Document text
Principal Investigator: John O'Shea
Organization: NATIONAL INSTITUTE OF ARTHRITIS AND MUSCULOSKELETAL AND SKIN DISEASES
Fiscal Year: 2024
Award: $2,543,374
Funding agency: National Institute of Arthritis and Musculoskeletal and Skin Diseases
Discovery of patients with mutations of Janus kinase (JAKs) and STATs reveals new disease mechanisms and insights into pathophysiology. STAT6 (signal transducer and activator of transcription 6) is a transcription factor that plays a central role in the pathophysiology of allergic inflammation. We identified multiple families spanning three continents with profound allergic immune dysregulation including: atopic dermatitis, gastrointestinal disease, asthma, food allergies, and anaphylaxis. All patients carried monoallelic rare variants in STAT6 and functional studies established their gain-of-function (GOF) phenotype with sustained STAT6 phosphorylation, increased STAT6 target gene expression, and TH2 skewing.
Allergic diseases are common, affecting more than 20% of the population. Genetic variants in the TGFβ pathway are strongly associated allergy and asthma. We examined patients and mice that harbor loss-of-function mutations in the kinase domain of TGFΒR1/2. We found that variants lead to elevated total and allergen-specific IgE and increase in type 2 follicular helper T cells (TFH2). Further investigation revealed that TGFβ plays a conserved, T cell-intrinsic, and nonredundant role in restraining TFH2 development via the PI3Kγ/mTOR pathway, thereby constraining allergic disease. Additionally, we previously discovered that archetype proprotein convertase subtilisin/kexins (PCSKs), furin, has essential regulatory functions in both innate and adaptive immune responses through its role in modifying TGFb. We demonstrated that deletion of furin in T cells promoted response against lymphocytic choriomeningitis virus by virtue of restricting viral burden and augmenting interferon gamma production.
Cytokine expression during T cell differentiation is a highly regulated process that involves long-range promoter-enhancer and CTCF-CTCF contacts at cytokine loci with the gene encoding interferon gamma (being an outstanding example. Here, we investigated the impact of dynamic chromatin loop formation within the topologically associating domain (TAD) in regulating the expression of interferon gamma and interleukin-22 (IL-22); these cytokine loci are closely located in the genome and are associated with complex enhancer landscapes, which are selectively active in type 1 and type 3 lymphocytes. In situ Hi-C analyses revealed inducible TADs that insulated Ifng and Il22 enhancers during Th1 cell differentiation. Targeted deletion of a 17 bp boundary motif of these TADs imbalanced Th1- and Th17-associated immunity, both in vitro and in vivo, upon Toxoplasma gondii infection. In contrast, this boundary element was dispensable for cytokine regulation in natural killer cells. Our findings suggest that precise cytokine regulation relies on lineage- and developmental stage-specific interactions of 3D chromatin architectures and enhancer landscapes.
To better understand mechanisms of disease we generate mice with STAT loss- and gain-of-function variants. Patients with loss-of-function mutations of STAT1 exhibit viral and bacterial susceptibility. Mice lacking STAT1 in T cells control parasitic infection. However, a consequence of infection is lethal immunopathology associated with a type 1 (interferon gamma) response, reduced IL-10 generation and increased IL-13 production. Conversely, STAT1 GOF mutations underlie heterogenous disorders. Such patients exhibit both immunodeficiency and autoimmunity (including SLE, scleroderma and related disorders). To better understand immunopathogenic mechanisms of disease, we developed a STAT1 GOF mouse. We found that these mice have increased susceptibility to viral infection, including SARS-CoV2. These mice exhibit a cytokine storm-like phenotype. We found that exogenous interferon gamma can reverse aspects of the pathology.
CD4+ T helper 17 (TH17) cells protect barrier tissues but also trigger autoimmunity. The mechanisms behind these opposing processes remain unclear. We identified that the transcription factor EGR2 controlled the transcriptional program of pathogenic TH17 cells in the central nervous system (CNS). EGR2 was significantly elevated in myelin-reactive CD4+ T cells from patients with multiple sclerosis and mice with autoimmune neuroinflammation. EGR2 enhanced TH17 cell differentiation and myeloid cell recruitment to the CNS.
STAT5 encompasses two genes, STAT5a and STAT5b. Our previous work established that STAT5 is a master regulator of energy and amino acid metabolism and is critical for hematopoiesis. We further established that STAT5 regulates the activity of key metabolically relevant biochemical pathways including AKT, mTOR and MYC. We demonstrated that NOD1, a cytosolic innate sensor of bacterial peptidoglycan, also functions in murine hematopoietic cells as a major regulator of both the generation and differentiation of lymphoid progenitors as well as peripheral T lymphocyte homeostasis. We further show that NOD1 mediates these functions by facilitating STAT5 signaling downstream of hematopoietic cytokines.
Disabling pansclerotic morphea (DPM) is a rare systemic inflammatory disorder, characterized by poor wound healing, fibrosis, cytopenias, hypogammaglobulinemia, and squamous-cell carcinoma. The cause is unknown, and mortality is high. We identified three unrelated families with an autosomal dominant pattern of inheritance of DPM. We identified three heterozygous variants in a specific region of the gene that encodes STAT4. In vitro, primary skin fibroblasts showed enhanced interleukin-6 secretion, with impaired wound healing, contraction of the collagen matrix, and matrix secretion. Inhibition of JAK-STAT signaling with ruxolitinib led to improvement in patients.
Discontinuous transcription is evolutionarily conserved and a fundamental feature of gene regulation; yet, the exact mechanisms underlying transcriptional bursting are unresolved. Analyses of bursting transcriptome-wide have focused on the role of cis-regulatory elements, but other factors that regulate this process remain elusive. We applied mathematical modeling to infer bursting dynamics transcriptome-wide and identified that Mediator complex subunit 26 (MED26) primarily regulates frequency, MYC regulates burst size, while cohesin and Bromodomain-containing protein 4 (BRD4) can modulate both. Currently, we are investigating STAT variants to dissect the impact on transcriptional bursting.
Terms: <2019 novel corona virus><2019 novel coronavirus><2019-nCoV><3-D><3-Dimensional><3D><AKT><Affect><Akt protein><Allergens><Allergic><Allergic Disease><Allergic inflammation><Allergic to food><Allergy><Allergy to food><Anaphylactic Reaction><Anaphylactic Shock><Anaphylaxis><Architecture><Asthma><Atopic Dermatitis><Atopic Eczema><Atopic Neurodermatitis><Atrophic Arthritis><Autoimmune><Autoimmune Diseases><Autoimmune Status><Autoimmunity><Autoregulation><B cell differentiation factor><B cell stimulating factor 2><B-Cell Differentiation Factor><B-Cell Differentiation Factor-2><B-Cell Stimulatory Factor-2><BCDF><BSF-2><BSF2><Bacteria><Basal Transcription Factor><Basal transcription factor genes><Biochemical Pathway><Body Tissues><Bone-Derived Transforming Growth Factor><Boundary Elements><Bromodomain><Bronchial Asthma><CD4 Cells><CD4 Positive T Lymphocytes><CD4 T cells><CD4 helper T cell><CD4 lymphocyte><CD4+ T-Lymphocyte><CD4-Positive Lymphocytes><CNS Nervous System><COVID-19 virus><COVID19 virus><CSIF><CSIF-10><Cell Body><Cell Communication and Signaling><Cell Differentiation><Cell Differentiation process><Cell Protection><Cell Signaling><Cells><Cellular Immune Function><Central Nervous System><Chromatin><Chromatin Loop><Chromatin Loop Domains><CoV-2><CoV2><Collagen><Complex><Cytokine Signal Transduction><Cytokine Signaling><Cytokine Synthesis Inhibitory Factor><Cytoprotection><Cytotoxic cell><DNA Loop><DNA-Binding Proteins><Development><Disabling><Disease><Disorder><Disseminated Neurodermatitis><Dysfunction><EGR2><EGR2 gene><Engineering / Architecture><Enhancers><Eosinophil-Mast Cell Growth-Factor><Epidermoid Carcinoma><Epigenetic><Epigenetic Change><Epigenetic Mechanism><Epigenetic Process><Erythrocyte Burst-Promoting Factor><Exhibits><FK506 Binding Protein 12-Rapamycin Associated Protein 1><FKBP12 Rapamycin Complex Associated Protein 1><FRAP1><FRAP1 gene><FRAP2><Family><Fibroblasts><Fibrosis><Food Allergy><Food Hypersensitivity><Frequencies><Functional disorder><Gastrointestinal Diseases><Gene Action Regulation><Gene Expression><Gene Expression Regulation><Gene Regulation><Gene Regulation Process><Gene Transcription><Gene variant><General Transcription Factor Gene><General Transcription Factors><Generations><Genes><Genetic Alteration><Genetic Change><Genetic Transcription><Genetic defect><Genome><Goals><Growth and Development><Growth and Development function><HPGF><Helper Cells><Helper T-Cells><Helper T-Lymphocytes><Helper-Inducer T-Cells><Helper-Inducer T-Lymphocyte><Hematopoiesis><Hematopoietic><Hematopoietic Cellular Control Mechanisms><Hematopoietic Cytokine><Hepatocyte-Stimulating Factor><Heterozygote><Hi-C><Homeostasis><Host Defense><Human><Hybridoma Growth Factor><Hypersensitivity><IFN-Gamma><IFN-beta 2><IFN-g><IFN-γ><IFNB2><IFNG><IFNγ><IL-10><IL-13><IL-22><IL-3><IL-3 Gene><IL-6><IL10><IL10A><IL13><IL3><IL3 Gene><IL3 Protein><IL6 Protein><IgE><Immune><Immune Interferon><Immune response><Immunes><Immunity><Immunoglobulin E><Immunological response><Immunomodulation><Impaired tissue repair><Impaired wound healing><In Situ><In Vitro><Inducer Cells><Inducer T-Lymphocytes><Infection><Inflammatory><Inflammatory Bowel Diseases><Inflammatory Bowel Disorder><Inflammatory Response><Inheritance Patterns><Innate Immune Response><Interferon Gamma><Interferon Type II><Interleukin 10 Precursor><Interleukin-10><Interleukin-13><Interleukin-6><Intermediary Metabolism><Intracellular Communication and Signaling><Investigation><JAK kinase><Janus kinase><K lymphocyte><KO mice><KROX20><Kinases><Knock-out Mice><Knockout Mice><LCM Viruses><LCMV><Lead><Link><Lupus Erythematosus Disseminatus><Lymphatic cell><Lymphocyte><Lymphocytic><Lymphocytic choriomeningitis virus><Lymphoid><MGF protein><MGI-2><MGSNF protein><MS patient><Mast-Cell Colony-Stimulating Factor><Math Models><Mechanistic Target of Rapamycin><Mediating><Mediator><Metabolic><Metabolic Networks><Metabolic Processes><Metabolism><Mice><Mice Mammals><Milk Growth Factor><Modern Man><Modification><Molecular><Morphea><Murein><Murine><Mus><Mutation><Myelin><Myeloid Cells><Myeloid Differentiation-Inducing Protein><NK Cells><Natural Killer Cells><Neuraxis><Null Mouse><P-CSF><P-Cell Stimulating Factor><PI 3-kinase gamma><PI3K gamma><PI3K γ><PI3Kγ><Parasitic infection><Pathogenesis><Pathogenicity><Pathology><Pathway interactions><Patients><Pb element><Peptidoglycan><Peripheral><Phenotype><Phosphorylation><Phosphotransferase Gene><Phosphotransferases><Physiological Homeostasis><Physiopathology><Planocellular Carcinoma><Plasmacytoma Growth Factor><Platelet Transforming Growth Factor><Play><Population><Predisposition><Process><Production><Proprotein Convertases><Protein Kinase B><Protein Phosphorylation><Protein Secretion><Proteins><Proto-Oncogene Proteins c-akt><Psoriasis><RAC-PK protein><RAFT1><RNA Expression><Regulation><Regulatory Element><Rheumatoid Arthritis><Role><SARS corona virus 2><SARS-CO-V2><SARS-COVID-2><SARS-CoV-2><SARS-CoV2><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><SLE><STAT protein><STAT1><STAT1 gene><STAT4><STAT4 gene><STAT5><STAT5A><STAT5A gene><STAT5a Transcription Factor><STAT6 Transcription Factor><STAT91><Scleroderma><Severe Acute Respiratory Coronavirus 2><Severe Acute Respiratory Distress Syndrome CoV 2><Severe Acute Respiratory Distress Syndrome Corona Virus 2><Severe Acute Respiratory Distress Syndrome Coronavirus 2><Severe Acute Respiratory Syndrome CoV 2><Severe Acute Respiratory Syndrome-associated coronavirus 2><Severe Acute Respiratory Syndrome-related coronavirus 2><Severe acute respiratory syndrome associated corona virus 2><Severe acute respiratory syndrome coronavirus 2><Severe acute respiratory syndrome related corona virus 2><Signal Transducer and Activator of Transcription><Signal Transducer and Activator of Transcription 5A><Signal Transducer and Activator of Transcription 6><Signal Transduction><Signal Transduction Systems><Signaling><Skin><Squamous Carcinoma><Squamous Cell Epithelioma><Squamous cell carcinoma><Stat5 protein><Stat5a protein><Stat5alpha protein><Stat6 protein><Subtilisins><Susceptibility><Systemic Lupus Erythematosus><Systemic Lupus Erythematous><Systemic Lupus Erythmatosus><T cell differentiation><T gondii infection><T-Cells><T-Lymphocyte><T. gondii infection><T4 Cells><T4 Lymphocytes><TGF B><TGF-beta><TGF-β><TGFbeta><TGFβ><Th-1 Cell><Th1 Cells><Therapeutic><Tissues><Toxoplasma gondii Infection><Toxoplasmosis><Transcription><Transcription Factor Proto-Oncogene><Transcription factor genes><Transforming Growth Factor beta><Transforming Growth Factor-Beta Family Gene><Transphosphorylases><Type 1 Helper Cell><Variant><Variation><Viral><Viral Burden><Viral Diseases><Viral Load><Viral Load result><Virus><Virus Diseases><Work><Wound Repair><Wuhan coronavirus><abnormal tissue repair><adaptive immune response><allele variant><allelic variant><allergic dermatitis><allergic eczema><amino acid metabolism><autoimmune attack><autoimmune condition><autoimmune destruction><autoimmune disorder><autoimmune pathogenesis><autoimmunity disease><autosome><biological signal transduction><blood cell formation><c-akt protein><cellular differentiation><cohesin><coronavirus disease 2019 virus><coronavirus disease-19 virus><cytokine><cytokine release syndrome><cytokine storm><cytopenia><cytoprotective><delayed wound healing><dermatosclerosis><developmental><disseminated lupus erythematosus><epigenetically><epigenomics><fungus><gain of function><gain of function mutation><gastrointestinal disorder><genetic variant><genome mutation><genomic variant><global gene expression><global transcription profile><hCoV19><heavy metal Pb><heavy metal lead><hemopoietic><heterozygosity><host response><human disease><hypogammaglobulinemia><hypoimmunity><immune deficiency><immune function><immune modulation><immune regulation><immune system response><immunodeficiency><immunologic reactivity control><immunomodulatory><immunopathology><immunoregulation><immunoregulatory><immunoresponse><improved><in vivo><infectious disease model><inflammatory disease of the intestine><inflammatory disorder of the intestine><insight><interferon beta 2><interleukin-22><intestinal autoinflammation><lFN-Gamma><loss of function><loss of function mutation><lymph cell><mTOR><mammalian target of rapamycin><mammary gland factor><mammary gland-specific nuclear factor><mathematic model><mathematical model><mathematical modeling><mortality><multiple sclerosis patient><nCoV2><neural inflammation><neuroinflammation><neuroinflammatory><new drug target><new druggable target><new pharmacotherapy target><new therapeutic target><new therapy target><novel drug target><novel druggable target><novel pharmacotherapy target><novel therapeutic target><novel therapy target><parasite infection><pathogen><pathophysiology><pathway><patients with MS><patients with multiple sclerosis><people with Multiple sclerosis><phosphatidylinositol 3-kinase gamma><phosphatidylinositol 3-kinase γ><progenitor><programs><promoter><promotor><proto-oncogene protein RAC><proto-oncogene protein akt><psoriasiform><psoriatic><rac protein kinase><rare allele><rare mutation><rare variant><recruit><related to A and C-protein><response><restraint><rheumatic arthritis><sensor><signal tranducer and activator of transcription 5><social role><systemic lupus erythematosis><three dimensional><thymus derived lymphocyte><tissue repair><transcription factor><transcriptome><viral infection><virus infection><virus-induced disease><wound healing><wound recovery><wound resolution>