Identification and characterization of novel molecular causes of primary immunodeficiency : RELA mutations are associated to common variable immunodeficiency and systemic lupus erythematosus Hicham Lamrini To cite this version: Hicham Lamrini. Identification and characterization of novel molecular causes of primary immunod- eficiency : RELA mutations are associated to common variable immunodeficiency and systemic lupus erythematosus. Hematology. Université Sorbonne Paris Cité, 2018. English. NNT : 2018USPCB211. tel-02466295 HAL Id: tel-02466295 https://tel.archives-ouvertes.fr/tel-02466295 Submitted on 4 Feb 2020 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. Université Paris Descartes Ecole doctorale HOB 561 Spécialité Hématologie Identification and characterization of novel molecular causes of primary immunodeficiency: RELA mutations are associated to common variable immunodeficiency and systemic lupus erythematosus Présenté par: Hicham LAMRINI Thèse de doctorat en Immunogénétique Dirigée par Pr. Marina CAVAZZANA Laboratoire de Lympho-Hématopoïese Humaine, Unité INSERM U1163 Soutenue publiquement à l’institut IMAGINE le 25 Juin 2018 à Paris Devant un jury composé de: Pr. Anne-Sophie KORGANOW Rapportrice Pr. Marc SCHMIDT-SUPPRIAN Rapporteur Pr. Catherine ALCAÏDE-LORIDAN Présidente du jury Dr. Sven KRACKER Examinateur Pr. Marina CAVAZZANA Directrice de thèse Table of Contents 1 Abstract ...................................................................................................................... 8 1.1 English abstract ........................................................................................................................................................ 8 1.2 French abstract ......................................................................................................................................................... 9 2 Abbreviations ........................................................................................................... 10 3 Introduction .............................................................................................................. 11 3.1 Overview of the adaptive immune response ............................................................................................ 11 3.2 Cell-mediated immunity .................................................................................................................................... 12 3.2.1 Antigen recognition and formation of the immune synapse ......................................................... 13 3.2.2 T cell development ............................................................................................................................................ 15 3.2.3 Effector T cells .................................................................................................................................................... 18 3.2.4 Memory T cells ................................................................................................................................................... 19 3.3 Humoral immunity ............................................................................................................................................... 22 3.3.1 ImmunogloBulin: structures and functions ........................................................................................... 22 3.3.2 B cell maturation and activation ............................................................................................................... 24 3.3.3 Memory B cells ................................................................................................................................................... 27 3.4 Overview on primary immunodeficiency diseases ................................................................................ 27 3.4.1 Brief history ......................................................................................................................................................... 27 3.4.2 Primary immunodeficiencies diagnosis and etiology ....................................................................... 28 3.4.3 Common variable immunodeficiency ....................................................................................................... 30 3.4.4 Systemic Lupus Erythematosus associated to PID ............................................................................. 32 3.5 NFκB ........................................................................................................................................................................... 35 3.5.1 Brief history ......................................................................................................................................................... 35 3.5.2 The NF-κB family proteins, their inhibitors (IκB proteins) and activators (IKK complex) 36 3.5.3 A complex pathway, nexus of cell signaling .......................................................................................... 44 3.5.4 NF-κB in immunology ..................................................................................................................................... 48 3.6 RELA in genetic diseases ................................................................................................................................... 54 3.6.1 RELA in chronic mucocutaneous ulcerations ....................................................................................... 54 3.6.2 RELA in CD4 lymphoproliferative disease with autoimmune cytopenias ................................ 55 3.7 Overview on whole exome sequencing and identification of deleterious single nucleotide variant detections ............................................................................................................................................................. 56 3.7.1 Concepts ................................................................................................................................................................ 56 3.7.2 WES and search for a candidate gene ..................................................................................................... 57 4 Aim of the project ..................................................................................................... 60 5 Results ...................................................................................................................... 61 5.1 Investigating RELA mutations ......................................................................................................................... 61 5.1.1 Patients clinical and immunological features ...................................................................................... 61 5.1.2 WES of CVID and SLE patients .................................................................................................................... 67 5.1.3 Mutated RELA expression ............................................................................................................................. 70 5.1.4 Cellular localiZation of mutant RELA proteins .................................................................................... 74 5.1.5 DNA Binding of mutant RELA proteins ................................................................................................... 80 5.1.6 Mutant RELA proteins interactions with partner proteins ............................................................ 85 5.1.7 Mutant RELA proteins transcriptional activity ................................................................................... 89 2 6 Discussion ................................................................................................................. 97 7 Perspectives ........................................................................................................... 103 8 Material and methods ............................................................................................ 105 9 Bibliography ........................................................................................................... 111 10 AcknowledGments ............................................................................................... 123 11 Publications ........................................................................................................ 130 3 Index of figures Figure 1. Overview of the immune response. ...................................................................................... 12 Figure 2. The immunological synapse. ................................................................................................... 15 Figure 3. Lymphocyte development. ....................................................................................................... 17 Figure 4. First and secondary exposure to antigen. .......................................................................... 21 Figure 5. The immunoglobulin, an antibody and a cell receptor. ............................................... 24 Figure 6. B cell maturation. ........................................................................................................................