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Short Course in Extracellular Vesicles — The Transition from Tissue to Liquid Biopsies 细胞外囊泡的短期病程-从组织活检到液体活检的转变
Q3 Medicine Pub Date : 2014-01-01 DOI: 10.5772/60053
J. Lötvall, J. Skog, A. Vlassov, A. Sacido, E. Rohde, J. Gere, W. Kuo
Extracellular vesicles (EVs), including exosomes and microvesicles, carry a variety of bio-macromolecules, including mRNA, microRNA, other non-coding RNAs, proteins and lipids. EVs have emerged as a promising, minimally invasive (liquid biopsies) and novel source of material for molecular diagnostics, and may provide a surrogate to tissue biopsy-based biomarkers for a variety of diseases. Although EVs can be easily identified and collected from biological fluids using commercial kits, further research and proper validation is needed in order for them to be useful in the clinical setting. Currently, several EV-based research and diagnostic companies have developed research-based kits and are in the process of working with clinical laboratories to develop and validate EV-based assays for a variety of diseases. The successful clinical application of EV-based diagnostic assays will require close collaboration between industry, academia, regulatory agencies and access to patient samples. We expect that international, integrative and interdisciplinary translational research teams, along with the emergence of FDA-approved platforms, will set the framework for EV-based diagnostics. We recognize that the EV field offers new promise for personalized/precision medicine and targeted treatment in a variety of diseases. A short course was held as a four-session webinar series in September and October 2014, presented by pioneers and experts in the EV domain, covering a broad range of topics from an overview of the field to its applications, and the current state and challenges of the commercialization of EVs for research and an introduction to the clinic. It was concluded with a panel discussion on the regulatory aspects and funding opportunities in this field. A summary of the short course is presented as a meeting dispatch.
细胞外囊泡(EVs),包括外泌体和微囊泡,携带多种生物大分子,包括mRNA、microRNA、其他非编码rna、蛋白质和脂质。电动汽车已经成为一种有前途的、微创的(液体活检)和分子诊断材料的新来源,并可能为各种疾病提供基于组织活检的生物标志物的替代品。虽然使用商业试剂盒可以很容易地从生物体液中识别和收集EVs,但为了使其在临床环境中有用,还需要进一步的研究和适当的验证。目前,几家基于ev的研究和诊断公司已经开发了基于研究的试剂盒,并正在与临床实验室合作,开发和验证基于ev的各种疾病检测方法。基于ev的诊断分析的成功临床应用将需要工业界、学术界、监管机构之间的密切合作,并获得患者样本。我们期待国际、综合和跨学科的转化研究团队,以及fda批准的平台的出现,将为基于ev的诊断设定框架。我们认识到,电动汽车领域为各种疾病的个性化/精准医疗和靶向治疗提供了新的希望。2014年9月和10月举办了一个短期课程,由电动汽车领域的先驱和专家介绍,包括从该领域的概述到其应用的广泛主题,以及电动汽车商业化研究和临床应用的现状和挑战。会议结束时,小组讨论了这一领域的监管方面和筹资机会。短期课程的摘要以会议简报的形式呈现。
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引用次数: 2
Uncovering the Role of Erythrocyte-Derived Extracellular Vesicles in Malaria: From Immune Regulation to Cell Communication 揭示红细胞来源的细胞外囊泡在疟疾中的作用:从免疫调节到细胞通讯
Q3 Medicine Pub Date : 2014-01-01 DOI: 10.5772/58596
Johan Ankarklev, D. Hjelmqvist, Pierre-Yves Mantel
Investigation of the involvement of extracellular vesicles (EVs) in parasite biology has burgeoned in recent years. Human infecting protozoan parasites, such as Trypanosoma cruzi, Lesihmania sp. and Trichomonas vaginalis, have all demonstrated the utilization of EVs as virulence factors in order to activate or hamper host immunity. Novel findings have provided evidence that the deployment of EVs by Plasmodium sp. has a major impact in disease outcomes and serves as an integral part in controlling stage switching in its life cycle. Clinical studies have highlighted elevated levels of EVs in patients with severe malaria disease and EVs have been linked to increased sequestration of infected red blood cells to the endothelium, causing obstruction of blood flow. It has also been found that EVs produced during malaria disease activate innate immunity. Intriguingly, recent discoveries indicate that Plasmodium sp. “highjack” the erythrocyte microvesiculation system in order to cross-communicate. Both the transfer of DNA and parasite density regulation has been suggested as key mechanisms of EVs in malaria biology.
近年来,关于细胞外囊泡(EVs)在寄生虫生物学中的作用的研究迅速发展。人类感染的原生动物寄生虫,如克氏锥虫、莱希曼原虫和阴道毛滴虫,都显示出利用EVs作为毒力因子来激活或阻碍宿主免疫。新的研究结果提供了证据,证明疟原虫的ev部署对疾病结局有重大影响,并在控制其生命周期的阶段转换中起着不可或缺的作用。临床研究强调,严重疟疾患者的EVs水平升高,并且EVs与受感染的红细胞与内皮细胞的隔离增加有关,从而导致血液流动受阻。还发现疟疾期间产生的ev可激活先天免疫。有趣的是,最近的发现表明,疟原虫“劫持”红细胞微泡系统以进行交叉交流。DNA转移和寄生虫密度调节被认为是EVs在疟疾生物学中的关键机制。
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引用次数: 8
Exosomal Heat Shock Proteins as New Players in Tumour Cell-to-Cell Communication 外泌体热休克蛋白在肿瘤细胞间通讯中的新作用
Q3 Medicine Pub Date : 2014-01-01 DOI: 10.5772/58721
C. Campanella, C. C. Bavisotto, A. M. Gammazza, D. Nikolić, F. Rappa, S. David, F. Cappello, F. Bucchieri, S. Fais
Exosomes have recently been proposed as novel elements in the study of intercellular communication in normal and pathological conditions. The biomolecular composition of exosomes reflects the specialized functions of the original cells. Heat shock proteins (Hsps) are a group of chaperone proteins with diverse biological roles. In recent years, many studies have focused on the extracellular roles played by Hsps that appear to be involved in cancer development and immune system stimulation. Hsps localized on the surface of exosomes, secreted by normal and tumour cells, could be key players in intercellular cross-talk, particularly during the course of different diseases, such as cancer. Exosomal Hsps offer significant opportunities for clinical applications, including their use as potential novel biomarkers for the diagnoses or prognoses of different diseases, or for therapeutic applications and drug delivery.
外泌体最近被认为是研究正常和病理条件下细胞间通讯的新元素。外泌体的生物分子组成反映了原始细胞的特化功能。热休克蛋白(Hsps)是一类具有多种生物学作用的伴侣蛋白。近年来,许多研究都集中在热休克蛋白在癌症发展和免疫系统刺激中所起的细胞外作用上。位于正常细胞和肿瘤细胞分泌的外泌体表面的热休克蛋白可能是细胞间串扰的关键角色,特别是在不同疾病(如癌症)的过程中。外泌体热休克蛋白为临床应用提供了重要的机会,包括它们作为潜在的新型生物标志物用于不同疾病的诊断或预后,或用于治疗应用和药物输送。
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引用次数: 45
Influence of Lung Parenchyma Surgical Manipulation on Circulating Free DNA 肺实质手术操作对循环游离DNA的影响
Q3 Medicine Pub Date : 2014-01-01 DOI: 10.5772/59875
M. Anile, C. Chiappetta, D. Diso, V. Liparulo, M. Leopizzi, C. Della Rocca, F. Venuta
Objectives: Metastatic recurrence is the most frequent cause of death after surgical resection of lung cancer. Manipulation during surgery has been advocated as one of the causes contributing to promotion of spreading. Methods: We investigated if the detection of plasma circulating free DNA (cfDNA) is influenced by surgical manipulation in 25 lung cancer patients (17 males and eight females) undergoing complete resection; 20 health subjects formed the control group. Bloodstream levels of cfDNA were detected before surgery, one week and one month after surgery. Results: CfDNA levels measured preoperatively and in the control group were 23 07 ± 7 4 ng/mL and 7 5 ± 3 4 ng/mL respectively (p=0 0002); levels at one week and one month were 68 2 ± 36 2 ng/mL and 9 6 ± 3 1 ng/mL respectively. The difference between the three time points were statistically significant (preop vs. one week p=0 0006; one week vs. one month p=0 0003) with an increase in the first week and a strong decrease after one month. CfDNA levels at one month were not statistically different from those recorded in the control group. There was no correlation between preoperative cfDNA levels, tumour stage, grading and histology and patient demographics. No correlation was found between postoperative cfDNA, type of surgical procedure, histology and stage. After a median follow-up of 16 months no recurrence was detected. Conclusions: Surgical manipulation determines increased cfDNA levels in the early postoperative period; however, after one month they decrease within the normal range, at levels that are statistically comparable with healthy subjects.
目的:转移性复发是肺癌手术切除后最常见的死亡原因。术中操作被认为是促进扩散的原因之一。方法:观察25例肺癌全切除术患者(男17例,女8例)手术操作对血浆循环游离DNA (cfDNA)检测的影响;健康受试者20人作为对照组。分别于术前、术后1周和1个月检测血液中cfDNA水平。结果:术前与对照组CfDNA水平分别为23 07±7 4 ng/mL、7 5±3 4 ng/mL (p=0 0002);1周和1个月的水平分别为68 2±36 2 ng/mL和9 6±31 ng/mL。三个时间点之间的差异具有统计学意义(术前vs.一周p= 0006;一周vs一个月p=0 0003),第一周增加,一个月后大幅下降。1个月时CfDNA水平与对照组无统计学差异。术前cfDNA水平与肿瘤分期、分级、组织学和患者人口统计学无相关性。术后cfDNA与手术方式、组织学及分期无相关性。中位随访16个月后未发现复发。结论:手术操作决定术后早期cfDNA水平升高;然而,一个月后,它们在正常范围内下降,在统计上与健康受试者相当。
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引用次数: 0
Identification of Immunoreactive Tumour Antigens Using Free and Exosome-Associated Humoral Responses 利用游离和外泌体相关体液反应鉴定免疫反应性肿瘤抗原
Q3 Medicine Pub Date : 2013-01-01 DOI: 10.5772/57524
Carolyn D. Roberson, Ç. Gerçel-Taylor, Ying Qi, K. Schey, D. Taylor
Altered tumour antigens can initiate cellular and humoral immune responses; however, they often fail to eliminate tumours. In humans, the presence of cancer is generally associated with the suppression of T cell activation and effector responses, characterized as a Th1 to Th2 biased response. This Th2 response leads to the production of tumour-reactive antibodies. Further, neoplastic lesions and biological fluids of cancer patients contain an abundance of tumour-derived exosomes (TDE) expressing tumour antigens. Expression of tumour antigens on TDE may represent an antibody target and serve to block antibody binding to the tumour, implicating a role for these nanovesicles in tumour survival. In this study, ovarian tumour cell proteins were separated by two-dimensional electrophoresis (2-DE) and patient-derived antibodies were used to analyse immunoreactivity. Common immunoreactive proteins among ovarian cancer patients were identified by mass spectrometry and six proteins were selected based on recognition and correlation with cancer pathogenesis. The identity of these proteins were confirmed by immunoreactivity of patient-derived antibodies with recombinant proteins and their presence on in vivo and in vitro-derived ovarian tumour exosomes was defined. Analysis of the TDE demonstrated bound tumour-reactive immunoglobulins, exhibiting immunoreactivity with specific antigens, suggesting that patient-derived antibodies recognize tumour antigens on circulating exosomes.
改变的肿瘤抗原可以启动细胞和体液免疫反应;然而,它们往往不能消除肿瘤。在人类中,癌症的存在通常与T细胞活化和效应反应的抑制有关,其特征是Th1对Th2的偏倚反应。这种Th2反应导致肿瘤反应性抗体的产生。此外,肿瘤病变和癌症患者的生物体液中含有大量表达肿瘤抗原的肿瘤源性外泌体(TDE)。肿瘤抗原在TDE上的表达可能代表抗体靶点,并阻断抗体与肿瘤的结合,这暗示了这些纳米囊泡在肿瘤存活中的作用。本研究采用双向电泳(2-DE)分离卵巢肿瘤细胞蛋白,并采用患者源性抗体分析免疫反应性。采用质谱法对卵巢癌患者中常见的免疫反应蛋白进行鉴定,并根据识别程度及与肿瘤发病机制的相关性筛选出6种免疫反应蛋白。这些蛋白的身份通过患者来源的抗体与重组蛋白的免疫反应性得到证实,并确定了它们在体内和体外来源的卵巢肿瘤外泌体上的存在。TDE分析显示结合肿瘤反应性免疫球蛋白,对特定抗原表现出免疫反应性,表明患者来源的抗体识别循环外泌体上的肿瘤抗原。
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引用次数: 3
Announcing Exosomes and Microvesicles, the Official Journal of the American Society for Exosomes and Microvesicles 宣布外泌体和微囊泡,美国外泌体和微囊泡学会官方期刊
Q3 Medicine Pub Date : 2013-01-01 DOI: 10.5772/56520
S. Gould, D. Taylor, A. Chiesi, W. Kuo
This editorial article introduces the new scientific journal Exosomes and Microvesicles (EXMV), the official journal of the American Society for Exosomes and Microvesicles (ASEMV), and describes its editorial line and mission in relation to the role of the Society, the state of the art of the study of exosomes and microvesicles, and the overall approach of the publication.
这篇社论文章介绍了美国外泌体和微囊泡学会(ASEMV)的官方期刊《外泌体和微囊泡》(EXMV)的新科学期刊,并描述了其编辑路线和任务,包括该学会的作用、外泌体和微囊泡研究的最新技术状况以及出版的总体方法。
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引用次数: 1
Rab27B-Mediated Metabolic Reprogramming Induces Secretome Acidification and Chemoresistance in Breast Cancer Cells rab27b介导的代谢重编程诱导乳腺癌细胞分泌组酸化和化疗耐药
Q3 Medicine Pub Date : 2013-01-01 DOI: 10.5772/56521
A. Hendrix, C. Ciccone, C. Gespach, M. Bracke, O. De Wever, W. Westbroek
The secretory Rab27B small GTPase promotes invasive growth, tumourigenicity and metastasis in oestrogen receptor (ER)-positive human breast cancer cells. Coherently, increased Rab27B expression in breast cancer patients is associated with a poor prognosis. In the present study, bio-energetic profiling revealed that oxidative phosphorylation is significantly reduced in ER-positive breast cancer cells engineered to overexpress Rab27B levels as observed in invasive clinical primary breast cancer. Rab27B-induced metabolic reprogramming to aerobic glycolysis was further evidenced by increased extracellular acidification followed by cathepsin B activation and doxorubicin resistance. Transient silencing of Rab27B and stable transfection of Rab27A, and Rab27B mutants in ER-positive breast cancer cells confirmed that this response was Rab27B-specific and dependent upon Rab27B-GTP activation and vesicle membrane attachment through the C-terminal geranylgeranyl group of this small GTPase. Rab27B-driven extracellular acidification is required and is sufficient to induce filopodia-like morphological changes, primarily involved in the process of cancer cell invasion. Our data demonstrate that a Rab27B-dependent switch from oxidative phosphorylation towards aerobic glycolysis in ER-positive breast cancer cells is accompanied by acidification of the tumour environment.
分泌Rab27B小GTPase促进雌激素受体(ER)阳性人乳腺癌细胞的侵袭性生长、致瘤性和转移。Rab27B在乳腺癌患者中的表达升高与预后不良相关。在本研究中,生物能量谱分析显示,在侵袭性临床原发性乳腺癌中观察到的er阳性乳腺癌细胞中,氧化磷酸化水平显著降低,从而过度表达Rab27B水平。rab27b诱导代谢重编程为有氧糖酵解的进一步证据是细胞外酸化增加,随后是组织蛋白酶B活化和阿霉素耐药性。在er阳性乳腺癌细胞中短暂沉默Rab27B并稳定转染Rab27A和Rab27B突变体证实了这种反应是Rab27B特异性的,并且依赖于Rab27B- gtp的激活和通过这种小GTPase的c端香叶基基基团附着在囊泡膜上。rab27b驱动的细胞外酸化是必需的,并且足以诱导丝状足样形态变化,主要参与癌细胞侵袭过程。我们的数据表明,在er阳性乳腺癌细胞中,依赖rab27b从氧化磷酸化到有氧糖酵解的转变伴随着肿瘤环境的酸化。
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引用次数: 6
Tumour Cells Incorporate Exosomes Derived from Dendritic Cells through a Mechanism Involving the Tetraspanin CD9 肿瘤细胞通过与四蛋白CD9相关的机制整合来自树突状细胞的外泌体
Q3 Medicine Pub Date : 2013-01-01 DOI: 10.5772/52069
G. Romagnoli, P. A. Toniolo, Isabela Katz Migliori, E. Caldini, M. A. Ferreira, C. R. Pizzo, P. C. Bergami-Santos, J. Barbuto
Exosomes (Exos) are secreted nanovesicles that contain membrane proteins and genetic material, which can be transferred between cells and contribute to their communication in the body. We show that Exos, obtained from mature human dendritic cells (DCs), are incorporated by tumour cells, which after Exos treatment, acquire the expression of HLA-class I, HLA-class II, CD86, CD11c, CD54 and CD18. This incorporation reaches its peak eight hours after treatment, can be observed in different cell tumour lines (SK-BR-3, U87 and K562) and could be a means to transform non-immunogenic into immunogenic tumour cells. Interestingly, tetraspanins, which are expressed by the tumour cells, have their surface level decreased after Exo treatment. Furthermore, the intensity of Exo incorporation by the different tumour cell lines was proportional to their CD9 expression levels and pre-treatment of Exos with anti-CD9 decreased their incorporation (by SK-BR-3 cells). This modification of tumour cells by DC-derived Exos may allow their use in new immunotherapeutic approaches to cancer. Furthermore, by showing the involvement of CD9 in this incorporation, we provide a possible selection criterion for tumours to be addressed by this strategy.
外泌体(Exos)是分泌的纳米囊泡,含有膜蛋白和遗传物质,可以在细胞之间转移,并有助于它们在体内的交流。我们发现,从成熟的人树突状细胞(dc)中获得的Exos与肿瘤细胞结合,经过Exos处理后,肿瘤细胞获得hla - I类、hla - II类、CD86、CD11c、CD54和CD18的表达。这种结合在治疗后8小时达到峰值,可以在不同的细胞肿瘤系(SK-BR-3、U87和K562)中观察到,可能是将非免疫原性肿瘤细胞转化为免疫原性肿瘤细胞的一种手段。有趣的是,肿瘤细胞表达的四联蛋白在Exo治疗后其表面水平降低。此外,不同肿瘤细胞系的Exo掺入强度与其CD9表达水平成正比,用抗CD9预处理Exo可以减少它们的掺入(SK-BR-3细胞)。这种由dc衍生的Exos修饰的肿瘤细胞可能允许它们在新的癌症免疫治疗方法中使用。此外,通过显示CD9参与这种结合,我们提供了一种可能的肿瘤选择标准,通过这种策略来解决。
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引用次数: 7
Therapeutic Uses of Exosomes 外泌体的治疗用途
Q3 Medicine Pub Date : 2013-01-01 DOI: 10.5772/56522
Z. Suntres, Milton G. Smith, F. Momen-Heravi, Jie Hu, Xin Zhang, Ying Wu, Hongguang Zhu, Jiping Wang, Jian Zhou, W. Kuo
Exosomes are membrane vesicles with a diameter of 40–100 nm that are secreted by many cell types into the extracellular milieu. Exosomes are found in cell culture supernatants and in different biological fluids and are known to be secreted by most cell types under normal and pathological conditions. Considerable research is focusing on the exploitation of exosomes in biological fluids for biomarkers in the diagnosis of disease. More recently, exosomes are being exploited for their therapeutic potential. Exosomes derived from dendritic cells, tumor cells, and malignant effusions demonstrate immunomodulatory functions and are able to present antigens to T-cells and stimulate antigen-specific T-cell responses. Exosomes have also been examined for their therapeutic potential in the treatment of infections such as toxoplasmosis, diphtheria, tuberculosis and atypical severe acute respiratory syndrome as well as autoimmune diseases. Attempts to find practical applications for exosomes continue to expand with the role of exosomes as a drug delivery system for the treatment of autoimmune/inflammatory diseases and cancers.
外泌体是一种直径为40 - 100nm的膜囊泡,由许多细胞分泌到细胞外环境中。外泌体存在于细胞培养上清和不同的生物液体中,已知在正常和病理条件下大多数细胞类型都会分泌外泌体。相当多的研究集中在利用生物液体中的外泌体作为疾病诊断的生物标志物。最近,外泌体因其治疗潜力而被开发。来源于树突状细胞、肿瘤细胞和恶性积液的外泌体显示出免疫调节功能,能够向t细胞呈递抗原并刺激抗原特异性t细胞反应。还研究了外泌体在治疗诸如弓形虫病、白喉、肺结核和非典型严重急性呼吸系统综合症以及自身免疫性疾病等感染方面的治疗潜力。随着外泌体作为治疗自身免疫性/炎症性疾病和癌症的药物传递系统的作用,寻找外泌体实际应用的尝试不断扩大。
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引用次数: 32
Exosome: A Novel and Safer Therapeutic Refinement of Mesenchymal Stem Cell 外泌体:一种新的、更安全的间充质干细胞治疗方法
Q3 Medicine Pub Date : 2013-01-01 DOI: 10.5772/57460
R. Yeo, R. C. Lai, K. Tan, S. Lim
Mesenchymal stem cell (MSC) has just been approved as the first “off-the-shelf” stem cell pharmaceutical drug with an anticipation of more approvals following completion of numerous rigorous clinical trials. Despite this progress, the rationale for MSC therapeutic efficacy remains tenuous and is increasingly rationalized on a secretion rather than differentiation mechanism. Recent studies identifying exosome as the secreted agent mediating MSC therapeutic efficacy could potentially reduce a cell-based drug to a safer biologic-based alternative. Here we review the development of MSC exosome as a potential first-in-class therapeutic, and the unique challenges in the manufacture and regulatory oversight of this new class of therapeutics.
间充质干细胞(MSC)刚刚被批准为第一个“现成的”干细胞药物,在完成大量严格的临床试验后,预计会有更多的批准。尽管取得了这些进展,但MSC治疗效果的理论基础仍然薄弱,并且越来越多地在分泌机制而不是分化机制上合理化。最近的研究发现外泌体作为介导MSC治疗效果的分泌剂可能会将基于细胞的药物减少到更安全的基于生物的替代药物。在这里,我们回顾了MSC外泌体作为一种潜在的一流治疗药物的发展,以及这种新型治疗药物在生产和监管方面的独特挑战。
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引用次数: 50
期刊
Journal of Circulating Biomarkers
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