The Analysis of the Human Megakaryocyte and Platelet Coding Transcriptome in Healthy and Diseased Subjects
Abstract
:1. Introduction to Megakaryopoiesis and Platelet Formation with a Focus on RNA
2. Bulk Transcriptomics of Healthy Megakaryocytes
3. Bulk Transcriptomics of Healthy Platelets
4. Single-Cell Transcriptomics of Healthy Megakaryocytes
Reference | Cell Type(S) | Summary of Results | Technology |
---|---|---|---|
Choudry et al. (2021) [73] | MKs and HSCs | MKs in lower ploidy states highly express platelet-specific genes. As polyploidization increases and the cell prepares for thrombopoiesis, gene expression is redirected towards transcriptional programs involved in translation and posttranslational processing. Two MK-biased HSC subpopulations were also observed and shown to originate from the BM. Finally, BM MKs from patients with recent myocardial infarction showed a specific gene expression signature that supports the modulation of MK differentiation in this thrombotic state. | G&T-seq |
Estevez et al. (2021) [98] | HSCs | The effect of germline monoallelic mutations in RUNX1, found in patients suffering from familial platelet disorder with a predisposition to myeloid malignancy, was studied by inserting the patient mutations into iPSC-derived hematopoietic progenitor cells (iHSCs) and performing scRNA-seq. There was found to be a marked deficiency of MK-biased iHSCs in mutated cultures, and gene sets that were upregulated included response to stress, regulation of signal transduction and immune signaling-related gene sets. An increased sensitivity to transforming growth factor β1 and an increase in the stress pathway through upregulation of c-jun N-terminal kinase-2 phosphorylation were observed. | 10X Chromium |
Lawrence et al. (2022) [94] | In vitro differentiating cells from iPSCs and HSCs up to MKs | Analysis of iPSC-derived MK differentiation and transcriptomic comparison with primary hematopoietic stem and progenitor cells. The in vitro cells do not pass through states resembling HSCs or MPPs as seen in vivo, but the further differentiated MK progenitor cells do exhibit a very similar transcriptome to their in vivo counterparts. A surface marker panel is described for MK progenitors, allowing for selection from culture and for insights into this intermediary state. | 10X Chromium and smart-seq |
Liu et al. (2021) [90] | MKs | Cellular heterogeneity within MKs was mapped, and an MK subpopulation with high enrichment of immune-associated genes was identified. The immune signature could be traced back to the progenitor stage, and two surface markers, CD148 and CD48, were identified. This type of MK can respond rapidly to immune stimuli both in vitro and in vivo, exhibiting high expression of immune receptors and mediators, which might act as immune-surveillance cells. | Smart-seq |
Lu et al. (2018) [96] | MEPs, common myeloid progenitors, and MK and erythroid progenitors | MEPs have a distinct gene expression signature that represents a continuous transition state from common myeloid progenitor cells to MK and erythroid progenitor cells. | Fluidigm C1 |
Psaila et al. (2017) [95] | CD34+ peripheral blood cells | Myelofibrosis causes an increased number of immature/low ploidy MKs with an altered transcriptome. Patient HSPCs have increased expression of MK-associated genes, including VWF and ITGA2B. Patient CD34+ progenitor cells showed increased expression of PF4 and TGFβ. | 10X Chromium |
Psaila et al. (2020) [97] | HSPCs | MK-biased hematopoiesis in myelofibrosis was observed, with heterogeneous MKp showing a highly expressed fibrosis signature and an aberrant metabolic and inflammatory signature. Targeting the aberrant expression of surface G6B may selectively ablate the myelofibrosis HSPC clone. | 10X Chromium |
Riemondy et al. (2019) [93] | Lymphocytes and MK mixture | The introduction of a method for resampling cell-type-wise, cell-wise or sample-wise from an existing complex scRNA library. | 10X Chromium |
Sun et al. (2021) [91] | Human and mouse MKs | Three distinct MK subpopulations were observed to possess gene signatures related to platelet generation, HSC niche interaction or inflammatory response. The first type of MK was mostly found near blood vessels, and the second was near HSCs. The third type, containing a gene signature related to the inflammatory response, was lower in ploidy, consisted of 5% of MKs and was capable of engulfing and digesting bacteria and stimulating T cells in vitro. | Smart-seq |
Wang et al. (2021) [92] | Human MKs | A comprehensive single-cell transcriptomic landscape of human MKs was constructed where MKs show cellular heterogeneity with distinct metabolic and cell cycle signatures. CD14+ MKs with immune characteristics were generated along a distinct trajectory, and THBS1 was identified as an early marker for MK-biased endothelial cells from human embryonic stem cells. | 10X Chromium |
5. Single-Cell Transcriptomics of Healthy Platelets
6. Platelet and Megakaryocyte Transcriptome to Decipher Inherited Platelet Disorders
7. Conclusions and Future Perspectives
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Plate-Based | Droplet-Based |
---|---|
Higher sensitivity | Higher cell throughput |
Better for large and fragile cells | Lower labor intensity |
Cheaper setup cost | Cheaper cost per cell |
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De Wispelaere, K.; Freson, K. The Analysis of the Human Megakaryocyte and Platelet Coding Transcriptome in Healthy and Diseased Subjects. Int. J. Mol. Sci. 2022, 23, 7647. https://doi.org/10.3390/ijms23147647
De Wispelaere K, Freson K. The Analysis of the Human Megakaryocyte and Platelet Coding Transcriptome in Healthy and Diseased Subjects. International Journal of Molecular Sciences. 2022; 23(14):7647. https://doi.org/10.3390/ijms23147647
Chicago/Turabian StyleDe Wispelaere, Koenraad, and Kathleen Freson. 2022. "The Analysis of the Human Megakaryocyte and Platelet Coding Transcriptome in Healthy and Diseased Subjects" International Journal of Molecular Sciences 23, no. 14: 7647. https://doi.org/10.3390/ijms23147647