Extracellular vesicle RNA as systemic modulators of the tumor microenvironment in B cell lymphoma

细胞外小泡(EV)是细胞间通讯的重要媒介，参与多种生理过程。它们几乎存在于我们身体的任何液体隔间中，甚至在它们起源的细胞的远处也能被检测到。在过去的十年里，它们在癌症的发生、发展和扩散中的关键作用变得越来越明显。EVS由一层脂质双层膜组成，显示其来源细胞的细胞表面蛋白，主要含有蛋白质和RNA分子。现在很清楚，电动汽车可以通过它们的表面分子与细胞相互作用，并可以将其内容转移到靶细胞。转移的物质，例如蛋白质、信使RNAs或microRNAs，被证明在受体细胞中是活跃的，导致它们的表型发生变化。在生理条件下，EVS作为免疫反应的介质，有迹象表明肿瘤来源的EVS有助于疾病相关的炎症和免疫抑制。迄今为止，肿瘤细胞来源的EVS主要用于实体癌的研究，而关于血液系统疾病中恶性细胞产生的EVS的数据很少。因此，我们提出的项目的目的是对EVS进行广泛的分析，重点是来自慢性淋巴细胞白血病(CLL)、多发性骨髓瘤(MM)和弥漫性大B细胞淋巴瘤(DLBCL)的恶性细胞的外体。这项研究包括：i)外切体的比较蛋白质组和RNA分析，重点放在Y RNAs和microRNAs及其分选机制；ii)在体外和小鼠模型中鉴定造血微环境中摄取肿瘤衍生的exosome的靶细胞；iii)描述靶细胞的转录体、分泌体和信号能力的下游变化，据此侧重于Toll样受体介导的炎症反应；以及iv)研究以评估exosome对合适的小鼠模型中B细胞淋巴瘤发展的影响。最后，我们将v)在小鼠的临床前研究中，研究外切体作为生物标记物和治疗靶点的潜力。

Extracellular vesicles (EVs) are important mediators of intercellular communication and are involved in many physiological processes. They are present in almost any fluid compartment of our bodies and can be detected even at distant sites of their cell of origin. Within the last decade, their critical involvement in cancer development, progression and dissemination has become more and more obvious. EVs consist of a lipid bilayer membrane displaying cell surface proteins of their cell of origin and contain mainly protein and RNA molecules. It is now clear that EVs can interact with cells via their surface molecules and can transfer their content to target cells. The transferred material, e.g. proteins, messengerRNAs or microRNAs, was shown to be active in recipient cells, leading to changes in their phenotypes. Under physiological conditions, EVs act as mediators of immune responses, and there are indications that tumor-derived EVs contribute to disease-associated inflammation and immune suppression.Tumor cell-derived EVs have so far been mainly studied in solid cancers, and there is only little data on EVs produced by malignant cells in hematological diseases. Therefore, the aim of our proposed project is an extensive analysis of EVs, with a focus on exosomes, derived from malignant cells of chronic lymphocytic leukemia (CLL), multiple myeloma (MM) and diffuse large B cell lymphoma (DLBCL). The study includes i) comparative proteome and RNA analyses of exosomes, with a focus on Y RNAs and microRNAs and their sorting mechanisms; ii) the identification of target cells within the hematopoietic microenvironment that take up tumor-derived exosomes in vitro and in mouse models; iii) a characterization of downstream changes within the transcriptome, secretome and signaling capacities of target cells, focusing hereby on inflammatory responses that are mediated by toll-like receptors; as well as iv) investigations to evaluate the impact of exosomes on B cell lymphoma development in suitable mouse models. Finally, we will v) investigate in a translational approach the potential of exosomes as biomarkers and as therapeutic target in preclinical studies in mice.