Single-Cell Transcriptome Analysis of CD34+ Stem Cell-Derived Myeloid Cells Infected With Human Cytomegalovirus
Myeloid cells are important sites of lytic and latent infection by human cytomegalovirus (CMV). We previously showed that only a small subset of myeloid cells differentiated from CD34+ hematopoietic stem cells is permissive to CMV replication, underscoring the heterogeneous nature of these populatio...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2019-03-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fmicb.2019.00577/full |
| _version_ | 1819151479269752832 |
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| author | Melissa Galinato Kristen Shimoda Alexis Aguiar Fiona Hennig Dario Boffelli Michael A. McVoy Laura Hertel |
| author_facet | Melissa Galinato Kristen Shimoda Alexis Aguiar Fiona Hennig Dario Boffelli Michael A. McVoy Laura Hertel |
| author_sort | Melissa Galinato |
| collection | DOAJ |
| description | Myeloid cells are important sites of lytic and latent infection by human cytomegalovirus (CMV). We previously showed that only a small subset of myeloid cells differentiated from CD34+ hematopoietic stem cells is permissive to CMV replication, underscoring the heterogeneous nature of these populations. The exact identity of resistant and permissive cell types, and the cellular features characterizing the latter, however, could not be dissected using averaging transcriptional analysis tools such as microarrays and, hence, remained enigmatic. Here, we profile the transcriptomes of ∼7000 individual cells at day 1 post-infection using the 10× genomics platform. We show that viral transcripts are detectable in the majority of the cells, suggesting that virion entry is unlikely to be the main target of cellular restriction mechanisms. We further show that viral replication occurs in a small but specific sub-group of cells transcriptionally related to, and likely derived from, a cluster of cells expressing markers of Colony Forming Unit – Granulocyte, Erythrocyte, Monocyte, Megakaryocyte (CFU-GEMM) oligopotent progenitors. Compared to the remainder of the population, CFU-GEMM cells are enriched in transcripts with functions in mitochondrial energy production, cell proliferation, RNA processing and protein synthesis, and express similar or higher levels of interferon-related genes. While expression levels of the former are maintained in infected cells, the latter are strongly down-regulated. We thus propose that the preferential infection of CFU-GEMM cells may be due to the presence of a pre-established pro-viral environment, requiring minimal optimization efforts from viral effectors, rather than to the absence of specific restriction factors. Together, these findings identify a potentially new population of myeloid cells permissive to CMV replication, and provide a possible rationale for their preferential infection. |
| first_indexed | 2024-12-22T14:34:03Z |
| format | Article |
| id | doaj.art-bc5153a894ae4ca19e0fc84b21e08644 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-22T14:34:03Z |
| publishDate | 2019-03-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-bc5153a894ae4ca19e0fc84b21e086442022-12-21T18:22:40ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2019-03-011010.3389/fmicb.2019.00577434560Single-Cell Transcriptome Analysis of CD34+ Stem Cell-Derived Myeloid Cells Infected With Human CytomegalovirusMelissa Galinato0Kristen Shimoda1Alexis Aguiar2Fiona Hennig3Dario Boffelli4Michael A. McVoy5Laura Hertel6Center for Immunobiology and Vaccine Development, Children’s Hospital Oakland Research Institute, Oakland, CA, United StatesCenter for Immunobiology and Vaccine Development, Children’s Hospital Oakland Research Institute, Oakland, CA, United StatesCenter for Immunobiology and Vaccine Development, Children’s Hospital Oakland Research Institute, Oakland, CA, United StatesCenter for Genetics, Children’s Hospital Oakland Research Institute, Oakland, CA, United StatesCenter for Genetics, Children’s Hospital Oakland Research Institute, Oakland, CA, United StatesDepartment of Pediatrics, Virginia Commonwealth University, Richmond, VA, United StatesCenter for Immunobiology and Vaccine Development, Children’s Hospital Oakland Research Institute, Oakland, CA, United StatesMyeloid cells are important sites of lytic and latent infection by human cytomegalovirus (CMV). We previously showed that only a small subset of myeloid cells differentiated from CD34+ hematopoietic stem cells is permissive to CMV replication, underscoring the heterogeneous nature of these populations. The exact identity of resistant and permissive cell types, and the cellular features characterizing the latter, however, could not be dissected using averaging transcriptional analysis tools such as microarrays and, hence, remained enigmatic. Here, we profile the transcriptomes of ∼7000 individual cells at day 1 post-infection using the 10× genomics platform. We show that viral transcripts are detectable in the majority of the cells, suggesting that virion entry is unlikely to be the main target of cellular restriction mechanisms. We further show that viral replication occurs in a small but specific sub-group of cells transcriptionally related to, and likely derived from, a cluster of cells expressing markers of Colony Forming Unit – Granulocyte, Erythrocyte, Monocyte, Megakaryocyte (CFU-GEMM) oligopotent progenitors. Compared to the remainder of the population, CFU-GEMM cells are enriched in transcripts with functions in mitochondrial energy production, cell proliferation, RNA processing and protein synthesis, and express similar or higher levels of interferon-related genes. While expression levels of the former are maintained in infected cells, the latter are strongly down-regulated. We thus propose that the preferential infection of CFU-GEMM cells may be due to the presence of a pre-established pro-viral environment, requiring minimal optimization efforts from viral effectors, rather than to the absence of specific restriction factors. Together, these findings identify a potentially new population of myeloid cells permissive to CMV replication, and provide a possible rationale for their preferential infection.https://www.frontiersin.org/article/10.3389/fmicb.2019.00577/fullsingle-celltropismcytomegalovirusmyeloid10× genomicsRNAseq |
| spellingShingle | Melissa Galinato Kristen Shimoda Alexis Aguiar Fiona Hennig Dario Boffelli Michael A. McVoy Laura Hertel Single-Cell Transcriptome Analysis of CD34+ Stem Cell-Derived Myeloid Cells Infected With Human Cytomegalovirus Frontiers in Microbiology single-cell tropism cytomegalovirus myeloid 10× genomics RNAseq |
| title | Single-Cell Transcriptome Analysis of CD34+ Stem Cell-Derived Myeloid Cells Infected With Human Cytomegalovirus |
| title_full | Single-Cell Transcriptome Analysis of CD34+ Stem Cell-Derived Myeloid Cells Infected With Human Cytomegalovirus |
| title_fullStr | Single-Cell Transcriptome Analysis of CD34+ Stem Cell-Derived Myeloid Cells Infected With Human Cytomegalovirus |
| title_full_unstemmed | Single-Cell Transcriptome Analysis of CD34+ Stem Cell-Derived Myeloid Cells Infected With Human Cytomegalovirus |
| title_short | Single-Cell Transcriptome Analysis of CD34+ Stem Cell-Derived Myeloid Cells Infected With Human Cytomegalovirus |
| title_sort | single cell transcriptome analysis of cd34 stem cell derived myeloid cells infected with human cytomegalovirus |
| topic | single-cell tropism cytomegalovirus myeloid 10× genomics RNAseq |
| url | https://www.frontiersin.org/article/10.3389/fmicb.2019.00577/full |
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