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| Status |
Public on Jun 06, 2018 |
| Title |
AmpliSeq Transcriptome Analysis of Human Alveolar and Monocyte-Derived Macrophages Over Time in Response to Mycobacterium tuberculosis infection |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
Non-coding RNA profiling by high throughput sequencing
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| Summary |
Human alveolar macrophages (HAM) are primary bacterial niche and immune response cells during Mycobacterium tuberculosis (M.tb) infection, and human blood monocyte-derived macrophages (MDM) are a model for investigating M.tb-macrophage interactions. Here, we use a targeted RNA-Seq method to measure transcriptome-wide changes in RNA expression patterns of freshly obtained HAM (used within 6 h) and 6 day cultured MDM upon M.tb infection over time (2, 24 and 72 h), in both uninfected and infected cells from three donors each. The Ion AmpliSeq™ Transcriptome Human Gene Expression Kit (AmpliSeq) uses primers targeting 18,574 mRNAs and 2,228 non-coding RNAs (ncRNAs) for a total of 20,802 transcripts. AmpliSeqTM yields highly precise and reproducible gene expression profiles (R2 >0.99). Taking advantage of AmpliSeq’s reproducibility, we establish well-defined quantitative RNA expression patterns of HAM versus MDM, including significant M.tb-inducible genes, in networks and pathways that differ in part between MDM and HAM. A similar number of expressed genes are detected at all time-points between uninfected MDM and HAM, in common pathways including inflammatory and immune functions, but canonical pathway differences also exist. In particular, at 2 h, multiple genes relevant to the immune response are preferentially expressed in either uninfected HAM or MDM, while the HAM RNA profiles approximate MDM profiles over time in culture, highlighting the unique RNA expression profile of freshly obtained HAM. MDM demonstrate a greater transcriptional response than HAM upon M.tb infection, with 2 to >10 times more genes up- or down-regulated. The results identify key genes involved in cellular responses to M.tb in two different human macrophage types. Follow-up bioinformatics analysis indicates that approximately 30% of response genes have expression quantitative trait loci (eQTLs in GTEx), common DNA variants that can influence host gene expression susceptibility or resistance to M.tb, illustrated with the TREM1 gene cluster and IL-10.
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| Overall design |
Assessment of transcriptome profiles from cells infected with Mycobacterium tuberculosis using AmpliSeq.
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| Contributor(s) |
Sadee W, Schlesinger LS, Papp A, Azad A, Pietrzak M |
| Citation(s) |
29847580 |
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| Submission date |
May 11, 2018 |
| Last update date |
Mar 01, 2019 |
| Contact name |
Maciej Pietrzak |
| E-mail(s) |
pietrzak.20@osu.edu
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| Phone |
+1(614) 688 9721
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| Organization name |
Ohio State University
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| Department |
Department of Biomedical Informatics College of Medicine
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| Street address |
1800 Cannon Drive, 340C Lincoln Tower
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| City |
Columbus |
| State/province |
OH |
| ZIP/Postal code |
43210 |
| Country |
USA |
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| Platforms (1) |
| GPL17303 |
Ion Torrent Proton (Homo sapiens) |
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| Samples (36)
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| Relations |
| BioProject |
PRJNA471095 |
| SRA |
SRP145502 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE114371_FeatureCounts.txt.gz |
860.5 Kb |
(ftp)(http) |
TXT |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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