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1.
Lupus Sci Med ; 11(1)2024 Jan 31.
Article in English | MEDLINE | ID: mdl-38302132

ABSTRACT

OBJECTIVES: To illuminate the poorly understood aetiology of SLE by comparing the gene expression profile of SLE neutrophils with that of neutrophils from patients infected by SARS-CoV-2, a disease (COVID-19) with well-defined antigens and a similar type I interferon response. METHODS: RNA sequencing of neutrophils from patients with SLE (n=15) and healthy controls (n=12) was analysed for differential gene expression and modulated pathways. The same analyses were performed on a similar neutrophil dataset from patients with SARS-CoV-2 infection (n=30) and healthy controls (n=8). Next, we carried out comparative analyses to identify common and unique transcriptional changes between the two disease contexts, emphasising genes regulated in opposite directions. RESULTS: We identified 372 differentially expressed genes in SLE neutrophils compared with healthy donor neutrophils (≥2 fold, p<0.05), 181 of which were concordant with transcriptional changes in SARS-CoV-2-infected individuals compared with their respective healthy controls. In contrast, 118 genes demonstrated statistically significant alterations exclusive to SLE, including 28 genes that were differentially expressed in opposite directions in the two diseases. CONCLUSIONS: The substantial overlap between neutrophil responses in SLE and COVID-19 suggests that the unknown cause of SLE is functionally similar to a viral infection and drives a similar immune activation and type I interferon response. Conversely, the genes regulated in the opposite direction represent responses unique to SLE. These include tyrosylprotein sulfotransferase-1 and nucleic acid deaminases of the APOBEC family, which can catalyse cytosine-to-uridine editing of both RNA and DNA, and other RNA-modifying enzymes.


Subject(s)
COVID-19 , Interferon Type I , Lupus Erythematosus, Systemic , Humans , Neutrophils , Transcriptome , COVID-19/genetics , SARS-CoV-2/genetics , SARS-CoV-2/metabolism , Lupus Erythematosus, Systemic/genetics , RNA/metabolism , Interferon Type I/genetics
2.
Sci Adv ; 7(22)2021 05.
Article in English | MEDLINE | ID: mdl-34039612

ABSTRACT

Chronic infectious diseases have a substantial impact on the human B cell compartment including a notable expansion of B cells here termed atypical B cells (ABCs). Using unbiased single-cell RNA sequencing (scRNA-seq), we uncovered and characterized heterogeneities in naïve B cell, classical memory B cells, and ABC subsets. We showed remarkably similar transcriptional profiles for ABC clusters in malaria, HIV, and autoimmune diseases and demonstrated that interferon-γ drove the expansion of ABCs in malaria. These observations suggest that ABCs represent a separate B cell lineage with a common inducer that further diversifies and acquires disease-specific characteristics and functions. In malaria, we identified ABC subsets based on isotype expression that differed in expansion in African children and in B cell receptor repertoire characteristics. Of particular interest, IgD+IgMlo and IgD-IgG+ ABCs acquired a high antigen affinity threshold for activation, suggesting that ABCs may limit autoimmune responses to low-affinity self-antigens in chronic malaria.

3.
J Immunol ; 203(7): 1820-1829, 2019 10 01.
Article in English | MEDLINE | ID: mdl-31451676

ABSTRACT

The clear role of autophagy in human inflammatory diseases such as Crohn disease was first identified by genome-wide association studies and subsequently dissected in multiple mechanistic studies. ATG16L1 has been particularly well studied in knockout and hypomorph settings as well as models recapitulating the Crohn disease-associated T300A polymorphism. Interestingly, ATG16L1 has a single homolog, ATG16L2, which is independently implicated in diseases, including Crohn disease and systemic lupus erythematosus. However, the contribution of ATG16L2 to canonical autophagy pathways and other cellular functions is poorly understood. To better understand its role, we generated and analyzed the first, to our knowledge, ATG16L2 knockout mouse. Our results show that ATG16L1 and ATG16L2 contribute very distinctly to autophagy and cellular ontogeny in myeloid, lymphoid, and epithelial lineages. Dysregulation of any of these lineages could contribute to complex diseases like Crohn disease and systemic lupus erythematosus, highlighting the value of examining cell-specific effects. We also identify a novel genetic interaction between ATG16L2 and epithelial ATG16L1. These findings are discussed in the context of how these genes may contribute distinctly to human disease.


Subject(s)
Autophagic Cell Death , Autophagy-Related Proteins , Carrier Proteins , Crohn Disease , Lupus Erythematosus, Systemic , Animals , Autophagic Cell Death/genetics , Autophagic Cell Death/immunology , Autophagy-Related Proteins/genetics , Autophagy-Related Proteins/immunology , Carrier Proteins/genetics , Carrier Proteins/immunology , Crohn Disease/genetics , Crohn Disease/immunology , Disease Models, Animal , Humans , Lupus Erythematosus, Systemic/genetics , Lupus Erythematosus, Systemic/immunology , Mice , Mice, Knockout , Organ Specificity/genetics , Organ Specificity/immunology
4.
Cell ; 178(3): 714-730.e22, 2019 07 25.
Article in English | MEDLINE | ID: mdl-31348891

ABSTRACT

Genome-wide association studies (GWAS) have revealed risk alleles for ulcerative colitis (UC). To understand their cell type specificities and pathways of action, we generate an atlas of 366,650 cells from the colon mucosa of 18 UC patients and 12 healthy individuals, revealing 51 epithelial, stromal, and immune cell subsets, including BEST4+ enterocytes, microfold-like cells, and IL13RA2+IL11+ inflammatory fibroblasts, which we associate with resistance to anti-TNF treatment. Inflammatory fibroblasts, inflammatory monocytes, microfold-like cells, and T cells that co-express CD8 and IL-17 expand with disease, forming intercellular interaction hubs. Many UC risk genes are cell type specific and co-regulated within relatively few gene modules, suggesting convergence onto limited sets of cell types and pathways. Using this observation, we nominate and infer functions for specific risk genes across GWAS loci. Our work provides a framework for interrogating complex human diseases and mapping risk variants to cell types and pathways.


Subject(s)
Colitis, Ulcerative/pathology , Colon/metabolism , Adult , Aged , Antibodies, Monoclonal/therapeutic use , Bestrophins/metabolism , CD8 Antigens/metabolism , Case-Control Studies , Colitis, Ulcerative/drug therapy , Colitis, Ulcerative/metabolism , Colon/pathology , Enterocytes/cytology , Enterocytes/metabolism , Female , Genetic Loci , Genome-Wide Association Study , Humans , Interleukin-17/metabolism , Male , Middle Aged , Risk Factors , T-Lymphocytes/cytology , T-Lymphocytes/metabolism , Thrombospondins/metabolism , Tumor Necrosis Factor-alpha/immunology , Tumor Necrosis Factor-alpha/metabolism , Young Adult
5.
Genome Res ; 29(1): 53-63, 2019 01.
Article in English | MEDLINE | ID: mdl-30552105

ABSTRACT

The evolutionary history of a gene helps predict its function and relationship to phenotypic traits. Although sequence conservation is commonly used to decipher gene function and assess medical relevance, methods for functional inference from comparative expression data are lacking. Here, we use RNA-seq across seven tissues from 17 mammalian species to show that expression evolution across mammals is accurately modeled by the Ornstein-Uhlenbeck process, a commonly proposed model of continuous trait evolution. We apply this model to identify expression pathways under neutral, stabilizing, and directional selection. We further demonstrate novel applications of this model to quantify the extent of stabilizing selection on a gene's expression, parameterize the distribution of each gene's optimal expression level, and detect deleterious expression levels in expression data from individual patients. Our work provides a statistical framework for interpreting expression data across species and in disease.


Subject(s)
Evolution, Molecular , Gene Expression Regulation/physiology , Models, Genetic , Animals , Dogs , Rabbits
6.
Cell Rep ; 25(11): 2992-3005.e5, 2018 12 11.
Article in English | MEDLINE | ID: mdl-30540934

ABSTRACT

Long-term hematopoietic stem cells (LT-HSCs) maintain hematopoietic output throughout an animal's lifespan. However, with age, the balance is disrupted, and LT-HSCs produce a myeloid-biased output, resulting in poor immune responses to infectious challenge and the development of myeloid leukemias. Here, we show that young and aged LT-HSCs respond differently to inflammatory stress, such that aged LT-HSCs produce a cell-intrinsic, myeloid-biased expression program. Using single-cell RNA sequencing (scRNA-seq), we identify a myeloid-biased subset within the LT-HSC population (mLT-HSCs) that is prevalent among aged LT-HSCs. We identify CD61 as a marker of mLT-HSCs and show that CD61-high LT-HSCs are uniquely primed to respond to acute inflammatory challenge. We predict that several transcription factors regulate the mLT-HSCs gene program and show that Klf5, Ikzf1, and Stat3 play an important role in age-related inflammatory myeloid bias. We have therefore identified and isolated an LT-HSC subset that regulates myeloid versus lymphoid balance under inflammatory challenge and with age.


Subject(s)
Aging/pathology , Hematopoietic Stem Cells/metabolism , Inflammation/pathology , Animals , Biomarkers/metabolism , Inflammation/genetics , Ligands , Mice, Inbred C57BL , Models, Biological , Myeloid Cells/metabolism , Toll-Like Receptors/metabolism , Transcription, Genetic
7.
Genome Res ; 28(12): 1812-1825, 2018 12.
Article in English | MEDLINE | ID: mdl-30446528

ABSTRACT

While genetic variants are known to be associated with overall gene abundance in stimulated immune cells, less is known about their effects on alternative isoform usage. By analyzing RNA-seq profiles of monocyte-derived dendritic cells from 243 individuals, we uncovered thousands of unannotated isoforms synthesized in response to influenza infection and type 1 interferon stimulation. We identified more than a thousand quantitative trait loci (QTLs) associated with alternate isoform usage (isoQTLs), many of which are independent of expression QTLs (eQTLs) for the same gene. Compared with eQTLs, isoQTLs are enriched for splice sites and untranslated regions, but depleted of sequences upstream of annotated transcription start sites. Both eQTLs and isoQTLs explain a significant proportion of the disease heritability attributed to common genetic variants. At the ERAP2 locus, we shed light on the function of the gene and how two frequent, highly differentiated haplotypes with intermediate frequencies could be maintained by balancing selection. At baseline and following type 1 interferon stimulation, the major haplotype is associated with low ERAP2 expression caused by nonsense-mediated decay, while the minor haplotype, known to increase Crohn's disease risk, is associated with high ERAP2 expression. In response to influenza infection, we found two uncharacterized isoforms expressed from the major haplotype, likely the result of multiple perfectly linked variants affecting the transcription and splicing at the locus. Thus, genetic variants at a single locus could modulate independent gene regulatory processes in innate immune responses and, in the case of ERAP2, may confer a historical fitness advantage in response to virus.


Subject(s)
Alternative Splicing , Aminopeptidases/genetics , Genetic Predisposition to Disease , Host-Pathogen Interactions/genetics , Influenza A virus , Influenza, Human/genetics , Influenza, Human/virology , Adolescent , Adult , Chromosome Mapping , Computational Biology/methods , Dendritic Cells/metabolism , Female , Gene Expression Profiling , Gene Expression Regulation , Gene Ontology , Genetic Testing , Genetic Variation , Humans , Interferon Type I/metabolism , Male , Middle Aged , Models, Biological , Molecular Sequence Annotation , Monocytes/metabolism , Quantitative Trait Loci , Transcriptome , Young Adult
8.
Cell ; 175(5): 1307-1320.e22, 2018 11 15.
Article in English | MEDLINE | ID: mdl-30392957

ABSTRACT

In the small intestine, a niche of accessory cell types supports the generation of mature epithelial cell types from intestinal stem cells (ISCs). It is unclear, however, if and how immune cells in the niche affect ISC fate or the balance between self-renewal and differentiation. Here, we use single-cell RNA sequencing (scRNA-seq) to identify MHC class II (MHCII) machinery enrichment in two subsets of Lgr5+ ISCs. We show that MHCII+ Lgr5+ ISCs are non-conventional antigen-presenting cells in co-cultures with CD4+ T helper (Th) cells. Stimulation of intestinal organoids with key Th cytokines affects Lgr5+ ISC renewal and differentiation in opposing ways: pro-inflammatory signals promote differentiation, while regulatory cells and cytokines reduce it. In vivo genetic perturbation of Th cells or MHCII expression on Lgr5+ ISCs impacts epithelial cell differentiation and IEC fate during infection. These interactions between Th cells and Lgr5+ ISCs, thus, orchestrate tissue-wide responses to external signals.


Subject(s)
Cell Differentiation , Cell Self Renewal , Interleukin-10/metabolism , Stem Cells/cytology , T-Lymphocytes, Helper-Inducer/metabolism , Animals , Cell Differentiation/drug effects , Cell Self Renewal/drug effects , Cytokines/pharmacology , Epithelial Cells/cytology , Epithelial Cells/metabolism , Female , Histocompatibility Antigens Class II/metabolism , Immune System/metabolism , Intestines/cytology , Intestines/microbiology , Male , Mice , Mice, Inbred C57BL , Organoids/cytology , Organoids/drug effects , Organoids/metabolism , Receptors, G-Protein-Coupled/metabolism , Salmonella enterica/pathogenicity , Stem Cells/metabolism , T-Lymphocytes, Helper-Inducer/cytology
9.
Nature ; 560(7718): 319-324, 2018 08.
Article in English | MEDLINE | ID: mdl-30069044

ABSTRACT

The airways of the lung are the primary sites of disease in asthma and cystic fibrosis. Here we study the cellular composition and hierarchy of the mouse tracheal epithelium by single-cell RNA-sequencing (scRNA-seq) and in vivo lineage tracing. We identify a rare cell type, the Foxi1+ pulmonary ionocyte; functional variations in club cells based on their location; a distinct cell type in high turnover squamous epithelial structures that we term 'hillocks'; and disease-relevant subsets of tuft and goblet cells. We developed 'pulse-seq', combining scRNA-seq and lineage tracing, to show that tuft, neuroendocrine and ionocyte cells are continually and directly replenished by basal progenitor cells. Ionocytes are the major source of transcripts of the cystic fibrosis transmembrane conductance regulator in both mouse (Cftr) and human (CFTR). Knockout of Foxi1 in mouse ionocytes causes loss of Cftr expression and disrupts airway fluid and mucus physiology, phenotypes that are characteristic of cystic fibrosis. By associating cell-type-specific expression programs with key disease genes, we establish a new cellular narrative for airways disease.


Subject(s)
Cell Differentiation/genetics , Cell Lineage/genetics , Cystic Fibrosis Transmembrane Conductance Regulator/genetics , Cystic Fibrosis/genetics , Epithelial Cells/metabolism , Animals , Asthma/genetics , Epithelial Cells/cytology , Female , Forkhead Transcription Factors/deficiency , Forkhead Transcription Factors/genetics , Gene Expression Profiling , Gene Expression Regulation , Goblet Cells/cytology , Goblet Cells/metabolism , Humans , Lung/cytology , Male , Mice , Sequence Analysis, RNA , Single-Cell Analysis , Trachea/cytology
10.
Nature ; 551(7680): 333-339, 2017 11 16.
Article in English | MEDLINE | ID: mdl-29144463

ABSTRACT

Intestinal epithelial cells absorb nutrients, respond to microbes, function as a barrier and help to coordinate immune responses. Here we report profiling of 53,193 individual epithelial cells from the small intestine and organoids of mice, which enabled the identification and characterization of previously unknown subtypes of intestinal epithelial cell and their gene signatures. We found unexpected diversity in hormone-secreting enteroendocrine cells and constructed the taxonomy of newly identified subtypes, and distinguished between two subtypes of tuft cell, one of which expresses the epithelial cytokine Tslp and the pan-immune marker CD45, which was not previously associated with non-haematopoietic cells. We also characterized the ways in which cell-intrinsic states and the proportions of different cell types respond to bacterial and helminth infections: Salmonella infection caused an increase in the abundance of Paneth cells and enterocytes, and broad activation of an antimicrobial program; Heligmosomoides polygyrus caused an increase in the abundance of goblet and tuft cells. Our survey highlights previously unidentified markers and programs, associates sensory molecules with cell types, and uncovers principles of gut homeostasis and response to pathogens.


Subject(s)
Epithelial Cells/cytology , Epithelium/metabolism , Intestine, Small/cytology , Single-Cell Analysis , Animals , Cell Differentiation , Cytokines/metabolism , Enterocytes/metabolism , Epithelial Cells/metabolism , Female , Gene Expression Profiling , Homeostasis , Leukocyte Common Antigens/metabolism , Male , Mice , Organoids/cytology , Organoids/metabolism , Paneth Cells/metabolism , Transcription, Genetic , Thymic Stromal Lymphopoietin
11.
Nanotoxicology ; 11(7): 846-856, 2017 Sep.
Article in English | MEDLINE | ID: mdl-28885066

ABSTRACT

In this work, we evaluated the impact of gold nanoparticles on endothelial cell behavior and function beyond the influence on cell viability. Five types of gold nanoparticles were studied: 5 nm and 20 nm bare gold nanoparticles, 5 nm and 20 nm gold nanoparticles with biocompatible polyethylene glycol (PEG) coating and 60 nm bare gold nanoparticles. We found that all tested gold nanoparticles did not affect cell viability significantly and reduced the reactive oxygen species (ROS) level in endothelial cells. Only 20 nm bare gold nanoparticles caused an over 50% increase in endothelial barrier permeability and slow recovery of barrier function was observed after the gold nanoparticles were removed. This impairment in endothelial barrier function was caused by unbalanced forces between intracellular tensions and paracellular forces, actin microfilament rearrangement, which occurred through a Rho/ROCK kinase-dependent pathway and broke the force balance between intracellular tensions and paracellular forces. The size-specific effect of gold nanoparticles on endothelial cells may have important implications regarding the behavior of nanoparticles in the biological system and provide valuable guidance in nanomaterial design and biomedical applications.


Subject(s)
Actin Cytoskeleton/metabolism , Gold/toxicity , Human Umbilical Vein Endothelial Cells/drug effects , Metal Nanoparticles/toxicity , Biocompatible Materials/chemistry , Biocompatible Materials/toxicity , Cell Culture Techniques , Cell Membrane Permeability/drug effects , Cell Survival/drug effects , Gold/chemistry , Human Umbilical Vein Endothelial Cells/metabolism , Humans , Metal Nanoparticles/chemistry , Microscopy, Fluorescence , Particle Size , Polyethylene Glycols/chemistry , Polyethylene Glycols/toxicity , Reactive Oxygen Species/metabolism , Surface Properties
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