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GEO help: Mouse over screen elements for information. |
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| Status |
Public on Sep 24, 2021 |
| Title |
Single cell RNA-seq profiling of murine endothelial cells in response to pulmonary hypertension |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
(1) Rationale: Endothelial cell dysfunction plays a critical role in the development and pathogenesis of pulmonary arterial hypertension (PAH). (2) Objectives: We aimed to characterise the endothelial cell dynamics in PAH at a single cell resolution. (3) Methods: We carried out single-cell RNA sequencing of lung endothelial cells isolated from an endothelial cell lineage tracing mouse model in control and SU5416/Hypoxia-induced PAH conditions. (4) Measurements and Main Results: Endothelial cell populations corresponding to the different lung vessel types could be identified in both control and PAH mice. Differential gene expression analysis revealed novel global and vessel-type specific responses in endothelial cells due to PAH. Global changes included the up-regulation of the major histocompatibility complex class II pathway, supporting a role for endothelial cells in the inflammatory response in PAH. We also identified a PAH response specific to the second capillary EC population, with the up-regulation of genes involved in cell localization and angiogenesis. Comparison with human genetics and transcriptomics data revealed the regulation of four genes with variants associated to PAH and five genes up-regulated in endothelial cells in both human and mouse scRNA-seq. Among them, Aqp1 and Adam15 genes represent promising new candidates to target endothelial dysfunction. Finally, we identified zonation-dependent changes across the arteriovenous axis in PAH using an in-silico cell ordering approach and showed the up-regulation of the Serine/threonine-protein kinase Sgk1 at the junction between the macro- and micro-vasculature. (5) Conclusions: This study uncovers the murine endothelial cell transcriptomics changes in PAH at a high resolution, revealing novel candidates relevant to PAH.
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| Overall design |
(1) Experiment1: To study ECs in murine lungs, we used a Cdh5-CreERT2-TdTomato mouse line, in which the EC-specific (Cdh5) expression of TdTomato is inducible with tamoxifen. After a two week wash out period, TdTomato-positive cells were obtained from the lung of two mice using flow cytometry and analysed using 10X Genomics scRNA-seq (ContA/ContB). (2) Experiment2: To examine EC changes due to pulmonary arterial hypertension (PAH), we used the SuHx model, exposing the Cdh5-CreERT2-TdTomato mice to chronic hypoxia for three weeks in combination with weekly subcutaneous injections of SU5416. ScRNA-seq was carried out for three SuHx mice (PAH1, PAH2, PAH3) and one control mouse that was maintained in normoxia (Cont1).
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| Contributor(s) |
Rodor J, Scanlon JP, Spiroski A, Monteiro JP, Caudrillier A, Stewart KR, Shmakova A, Dobie R, Henderson BE, Chen S, Henderson NC, Baker AH |
| Citation(s) |
34528097 |
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| Submission date |
Jul 23, 2020 |
| Last update date |
Sep 26, 2021 |
| Contact name |
Julie Rodor |
| E-mail(s) |
Julie.Rodor@ed.ac.uk
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| Phone |
+44 (0)7580200884
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| Organization name |
UNIVERSITY OF EDINBURGH
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| Department |
Centre for Cardiovascular Science
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| Street address |
47 Little France Crescent
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| City |
Edinburgh |
| State/province |
NA |
| ZIP/Postal code |
EH16 4TJ |
| Country |
United Kingdom |
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| Platforms (1) |
| GPL24247 |
Illumina NovaSeq 6000 (Mus musculus) |
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| Samples (6)
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| GSM4684977 |
scRNA-seq of TdTomato+ cells from ContA mouse |
| GSM4684978 |
scRNA-seq of TdTomato+ cells from ContB mouse |
| GSM4684979 |
scRNA-seq of TdTomato+ cells from Cont1 mouse |
| GSM4684980 |
scRNA-seq of TdTomato+ cells from PAH1 mouse |
| GSM4684981 |
scRNA-seq of TdTomato+ cells from PAH2 mouse |
| GSM4684982 |
scRNA-seq of TdTomato+ cells from PAH3 mouse |
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| Relations |
| BioProject |
PRJNA648057 |
| SRA |
SRP273247 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE154959_RAW.tar |
55.7 Mb |
(http)(custom) |
TAR (of H5) |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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