Academy of Military Medical Sciences, Academy of Military Sciences, Beijing 100850, PR China; Department of Hematopoietic Stem Cell Transplantation, the Fifth Medical Center of Chinese PLA General Hospital, Beijing 100071, PR China; Beijing Key Laboratory of Stem Cell Therapy and Transformation Research, Beijing 100071, PR China.
Department of Hematopoietic Stem Cell Transplantation, the Fifth Medical Center of Chinese PLA General Hospital, Beijing 100071, PR China; Beijing Key Laboratory of Stem Cell Therapy and Transformation Research, Beijing 100071, PR China.
Int Immunopharmacol. 2020 Apr;81:106266. doi: 10.1016/j.intimp.2020.106266. Epub 2020 Feb 12.
Autologous hematopoietic stem cell transplantation is an effective therapeutic strategy for lymphoma patients. However, some patients have to give up receiving transplantation because of failing to obtain sufficient CD34 cells yields. Therefore, we ex vivo expanded HSCs of lymphoma patients using UM171 to solve the problem of HSCs deficiency.
Mobilized peripheral blood-derived CD34 cells from lymphoma patients were cultured for 10 days with or without UM171. The fold of cell expansion and the immunophenotype of expanded cells were assessed by flow cytometry. RNA-seq experiment was performed to identify the mechanism by which UM171 promoted HSCs expansion.
UM171 treatment increased the proportion of CD34 (68.97 ± 6.91%), CD34 CD38 cells (44.10 ± 9.20%) and CD34CD38CD45RACD90 LT-HSCs (3.05 ± 2.08%) compared to vehicle treatment (36.08 ± 11.14%, 18.30 ± 9.49% and 0.56 ± 0.45%, respectively). UM171 treatment led to an 85.08-fold increase in LT-HSC numbers relative to initial cells. Importantly, UM171 promoted expansion of LT-HSCs achieved 138.57-fold in patients with poor mobilization. RNA-seq data showed that UM171 upregulated expression of HSC-, mast cell-specific genes and non-canonical Wnt signaling related genes, and inhibited genes expression of erythroid, megakaryocyte and inflammatory mediated chemokine.
Our study shows that UM171 can efficiently promote ex vivo expansion of HSCs from lymphoma patients, especially for poorly mobilizing patients. In terms of mechanism, UM171 upregulate HSC-specific genes expression and suppress erythroid and megakaryocytic differentiation, as well as activate non-classical Wnt signaling.
自体造血干细胞移植是治疗淋巴瘤患者的有效治疗策略。然而，由于无法获得足够的 CD34 细胞产量，一些患者不得不放弃接受移植。因此，我们使用 UM171 体外扩增淋巴瘤患者的 HSCs 以解决 HSCs 缺乏的问题。
从淋巴瘤患者的动员外周血中分离出 CD34 细胞，用或不用 UM171 培养 10 天。通过流式细胞术评估细胞扩增的倍数和扩增细胞的免疫表型。进行 RNA-seq 实验以确定 UM171 促进 HSCs 扩增的机制。
与对照组相比，UM171 处理增加了 CD34（68.97±6.91%）、CD34 CD38 细胞（44.10±9.20%）和 CD34 CD38 CD45RACD90 LT-HSCs（3.05±2.08%）的比例。UM171 处理使 LT-HSC 的数量相对于初始细胞增加了 85.08 倍。重要的是，UM171 促进了动员不良患者的 LT-HSC 扩增，达到了 138.57 倍。RNA-seq 数据显示，UM171 上调了 HSC、肥大细胞特异性基因和非经典 Wnt 信号相关基因的表达，并抑制了红系、巨核细胞和炎症介导趋化因子的基因表达。
我们的研究表明，UM171 可以有效地促进淋巴瘤患者体外扩增 HSCs，特别是对动员不良的患者。就机制而言，UM171 上调 HSC 特异性基因的表达，抑制红系和巨核细胞的分化，并激活非经典 Wnt 信号。
