 119 6.2.4 Definition of hematopoietic cell types for FACS analysis and FACS sorting ...... 120 6.2.5 Envelope protein and Fcγr2b staining ................................................................... 121 6.2.6 Bone marrow transplantation ................................................................................ 121 6.2.7 Immunohistochemistry .......................................................................................... 121 6.2.8 Autoantibodies detection test on MEFs ................................................................ 122 6.2.9 ELISA ................................................................................................................... 122 6.2.10 B cell proliferation assay ....................................................................................... 123 6.2.11 Colony forming assay and B cell differentiation on OP9 cells ............................
Load more

Recommended publications
  • Universidad Autónoma De Madrid Regulatory Mechanisms of Germinal Centers
    Universidad Autónoma de Madrid Departamento de Biología Molecular Regulatory mechanisms of Germinal Centers PhD Thesis Arantxa Pérez García Madrid, 2016 Regulatory mechanisms of Germinal Centers Memoria presentada por la licenciada en Biología Arantxa Pérez García para optar al título de doctor por la Universidad Autónoma de Madrid Directora de tesis: Almudena R. Ramiro Este trabajo ha sido realizado en el laboratorio de Biología de linfocitos B, en el Centro Nacional de Investigaciones Cardiovasculares (CNIC) Madrid, 2016 Memoria presentada por Arantxa Pérez García, licenciada en Biología, para optar al grado de doctor por la Universidad Autónoma de Madrid. Esta tesis ha sido realizada en el laboratorio de Biología de Linfocitos B del Centro Nacional de Investigaciones Cardiovasculares (CNIC), bajo la dirección de la Doctora Almudena R. Ramiro, y para que así conste y a los efectos oportunos, firma el siguiente certificado; En Madrid, a 21 de Abril de 2016 Almudena R. Ramiro RESUMEN Tras el reconocimiento del antígeno, los linfocitos B pueden iniciar la reacción de centro germinal (GC), en la cual diversifican sus genes de inmunoglobulinas, mediante las reacciones de hipermutación somática (SHM) y cambio de isotipo (CSR), dando lugar a células plasmáticas o B memoria. La transición a través de los diferentes estadios de esta reacción implica la expresión coordinada de redes de genes que permiten una correcta diversificación de los linfocitos B. A nivel molecular, las reacciones de SHM y CSR se desencadenan por la desaminación de citosinas en los genes de las inmunoglobulinas, mediada por AID. La actividad de AID en linfocitos B no está restringida a los genes de las inmunoglobulinas, pudiendo introducir mutaciones en otros genes y mediar translocaciones cromosómicas con potencial linfomagénico.
    [Show full text]
  • A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus
    Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated.
    [Show full text]
  • Expression of Human Endogenous Retrovirus K Envelope Transcripts in Human Breast Cancer1
    Vol. 7, 1553–1560, June 2001 Clinical Cancer Research 1553 Advances in Brief Expression of Human Endogenous Retrovirus K Envelope Transcripts in Human Breast Cancer1 Feng Wang-Johanning, Andra R. Frost, breast tumor sample showed nucleotide sequence differ- Gary L. Johanning, M. B. Khazaeli, ences, suggesting that multiple loci may be transcribed. Conclusions: These data indicate that HERV-K tran- Albert F. LoBuglio, Denise R. Shaw, and scripts with coding potential for the envelope region are 2 Theresa V. Strong expressed frequently in human breast cancer. The Comprehensive Cancer Center and Division of Hematology Oncology, Departments of Medicine [F. W-J., M. B. K., A. F. L., Introduction D. R. S., T. V. S.], Pathology [A. R. F.], and Nutrition Sciences 3 [G. L. J.], University of Alabama at Birmingham, Birmingham, HERVs are stably inherited sequences thought to have Alabama 35294-3300 entered the germ line of their host more than a million years ago (1, 2). These elements are widely dispersed throughout the genome and are estimated to comprise Ͼ1% of the entire human Abstract genome. Most HERVs are defective because of multiple termi- Purpose: We investigated the expression of human en- nation codons and deletions (3), but some appear to contain all dogenous retroviral (HERV) sequences in breast cancer. structural features necessary for viral replication (4). HERVs are Experimental Design: Reverse transcription-PCR (RT- grouped into single- and multiple-copy families, usually classi- PCR) was used to examine expression of the envelope (env) fied according to the tRNA used for reverse transcription (2). region of ERV3, HERV-E4-1, and HERV-K in breast cancer The type K family (HERV-K) is present in an estimated 30–50 cell lines, human breast tumor samples, adjacent uninvolved copies/human genome and includes some elements with long breast tissues, nonmalignant breast tissues, and placenta.
    [Show full text]
  • Supp Material.Pdf
    Simon et al. Supplementary information: Table of contents p.1 Supplementary material and methods p.2-4 • PoIy(I)-poly(C) Treatment • Flow Cytometry and Immunohistochemistry • Western Blotting • Quantitative RT-PCR • Fluorescence In Situ Hybridization • RNA-Seq • Exome capture • Sequencing Supplementary Figures and Tables Suppl. items Description pages Figure 1 Inactivation of Ezh2 affects normal thymocyte development 5 Figure 2 Ezh2 mouse leukemias express cell surface T cell receptor 6 Figure 3 Expression of EZH2 and Hox genes in T-ALL 7 Figure 4 Additional mutation et deletion of chromatin modifiers in T-ALL 8 Figure 5 PRC2 expression and activity in human lymphoproliferative disease 9 Figure 6 PRC2 regulatory network (String analysis) 10 Table 1 Primers and probes for detection of PRC2 genes 11 Table 2 Patient and T-ALL characteristics 12 Table 3 Statistics of RNA and DNA sequencing 13 Table 4 Mutations found in human T-ALLs (see Fig. 3D and Suppl. Fig. 4) 14 Table 5 SNP populations in analyzed human T-ALL samples 15 Table 6 List of altered genes in T-ALL for DAVID analysis 20 Table 7 List of David functional clusters 31 Table 8 List of acquired SNP tested in normal non leukemic DNA 32 1 Simon et al. Supplementary Material and Methods PoIy(I)-poly(C) Treatment. pIpC (GE Healthcare Lifesciences) was dissolved in endotoxin-free D-PBS (Gibco) at a concentration of 2 mg/ml. Mice received four consecutive injections of 150 μg pIpC every other day. The day of the last pIpC injection was designated as day 0 of experiment.
    [Show full text]
  • How Relevant Are Bone Marrow-Derived Mast Cells (Bmmcs) As Models for Tissue Mast Cells? a Comparative Transcriptome Analysis of Bmmcs and Peritoneal Mast Cells
    cells Article How Relevant Are Bone Marrow-Derived Mast Cells (BMMCs) as Models for Tissue Mast Cells? A Comparative Transcriptome Analysis of BMMCs and Peritoneal Mast Cells 1, 2, 1 1 2,3 Srinivas Akula y , Aida Paivandy y, Zhirong Fu , Michael Thorpe , Gunnar Pejler and Lars Hellman 1,* 1 Department of Cell and Molecular Biology, Uppsala University, The Biomedical Center, Box 596, SE-751 24 Uppsala, Sweden; [email protected] (S.A.); [email protected] (Z.F.); [email protected] (M.T.) 2 Department of Medical Biochemistry and Microbiology, Uppsala University, The Biomedical Center, Box 589, SE-751 23 Uppsala, Sweden; [email protected] (A.P.); [email protected] (G.P.) 3 Department of Anatomy, Physiology and Biochemistry, Swedish University of Agricultural Sciences, Box 7011, SE-75007 Uppsala, Sweden * Correspondence: [email protected]; Tel.: +46-(0)18-471-4532; Fax: +46-(0)18-471-4862 These authors contributed equally to this work. y Received: 29 July 2020; Accepted: 16 September 2020; Published: 17 September 2020 Abstract: Bone marrow-derived mast cells (BMMCs) are often used as a model system for studies of the role of MCs in health and disease. These cells are relatively easy to obtain from total bone marrow cells by culturing under the influence of IL-3 or stem cell factor (SCF). After 3 to 4 weeks in culture, a nearly homogenous cell population of toluidine blue-positive cells are often obtained. However, the question is how relevant equivalents these cells are to normal tissue MCs. By comparing the total transcriptome of purified peritoneal MCs with BMMCs, here we obtained a comparative view of these cells.
    [Show full text]
  • Indicators of the Molecular Pathogenesis of Virulent Newcastle
    www.nature.com/scientificreports OPEN Indicators of the molecular pathogenesis of virulent Newcastle disease virus in chickens revealed by transcriptomic profling of spleen Mohammad Rabiei1*, Wai Yee Low2, Yan Ren2, Mohamad Indro Cahyono3, Phuong Thi Kim Doan1,4, Indi Dharmayanti3, Eleonora Dal Grande1 & Farhid Hemmatzadeh1,2 Newcastle disease virus (NDV) has caused signifcant outbreaks in South-East Asia, particularly in Indonesia in recent years. Recently emerged genotype VII NDVs (NDV-GVII) have shifted their tropism from gastrointestinal/respiratory tropism to a lymphotropic virus, invading lymphoid organs including spleen and bursa of Fabricius to cause profound lymphoid depletion. In this study, we aimed to identify candidate genes and biological pathways that contribute to the disease caused by this velogenic NDV-GVII. A transcriptomic analysis based on RNA-Seq of spleen was performed in chickens challenged with NDV-GVII and a control group. In total, 6361 genes were diferentially expressed that included 3506 up-regulated genes and 2855 down-regulated genes. Real-Time PCR of ten selected genes validated the RNA-Seq results as the correlation between them is 0.98. Functional and network analysis of Diferentially Expressed Genes (DEGs) showed altered regulation of ElF2 signalling, mTOR signalling, proliferation of cells of the lymphoid system, signalling by Rho family GTPases and synaptogenesis signalling in spleen. We have also identifed modifed expression of IFIT5, PI3K, AGT and PLP1 genes in NDV-GVII infected chickens. Our fndings in activation of autophagy-mediated cell death, lymphotropic and synaptogenesis signalling pathways provide new insights into the molecular pathogenesis of this newly emerged NDV-GVII. Newcastle disease virus (NDV) has a worldwide distribution.
    [Show full text]
  • Figure S1. Reverse Transcription‑Quantitative PCR Analysis of ETV5 Mrna Expression Levels in Parental and ETV5 Stable Transfectants
    Figure S1. Reverse transcription‑quantitative PCR analysis of ETV5 mRNA expression levels in parental and ETV5 stable transfectants. (A) Hec1a and Hec1a‑ETV5 EC cell lines; (B) Ishikawa and Ishikawa‑ETV5 EC cell lines. **P<0.005, unpaired Student's t‑test. EC, endometrial cancer; ETV5, ETS variant transcription factor 5. Figure S2. Survival analysis of sample clusters 1‑4. Kaplan Meier graphs for (A) recurrence‑free and (B) overall survival. Survival curves were constructed using the Kaplan‑Meier method, and differences between sample cluster curves were analyzed by log‑rank test. Figure S3. ROC analysis of hub genes. For each gene, ROC curve (left) and mRNA expression levels (right) in control (n=35) and tumor (n=545) samples from The Cancer Genome Atlas Uterine Corpus Endometrioid Cancer cohort are shown. mRNA levels are expressed as Log2(x+1), where ‘x’ is the RSEM normalized expression value. ROC, receiver operating characteristic. Table SI. Clinicopathological characteristics of the GSE17025 dataset. Characteristic n % Atrophic endometrium 12 (postmenopausal) (Control group) Tumor stage I 91 100 Histology Endometrioid adenocarcinoma 79 86.81 Papillary serous 12 13.19 Histological grade Grade 1 30 32.97 Grade 2 36 39.56 Grade 3 25 27.47 Myometrial invasiona Superficial (<50%) 67 74.44 Deep (>50%) 23 25.56 aMyometrial invasion information was available for 90 of 91 tumor samples. Table SII. Clinicopathological characteristics of The Cancer Genome Atlas Uterine Corpus Endometrioid Cancer dataset. Characteristic n % Solid tissue normal 16 Tumor samples Stagea I 226 68.278 II 19 5.740 III 70 21.148 IV 16 4.834 Histology Endometrioid 271 81.381 Mixed 10 3.003 Serous 52 15.616 Histological grade Grade 1 78 23.423 Grade 2 91 27.327 Grade 3 164 49.249 Molecular subtypeb POLE 17 7.328 MSI 65 28.017 CN Low 90 38.793 CN High 60 25.862 CN, copy number; MSI, microsatellite instability; POLE, DNA polymerase ε.
    [Show full text]
  • University of California, San Diego
    UC San Diego UC San Diego Electronic Theses and Dissertations Title The post-terminal differentiation fate of RNAs revealed by next-generation sequencing Permalink https://escholarship.org/uc/item/7324r1rj Author Lefkowitz, Gloria Kuo Publication Date 2012 Peer reviewed|Thesis/dissertation eScholarship.org Powered by the California Digital Library University of California UNIVERSITY OF CALIFORNIA, SAN DIEGO The post-terminal differentiation fate of RNAs revealed by next-generation sequencing A dissertation submitted in partial satisfaction of the requirements for the degree Doctor of Philosophy in Biomedical Sciences by Gloria Kuo Lefkowitz Committee in Charge: Professor Benjamin D. Yu, Chair Professor Richard Gallo Professor Bruce A. Hamilton Professor Miles F. Wilkinson Professor Eugene Yeo 2012 Copyright Gloria Kuo Lefkowitz, 2012 All rights reserved. The Dissertation of Gloria Kuo Lefkowitz is approved, and it is acceptable in quality and form for publication on microfilm and electronically: __________________________________________________________________ __________________________________________________________________ __________________________________________________________________ __________________________________________________________________ __________________________________________________________________ Chair University of California, San Diego 2012 iii DEDICATION Ma and Ba, for your early indulgence and support. Matt and James, for choosing more practical callings. Roy, my love, for patiently sharing the ups and downs
    [Show full text]
  • Epigenetic Mechanisms Are Involved in the Oncogenic Properties of ZNF518B in Colorectal Cancer
    Epigenetic mechanisms are involved in the oncogenic properties of ZNF518B in colorectal cancer Francisco Gimeno-Valiente, Ángela L. Riffo-Campos, Luis Torres, Noelia Tarazona, Valentina Gambardella, Andrés Cervantes, Gerardo López-Rodas, Luis Franco and Josefa Castillo SUPPLEMENTARY METHODS 1. Selection of genomic sequences for ChIP analysis To select the sequences for ChIP analysis in the five putative target genes, namely, PADI3, ZDHHC2, RGS4, EFNA5 and KAT2B, the genomic region corresponding to the gene was downloaded from Ensembl. Then, zoom was applied to see in detail the promoter, enhancers and regulatory sequences. The details for HCT116 cells were then recovered and the target sequences for factor binding examined. Obviously, there are not data for ZNF518B, but special attention was paid to the target sequences of other zinc-finger containing factors. Finally, the regions that may putatively bind ZNF518B were selected and primers defining amplicons spanning such sequences were searched out. Supplementary Figure S3 gives the location of the amplicons used in each gene. 2. Obtaining the raw data and generating the BAM files for in silico analysis of the effects of EHMT2 and EZH2 silencing The data of siEZH2 (SRR6384524), siG9a (SRR6384526) and siNon-target (SRR6384521) in HCT116 cell line, were downloaded from SRA (Bioproject PRJNA422822, https://www.ncbi. nlm.nih.gov/bioproject/), using SRA-tolkit (https://ncbi.github.io/sra-tools/). All data correspond to RNAseq single end. doBasics = TRUE doAll = FALSE $ fastq-dump -I --split-files SRR6384524 Data quality was checked using the software fastqc (https://www.bioinformatics.babraham. ac.uk /projects/fastqc/). The first low quality removing nucleotides were removed using FASTX- Toolkit (http://hannonlab.cshl.edu/fastxtoolkit/).
    [Show full text]
  • Engineered Type 1 Regulatory T Cells Designed for Clinical Use Kill Primary
    ARTICLE Acute Myeloid Leukemia Engineered type 1 regulatory T cells designed Ferrata Storti Foundation for clinical use kill primary pediatric acute myeloid leukemia cells Brandon Cieniewicz,1* Molly Javier Uyeda,1,2* Ping (Pauline) Chen,1 Ece Canan Sayitoglu,1 Jeffrey Mao-Hwa Liu,1 Grazia Andolfi,3 Katharine Greenthal,1 Alice Bertaina,1,4 Silvia Gregori,3 Rosa Bacchetta,1,4 Norman James Lacayo,1 Alma-Martina Cepika1,4# and Maria Grazia Roncarolo1,2,4# Haematologica 2021 Volume 106(10):2588-2597 1Department of Pediatrics, Division of Stem Cell Transplantation and Regenerative Medicine, Stanford School of Medicine, Stanford, CA, USA; 2Stanford Institute for Stem Cell Biology and Regenerative Medicine, Stanford School of Medicine, Stanford, CA, USA; 3San Raffaele Telethon Institute for Gene Therapy, Milan, Italy and 4Center for Definitive and Curative Medicine, Stanford School of Medicine, Stanford, CA, USA *BC and MJU contributed equally as co-first authors #AMC and MGR contributed equally as co-senior authors ABSTRACT ype 1 regulatory (Tr1) T cells induced by enforced expression of interleukin-10 (LV-10) are being developed as a novel treatment for Tchemotherapy-resistant myeloid leukemias. In vivo, LV-10 cells do not cause graft-versus-host disease while mediating graft-versus-leukemia effect against adult acute myeloid leukemia (AML). Since pediatric AML (pAML) and adult AML are different on a genetic and epigenetic level, we investigate herein whether LV-10 cells also efficiently kill pAML cells. We show that the majority of primary pAML are killed by LV-10 cells, with different levels of sensitivity to killing. Transcriptionally, pAML sensitive to LV-10 killing expressed a myeloid maturation signature.
    [Show full text]
  • 1 Novel Expression Signatures Identified by Transcriptional Analysis
    ARD Online First, published on October 7, 2009 as 10.1136/ard.2009.108043 Ann Rheum Dis: first published as 10.1136/ard.2009.108043 on 7 October 2009. Downloaded from Novel expression signatures identified by transcriptional analysis of separated leukocyte subsets in SLE and vasculitis 1Paul A Lyons, 1Eoin F McKinney, 1Tim F Rayner, 1Alexander Hatton, 1Hayley B Woffendin, 1Maria Koukoulaki, 2Thomas C Freeman, 1David RW Jayne, 1Afzal N Chaudhry, and 1Kenneth GC Smith. 1Cambridge Institute for Medical Research and Department of Medicine, Addenbrooke’s Hospital, Hills Road, Cambridge, CB2 0XY, UK 2Roslin Institute, University of Edinburgh, Roslin, Midlothian, EH25 9PS, UK Correspondence should be addressed to Dr Paul Lyons or Prof Kenneth Smith, Department of Medicine, Cambridge Institute for Medical Research, Addenbrooke’s Hospital, Hills Road, Cambridge, CB2 0XY, UK. Telephone: +44 1223 762642, Fax: +44 1223 762640, E-mail: [email protected] or [email protected] Key words: Gene expression, autoimmune disease, SLE, vasculitis Word count: 2,906 The Corresponding Author has the right to grant on behalf of all authors and does grant on behalf of all authors, an exclusive licence (or non-exclusive for government employees) on a worldwide basis to the BMJ Publishing Group Ltd and its Licensees to permit this article (if accepted) to be published in Annals of the Rheumatic Diseases and any other BMJPGL products to exploit all subsidiary rights, as set out in their licence (http://ard.bmj.com/ifora/licence.pdf). http://ard.bmj.com/ on September 29, 2021 by guest. Protected copyright. 1 Copyright Article author (or their employer) 2009.
    [Show full text]
  • Supplemental Materials
    The infection-tolerant mammalian reservoir of Lyme disease and other zoonoses broadly counters the inflammatory effects of endotoxin Supplemental Materials Figures S1-S5 Tables S1-S20 Figure S1. Digital photograph of two adult Peromyscus leucopus with exudative conjunctivitis and huddled together. The animals had received 10 mg/gm body of Escherichia coli lipopolysaccharide intraperitoneally the day before. Figure S2. Species- and tissue-specific responses to LPS. Independent differential gene expression analysis of RNA-seq data were performed for blood, spleen, and liver tissues of P. leucopus and M. musculus collected 4 h after injection with LPS or buffer alsone as control. These are represented as volcano plots with range- adjusted scales for the log2-transformed fold-changes on x-axes and log10-transformed FDR p values on y- axes. Colors of symbols denote the following: red, up-regulated gene with absolute fold-change > 4.0 and p value < 0.05; purple, down-regulated gene with absolute fold-change > 4.0 and p value < 0.05; and gray, all others. Numbers at the top left and right corners in each plot respresent numbers of down- and up-regulated genes, respectively. Figure 3 is same data with constant scales for x- and y-axes across the plots. Numerical values for each gene in the 6 datasets are provided in Tables S4-S9. Figure S3. Correlation of IL-10 and IL-10 P. leucopus and M. musculus from RNA-seq of spleen and of Figure 6B and TaBle S14. The scatter plot is log10 values of normalized unique reads of one coding sequence against another for each of the four groups, as defined in the legend for Figure 6 and indicated By different symBols.
    [Show full text]