Ruiting Wen1, Chen Dong1, Chen Xu1, Long Zhao2, Yang Yang2, Zhihua Zhang2, Yanfu Chen2, Lian Duan3, Hu Chen2, Zhigang Yang4, Bin Zhang5. 1. 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. 2. 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. 3. The Fifth Medical Center of Chinese PLA General Hospital, Beijing 100071, PR China. 4. Department of Hematology and Rheumatology, Zhanjiang Central Hospital, Guangdong Medical University, Zhanjiang 524045, PR China. Electronic address: yangzg@gdmu.edu.cn. 5. 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. Electronic address: zb307ctc@163.com.
Abstract
BACKGROUND: 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. METHODS: 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. RESULTS: UM171 treatment increased the proportion of CD34+ (68.97 ± 6.91%), CD34+ CD38- cells (44.10 ± 9.20%) and CD34+CD38-CD45RA-CD90+ 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. CONCLUSIONS: 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.
BACKGROUND: Autologous hematopoietic stem cell transplantation is an effective therapeutic strategy for lymphomapatients. 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 lymphomapatients using UM171 to solve the problem of HSCs deficiency. METHODS: Mobilized peripheral blood-derived CD34+ cells from lymphomapatients 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. RESULTS: UM171 treatment increased the proportion of CD34+ (68.97 ± 6.91%), CD34+ CD38- cells (44.10 ± 9.20%) and CD34+CD38-CD45RA-CD90+ 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. CONCLUSIONS: Our study shows that UM171 can efficiently promote ex vivo expansion of HSCs from lymphomapatients, 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.
Authors: James L C Che; Daniel Bode; Iwo Kucinski; Alyssa H Cull; Fiona Bain; Hans J Becker; Maria Jassinskaja; Melania Barile; Grace Boyd; Miriam Belmonte; Andy G X Zeng; Kyomi J Igarashi; Juan Rubio-Lara; Mairi S Shepherd; Anna Clay; John E Dick; Adam C Wilkinson; Hiromitsu Nakauchi; Satoshi Yamazaki; Berthold Göttgens; David G Kent Journal: EMBO Rep Date: 2022-08-16 Impact factor: 9.071