Differental Distribution of Hematopoietic Cells in Bone Marrow of PNH Patients Were Showed By Single-Cell Sequencing

Background：Paroxysmal nocturnal hemoglobinuria (PNH) is a rare clonogenic disease of hematopoietic stem cells. However, the mechanism of proliferative advantage of PNH clones remains unclear. Single-cell sequencing has been widely implemented to explore the cellular and molecular features in hematopoiesis.

Methods: Bone marrow mononuclear cells were acquired from 3 PNH patients and 4 healthy controls. CD59+ and CD59- cells were sorted by flow cytometry in PNH patients. Then we performed deep single-cell RNA sequencing for all cell lines of hematopoietic cells. We preliminarily analyzed the distribution and changes of hematopoietic cells in bone marrow of PNH patients.

Results: Fourteen types of hematopietic cells were identified in our study, including myeloid cells (CMP, GMP, neutrophils and monocytes), lymphocytes (CLP, B cells, T cells and NK cells), megakaryocyte-erythroid progenitor cell (MEP) and erythrocytes. We observed that AVP, CD79B, GZMH, CCR7,SPI1 and GATA1, respected specifically for HSPCs, B cells, NK cells, T cells, Neutrophil/Monocytes and Erythrocytes. Compared with normal controls, we found increased erythroid, significantly reduced B cells and slightly reduced neutrophils in CD59- cells, and sharply reduced T/NK cells in CD59+ cells of PNH patients. Furthermore, we analyzed deeply the changes of T/NK cells in PNH patients. We found that T cells decreased and NK cells increased in CD59- cells, while the ratio of T/NK cells in CD59+ cells of PNH patients were no significant difference. For the subtype of NK cells, CD56dim NK cells increased in CD59- cells, while Cytokine-inducedmemory-like (CIML) NK cells significantly increased in CD59+ cells in PNH patients. Interestingly, we observed one type of CD56dim NK cells (10.0 in Fig 1D1), which is sharply increased in CD59- cells. More analysis in differential genes and pathway analysis should be done further.

Conclusion: Differental distribution of hematopoietic cells in bone marrow of PNH patients were showed by single-cell RNA sequencing, especially subtypes of NK cells in CD59- and CD59+ cells.

Keywords: PNH, single-cell RNA sequencing, NK cells, CIML NK cells

No relevant conflicts of interest to declare.