异基因造血干细胞移植后发生移植物抗宿主病患者血清外泌体miRNAs敏感生物标志物的表达分析
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摘要
背景:当前国内外关于人类造血干细胞移植后外泌体miRNA和移植物抗宿主病发生的相关性研究甚少。国内研究匮乏,外泌体来源的核酸分子有望成为新型生物标志物预测急性移植物抗宿主病的发生。目的:探讨异基因造血干细胞移植后发生急性移植物抗宿主病患者外周血血清外泌体中候选基因miRNAs的差异表达。方法:收集2017年8月至2018年5月异基因造血干细胞移植后发生急性移植物抗宿主病以及未发生急性移植物抗宿主病患者共30例,其中急性移植物抗宿主病组20例,非急性移植物抗宿主病组10例。提取移植前后血清外泌体及mi RNA,基因芯片杂交后图像采集和数据分析。结果与结论:与对照组相比,移植前后呈上升或下降趋势且信号差异倍数超过10倍的mi RNAs在急性移植物抗宿主病组有11个mi RNAs上升,26个下降。hsa-mi R-3976、hsa-mi R-122-5p、hsa-miR-3125明显上调,hsa-mi R-941、hsa-mi R-4687-5P和hsa-mi R-4769-5p明显下调。
BACKGROUND: There are few studies, especially in China, addressing the correlation between exosome miRNAs and graft-versus-host disease(GVHD) following human hematopoietic stem cell transplantation. Exosome-derived nucleic acid molecules are expected to become new biomarkers for predicting the occurrence of acute GVHD(aGVHD).OBJECTIVE: To investigate the differential expression of candidate gene microRNAs in peripheral blood serum exosomes of aGVHD patients with after allogeneic hematopoietic stem cell transplantation.METHODS: A total of 30 patients with aGVHD and without aGVHD after allogeneic hematopoietic stem cell transplantation from August 2017 to May 2018 were enrolled, including 20 in aGVHD group and 10 in non-aGVHD group. Serum exosomes and miRNAs were extracted before and after transplantation. Image acquisition and data analysis were performed after gene chip hybridization.RESULTS AND CONCLUSION: After microarray hybridization microRNAs showed an over 10-fold upward or downward trend before and after transplantation. Compared with the control group, 11 microRNAs increased and 26 decreased in the aGVHD group. hsa-miR-3976,hsa-miR-122-5 p, and hsa-miR-3125 were significantly up-regulated, while hsa-miR-941, hsa-miR-4687-5 P and hsa-miR-4769-5 p were significantly down-regulated.
BACKGROUND: There are few studies, especially in China, addressing the correlation between exosome miRNAs and graft-versus-host disease(GVHD) following human hematopoietic stem cell transplantation. Exosome-derived nucleic acid molecules are expected to become new biomarkers for predicting the occurrence of acute GVHD(aGVHD).OBJECTIVE: To investigate the differential expression of candidate gene microRNAs in peripheral blood serum exosomes of aGVHD patients with after allogeneic hematopoietic stem cell transplantation.METHODS: A total of 30 patients with aGVHD and without aGVHD after allogeneic hematopoietic stem cell transplantation from August 2017 to May 2018 were enrolled, including 20 in aGVHD group and 10 in non-aGVHD group. Serum exosomes and miRNAs were extracted before and after transplantation. Image acquisition and data analysis were performed after gene chip hybridization.RESULTS AND CONCLUSION: After microarray hybridization microRNAs showed an over 10-fold upward or downward trend before and after transplantation. Compared with the control group, 11 microRNAs increased and 26 decreased in the aGVHD group. hsa-miR-3976,hsa-miR-122-5 p, and hsa-miR-3125 were significantly up-regulated, while hsa-miR-941, hsa-miR-4687-5 P and hsa-miR-4769-5 p were significantly down-regulated.
引文
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[2]Báez A, Martín-Antonio B, Piruat JI, et al. Gene and miRNA expression profiles of hematopoietic progenitor cells vary depending on their origin. Biol Blood Marrow Transplant.2014;20(5):630-639.
[3]Dickinson AM, Norden J. Non-HLA genomics:does it have a role in predicting haematopoietic stem cell transplantation outcome. Int J Immunogenet. 2015;42(4):229-238.
[4]Paczesny S, Raiker N, Brooks S, et al. Graft-versus-host disease biomarkers:omics and personalized medicine. Int J Hematol. 2013;98(3):275-292.
[5]Sanghvi VR, Mavrakis KJ, Van der Meulen J, et al.Characterization of a set of tumor suppressor microRNAs in T cell acute lymphoblastic leukemia. Sci Signal. 2014;7(352):ra111.
[6]Gentner B, Pochert N, Rouhi A, et al. MicroRNA-223 dose levels fine tune proliferation and differentiation in human cord blood progenitors and acute myeloid leukemia. Exp Hematol.2015;43(10):858-868.e7.
[7]An T, Qin S, Xu Y, et al. Exosomes serve as tumour markers for personalized diagnostics owing to their important role in cancer metastasis. J Extracell Vesicles. 2015;4:27522.
[8]黄依瑶,郑磊.重视外泌体的实验诊断价值[J].中华检验医学杂志,2015, 38(11):724-726.
[9]Muntión S, Ramos TL, Diez-Campelo M, et al. Microvesicles from Mesenchymal Stromal Cells Are Involved in HPC-Microenvironment Crosstalk in Myelodysplastic Patients.PLoS One. 2016;11(2):e0146722.
[10]黄依瑶,唐月汀,覃思华,等.血清中外泌体及外泌体RNA提取方法的比较[J].中华检验医学杂志,2016, 39(6):427-432.
[11]赵晓琦,程敏.外泌体相关微小RNA的生物学特性及其在多种心血管疾病中的诊断价值[J].临床心血管病杂志,2015, 31(9):924-928.
[12]杨雅芝,张小燕,万倩.外泌小体内miRNA在多发性骨髓瘤进展中的作用及可能机制[J].中国实验血液学杂志, 2017,25(1):301-305.
[13] Wang L, Romero M, Ratajczak P, et al. Increased apoptosis is linked to severe acute GVHD in patients with Fanconi anemia.Bone Marrow Transplant. 2013;48(6):849-853.
[14] De Tullio G, De Fazio V, Sgherza N, et al. Challenges and opportunities of microRNAs in lymphomas. Molecules. 2014;19(9):14723-14781.
[15]简雯.帕金森病患者血浆外泌体的分离鉴定及miRNA表达谱分析[D].南昌:南昌大学, 2016.
[16]胡彬,付伟,黄斯勇,等.异基因造血干细胞移植后血浆microRNA的表达与急性移植物抗宿主病的相关性研究[J].中国实验血液学杂志,2016,24(3):827-832.
[17]郭智,陈惠仁.异基因造血干细胞移植后的相关并发症及危险因素[J].中国组织工程研究,2013,17(40):7163-7168.
[18]田淼,谭三勤,刘静茹,等.高通量测序技术分析急性髓系白血病血浆外泌体微RNA表达谱差异[J].中国生物化学与分子生物学报,2017, 33(2):138-149.
[19] Wojtowicz EE, Walasek MA, Broekhuis MJ, et al.MicroRNA-125 family members exert a similar role in the regulation of murine hematopoiesis. Exp Hematol. 2014;42(10):909-918.e1.
[20] Zhang J, Li S, Li L, et al. Exosome and exosomal microRNA:trafficking, sorting, and function. Genomics Proteomics Bioinformatics. 2015;13(1):17-24.
[21] Lia G, Brunello L, Bruno S, et al. Extracellular vesicles as potential biomarkers of acute graft-vs-host disease. Leukemia.2018;32(3):765-773.
[22] Xiao B, Wang Y, Li W, et al. Plasma microRNA signature as a noninvasive biomarker for acute graft-versus-host disease.Blood. 2013;122(19):3365-3375.
[23] Boon RA, Vickers KC. Intercellular transport of microRNAs.Arterioscler Thromb Vasc Biol. 2013;33(2):186-192.
[24]田淼,谭三勤,刘静茹,等.高通量测序技术分析急性髓系白血病血浆外泌体微RNA表达谱差异[J].中国生物化学与分子生物学报,2017, 33(2):138-149.
[25]陈瑶,陆道培,刘开彦,等.异基因造血干细胞移植后急性移植物抗宿主病151例危险因素分析[J].中华血液学杂志, 2005, 26(2):74-77.
[26] Lechman ER, Gentner B, Ng SW, et al. miR-126 Regulates Distinct Self-Renewal Outcomes in Normal and Malignant Hematopoietic Stem Cells. Cancer Cell. 2016;29(2):214-228.
[27]邓雪蓉,任汉云,岑溪南,等.100例异基因造血干细胞移植后移植物抗宿主病的发生及其对患者复发和生存的影响[J].中国实验血液学杂志, 2009, 17(4):994-998.
[28]胡国文,汪泱,沈晓黎,等.外泌体及其在干细胞中的研究进展[J].上海交通大学学报(医学版),2013, 33(6):868-873.
[29] Koutova L, Sterbova M, Pazourkova E, et al. The impact of standard chemotherapy on miRNA signature in plasma in AML patients. Leuk Res. 2015;39(12):1389-1395.
[2]Báez A, Martín-Antonio B, Piruat JI, et al. Gene and miRNA expression profiles of hematopoietic progenitor cells vary depending on their origin. Biol Blood Marrow Transplant.2014;20(5):630-639.
[3]Dickinson AM, Norden J. Non-HLA genomics:does it have a role in predicting haematopoietic stem cell transplantation outcome. Int J Immunogenet. 2015;42(4):229-238.
[4]Paczesny S, Raiker N, Brooks S, et al. Graft-versus-host disease biomarkers:omics and personalized medicine. Int J Hematol. 2013;98(3):275-292.
[5]Sanghvi VR, Mavrakis KJ, Van der Meulen J, et al.Characterization of a set of tumor suppressor microRNAs in T cell acute lymphoblastic leukemia. Sci Signal. 2014;7(352):ra111.
[6]Gentner B, Pochert N, Rouhi A, et al. MicroRNA-223 dose levels fine tune proliferation and differentiation in human cord blood progenitors and acute myeloid leukemia. Exp Hematol.2015;43(10):858-868.e7.
[7]An T, Qin S, Xu Y, et al. Exosomes serve as tumour markers for personalized diagnostics owing to their important role in cancer metastasis. J Extracell Vesicles. 2015;4:27522.
[8]黄依瑶,郑磊.重视外泌体的实验诊断价值[J].中华检验医学杂志,2015, 38(11):724-726.
[9]Muntión S, Ramos TL, Diez-Campelo M, et al. Microvesicles from Mesenchymal Stromal Cells Are Involved in HPC-Microenvironment Crosstalk in Myelodysplastic Patients.PLoS One. 2016;11(2):e0146722.
[10]黄依瑶,唐月汀,覃思华,等.血清中外泌体及外泌体RNA提取方法的比较[J].中华检验医学杂志,2016, 39(6):427-432.
[11]赵晓琦,程敏.外泌体相关微小RNA的生物学特性及其在多种心血管疾病中的诊断价值[J].临床心血管病杂志,2015, 31(9):924-928.
[12]杨雅芝,张小燕,万倩.外泌小体内miRNA在多发性骨髓瘤进展中的作用及可能机制[J].中国实验血液学杂志, 2017,25(1):301-305.
[13] Wang L, Romero M, Ratajczak P, et al. Increased apoptosis is linked to severe acute GVHD in patients with Fanconi anemia.Bone Marrow Transplant. 2013;48(6):849-853.
[14] De Tullio G, De Fazio V, Sgherza N, et al. Challenges and opportunities of microRNAs in lymphomas. Molecules. 2014;19(9):14723-14781.
[15]简雯.帕金森病患者血浆外泌体的分离鉴定及miRNA表达谱分析[D].南昌:南昌大学, 2016.
[16]胡彬,付伟,黄斯勇,等.异基因造血干细胞移植后血浆microRNA的表达与急性移植物抗宿主病的相关性研究[J].中国实验血液学杂志,2016,24(3):827-832.
[17]郭智,陈惠仁.异基因造血干细胞移植后的相关并发症及危险因素[J].中国组织工程研究,2013,17(40):7163-7168.
[18]田淼,谭三勤,刘静茹,等.高通量测序技术分析急性髓系白血病血浆外泌体微RNA表达谱差异[J].中国生物化学与分子生物学报,2017, 33(2):138-149.
[19] Wojtowicz EE, Walasek MA, Broekhuis MJ, et al.MicroRNA-125 family members exert a similar role in the regulation of murine hematopoiesis. Exp Hematol. 2014;42(10):909-918.e1.
[20] Zhang J, Li S, Li L, et al. Exosome and exosomal microRNA:trafficking, sorting, and function. Genomics Proteomics Bioinformatics. 2015;13(1):17-24.
[21] Lia G, Brunello L, Bruno S, et al. Extracellular vesicles as potential biomarkers of acute graft-vs-host disease. Leukemia.2018;32(3):765-773.
[22] Xiao B, Wang Y, Li W, et al. Plasma microRNA signature as a noninvasive biomarker for acute graft-versus-host disease.Blood. 2013;122(19):3365-3375.
[23] Boon RA, Vickers KC. Intercellular transport of microRNAs.Arterioscler Thromb Vasc Biol. 2013;33(2):186-192.
[24]田淼,谭三勤,刘静茹,等.高通量测序技术分析急性髓系白血病血浆外泌体微RNA表达谱差异[J].中国生物化学与分子生物学报,2017, 33(2):138-149.
[25]陈瑶,陆道培,刘开彦,等.异基因造血干细胞移植后急性移植物抗宿主病151例危险因素分析[J].中华血液学杂志, 2005, 26(2):74-77.
[26] Lechman ER, Gentner B, Ng SW, et al. miR-126 Regulates Distinct Self-Renewal Outcomes in Normal and Malignant Hematopoietic Stem Cells. Cancer Cell. 2016;29(2):214-228.
[27]邓雪蓉,任汉云,岑溪南,等.100例异基因造血干细胞移植后移植物抗宿主病的发生及其对患者复发和生存的影响[J].中国实验血液学杂志, 2009, 17(4):994-998.
[28]胡国文,汪泱,沈晓黎,等.外泌体及其在干细胞中的研究进展[J].上海交通大学学报(医学版),2013, 33(6):868-873.
[29] Koutova L, Sterbova M, Pazourkova E, et al. The impact of standard chemotherapy on miRNA signature in plasma in AML patients. Leuk Res. 2015;39(12):1389-1395.
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