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RUNX1-205, a novel splice variant of the human RUNX1 gene, has blockage effect on mesoderm-hemogenesis transition and promotion effect during the late stage of hematopoiesis

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Journal of Molecular Cell Biology 2020年第5期12卷 386-396页

作者： Wencui Sun Jiahui Zeng Jing Chang Yuan Xue Yonggang Zhang Xu Pan Ya Zhou mowen lai Guohui Bian Qiongxiu Zhou Jiaxing Liu Bo Chen Feng Ma Institute of Blood Transfusion Chinese Academy of Medical Sciences&Peking Union Medical College(CAMS&PUMC)Chengdu 610052China State Key Laboratory of Biotherapy Sichuan UniversityChengdu 61006China State Key Laboratory of Experimental Hematology CAMS&PUMCTianjin 300020China

Runt-related transcription factor 1(RUNX1)is required for definitive hematopoiesis;however,the functions of most human RUNX1 isoforms are unclear.In particular,the effects of RUNX1-205(a novel splice variant that lacks exon 6 in comparison with RUNX1b)on human hematopoiesis are not clear.In this study,a human embryonic stem cell(hESC)line with inducible RUNX1-205 overexpression was established.Analyses of these cells revealed that induction of RUNX1-205 overexpression at early stage did not influence the induction of mesoderm but blocked the emergence of CD34+cells,and the production of hematopoietic stem/progenitor cells was significantly reduced.In addition,the expression of hematopoiesis-related factors was downregulated.However,these effects were abolished when RUNX1-205 overexpression was induced after Day 6 in co-cultures of hESCs and AGM-S3 cells,indicating that the inhibitory effect occurred prior to generation of hemogenic endothelial cells,while the promotive effect could be observed during the late stage of hematopoiesis.This is very similar to that of RUNX1b.Interestingly,the mRNA expression profile of RUNX1-205 during hematopoiesis was distinct from that of RUNX1b,and the protein stability of RUNX1-205 was much higher than that of RUNX1b.Thus,the function of RUNX1-205 in normal and diseased models should be further explored.

关键词： mesoderm-hemogenesis transition hematopoiesis hematopoietic stem/progenitor cells (HSPCs) RUNX1 mesoderm embryonic stem cells (ESCs)

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Inducible overexpression of RUNX1b/c in human embryonic stem cells blocks early hematopoiesis from mesoderm

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Journal of Molecular Cell Biology 2017年第4期9卷 262-273页

作者： Bo Chen Jiawen Teng Hongwei Liu Xu Pan Ya Zhou Shu Huang mowen lai Guohui Bian Bin Mao Wencui Sun Qiongxiu Zhou Shengyong Yang Tatsutoshi Nakahata Feng Ma Institute of Blood Transfusion Chinese Academy of Medical Sciences & Peking Union Medical College (CAMS & PUMC) Chengdu 610052 China State Key Laboratory of Biotherapy Sichuan University Chengdu 610065 China Center for iPS Cell Research and Application (CiRA) Kyoto University Kyoto 606-8507 Japan State Key Laboratory of Experimental Hematology CAMS & PUMC Tianjin 300020 China

RUNX1 绝对为 RUNX1b/c 的权威的造血作用，而是功能被要求，人的 RUNX1 的二 isoforms，是不清楚的。我们建立了可诱导的 RUNX1b/c-overexpressing 人的胚胎的干细胞(hESC ) 线，在 RUNX1b/c， overexpression 阻止了 CD34+ 房间的出... 详细信息

RUNX1 绝对为 RUNX1b/c 的权威的造血作用，而是功能被要求，人的 RUNX1 的二 isoforms，是不清楚的。我们建立了可诱导的 RUNX1b/c-overexpressing 人的胚胎的干细胞(hESC ) 线，在 RUNX1b/c， overexpression 阻止了 CD34+ 房间的出现早舞台，急速地从而减少造血的干细胞的生产。同时，造血作用相关的因素的表示是 downregulated。然而，如此的阻塞效果从在 hESC/AGM-S3 房间合作文化的白天 6 消失了，证明阻塞发生在 hemogenic endothelial 房间的产生前。这阻塞被 RepSox 部分救，转变生长因素(TGF ) 的一个禁止者 - 发信号的小径，显示在 RUNX1b/c 和 TGF- 小径之间的一种靠近的关系。我们的结果在早造血作用的开发建议 RUNX1b/c 的一个唯一的禁止的函数并且可以在正常和 diseased 模型帮助它的生物函数的进一步的理解。

关键词：人类胚胎干细胞造血功能诱导表达早期中胚层 TGF-β 堵塞效应转化生长因子

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Early development and functional properties of tryptase/chymase double-positive mast cells from human pluripotent stem cells

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Journal of Molecular Cell Biology 2021年第2期13卷 104-115页

作者： Guohui Bian Yanzheng Gu Changlu Xu Wenyu Yang Xu Pan Yijin Chen mowen lai Ya Zhou Yong Dong Bin Mao Qiongxiu Zhou Bo Chen Tatsutoshi Nakathata Lihong Shi Min Wu Yonggang Zhang Feng Ma Center for Stem Cell Research and Application Institute of Blood TransfusionChinese Academy of Medical Sciences&Peking Union Medical College(CAMS&PUMC)Chengdu 610052China State Key Laboratory of Experimental Hematology National Clinical Research Center for Blood DiseasesInstitute of Hematology&Blood Diseases HospitalCAMS&PUMCTianjin 300020China Stem Cell Key Laboratory of Jiangsu Province Institute of Medical BiotechnologySuzhou UniversitySuzhou 215123China Center for iPS Cell Research and Application(CiRA) Kyoto UniversityKyoto 606-8507Japan Department of Biomedical Sciences School of Medicine and Health SciencesUniversity of North DakotaGrand ForksND 58203US

Mast cells (MCs) play a pivotal role in the hypersensitivity reaction by regulating the innate and adaptive immune responses. Humans have two types of MCs. The first type, termed MCTC, is found in the skin and other connective tissues and expresses both tryptase and chymase, while the second, termed MCT, which only expresses tryptase, is found primarily in the mucosa. MCs induced from human adult-type CD34+ cells are reported to be of the MCT type, but the development of MCs during embryonic/fetal stages is largely unknown. Using an efficient coculture system, we identified that a CD34+c-kit+ cell population, which appeared prior to the emergence of CD34+CD45+ hematopoietic stem and progenitor cells (HSPCs), stimulated robust production of pure Tryptase+Chymase+ MCs (MCTCs). Single-cell analysis revealed dual development directions of CD34+c-kit+ progenitors, with one lineage developing into erythro-myeloid progenitors (EMP) and the other lineage developing into HSPC. Interestingly, MCTCs derived from early CD34+c-kit+ cells exhibited strong histamine release and immune response functions. Particularly, robust release of IL-17 suggested that these early developing tissue-type MCTCs could play a central role in tumor immunity. These findings could help elucidate the mechanisms controlling early development of MCTCs and have significant therapeutic implications.

关键词： mast cells human pluripotent stem cells(hPSCs) development tryptase chymase

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HOXC4 up-regulates NF-κB signaling and promotes the cell proliferation to drive development of human hematopoiesis, especially CD43+ cells

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Blood Science 2020年第4期2卷 117-128页

作者： Jiahui Zeng Wencui Sun Jing Chang Danying Yia Lijiao Zhu Yonggang Zhang Xu Pan Ya Zhou mowen lai Guohui Bian Qiongxiu Zhou Jiaxin Liu Bo Chen FengMa Institute of Blood Transfusion Chinese Academy of Medical Sciences&Peking Union Medical College(CAMS&PUMC)Chengdu 610052China State Key Laboratory of Biotherapy Sichuan UniversityChengdu 61006China State Key Laboratory of Experimental Hematology CAMS&PUMCTianjin 300020China

The hematopoietic function of HOXC4 has not been extensively investigated.Our research indicated that induction of HOXC4 in co-culture system from D10 significantly promoted productions of most hematopoietic progenitor cells.CD34−CD43+cells could be clearly classified into CD34−CD43^(low) and CD34−CD43^(high) sub-populations at D14.The former cells had greater myelogenic potential,and their production was not significantly influenced by induction of HOXC4.By contrast,the latter cells had greater potential to differentiate into megakaryocytes and erythroid cells,and thus had properties of erythroid–megakaryocyte common progenitors,which abundance was increased by∼2-fold when HOXC4 was induced from D10.For CD34−CD43^(low),CD34+CD43+,and CD34−CD43^(high) sub-populations,CD43 level served as a natural index for the tendency to undergo hematopoiesis.Induction of HOXC4 from D10 caused more CD43+cells sustain in S-phase with up-regulation of NF-κB signaling,which could be counteracted by inhibition of NF-κB signaling.These observations suggested that promotion of hematopoiesis by HOXC4 is closely related to NF-κB signaling and a change in cell-cycle status,which containing potential of clinical applications.

关键词： CD43 Erythroid–megakaryocyte common progenitor(EMkP) Hematopoiesis HOXC4 Human embryonic stem cells(hESCs) Inducible expression

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Overexpression of HOXA9 upregulates NF-κB signaling to promote human hematopoiesis and alter the hematopoietic differentiation potentials

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Cell Regeneration 2021年第1期10卷 95-106页

作者： Jiahui Zeng Danying Yi Wencui Sun Yuanlin Liu Jing Chang Lijiao Zhu Yonggang Zhang Xu Pan Yong Dong Ya Zhou mowen lai Guohui Bian Qiongxiu Zhou Jiaxin Liu Bo Chen Feng Ma Chinese Academy of Medical Sciences&Peking Union Medical College(CAMS&PUMC) Institute of Blood TransfusionNo.26Huacai RoadLongtan Industry ParkChenghua DistrictChengdu 610052China State Key Laboratory of Biotherapy Sichuan UniversityChengdu 610065China State Key Laboratory of Experimental Hematology CAMS&PUMCTianjin 300020China

Background: The HOX genes are master regulators of embryogenesis that are also involved inhematopoiesis. HOXA9 belongs to a cluster of HOX genes that play extensively studied roles inhematopoiesis and leukemogenesis.Methods: We established HOXA9-inducible human embryonic stem cells (HOXA9/hESCs) with normalpluripotency and potential for hematopoiesis, which could be used to analyze gene function with highaccuracy. HOXA9/hESCs co-cultured with aorta–gonad–mesonephros-derived stromal cells (AGM-S3) wereinduced to overexpress HOXA9 with doxycycline (DOX) at various times after hematopoiesis started andthen subjected to flow cytometry.Results: Induction of HOXA9 from Day 4 (D4) or later notably promoted hematopoiesis and also increasedthe production of CD34+ cells and derived populations. The potential for myelogenesis was significantlyelevated while the potential for erythrogenesis was significantly reduced. At D14, a significant promotion ofS phase was observed in green fluorescent protein positive (GFP+) cells overexpressing HOXA9. NF-κBsignaling was also up-regulated at D14 following induction of HOXA9 on D4. All of these effects could becounteracted by addition of an NF-κB inhibitor or siRNA against NFKB1 along with DOX.Conclusions: Overexpression of HOXA9 starting at D4 or later during hematopoiesis significantly promotedhematopoiesis and the production of myeloid progenitors while reduced the production of erythroidprogenitors, indicating that HOXA9 plays a key role in hematopoiesis and differentiation of hematopoieticlineages.

关键词： Hematopoiesis Differentiation of hematopoietic lineages HOXA9 Mesoderm Human embryonic stem cells(hESCs) Tet-on Inducible expression system

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Inhibition of aryl hydrocarbon receptor signaling promotes the terminal differentiation of human erythroblasts

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Journal of Molecular Cell Biology 2022年第2期14卷 18-31页

作者： Yĳin Chen Yong Dong Xulin Lu Wanjing Li Yimeng Zhang Bin Mao Xu Pan Xiaohong Li Ya Zhou Quanming An Fangxin Xie Shihui Wang Yuan Xue Xinping Cai mowen lai Qiongxiu Zhou Yan Yan Ruohan Fu Hong Wang Tatsutoshi Nakahata Xiuli An Lihong Shi Yonggang Zhang Feng Ma Institute of Blood Transfusion Chinese Academy of Medical Sciences&Peking Union Medical College(CAMS&PUMC)Chengdu 610052China State Key Laboratory of Experimental Hematology Institute of Hematology and Blood Diseases HospitalCAMS&PUMCTianjin 300020China School of Life Sciences Zhengzhou UniversityZhengzhou 450001China Department of Obstetrics and Gynecology Jinjiang Maternity and Child Health HospitalChengdu 610065China Center for iPS Cell Research and Application(CiRA) Kyoto UniversityKyoto 606-8507Japan Laboratory of Membrane Biology New York Blood CenterNew YorkNY 10065USA

The aryl hydrocarbon receptor(AHR)plays an important role during mammalian embryo development.Inhibition of AHR signaling promotes the development of hematopoietic stem/progenitor cells.AHR also regulates the functional maturation of blood cells,such as T cells and megakaryocytes.However,little is known about the role of AHR modulation during the development of erythroid cells.In this study,we used the AHR antagonist StemRegenin 1(SR1)and the AHR agonist 2,3,7,8-tetrachlorodibenzo-p-dioxin during different stages of human erythropoiesis to elucidate the function of AHR.We found that antagonizing AHR signaling improved the production of human embryonic stem cell derived erythrocytes and enhanced erythroid terminal differentiation.RNA sequencing showed that SR1 treatment of proerythroblasts upregulated the expression of erythrocyte differentiation-related genes and downregulated actin organization-associated genes.We found that SR1 accelerated F-actin remodeling in terminally differentiated erythrocytes,favoring their maturation of the cytoskeleton and enucleation.We demonstrated that the effects of AHR inhibition on erythroid maturation were associated with F-actin remodeling.Our findings help uncover the mechanism for AHRmediated human erythroid cell differentiation.We also provide a new approach toward the large-scale production of functionally mature human pluripotent stem cell-derived erythrocytes for use in translational applications.

关键词： erythroblast AHR SR-1 human pluripotent stem cells differentiation

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未成熟CD71 +红细胞免疫功能研究进展

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中华微生物学和免疫学杂志 2023年第9期43卷 671-676页

作者：李晓红周涯赖默温张勇刚马峰中国医学科学院输血研究所/干细胞与再生医学研究中心成都610052

红细胞的主要功能是氧气运输,但其与机体免疫也存在密切关系。越来越多的证据表明不成熟的红细胞在疾病,特别是肿瘤的发展过程中,具有重要作用。本文就未成熟CD71+红细胞在生理和病理等状态下发挥的免疫调控作用研究进展作一综述。

关键词： CD71+红细胞异质性红细胞免疫调控生理和病理

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人脑类器官来源间充质干细胞促进脐带血造血干祖细胞向巨噬细胞分化和功能极化

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生命的化学 2022年第6期42卷 1161-1170页

作者：蔡信平周涯张秀秀潘旭李晓红赖默温张勇刚马峰中国医学科学院北京协和医学院输血研究所成都610052

本文探讨了人脑类器官来源神经源性间充质干细胞(neural mesenchymal stem cells,nMSCs)对脐带血来源CD34(umbilical cord blood CD34,UCB-CD34)细胞向各系分化的作用,尤其是对巨噬细胞(macrophage,Mφ)诱导分化和极化的影响。本文通过U... 详细信息

本文探讨了人脑类器官来源神经源性间充质干细胞(neural mesenchymal stem cells,nMSCs)对脐带血来源CD34(umbilical cord blood CD34,UCB-CD34)细胞向各系分化的作用,尤其是对巨噬细胞(macrophage,Mφ)诱导分化和极化的影响。本文通过UCB-CD34细胞与H1-nMSCs或hi PSC-nMSCs贴壁共培养,无MSCs组作为对照(control),计数共培养组和对照组造血细胞集落数、对集落细胞进行MGG染色、流式检测Mφ相关表面分子、转录组测序检测共培养7 d后CD34细胞红系和髓系基因表达的差异。将nMSCs通过trans-well与UCB-CD34来源的Mφ(UCB-Mφ)共培养,经IL-4刺激后,流式检测Mφ表面分子CD206的表达情况,并用U-PLEX平台测定培养基上清中IL-10的浓度。相较于对照组,UCB-CD34细胞分别与H1-nMSCs、hi PSC-nMSCs共培养时可获得更多的造血细胞集落(P<0.05)以及更多的M集落(P<0.01);UCB-CD34细胞分别与H1-nMSCs、hi PSC-nMSCs共培养第7天,可产生更多GMP细胞,而CFU-E产量减少。UCB-CD34细胞分别与H1-nMSCs、hi PSC-nMSCs在Mφ诱导分化培养基或无细胞因子分化培养基中共培养21 d后,均可获得更高比例和数量的Mφ。共培养7 d后CD34细胞转录组测序显示,与H1-nMSCs、hi PSC-nMSCs共培养组高表达GMP相关基因:NAPSB、HCK、FCGR1A、MS4A6A、PRTN3等和Mφ相关基因CD14、CSF1R、TRIB1和APP等;与nMSCs共培养组的UCB-Mφ在IL-4刺激时,高表达CD206表面标记分子,且分泌更多IL-10,即向M2型极化。本研究初步证明,H1-nMSCs、hi PSC-nMSCs能较好地促进UCB-CD34向Mφ分化及向M2型极化。

关键词：人多能干细胞脐带血造血干细胞间充质干细胞巨噬细胞极化

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Quantitative analysis of biotechnical reinforcement for a steep slope consisting of composite coal-gangue-soil medium adjacent to a mined-out area

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Journal of University of Science and Technology Beijing 2005年第6期12卷 489-494页

作者： Xingping lai mowen Xie Fenhua Ren Meifeng Cai Civil and Environmental Engineering School University of Science and Technology Beijing Beijing 100083 China Energy School Xi＇an University of Science and Technology Xi＇an 710054 China West Mining and Hazard Control Key Laboratory of the Ministry of Education of China Xi＇an 710054 China

The engineering and geological characteristics of a steep slope consisting of coal gangue, rock and soil medium in Huating coal mine have been comprehensively investigated. Owing to humid weather, heavy rainfall, vegetation and porous characteristics of the soil and rock mass, the steep slope will be destabilized and induce mud-rock flow or derive hazard easily. Firstly, based on the classical slope reinforcement theory, some regularity between the shear and displacement in the destabilized zone of the slope with or without root strength contribution is presented. Then, based on the experimental and statistical analysis of root strength, hydrological characteristics and stability status, etc., some possible biotechnical techniques for reinforcement of the steep slope have been suggested. These methods are important for quantitative analysis of destabilization of the slope and design of the biotechnical reinforcement.

关键词： mined-out area coal-gangue-soil composite medium steep slope root-soil effect biotechnical reinforcement

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人胚胎干细胞来源红系细胞膜蛋白表达的研究

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中国输血杂志 2017年第3期30卷 235-241页

作者：李晴邹庆毛斌黄淑赖默温潘旭孙文翠陈谊金刘嘉馨马峰中国医学科学院北京协和医学院输血研究所四川成都610052 中国医学科学院输血研究所四川新生命干细胞科技股份有限公司联合实验室

目的探讨人胚胎干细胞(h ESCs)来源红系细胞在发育过程中其膜蛋白,包括膜表型分子、膜骨架蛋白和跨膜蛋白的表达情况。方法 h ESCs与小鼠主动脉-性腺-中肾(m AGM-S3)基质细胞共培养产生造血干祖细胞(CD34+CD45+)和GPA+细胞,挑取共培养1... 详细信息

目的探讨人胚胎干细胞(h ESCs)来源红系细胞在发育过程中其膜蛋白,包括膜表型分子、膜骨架蛋白和跨膜蛋白的表达情况。方法 h ESCs与小鼠主动脉-性腺-中肾(m AGM-S3)基质细胞共培养产生造血干祖细胞(CD34+CD45+)和GPA+细胞,挑取共培养12 d的细胞,同时磁珠分选人脐血中的CD34^+细胞,在多种特定细胞因子SCF、IL-6、IL-3、FLT3-L、TPO、EPO等诱导下,分别进行集落培养12-14 d产生红系爆发式集落形成单位(BFU-E),运用流式细胞术(FACs)、qRT-PCR和免疫荧光染色等方法检测并比较hESCs和人脐血CD34^+细胞2种来源的红系细胞膜蛋白的表达情况,并以后者作为对照组。结果 hESCs来源BFU-E的膜蛋白的表达趋势与对照组相近:其膜骨架蛋白和跨膜蛋白的基因,如SPTA、SPTB、ANK1、EPB41等基因转录水平都在逐渐增强,而一些膜蛋白如黏附分子CD49d、CD29、CD71等的表达逐渐降低,CD47基本不表达;带3蛋白(Band 3)表达量较高且与成体型血红蛋白β-globin的表达呈正相关。结论在hESCs向红系诱导生成过程中,一些膜表型分子表达在逐渐降低,而膜蛋白表达逐渐增强。

关键词：人胚胎干细胞红系细胞膜蛋白表达红系爆发式集落形成单位带3蛋白

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