scholarly journals Single-cell analysis reveals the function of lung progenitor cells in COVID-19 patients

2020 ◽  
Author(s):  
Zixian Zhao ◽  
Yu Zhao ◽  
Yueqing Zhou ◽  
Xiaofan Wang ◽  
Ting Zhang ◽  
...  
Keyword(s):  
Single Cell ◽  
Progenitor Cells ◽  
Time Course ◽  
Single Cell Analysis ◽  
Lung Damage ◽  
Mouse Lung ◽  
Epithelial Barrier ◽  
Epithelial Barriers ◽  
Lung Progenitor

AbstractThe high mortality of severe 2019 novel coronavirus disease (COVID-19) cases is mainly caused by acute respiratory distress syndrome (ARDS), which is characterized by increased permeability of the alveolar epithelial barriers, pulmonary edema and consequently inflammatory tissue damage. Some but not all patients showed full functional recovery after the devastating lung damage, and so far there is little knowledge about the lung repair process1. Here by analyzing the bronchoalveolar lavage fluid (BALF) of COVID-19 patients through single cell RNA-sequencing (scRNA-Seq), we found that in severe (or critical) cases, there is remarkable expansion of TM4SF1+ and KRT5+ lung progenitor cells. The two distinct populations of progenitor cells could play crucial roles in alveolar cell regeneration and epithelial barrier re-establishment, respectively. In order to understand the function of KRT5+ progenitors in vivo, we transplanted a single KRT5+ cell-derived cell population into damaged mouse lung. Time-course single-cell transcriptomic analysis showed that the transplanted KRT5+ progenitors could long-term engrafted into host lung and differentiate into HOPX+ OCLN+ alveolar barrier cell which restored the epithelial barrier and efficiently prevented inflammatory cell infiltration. Similar barrier cells were also identified in some COVID-19 patients with massive leukocyte infiltration. Altogether this work uncovered the mechanism that how various lung progenitor cells work in concert to prevent and replenish alveoli loss post severe SARS-CoV-2 infection.

scholarly journals Single‐cell analysis identified lung progenitor cells in COVID‐19 patients

2020 ◽  
Vol 53 (12) ◽  
Author(s):  
Zixian Zhao ◽  
Yu Zhao ◽  
Yueqing Zhou ◽  
Xiaofan Wang ◽  
Ting Zhang ◽  
...  
Keyword(s):  
Single Cell ◽  
Progenitor Cells ◽  
Single Cell Analysis ◽  
Cell Analysis ◽  
Lung Progenitor

scholarly journals Single-cell analysis of RORα tracer mouse lung reveals ILC progenitors and effector ILC2 subsets

2019 ◽  
Vol 217 (3) ◽  
Author(s):  
Maryam Ghaedi ◽  
Zi Yi Shen ◽  
Mona Orangi ◽  
Itziar Martinez-Gonzalez ◽  
Lisa Wei ◽  
...  
Keyword(s):  
Single Cell ◽  
Single Cell Analysis ◽  
Retinoic Acid Receptor ◽  
Allergic Inflammation ◽  
Lymphoid Cells ◽  
Orphan Receptor ◽  
Mouse Lung ◽  
Mouse Lungs

Lung group 2 innate lymphoid cells (ILC2s) drive allergic inflammation and promote tissue repair. ILC2 development is dependent on the transcription factor retinoic acid receptor–related orphan receptor (RORα), which is also expressed in common ILC progenitors. To elucidate the developmental pathways of lung ILC2s, we generated RORα lineage tracer mice and performed single-cell RNA sequencing, flow cytometry, and functional analyses. In adult mouse lungs, we found an IL-18Rα+ST2− population different from conventional IL-18Rα−ST2+ ILC2s. The former was GATA-3intTcf7EGFP+Kit+, produced few cytokines, and differentiated into multiple ILC lineages in vivo and in vitro. In neonatal mouse lungs, three ILC populations were identified, namely an ILC progenitor population similar to that in adult lungs and two distinct effector ILC2 subsets that differentially produced type 2 cytokines and amphiregulin. Lung ILC progenitors might actively contribute to ILC-poiesis in neonatal and inflamed adult lungs. In addition, neonatal lung ILC2s include distinct proinflammatory and tissue-repairing subsets.

scholarly journals Single-cell analysis of RORα tracer mouse lung reveals ILC progenitors and effector ILC2 subsets

2019 ◽  
Author(s):  
Maryam Ghaedi ◽  
Mona Orangi ◽  
Itziar Martinez-Gonzalez ◽  
Lisa Wei ◽  
Xiaoxiao Lu ◽  
...  
Keyword(s):  
Single Cell ◽  
Single Cell Analysis ◽  
Retinoic Acid Receptor ◽  
Allergic Inflammation ◽  
Lymphoid Cells ◽  
Orphan Receptor ◽  
Mouse Lung ◽  
Mouse Lungs

Lung group 2 innate lymphoid cells (ILC2s) drive allergic inflammation and promote tissue repair. ILC2 development is dependent on the transcription factor retinoic acid receptor–related orphan receptor (RORα), which is also expressed in common ILC progenitors. To elucidate the developmental pathways of lung ILC2s, we generated RORα lineage tracer mice and performed single-cell RNA sequencing, flow cytometry, and functional analyses. In adult mouse lungs, we found an IL-18Rα+ST2− population different from conventional IL-18Rα−ST2+ ILC2s. The former was GATA-3intTcf7EGFP+Kit+, produced few cytokines, and differentiated into multiple ILC lineages in vivo and in vitro. In neonatal mouse lungs, three ILC populations were identified, namely an ILC progenitor population similar to that in adult lungs and two distinct effector ILC2 subsets that differentially produced type 2 cytokines and amphiregulin. Lung ILC progenitors might actively contribute to ILC-poiesis in neonatal and inflamed adult lungs. In addition, neonatal lung ILC2s include distinct proinflammatory and tissue-repairing subsets.

scholarly journals Single Cell Analysis Reveals That IL-4 Receptor/Stat6 Signaling Is Not Required for the In Vivo or In Vitro Development of CD4+Lymphocytes with a Th2 Cytokine Profile

2000 ◽  
Vol 164 (6) ◽  
pp. 3047-3055 ◽  
Author(s):  
Dragana Jankovic ◽  
Marika C. Kullberg ◽  
Nancy Noben-Trauth ◽  
Patricia Caspar ◽  
William E. Paul ◽  
...  
Keyword(s):  
Single Cell ◽  
Single Cell Analysis ◽  
Cytokine Profile ◽  
Cell Analysis ◽  
In Vitro Development ◽  
Th2 Cytokine ◽  
Vitro Development ◽  
Cd4 Lymphocytes

scholarly journals P-165 Cytometry-Based Single Cell Analysis of Intact Epithelial Signaling Reveals MAPK Activation Divergent from TNF-α-Induced Apoptosis in Vivo

2016 ◽  
Vol 22 ◽  
pp. S59-S60
Author(s):  
Alan Simmons ◽  
Amrita Banerjee ◽  
Eliot McKinley ◽  
Cherieʼ Scurrah ◽  
Jeffrey Franklin ◽  
...  
Keyword(s):  
Single Cell ◽  
Single Cell Analysis ◽  
Cell Analysis ◽  
Mapk Activation ◽  
Tnf Α ◽  
Induced Apoptosis

scholarly journals Receptor-mediated Immunoglobulin G Transport Across Mucosal Barriers in Adult Life

2002 ◽  
Vol 196 (3) ◽  
pp. 303-310 ◽  
Author(s):  
Gerburg M. Spiekermann ◽  
Patricia W. Finn ◽  
E. Sally Ward ◽  
Jennifer Dumont ◽  
Bonny L. Dickinson ◽  
...  
Keyword(s):  
Immune Surveillance ◽  
Adult Life ◽  
Secretory Iga ◽  
Mouse Lung ◽  
Bronchial Epithelial ◽  
Mucosal Surfaces ◽  
Dependent Absorption ◽  
Epithelial Barriers ◽  
Mucosal Secretions

Mucosal secretions of the human gastrointestinal, respiratory, and genital tracts contain the immunoglobulins (Ig)G and secretory IgA (sIgA) that function together in host defense. Exactly how IgG crosses epithelial barriers to function in mucosal immunity remains unknown. Here, we test the idea that the MHC class I–related Fc-receptor, FcRn, transports IgG across the mucosal surface of the human and mouse lung from lumen to serosa. We find that bronchial epithelial cells of the human, nonhuman primate, and mouse, express FcRn in adult-life, and demonstrate FcRn-dependent absorption of a bioactive Fc-fusion protein across the respiratory epithelium of the mouse in vivo. Thus, IgG, like dimeric IgA, can cross epithelial barriers by receptor-mediated transcytosis in adult animals. These data show that mucosal surfaces that express FcRn reabsorb IgG and explain a mechanism by which IgG may act in immune surveillance to retrieve lumenal antigens for processing in the lamina propria or systemically.

scholarly journals Single-cell analysis of circadian dynamics in tissue explants

2015 ◽  
Vol 26 (22) ◽  
pp. 3940-3945 ◽  
Author(s):  
Laura Lande-Diner ◽  
Jacob Stewart-Ornstein ◽  
Charles J. Weitz ◽  
Galit Lahav
Keyword(s):  
Circadian Clock ◽  
Single Cell ◽  
Cell Tracking ◽  
Single Cell Analysis ◽  
Single Cells ◽  
Tissue Imaging ◽  
Wide Field ◽  
Isolated Cells ◽  
Peripheral Tissues

Tracking molecular dynamics in single cells in vivo is instrumental to understanding how cells act and interact in tissues. Current tissue imaging approaches focus on short-term observation and typically nonendogenous or implanted samples. Here we develop an experimental and computational setup that allows for single-cell tracking of a transcriptional reporter over a period of >1 wk in the context of an intact tissue. We focus on the peripheral circadian clock as a model system and measure the circadian signaling of hundreds of cells from two tissues. The circadian clock is an autonomous oscillator whose behavior is well described in isolated cells, but in situ analysis of circadian signaling in single cells of peripheral tissues is as-yet uncharacterized. Our approach allowed us to investigate the oscillatory properties of individual clocks, determine how these properties are maintained among different cells, and assess how they compare to the population rhythm. These experiments, using a wide-field microscope, a previously generated reporter mouse, and custom software to track cells over days, suggest how many signaling pathways might be quantitatively characterized in explant models.

scholarly journals Using Flow Cytometry and Multistage Machine Learning to Discover Label-Free Signatures of Algal Lipid Accumulation

2018 ◽  
Author(s):  
Mohammad Tanhaemami ◽  
Elaheh Alizadeh ◽  
Claire Sanders ◽  
Babetta L. Marrone ◽  
Brian Munsky’
Keyword(s):  
Machine Learning ◽  
Flow Cytometry ◽  
Single Cell ◽  
Lipid Accumulation ◽  
Time Course ◽  
Fluorescent Dyes ◽  
Single Cell Analysis ◽  
Learning Approaches ◽  
Label Free ◽  
Unlabeled Cell

Abstract—Most applications of flow cytometry or cell sorting rely on the conjugation of fluorescent dyes to specific biomarkers. However, labeled biomarkers are not always available, they can be costly, and they may disrupt natural cell behavior. Label-free quantification based upon machine learning approaches could help correct these issues, but label replacement strategies can be very difficult to discover when applied labels or other modifications in measurements inadvertently modify intrinsic cell properties. Here we demonstrate a new, but simple approach based upon feature selection and linear regression analyses to integrate statistical information collected from both labeled and unlabeled cell populations and to identify models for accurate label-free single-cell quantification. We verify the method’s accuracy to predict lipid content in algal cells(Picochlorum soloecismus)during a nitrogen starvation and lipid accumulation time course. Our general approach is expected to improve label-free single-cell analysis for other organisms or pathways, where biomarkers are inconvenient, expensive, or disruptive to downstream cellular processes.

scholarly journals Long Noncoding RNA SNHG4 Remits Lipopolysaccharide‐Engendered Inflammatory Lung Damage by Inhibiting METTL3 - Mediated m6A Level of STAT2 mRNA

2021 ◽  
Author(s):  
Si-Xiu Li ◽  
Wen Yan ◽  
Jian-Ping Liu ◽  
Yu-Juan Zhao ◽  
Lu Chen
Keyword(s):  
Cell Proliferation ◽  
Noncoding Rna ◽  
Lung Damage ◽  
Lung Fibroblasts ◽  
Mouse Lung ◽  
Treated Cell ◽  
Cell Functions ◽  
Tnf Α

Abstract Background: Emerging evidence suggests that long non coding RNA (lncRNA) small nucleolar RNA host gene 4 (SNHG4) has become a new insight into lipopolysaccharide (LPS) - induced microglia inflammation, its role in neonatal pneumonia (NP) remains to be largely unrevealed.Methods: RT-qPCR was used to determine SNHG4 and METTL3 expression in the serum from NP patients and normal volunteers, as well as in WI-38 cells treated with LPS. The SNHG4 overexpression vector (pcDNA-SNHG4) was transfected into LPS - treated cells. CCK-8, Transwell, annexin V-FITC/PI and ELISA assays were used to determine cell proliferation, migration, apoptosis and contents of IL-6, TNF-α, SOD and MDA, respectively. The level of SNHG4 in the promoter region of METTL3 was assessed with RIP assay. m6A quantitative analysis illustrated the m6A level with or without SNHG4 overexpression or METTL3 silencing. Bioinformatics analysis and RIP-PCR were used to predict and validate YTHDF1 - mediated m6A levels on signal transducer and activator of transcription 2 (STAT2) mRNA in METTL3 inhibited cells. Then rescue experiments were performed to explore effects of SNHG4 and METTL3 or STAT2 on LPS-treated cell functions. Subsequently, in vivo functional experiments were performed to investigate the role of SNHG4 in LPS induced pneumonia in mice. Results: SNHG4 was downregulated and METTL3 was upregulated in NP patients and LPS-treated cells. SNHG4 overexpression facilitated cell proliferation, migration and SOD concentration, and inhibited apoptosis and IL-6, TNF-α and MDA contents. Mechanistically, SNHG4 bound with METTL3 and downregulated METTL3 expression. Besides, total m6A modification level was lower in the SNHG4 overexpressed or METTL3 inhibited cells. METTL3 interference reduced m6A levels of STAT2 mRNA, decreased STAT2 mRNA stability and promoted STAT2 translation level. METTL3 or STAT2 upregulated reversed the effects of SNHG4 overexpression on LPS - treated cell functions. Conclusions: This study reveals that SNHG4 promotes LPS induced inflammation in human lung fibroblasts and mouse lung tissues in vitro and in vivo by inhibiting METTL3 - mediated m6A level of STAT2 mRNA, which may provide a potential therapeutic mechanism for NP.

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