The embryonic mesoderm comprises heterogeneous cell subpopulations with distinct lineage biases.It is unclear whether a bias for the human hematopoietic lineage emerges at this early developmental stage.In this study,...The embryonic mesoderm comprises heterogeneous cell subpopulations with distinct lineage biases.It is unclear whether a bias for the human hematopoietic lineage emerges at this early developmental stage.In this study,we integrated single-cell transcriptomic analyses of human mesoderm cells from embryonic stem cells and embryos,enabling us to identify and define the molecular features of human hematopoietic mesoderm(HM)cells biased towards hematopoietic lineages.We discovered that BMP4 plays an essential role in HM specification and can serve as a marker for HM cells.Mechanistically,BMP4 acts as a downstream target of HDAC1,which modulates the expression of BMP4 by deacetylating its enhancer.Inhibition of HDAC significantly enhances HM specification and promotes subsequent hematopoietic cell differentiation.In conclusion,our study identifies human HM cells and describes new mechanisms for human hematopoietic development.展开更多
Development of animal embryos before zygotic genome activation at the mid blastula transition （MBT） is essentially supported by eggderived maternal products. Nodal proteins are crucial signals for mesoderm and endod...Development of animal embryos before zygotic genome activation at the mid blastula transition （MBT） is essentially supported by eggderived maternal products. Nodal proteins are crucial signals for mesoderm and endoderm induction after the MBT. It remains unclear which maternal factors activate zygotic expression of nodal genes in the ventrotateral blastodermal margin of the zebrafish blastulas. In this study, we show that loss of maternal Eomesodermin a （Eomesa）, a T-box transcription factor, impairs zygotic expression of the nodal genes ndr1 and ndr2 as well as mesodermal and endodermal markers, indicating an involvement in mesendoderm induction. Maternal Eomesa is also required for timely zygotic expression of the transcription factor gene mxtx2, a regulator of nodal gene expression. Eomesa directly binds to the Eomes-binding sites in the promoter or enhancer of ndr1, ndr2, and rnxtx2 to activate their transcrip- tion. Furthermore, human and mouse Nodal genes are also regulated by Eomes. Transfection of zebrafish eomesa into murine embryonic stem cells promotes mesendodermal differentiation with constant higher levels of endogenous Nodal expression, suggesting a conserved function of Eomes. Taken together, our findings reveal a conserved rote of maternal T-box transcription factors in regulating nodal gene expression and mesendoderm induction in vertebrate embryos.展开更多
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 lack...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.展开更多
RUNXI is absolutely required for definitive hematopoiesis, but the function of RUNXlb/c, two isoforms of human RUNX1, is unclear. We established inducible RUNXlb/c-overexpressing human embryonic stem cell （hESC） lin...RUNXI is absolutely required for definitive hematopoiesis, but the function of RUNXlb/c, two isoforms of human RUNX1, is unclear. We established inducible RUNXlb/c-overexpressing human embryonic stem cell （hESC） lines, in which RUNXlb/c overexpression prevented the emergence of CD34＋ cells from early stage, thereby drastically reducing the production of hematopoi- etic stem/prognnitor cells. Simultaneously, the expression of hematopoiesis-related factors was downregulated. However, such blockage effect disappeared from day 6 in hESC/AGM-S3 ceU co-cultures, proving that the blockage occurred before the generation of hemogenic endothelial cells. This blockage was partially rescued by RepSox, an inhibitor of the transforming growth factor （TGF）-β signaling pathway, indicating a close relationship between RUNX1b/c and TGF-β pathway. Our results suggest a unique inhibitory function of RUNX1b/c in the development of early hematopoiesis and may aid further understanding of its biological function in normal and diseased models.展开更多
本研究通过整合生物信息学分析与实验验证,系统探讨中胚层发育关键基因在黑色素瘤发生发展中的分子机制。基于基因集变异分析(gene set variation analysis,GSVA)算法对406例皮肤黑色素瘤(skin cutaneous melanoma,SKCM)患者的7752项生...本研究通过整合生物信息学分析与实验验证,系统探讨中胚层发育关键基因在黑色素瘤发生发展中的分子机制。基于基因集变异分析(gene set variation analysis,GSVA)算法对406例皮肤黑色素瘤(skin cutaneous melanoma,SKCM)患者的7752项生物功能进行富集分析,发现中胚层发育在黑色素瘤的发生发展中具有重要作用。通过LASSO-COX算法筛选出SMAD4、NODAL、BMPR1A和ZFP36L 1共4个核心调控基因,并构建预后相关风险评分体系。基因本体论(Gene Ontology,GO)和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路分析显示,这些基因主要富集于mRNA代谢过程和TGF-β信号通路相关通路。通过定量聚合酶链式反应、蛋白质印迹及免疫组化证实:SMAD4和BMPR1A在肿瘤组织中的表达下调与不良预后显著相关(P<0.05);NODAL通过调控上皮-间质转化(EMT)过程促进肿瘤侵袭转移;ZFP36L1高表达组则表现出更好的化疗敏感性。进一步分析发现,核心基因表达水平与肿瘤免疫浸润特征及免疫检查点分子存在显著相关性。本研究通过多组学分析结合实验验证,揭示了中胚层发育相关基因在黑色素瘤进展中的关键作用,阐明了SMAD4/NODAL/BMPR1A/ZFP36L1通过免疫微环境调控和EMT过程影响肿瘤生物学行为的分子机制,为黑色素瘤的分子分型和靶向治疗提供了新的理论依据。展开更多
The CCCTC-binding factor(CTCF)protein and its modified forms regulate gene expression and genome organization.However,information on CTCF acetylation and its biological function is still lacking.Here,we show that CTCF...The CCCTC-binding factor(CTCF)protein and its modified forms regulate gene expression and genome organization.However,information on CTCF acetylation and its biological function is still lacking.Here,we show that CTCF can be acetylated at lysine 20(CTCF-K20)by CREB-binding protein(CBP)and deacetylated by histone deacetylase 6(HDAC6).CTCF-K20 is required for the CTCF interaction with CBP.A CTCF point mutation at lysine 20 had no effect on self-renewal but blocked the mesoderm differentiation of mouse embryonic stem cells(mESCs).The CTCF-K20 mutation reduced CTCF binding to the promoters and enhancers of genes associated with early cardiac mesoderm differentia-tion,resulting in diminished chromatin accessibility and decreased enhancer-promoter interactions,impairing gene expression.In summary,this study reveals the important roles of CTCF-K20 in regulating CTCF genomic functions and mESC differentiation into mesoderm.展开更多
背景与目的:子宫恶性中胚叶混合瘤(malignant mixed mesodermal tumor,MMMT)是罕见的妇科恶性肿瘤,预后极差,是临床诊治的难点。本研究旨在探讨本病的临床、病理特征及孕激素受体亚型(progesterone receptor subtype A and B,PRA and P...背景与目的:子宫恶性中胚叶混合瘤(malignant mixed mesodermal tumor,MMMT)是罕见的妇科恶性肿瘤,预后极差,是临床诊治的难点。本研究旨在探讨本病的临床、病理特征及孕激素受体亚型(progesterone receptor subtype A and B,PRA and PRB)蛋白在子宫MMMT的表达及其意义。方法:回顾性分析17例患者的临床资料,并对其病理切片行光镜观察并应用免疫组化法测定PRA、PRB的表达情况,随访其中的11例患者。结果:①子宫MMMT表现缺乏特异性,主要表现为阴道出血。②病理上肿瘤成分复杂,形态多样,有上皮和间叶两种成分组成,相互间有穿插和移行变化。③同源性PRA阳性占55.6%,PRB阳性占33.3%;异源性PRA阳性占37.5%,PRB阳性占37.5%,两种亚型间差异无显著性(P>0.05)。PRA在Ⅰ期和Ⅱ期患者的表达率分别为66.7%和40%;PRB患者分别为55.6%和20%。④Ⅰ期患者平均存活43.8个月(32～59个月),Ⅱ期平均存活34.25个月(19～41个月);Ⅲ期1例,存活5个月。结论:子宫MMMT的诊断主要依赖组织形态学,疾病进展可能与PRA、PRB的丢失有关,PRA、PRB的表达可能与病理类型无关。预后可能与临床分期及PRA、PRB的表达有关。展开更多
基金supported by the CAMS Innovation Fund for Medical Sciences(2021-I2M-1-073,2021-I2M-1-040,2022-I2M-JB-015)the National Key Research and Development Program of China(2021YFA1100703,2021YFA1103000)+2 种基金Haihe Laboratory of Cell Ecosystem Innovation Fund(22HHXBSS00031)the National Natural Science Foundation of China(82125003,32271161,82200141)Tianjin Municipal Science and Technology Commission Grant(20JCYBJC00240,22ZXSYSY00010,22JCQNJC01270)。
文摘The embryonic mesoderm comprises heterogeneous cell subpopulations with distinct lineage biases.It is unclear whether a bias for the human hematopoietic lineage emerges at this early developmental stage.In this study,we integrated single-cell transcriptomic analyses of human mesoderm cells from embryonic stem cells and embryos,enabling us to identify and define the molecular features of human hematopoietic mesoderm(HM)cells biased towards hematopoietic lineages.We discovered that BMP4 plays an essential role in HM specification and can serve as a marker for HM cells.Mechanistically,BMP4 acts as a downstream target of HDAC1,which modulates the expression of BMP4 by deacetylating its enhancer.Inhibition of HDAC significantly enhances HM specification and promotes subsequent hematopoietic cell differentiation.In conclusion,our study identifies human HM cells and describes new mechanisms for human hematopoietic development.
基金Acknowledgements We thank Drs Alex Schier and Susan Mango （Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA, USA） for discussion and suggestions, Dr David Kimelman （Department of Biochemistry, University of Washington, Seattle, WA, USA） for myc-eomesa construct, and members of the Meng lab for discussion and technical assistance. This work was financially supported by grants from the Major Science Research Programs of China （2011CB943800） and the National Natural Science Foundation of China （31221064）.
文摘Development of animal embryos before zygotic genome activation at the mid blastula transition （MBT） is essentially supported by eggderived maternal products. Nodal proteins are crucial signals for mesoderm and endoderm induction after the MBT. It remains unclear which maternal factors activate zygotic expression of nodal genes in the ventrotateral blastodermal margin of the zebrafish blastulas. In this study, we show that loss of maternal Eomesodermin a （Eomesa）, a T-box transcription factor, impairs zygotic expression of the nodal genes ndr1 and ndr2 as well as mesodermal and endodermal markers, indicating an involvement in mesendoderm induction. Maternal Eomesa is also required for timely zygotic expression of the transcription factor gene mxtx2, a regulator of nodal gene expression. Eomesa directly binds to the Eomes-binding sites in the promoter or enhancer of ndr1, ndr2, and rnxtx2 to activate their transcrip- tion. Furthermore, human and mouse Nodal genes are also regulated by Eomes. Transfection of zebrafish eomesa into murine embryonic stem cells promotes mesendodermal differentiation with constant higher levels of endogenous Nodal expression, suggesting a conserved function of Eomes. Taken together, our findings reveal a conserved rote of maternal T-box transcription factors in regulating nodal gene expression and mesendoderm induction in vertebrate embryos.
基金supported by grants from the CAMS Initiatives for Innovative Medicine(2016-I2M-1-018 to F.M.and 2017-I2M-3-021 to J.L.)the Sichuan Provincial Health and Family Planning Commissi on research project(17PJ489 to B.C.)Chengdu Science and Technology Project-Technology Innovation R&D(2018-YF05-01341-SN to B.C.).
文摘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.
基金This work was supported by the National Program on Key Basic Research Project of China （973 Program 2015CB964902）, the National Natural Science Foundation of China （NSFC H81170466 and H81370597）, and the CAMS Initiatives for Innovative Medicine （2016-12M-1-018） awarded to F.M.
文摘RUNXI is absolutely required for definitive hematopoiesis, but the function of RUNXlb/c, two isoforms of human RUNX1, is unclear. We established inducible RUNXlb/c-overexpressing human embryonic stem cell （hESC） lines, in which RUNXlb/c overexpression prevented the emergence of CD34＋ cells from early stage, thereby drastically reducing the production of hematopoi- etic stem/prognnitor cells. Simultaneously, the expression of hematopoiesis-related factors was downregulated. However, such blockage effect disappeared from day 6 in hESC/AGM-S3 ceU co-cultures, proving that the blockage occurred before the generation of hemogenic endothelial cells. This blockage was partially rescued by RepSox, an inhibitor of the transforming growth factor （TGF）-β signaling pathway, indicating a close relationship between RUNX1b/c and TGF-β pathway. Our results suggest a unique inhibitory function of RUNX1b/c in the development of early hematopoiesis and may aid further understanding of its biological function in normal and diseased models.
文摘本研究通过整合生物信息学分析与实验验证,系统探讨中胚层发育关键基因在黑色素瘤发生发展中的分子机制。基于基因集变异分析(gene set variation analysis,GSVA)算法对406例皮肤黑色素瘤(skin cutaneous melanoma,SKCM)患者的7752项生物功能进行富集分析,发现中胚层发育在黑色素瘤的发生发展中具有重要作用。通过LASSO-COX算法筛选出SMAD4、NODAL、BMPR1A和ZFP36L 1共4个核心调控基因,并构建预后相关风险评分体系。基因本体论(Gene Ontology,GO)和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路分析显示,这些基因主要富集于mRNA代谢过程和TGF-β信号通路相关通路。通过定量聚合酶链式反应、蛋白质印迹及免疫组化证实:SMAD4和BMPR1A在肿瘤组织中的表达下调与不良预后显著相关(P<0.05);NODAL通过调控上皮-间质转化(EMT)过程促进肿瘤侵袭转移;ZFP36L1高表达组则表现出更好的化疗敏感性。进一步分析发现,核心基因表达水平与肿瘤免疫浸润特征及免疫检查点分子存在显著相关性。本研究通过多组学分析结合实验验证,揭示了中胚层发育相关基因在黑色素瘤进展中的关键作用,阐明了SMAD4/NODAL/BMPR1A/ZFP36L1通过免疫微环境调控和EMT过程影响肿瘤生物学行为的分子机制,为黑色素瘤的分子分型和靶向治疗提供了新的理论依据。
基金This work was supported in part by grants from the National Key R&D Program of China(2021YFA1100300)Strategic Priority Research Program of the Chinese Academy of Sciences(XDA16010502)+2 种基金National Natural Science Foundation of China(31925009,U21A20195,32000424,32100462,32100463,and 81902885)Science and Technology Planning Project of Guangdong Province,China(2019B020234004,2019A050510004 and 2020B1212060052)Macao Science and Technology Development Fund(FDCT0107/2019/A2).
文摘The CCCTC-binding factor(CTCF)protein and its modified forms regulate gene expression and genome organization.However,information on CTCF acetylation and its biological function is still lacking.Here,we show that CTCF can be acetylated at lysine 20(CTCF-K20)by CREB-binding protein(CBP)and deacetylated by histone deacetylase 6(HDAC6).CTCF-K20 is required for the CTCF interaction with CBP.A CTCF point mutation at lysine 20 had no effect on self-renewal but blocked the mesoderm differentiation of mouse embryonic stem cells(mESCs).The CTCF-K20 mutation reduced CTCF binding to the promoters and enhancers of genes associated with early cardiac mesoderm differentia-tion,resulting in diminished chromatin accessibility and decreased enhancer-promoter interactions,impairing gene expression.In summary,this study reveals the important roles of CTCF-K20 in regulating CTCF genomic functions and mESC differentiation into mesoderm.
文摘背景与目的:子宫恶性中胚叶混合瘤(malignant mixed mesodermal tumor,MMMT)是罕见的妇科恶性肿瘤,预后极差,是临床诊治的难点。本研究旨在探讨本病的临床、病理特征及孕激素受体亚型(progesterone receptor subtype A and B,PRA and PRB)蛋白在子宫MMMT的表达及其意义。方法:回顾性分析17例患者的临床资料,并对其病理切片行光镜观察并应用免疫组化法测定PRA、PRB的表达情况,随访其中的11例患者。结果:①子宫MMMT表现缺乏特异性,主要表现为阴道出血。②病理上肿瘤成分复杂,形态多样,有上皮和间叶两种成分组成,相互间有穿插和移行变化。③同源性PRA阳性占55.6%,PRB阳性占33.3%;异源性PRA阳性占37.5%,PRB阳性占37.5%,两种亚型间差异无显著性(P>0.05)。PRA在Ⅰ期和Ⅱ期患者的表达率分别为66.7%和40%;PRB患者分别为55.6%和20%。④Ⅰ期患者平均存活43.8个月(32～59个月),Ⅱ期平均存活34.25个月(19～41个月);Ⅲ期1例,存活5个月。结论:子宫MMMT的诊断主要依赖组织形态学,疾病进展可能与PRA、PRB的丢失有关,PRA、PRB的表达可能与病理类型无关。预后可能与临床分期及PRA、PRB的表达有关。
