人类胚胎期和成年期巨核细胞的分子特征比较

doi:10.19586/j.2095-2341.2021.0194

生物技术进展 ›› 2022, Vol. 12 ›› Issue (4): 577-583.DOI: 10.19586/j.2095-2341.2021.0194

人类胚胎期和成年期巨核细胞的分子特征比较

李敏敏(), 夏美娟, 赵晶晶, 陈肖源, 刘翠翠, 苏培, 王洪涛, 周家喜()

中国医学科学院血液病医院（中国医学科学院血液学研究所），北京协和医学院，实验血液学国家重点实验室；国家血液系统疾病临床医学研究中心；细胞生态海河实验室，天津 300020

收稿日期:2021-12-16 接受日期:2022-01-12 出版日期:2022-07-25 发布日期:2022-08-10
通讯作者: 周家喜
作者简介:李敏敏 E-mail：liminmin@ihcams.ac.cn；
基金资助:
国家自然科学基金项目(81870099);京津冀专项项目(19JCZDJC65700);北京协和医学院学科建设项目(201920101401);中央级公益性科研院所基本科研业务费项目(2021-RC310-019)

Comparison of the Molecular Characteristics of Human Megakaryocytes Between Embryonic Stage and Adult Stage

Minmin LI(), Meijuan XIA, Jingjing ZHAO, Xiaoyuan CHEN, Cuicui LIU, Pei SU, Hongtao WANG, Jiaxi ZHOU()

State Key Laboratory of Experimental Hematology； National Clinical Research Center for Blood Diseases； Haihe Laboratory of Cell Ecosystem，Institute of Hematology & Blood Diseases Hospital，Chinese Academy of Medical Sciences & Peking Union Medical College，Tianjin 300020，China

Received:2021-12-16 Accepted:2022-01-12 Online:2022-07-25 Published:2022-08-10
Contact: Jiaxi ZHOU

摘要/Abstract

摘要：

为探究人类不同发育时期巨核细胞的分子特征，基于人类胚胎期卵黄囊、胎肝和成年骨髓巨核细胞的单细胞转录组测序数据，从分子特征、基因调控网络等方面分别对其分子差异进行生物信息学分析。结果表明，胚胎期巨核细胞具有较强的增殖特征，高表达细胞增殖相关的转录因子；而成年期巨核细胞具有较强的血小板生成特征，高表达与巨核细胞分化成熟相关的转录因子。研究结果为研究不同发育阶段巨核细胞及其子代血小板的功能差异提供了理论依据。

关键词: 胚胎期, 成年期, 巨核细胞, 单细胞转录组测序, 分子特征

Abstract:

To explore the molecular characteristics of human megakaryocytes at different development stages， based on the single-cell RNA sequence data of the human embryonic yolk sac， fetal liver， and adult bone marrow megakaryocytes， the molecular differences were analyzed by bioinformatics from the aspects of molecular characteristics and gene regulation network. The results showed that， embryonic stage megakaryocytes had higher proliferation characteristics and highly expressed transcription factors related to cell proliferation， while adult stage megakaryocytes had strongly characteristics of platelet-producing and highly expressed transcription factors related to megakaryocyte differentiation and maturation. This paper provided a theoretical basis for studying the functional differences of megakaryocytes and their progeny platelets at different developmental stages.

Key words: embryonic stage, adult stage, megakaryocyte, single-cell RNA sequence, molecular characteristics

中图分类号:

Q291

李敏敏, 夏美娟, 赵晶晶, 陈肖源, 刘翠翠, 苏培, 王洪涛, 周家喜. 人类胚胎期和成年期巨核细胞的分子特征比较[J]. 生物技术进展, 2022, 12(4): 577-583.

Minmin LI, Meijuan XIA, Jingjing ZHAO, Xiaoyuan CHEN, Cuicui LIU, Pei SU, Hongtao WANG, Jiaxi ZHOU. Comparison of the Molecular Characteristics of Human Megakaryocytes Between Embryonic Stage and Adult Stage[J]. Current Biotechnology, 2022, 12(4): 577-583.

图/表 5

表1 数据来源信息

Table 1 Data source information

序号	来源	细胞数目	物种	器官	时期	测序方法	流式分选策略
1	Wang等^[5]	242	人	卵黄囊	胚胎（孕4~5周）	Smart⁃seq2	CD41a⁺CD42b⁺
2	Wang等^[5]	256	人	胎肝	胚胎（孕7~9周）	Smart⁃seq2	CD41a⁺CD42b⁺
3	Liu等^[6]	860	人	骨髓	成年	Smart⁃seq2	CD41a⁺CD42b⁺

图1 人YS、FL和BM来源的巨核细胞转录组水平PAGA图注：点代表不同来源的巨核细胞，线代表不同来源的巨核细胞的相关性，线越粗代表相关性越高。

Fig.1 Transcriptome?level PAGA picture of human YS, FL, and BM?derived megakaryocytes

图2 人胚胎期和成年期巨核细胞分子特征的差异分析A：热图展示不同组织来源巨核细胞的差异基因表达。热图上不同颜色的矩形方块代表不同组织来源的巨核细胞；左边数字代表不同组织来源的巨核细胞上调的基因数目；热图右侧的图例表示基因的表达情况，蓝色代表表达丰度较低，红色代表表达丰度较高。B：不同组织来源巨核细胞的代表性生物学过程（GO）。C：GSEA图展示了胚胎期组织（YS和FL）富集的生物学过程。D：GSEA图展示了成年期组织（BM）富集的生物学过程，绿色折线代表基因ES折线图，折线图的峰值是该基因集的ES得分，横轴代表该基因集的每个基因，峰值之前的基因为该基因集的核心基因。NES代表标准化后的ES，P<0.05为差异有统计学意义。E：点图展示不同组织来源巨核细胞的细胞周期和血小板产生相关基因的表达比例和表达水平。F：箱线图展示不同组织来源巨核细胞的细胞周期S期基因集（上）和血小板产生基因集（下）的评分。P值由Wilcoxon秩和检验确定，****代表差异在P<0.000 1水平有统计学意义；***代表差异在P<0.001水平有统计学意义；*代表差异在P<0.05水平有统计学意义。

Fig.2 Differential analysis of molecular characteristics of human embryonic and adult megakaryocytes

图3 人胚胎期和成年期巨核细胞的关键转录因子A：不同组织来源巨核细胞的转录子活性热图，热图左侧代表不同组织来源的巨核细胞高活性的转录因子，标红代表与不同组织来源的巨核细胞特征相符的转录因子；B：不同组织来源巨核细胞的转录因子调控网络；C：小提琴图展示不同组织来源巨核细胞代表性的转录因子的表达水平。P值由Wilcoxon秩和检验确定，****代表差异在P<0.000 1水平有统计学意义；***代表差异在P<0.001水平有统计学意义；*代表差异在P≤0.05水平有统计学意义。

Fig.3 The bey transcription factors of megakaryocytes in human embryonic stage and adult stage

图4 人胚胎期和成年期巨核细胞免疫亚群特征注：热图展示胚胎期巨核细胞免疫亚群和成年期巨核细胞免疫亚群的差异基因。热图上不同颜色的矩形方块代表不同时期的巨核细胞免疫亚群；热图左侧标注为不同时期的巨核细胞免疫亚群的特征性差异基因；热图右侧的图例表示基因的表达情况，蓝色代表表达丰度较低，红色代表表达丰度较高。

Fig.4 The characteristics of immune subsets megakaryocyte in human embryo stage and adult stage

参考文献 21

1	MACHLUS K R, ITALIANO J E. The incredible journey: From megakaryocyte development to platelet formation[J]. J. Cell Biol., 2013, 201(6): 785-796.
2	WOOLTHUIS C M, PARK C Y. Hematopoietic stem/progenitor cell commitment to the megakaryocyte lineage[J]. Blood. 2016, 127(10): 1242-1248.
3	IVANOVS A, RYBTSOV S, NG E S, et al.. Human haematopoietic stem cell development: from the embryo to the dish[J]. Development, 2017, 144(13): 2323-2337.
4	TAVIAN M, PEAULT B. Embryonic development of the human hematopoietic system[J]. Int. J. Dev. Biol., 2005, 49(2-3): 243-250.
5	WANG H, HE J, XU C, et al.. Decoding human megakaryocyte development[J]. Cell Stem Cell, 2021, 28(3): 535-549.
6	LIU C, WU D, XIA M, et al.. Characterization of cellular heterogeneity and an immune subpopulation of human megakaryocytes[J/OL]. Adv. Sci., 2021, 8(15): 2100921[2022-03-18]. .
7	BLUTEAU O, LANGLOIS T, RIVERA-MUNOZ P, et al.. Developmental changes in human megakaryopoiesis[J]. J. Thromb. Haemost., 2013, 11(9): 1730-1741.
8	BUTLER A, HOFFMAN P, SMIBERT P, et al.. Integrating single-cell transcriptomic data across different conditions, technologies, and species[J]. Nat. Biotechnol., 2018, 36(5): 411-420.
9	YU G, WANG L, HAN Y, et al.. Cluster profiler: an R package for comparing biological themes among gene clusters[J]. OMICS, 2012, 16(5): 284-287.
10	SUBRAMANIAN A, TAMAYO P, MOOTHA V K, et al.. Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles[J]. Proc. Natl. Acad. Sci. USA, 2005, 102(43): 15545-15550.
11	AIBAR S, GONZÁLEZ-BLAS C B, MOERMAN T, et al.. SCENIC: single-cell regulatory network inference and clustering[J]. Nature Methods, 2017, 14(11): 1083-1086.
12	JURIKOVA M, DANIHEL L, POLAK S, et al.. Ki67, PCNA, and MCM proteins: markers of proliferation in the diagnosis of breast cancer[J]. Acta Histochem., 2016, 118(5): 544-552.
13	ETO K, KUNISHIMA S. Linkage between the mechanisms of thrombocytopenia and thrombopoiesis[J]. Blood, 2016, 127(10): 1234-1241.
14	KANG Y, CUI Y, TAN M. MicroRNA-212 suppresses cell proliferation in nasopharyngeal carcinoma by targeting ELF3[J]. Oncol. Lett., 2020, 19(4): 2902-2908.
15	ZHANG S, ZHU X, LI G. E2F1/SNHG7/miR-186-5p/MMP2 axis modulates the proliferation and migration of vascular endothelial cell in atherosclerosis[J/OL]. Life Sci., 2020, 257: 118013[2022-03-18]. .
16	WANG H, LIU C, LIU X, et al.. MEIS1 regulates hemogenic endothelial generation, megakaryopoiesis, and thrombopoiesis in human pluripotent stem cells by targeting TAL1 and FLI1[J]. Stem Cell Rep., 2018, 10(2): 447-460.
17	IGNATZ M, SOLA-VISNER M, RIMSZA L M, et al.. Umbilical cord blood produces small megakaryocytes after transplantation[J]. Biol. Blood Marrow Transplant., 2007, 13(2): 145-150.
18	LIU Z, ITALIANO J, FERRER-MARIN F, et al.. Developmental differences in megakaryocytopoiesis are associated with up-regulated TPO signaling through mTOR and elevated GATA-1 levels in neonatal megakaryocytes[J]. Blood, 2011, 117(15): 4106-4117.
19	PARISER D N, HILT Z T, TURE S K, et al.. Lung megakaryocytes are immune modulatory cells[J/OL]. J. Clin. Invest., 2021, 131(1): e137377[2022-03-18]. .
20	NAKAMURA S, TAKAYAMA N, HIRATA S, et al.. Expandable megakaryocyte cell lines enable clinically applicable generation of platelets from human induced pluripotent stem cells[J]. Cell Stem Cell, 2014, 14(4): 535-548.
21	MOREAU T, EVANS A L, VASQUEZ L, et al.. Large-scale production of megakaryocytes from human pluripotent stem cells by chemically defined forward programming[J/OL]. Nat. Commun., 2016, 7: 11208[2022-03-18]. .

人类胚胎期和成年期巨核细胞的分子特征比较

Comparison of the Molecular Characteristics of Human Megakaryocytes Between Embryonic Stage and Adult Stage

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