Nanoscale flow cytometry to distinguish subpopulations of prostate extracellular vesicles in patient plasma
Nanoscale flow cytometry to distinguish subpopulations of prostate extracellular vesicles in patient plasma
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DOI：
10.1002/pros.23764
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发表时间：
2019-05-01
期刊：
影响因子：
2.8
通讯作者：
Leong, Hon S.
中科院分区：
文献类型：
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作者：
Padda, Ranjit S.;Deng, Florence K.;Leong, Hon S.
目的：确定患者血浆中存在的前列腺源性细胞外小泡(EV)是胞吐来源(Exosome)还是由细胞膜释放(微粒/微囊)。恶性和正常前列腺细胞都会释放两种类型的EV进入循环，即外体和微粒子/微囊，它们的大小、来源和释放方式都不同。确定前列腺源性EV外体型与微粒子/微囊型EV亚型的比例具有潜在的诊断意义。材料和方法：采用纳米级流式细胞仪(NFC)等多参数分析平台对前列腺源性胞外囊泡进行分析。分析了前列腺癌(Pca)患者血浆中各种外体标志物(CD9、CD63、CD81)和前列腺特异性组织标志物(前列腺特异性膜抗原/PSMA)。结果：通过纳米级流式细胞术，我们确定前列腺源性EV主要来源于细胞膜来源、微粒/微囊，并不是所有PSMA表达EVS的EV共表达外体标志物CD9、CD63和CD81。CD9是前列腺源性EVS中含量最丰富的外体标记物(12-19%)。随着前列腺癌分级的增加，PSMA+CD9或PSMA+CD63共表达EVS的比例没有增加的趋势。结论：前列腺癌组织中存在的EVS大多来自细胞膜，其大小和最重要的是缺乏CD9/CD63/CD81等外源标志物的共表达。事实上，CD81在患者血浆中的任何前列腺来源的EV上都不存在，而CD9在少数前列腺来源的EV上存在。增加用于检测前列腺源性EVS的胞外标记物并不能更清楚地区分前列腺释放的EVS。
Objective: To determine if prostate-derived extracellular vesicles (EVs) present in patient plasma samples are of exocytotic origin (exosomes) or released by the cell membrane (microparticles/microvesicles). Both malignant and normal prostate cells release two types of EVs into the circulation, exosomes, and microparticles/ microvesicles which differ in size, origin, and mode of release. Determining what proportion of prostate-derived EVs are of exosomal versus microparticle/microvesicle EV subtype is of potential diagnostic significance.Materials and Methods: Multi-parametric analytical platforms such as nanoscale flow cytometry (nFC) were used to analyze prostate derived extracellular vesicles. Plasmas from prostate cancer (PCa) patient plasmas representing benign prostatic hyperplasia (BPH), low grade prostate cancer (Gleason Score 3 + 3) and high grade prostate cancer (Gleason Score >= 4 + 4) were analyzed for various exosome markers (CD9, CD63, CD81) and a prostate specific tissue marker (prostate specific membrane antigen/ PSMA).Results: By using nanoscale flow cytometry, we determine that prostate derived EVs are primarily of cell membrane origin, microparticles/microvesicles, and not all PSMA expressing EVs co-express exosomal markers such as CD9, CD63, and CD81. CD9 was the most abundant exosomal marker on prostate derived EVs (12-19%). There was no trend observed in terms of more PSMA + CD9 or PSMA + CD63 co-expressing EVs versus increasing grade of prostate cancer.Conclusion: The majority of prostate derived EVs present in plasmas are from the cell membrane as evidenced by their size and most importantly, lack of co-expression of exosomal markers such as CD9/CD63/CD81. In fact, CD81 was not present on any prostate derived EVs in patient plasmas whereas CD9 was present on a minority of prostate derived EVs. The addition of an exosomal marker for detection of prostate-derived EVs does not provide greater clarity in distinguishing EVs released by the prostate.