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

Discovery and Optimization of a Series of Sulfonamide Inverse Agonists for the Retinoic Acid Receptor-Related Orphan Receptor-α

2019 ◽  
Vol 15 (6) ◽  
pp. 676-684
Author(s):  
Christelle Doebelin ◽  
Yuanjun He ◽  
Sean Campbell ◽  
Philippe Nuhant ◽  
Naresh Kumar ◽  
...  
Keyword(s):  
Retinoic Acid Receptor ◽  
Research Effort ◽  
Pharmacokinetic Profile ◽  
Orphan Receptor ◽  
Orphan Nuclear Receptor ◽  
Inverse Agonists ◽  
Close Family Member ◽  
Per Oral

Background: Despite a massive industry endeavor to develop RORγ-modulators for autoimmune disorders, there has been no indication of efforts to target the close family member RORα for similar indications. This may be due to the misconception that RORα is redundant to RORγ, or the inherent difficulty in cultivating tractable starting points for RORα. RORα-selective modulators would be useful tools to interrogate the biology of this understudied orphan nuclear receptor. Objectives: he goal of this research effort was to identify and optimize synthetic ligands for RORα starting from the known LXR agonist T0901317. Methods: Fourty-five analogs of the sulfonamide lead (1) were synthesized and evaluated for their ability to suppress the transcriptional activity of RORα, RORγ, and LXRα in cell-based assays. Analogs were characterized by 1H-NMR, 13C-NMR, and LC-MS analysis. The pharmacokinetic profile of the most selective RORα inverse agonist was evaluated in rats with intraperitoneal (i.p.) and per oral (p.o.)dosing. Results: Structure-activity relationship studies led to potent dual RORα/RORγ inverse agonists as well as RORα-selective inverse agonists (20, 28). LXR activity could be reduced by removing the sulfonamide nitrogen substituent. Attempts to improve the potency of these selective leads by varying substitution patterns throughout the molecule proved challenging. Conclusion: The synthetic RORα-selective inverse agonists identified (20, 28) can be utilized as chemical tools to probe the function of RORα in vitro and in vivo.

scholarly journals Recurrent pregnancy loss is associated with a pro-senescent decidual response during the peri-implantation window

2018 ◽  
Author(s):  
Emma S Lucas ◽  
Pavle Vrljicak ◽  
Joanne Muter ◽  
Maria M Diniz-da-Costa ◽  
Paul J Brighton ◽  
...  
Keyword(s):  
Single Cell ◽  
Pregnancy Loss ◽  
Recurrent Pregnancy Loss ◽  
Single Cell Analysis ◽  
Immune Surveillance ◽  
Marker Genes ◽  
Implantation Window ◽  
Decidual Cells

AbstractBreakdown of the feto-maternal interface in early pregnancy causes miscarriage. The cycling endometrium becomes poised to transition to a pregnant state during the midluteal implantation window, coinciding with differentiation of stromal cells into decidual cells (DC) and emergence of senescent decidual cells (snDC). Emerging evidence suggests that DC engage uterine natural killer cells to eliminate their senescent counterparts, thus enabling formation of a robust decidual matrix in pregnancy. To examine if failure to constrain snDC during the peri-implantation window increases the risk of miscarriage, we reconstructed the decidual pathway at single-cell levelin vitroand demonstrated that, without immune surveillance, secondary senescence rapidly transforms DC into progesterone-resistant cells that abundantly express extracellular matrix remodelling factors. Additional single-cell analysis of midluteal endometrium identifiedDIO2andSCARA5as marker genes of a diverging decidual responsein vivo. Finally, we report a conspicuous link between a pro-senescent decidual response in luteal phase endometrium and recurrent pregnancy loss, suggesting that pre-pregnancy screening and intervention may reduce the burden of miscarriage.

scholarly journals Interleukin (IL)-12 and IL-18 Synergize to Promote MAIT Cell IL-17A and IL-17F Production Independently of IL-23 Signaling

2020 ◽  
Vol 11 ◽  
Author(s):  
Suzanne Cole ◽  
Janine Murray ◽  
Catherine Simpson ◽  
Remi Okoye ◽  
Kerry Tyson ◽  
...  
Keyword(s):  
Th17 Cells ◽  
Cell Activation ◽  
Retinoic Acid Receptor ◽  
Γδ T Cells ◽  
Lymphoid Cells ◽  
Orphan Receptor ◽  
Mait Cells ◽  
Activation Assay ◽  
Mait Cell

IL-23 is considered a critical regulator of IL-17 in Th17 cells; however, its requirement for inducing IL-17 production in other human immune subsets remains incompletely understood. Mucosal associated invariant T (MAIT) cells uniformly express retinoic acid receptor-related orphan receptor gamma t (RORγt) but only a minor population have been shown to produce IL-17A. Here we show that IL-17F is the dominant IL-17 isoform produced by MAIT cells, not IL-17A. For optimal MAIT cell derived IL-17A and IL-17F production, T cell receptor (TCR) triggering, IL-18 and monocyte derived IL-12 signaling is required. Unlike Th17 cells, this process is independent of IL-23 signaling. Using an in vitro skin cell activation assay, we demonstrate that dual neutralization of both IL-17A and IL-17F resulted in greater suppression of inflammatory proteins than inhibition of IL-17A alone. Finally, we extend our findings by showing that other innate-like lymphocytes such as group 3 innate lymphoid cells (ILC3) and gamma delta (γδ) T cells are also capable of IL-23 independent IL-17A and IL-17F production. These data indicate both IL-17F and IL-17A production from MAIT cells may contribute to tissue inflammation independently of IL-23, in part explaining the therapeutic disconnect between targeting IL-17 or IL-23 in certain inflammatory diseases.

scholarly journals Single-cell analysis reveals a nestin+ tendon stem/progenitor cell population with strong tenogenic potentiality

2016 ◽  
Vol 2 (11) ◽  
pp. e1600874 ◽  
Author(s):  
Zi Yin ◽  
Jia-jie Hu ◽  
Long Yang ◽  
Ze-Feng Zheng ◽  
Cheng-rui An ◽  
...  
Keyword(s):  
Gene Expression ◽  
Single Cell ◽  
Cell Population ◽  
Single Cell Analysis ◽  
Expression Profiles ◽  
Gene Expression Profiles ◽  
Cell Analysis ◽  
Nestin Expression

The repair of injured tendons remains a formidable clinical challenge because of our limited understanding of tendon stem cells and the regulation of tenogenesis. With single-cell analysis to characterize the gene expression profiles of individual cells isolated from tendon tissue, a subpopulation of nestin+ tendon stem/progenitor cells (TSPCs) was identified within the tendon cell population. Using Gene Expression Omnibus datasets and immunofluorescence assays, we found that nestin expression was activated at specific stages of tendon development. Moreover, isolated nestin+ TSPCs exhibited superior tenogenic capacity compared to nestin− TSPCs. Knockdown of nestin expression in TSPCs suppressed their clonogenic capacity and reduced their tenogenic potential significantly both in vitro and in vivo. Hence, these findings provide new insights into the identification of subpopulations of TSPCs and illustrate the crucial roles of nestin in TSPC fate decisions and phenotype maintenance, which may assist in future therapeutic strategies to treat tendon disease.

scholarly journals The Trends of Single-Cell Analysis: A Global Study

2020 ◽  
Vol 2020 ◽  
pp. 1-10
Author(s):  
Hua Tian ◽  
Haifeng Liu ◽  
Yuanyuan Zhu ◽  
Dan Xing ◽  
Bin Wang
Keyword(s):  
Single Cell ◽  
Single Cell Analysis ◽  
Bibliographic Coupling ◽  
In Vivo Studies ◽  
Cell Analysis ◽  
The Usa ◽  
Visualization Of Data ◽  
Number Of Publications

Objective. The field of single-cell analysis has rapidly grown worldwide, and a bibliometric analysis and visualization of data and publications pertaining to such single-cell research has the potential to offer insights into the development of this field over the past two decades while also highlighting future avenues of research. Methods. Single-cell analysis-related studies published from 2000-2019 were identified through searches of the Web of Science, Scopus, and PubMed databases, and corresponding bibliometric data were systematically compiled. Extracted data from each study included author names, country of origin, and affiliations. GraphPad Prism was used to analyze these data, while VOSviewer was used to perform global analyses of bibliographic coupling, coauthorship, cocitation, and co-occurrence. Results. In total, 4,071 relevant studies were included in this analysis. The number of publications increased substantially with time, suggesting that single-cell analyses are becoming increasingly more prevalent in recent years. Studies from the USA had the greatest impact in this field, with higher H -index values and numbers of citations relative to other countries, whereas Israel exhibited the highest average number of citations per publication. Bibliographic coupling, coauthorship, cocitation, and co-occurrence analyses revealed that Analytical Chemistry was associated with the highest number of publications in this field, and the University of Stanford contributed the most to this field. The most cited study included in this analysis was published by Macosko et al. in 2015 in Cell. Co-occurrence analyses revealed that the most common single-cell research topics included “mechanistic studies,” “in vitro studies,” “in vivo studies,” and “fabrication studies.” Conclusions. Single-cell analyses are a rapidly growing area of scientific interest, and higher volumes of publications in this field are expected in the coming years, particularly for studies conducting fabrication and in vivo single-cell analyses.

scholarly journals Assessing the Heterogeneity of Cardiac Non-myocytes and the Effect of Cell Culture with Integrative Single Cell Analysis

2020 ◽  
Author(s):  
Brian S. Iskra ◽  
Logan Davis ◽  
Henry E. Miller ◽  
Yu-Chiao Chiu ◽  
Alexander R. Bishop ◽  
...  
Keyword(s):  
Cell Culture ◽  
Single Cell ◽  
Single Cell Analysis ◽  
Cell Types ◽  
Cell Analysis ◽  
Mass Cytometry ◽  
Single Cell Rna Sequencing ◽  
High Depth

AbstractCardiac non-myocytes comprise a diverse and crucial cell population in the heart that plays dynamic roles in cardiac wound healing and growth. Non-myocytes broadly fall into four cell types: endothelium, fibroblasts, leukocytes, and pericytes. Here we characterize the diversity of the non-myocytes in vivo and in vitro using mass cytometry. By leveraging single-cell RNA sequencing we inform the design of a mass cytometry panel. To aid in annotation of the mass cytometry datasets, we utilize data integration with a neural network. We introduce approximately 460,000∼ single cell proteomes of non-myocytes as well as 5,000∼ CD31 negative single cell transcriptomes. Using our data, as well as previously reported datasets, we characterize cardiac non-myocytes with high depth in six mice, characterizing novel surface markers (CD9, CD200, Notch3, and FolR2). Further, we find that extended cell culture promotes the proliferation of CD45+CD11b+FolR2+IAIE- myeloid cells in addition to fibroblasts.

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.

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