Deciphering transcriptome alterations in bone marrow hematopoiesis at single-cell resolution in immune thrombocytopenia

Abstract

Immune thrombocytopenia (ITP) is an autoimmune disorder, in which megakaryocyte dysfunction caused by an autoimmune reaction can lead to thrombocytopenia, although the underlying mechanisms remain unclear. Here, we performed single-cell transcriptome profiling of bone marrow CD34+ hematopoietic stem and progenitor cells (HSPCs) to determine defects in megakaryopoiesis in ITP. Gene expression, cell-cell interactions, and transcriptional regulatory networks varied in HSPCs of ITP, particularly in immune cell progenitors. Differentially expressed gene (DEG) analysis indicated that there was an impaired megakaryopoiesis of ITP. Flow cytometry confirmed that the number of CD9+ and HES1+ cells from Lin−CD34+CD45RA− HSPCs decreased in ITP. Liquid culture assays demonstrated that CD9+Lin−CD34+CD45RA− HSPCs tended to differentiate into megakaryocytes; however, this tendency was not observed in ITP patients and more erythrocytes were produced. The percentage of megakaryocytes differentiated from CD9+Lin−CD34+CD45RA− HSPCs was 3-fold higher than that of the CD9− counterparts from healthy controls (HCs), whereas, in ITP patients, the percentage decreased to only 1/4th of that in the HCs and was comparable to that from the CD9− HSPCs. Additionally, when co-cultured with pre-B cells from ITP patients, the differentiation of CD9+Lin−CD34+CD45RA− HSPCs toward the megakaryopoietic lineage was impaired. Further analysis revealed that megakaryocytic progenitors (MkP) can be divided into seven subclusters with different gene expression patterns and functions. The ITP-associated DEGs were MkP subtype-specific, with most DEGs concentrated in the subcluster possessing dual functions of immunomodulation and platelet generation. This study comprehensively dissects defective hematopoiesis and provides novel insights regarding the pathogenesis of ITP.

Single-Cell RNA Sequencing Profiling Cellular Heterogeneity and Specific Responses of Fish Gills to Microplastics and Nanoplastics

“…The cell suspensions were filtered through a 40 μm nylon cell strainer and the red blood cells were removed using red blood cell lysis buffer (Thermo Fisher Scientific, USA). This method was reported to exert no impact on thrombocytes . Cell viability of each sample was checked on a Countstar fluorescence cell analyzer (Aber Instruments Ltd., UK).…”

“…This method was reported to exert no impact on thrombocytes. 38 Cell viability of each sample was checked on a Countstar fluorescence cell analyzer (Aber Instruments Ltd., UK). Before the samples were sequenced, the cell viability for each sample was confirmed to be above 80%.…”

Single-cell transcriptomics dissects the transcriptome alterations of hematopoietic stem cells in myelodysplastic neoplasms

“…We used Monocle 2(version 2.28.0) to construct the trajectory tree of all groups of cells, which indicated the distinct lineage differentiation potential for each group of cells. For identification of major patterns along the pseudotime, the top 1500 pseudotime-dependent genes were selected by the differentialGeneTest function with setting the parameters “fullModelFormulaStr” as “ sm.ns(Pseudotime)” and were clustered into three distinct patterns by k-means clustering [ 61 ]. BEAM analysis was used to get the expression patterns in branches during development [ 62 ].…”

<strong></strong>Type 2 Hypersensitivity Disorders including Systemic Lupus Erythematosus, Sjogren’s Syndrome, Grave’s Disease, Myasthenia Gravis, Immune Thrombocytopenia, Autoimmune Hemolytic Anemia, Dermatomyositis, and Graft versus Host Disease Are THab Dominant A

“…The toll-like receptor relating to anti-viral immunity, TLR7, is up-regulated in immune thrombocytopenia [110]. Type 1 interferon signaling related transcription factor, IRF3, adaptive immunity related transcription factor, NFkB, are both over-represented in immune thrombocytopenia [112]. The master THαβ immune transcription factors, STAT1 and STAT3, are also over-represented in immune thrombocytopenia [113,114].…”