PRJNA661222: Next Generation Sequencing Quantitative Analysis on COVID-19 Kidney tissue
概要: Purpose: The goals of this study are to compare NGS-derived transcriptome profiling (RNA-seq) of COVID-19 kidney to normal controls. Methods/Results: Kidney mRNA profile of human COVID-19 tissue was generated by deep sequencing using Illumina Novaseq6000 Paired-end 150. After filtering reads mapped to contamination database, the reads that were uniquely aligned to the exon and splicing-junction segments with a maximal 2 mismatches for each transcript were then counted as expression level for corresponding transcript. Next, RNA-seq reads count data were downloaded from public resource GTEx project (https://www.gtexportal.org/home/datasets) and 12 normal kidney tissue samples were extracted as controls. The differential analysis by fold change difference was carried out to identify dysregulated genes at 1.5 fold change. The differential expressed genes were then subjected to Gene Ontology function and Pathway (KEGG, Ingenuity IPA, BIOCARTA, NABA, Panther, PID, REACTOME, Wiki-pathway) enrichment analysis by Fisher-exact test. Conclusions: RNA sequencing data revealed that biological processes from upregulated genes were enriched for cell cycle, chromosome segregation, response to wounding, humoral immune response, and blood coagulation, suggesting that cell injury/regeneration, inflammatory response, and endothelial injury were the major disease processes involved. The biological processes from downregulated genes were enriched for ion transport, metabolic processes, and oxidation, likely secondary to severe tubular cell injury. Pathway analysis from both up- and downregulated genes showed enrichment of transmembrane transport, oxidation, and blood coagulation consistent with the GO terms analysis. Upregulated genes were enriched only for the FOXM1 pathway, which was recently reported to promote tubular cell proliferation during injury repair. Additionally, genes related to the renin-angiotensin system were downregulated, but ACE2 expression did not differ from normal controls.
项目整体设计: NGS-derived transcriptome profiling (RNA-seq) of COVID-19 kidney
| 生长方案: |
- |
| 处理方案: |
- |
| 提取方案: |
RNA was extracted from frozen kidney biopsy tissue using a Qiagen miRNeasy Mini Kit (217004) and eluted in water. |
| 建库方案: |
RNA libraries were prepared for sequencing using standard Illumina protocols |
| 分子类型: |
polyA(+) RNA |
| 库的片段类型: |
PAIRED |
| 库的链类型: |
Forward |
| 测序平台: |
ILLUMINA |
| 测序仪型号: |
Illumina NovaSeq 6000 |
| 链特异性: |
Specific |
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物种 |
种族 |
族裔 |
年龄 |
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组织 |
细胞类型 |
细胞亚型 |
细胞系 |
疾病 |
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突变/变异 |
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Condition Detail |
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提取方案 |
建库方案 |
分子类型 |
库的片段类型 |
链特异性 |
库的链类型 |
加标(Spike-In) |
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细胞数 |
测序片段数 |
碱基数 |
平均测序片段长度_1 |
平均测序片段长度_2 |
唯一比对率 |
多重比对率 |
覆盖度 |
Molecular Analysis of the Kidney From a Patient With COVID-19-Associated Collapsing Glomerulopathy.
Kidney medicine . 2021-04-28 [PMID:
33942030]