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GEO help: Mouse over screen elements for information. |
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| Status |
Public on May 19, 2023 |
| Title |
RNA sequencing of isolated brain endothelial cells from Smad4, Alk1, or Eng endothelial cell-specific knock-out mutants, respective wildtypes (WTs), and Angiopoietin-2 inhibition transcriptional changes. |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Purpose: Hereditary Hemorrhagic Telangiectasia (HHT) is an autosomal dominant vascular disorder characterized by arteriovenous malformations (AVMs). Over 90% of HHT patients carry heterozygous loss-of-function mutations in Transforming Growth Factor-beta (TGFb) signaling components activin receptor-like kinase 1 (ACVRL1/ALK1), endoglin (ENG), or mothers against decapentaplegic homolog 4 (SMAD4). Brain AVMs can be lethal for HHT patients. Here, we use an inducible endothelial (EC)-specific mouse line to investigate brain AVMs characteristics in Alk1-, Eng-, and Smad4-HHT mouse models. Angiopientin-2 (Ang2) inhibition diminishes HHT associated cerebral vascular defects. We aim to identify the shared transcriptional changes in brain ECs between these three HHT types, and provide insights for transcriptional changes upon Ang2 inhibition.
Methods: ECs were isolated from brains of both WT and HHT mutant mice at postnatal day 7 (P7) for Smad4 and Eng models, P6 for Alk1 model. Total RNA was extracted from isolated brain ECs and quantified using Qubit RNA High Sensitivity Assay Kit. RNA integrity was determined using the Bioanalyzer RNA 6000 Nano assay kit. RNA library construction was performed with the TruSeq RNA Library Prep Kit v2 from Illumina. The resulting mRNA library was quantified using Qubit dsDNA High Sensitivity Assay Kit and verified using the Bioanalyzer DNA1000 assay kit. Verified samples were sequenced using the NextSeq 500/550 High Output Kit v2.5 (150 Cycles) on a Nextseq 550 system. Sequenced reads were aligned to the mouse (mm10) reference genome with RNA-seq alignment tool (version 2.0.1). The aligned reads were used to quantify mRNA expression and determine differentially expressed genes using the RNA-seq Differential Expression tool (version 1.0.1). Both alignment and differential expression analysis were performed using the tools in the BaseSpace Sequence Hub.
Results: We identified 5509, 5381, and 2663 differentially expressed genes in Smad4, Alk1, and Eng HHT models respectively. Also, Ang2 inhibition mainly funcioned through angiogenesis and cell migration processes to normalize cerebrovascular defects in Smad4-HHT model.
Conclusions: A common but unique pro-angiogenic program among all HHT models included 14 consistent upregulated and 5 consistent downregulated angiogic genes. Also, Ang2 inhibition mainly funcioned through angiogenesis and cell migration processes to normalize cerebrovascular defects in Smad4-HHT model.
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| Overall design |
Isolated brain ECs mRNA profiles of postnatal day 7 (P7) for Smad4 and Eng models, P6 for Alk1 model, 5 samples in each WT/mutant group. IgG/LC10 treatments were administered at P2 and P4 for Smad4-HHT model.
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| Web link |
https://pubmed.ncbi.nlm.nih.gov/37288572/
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| Contributor(s) |
Zhou X, Meadows SM |
| Citation(s) |
37288572 |
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| Submission date |
Feb 21, 2022 |
| Last update date |
Sep 16, 2023 |
| Contact name |
Xingyan Zhou |
| Organization name |
Tulane University
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| Street address |
6823 St Charles Ave
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| City |
New Orleans |
| ZIP/Postal code |
70118 |
| Country |
USA |
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| Platforms (1) |
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| Samples (39)
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| Relations |
| BioProject |
PRJNA808990 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE197105_Alk1_Gene_counts.csv.gz |
1.5 Mb |
(ftp)(http) |
CSV |
| GSE197105_Eng_Gene_counts.csv.gz |
1.1 Mb |
(ftp)(http) |
CSV |
| GSE197105_Genecounts_IgGWT_vs_IgGMUT.csv.gz |
687.1 Kb |
(ftp)(http) |
CSV |
| GSE197105_Genecounts_LC10WT_vs_LC10MUT.csv.gz |
934.1 Kb |
(ftp)(http) |
CSV |
| GSE197105_Smad4_Gene_counts.csv.gz |
1.1 Mb |
(ftp)(http) |
CSV |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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