Effects of extracellular vesicles on miRNA activity in the skin

项目总结/摘要 每年诊断出的新病例超过100万，皮肤癌占世界所有癌症的近一半。 美国鳞状细胞癌（SCC）是一种具有常见遗传特征的主要皮肤癌，但 没有明确的肿瘤发生机制开发SCC的靶向疗法一直具有挑战性， 该疾病的分子病因学描绘了一幅异质的画面。一个新兴的概念是， 或癌细胞和肿瘤微环境（TME）之间的串扰对于肿瘤起始是至关重要的， 进展这种细胞间质串扰的关键是膜衍生的纳米级细胞外基质的作用。 由肿瘤细胞分泌并释放到血液和其他体液中的囊泡（EV）携带它们的细胞， 分子大分子货物调节局部和远端TME。除了促有丝分裂信号蛋白 和mRNA，EV携带miRNA，短的非编码RNA，其抑制对细胞增殖至关重要的互补mRNA。 周期调节和细胞增殖和分化。我们研究的长期目标是了解 在分子和细胞水平上，发育过程中的正常信号事件是如何被破坏的， SCC进展过程中的致病信号传导。在恶性转化过程中，异常表达或 观察到桥粒钙粘蛋白桥粒芯糖蛋白2（Dsg 2）的定位。Dsg 2在正常的 细胞生长和发育，这是强调的事实，基因消融导致胚胎细胞 小鼠的生长缺陷和致死性。在人类中，Dsg 2的突变与心肌病有关， 呼吸道和泌尿道感染以及阿尔茨海默病。Dsg 2的过表达也发生在 各种其他恶性肿瘤，包括前列腺，结肠和皮肤，表明Dsg 2在肿瘤中的作用更普遍。 肿瘤发生Dsg 2促进细胞增殖、迁移和过度增殖的机制 疾病尚未完全了解。该提案的目标是证明Dsg 2促进肿瘤发生 通过增强携带能够引发角质形成细胞的癌细胞状态特异性货物的EV的释放 和成纤维细胞。我们的假设是Dsg 2的细胞表面呈递调节了EV相关细胞的输出。 可作为SCC癌症进展和/或侵袭的生物标志物和关键调节因子的miRNA 使它们成为新的治疗靶点。目的1将确定Dsg 2调节细胞凋亡的机制。 EV的生物合成。目的2研究EV相关miR 146 a和miR 155的细胞效应。目标3将 评估血清EV miRNA水平作为SCC预后危险因素的效用。这里得到的结果将 阐明Dsg 2的新功能，并阐明Dsg 2如何影响细胞生长和迁移： 生物学步骤对于适当的组织形态发生和肿瘤发生至关重要。一旦成功完成 在这项工作中，我们将定义控制细胞EV输出的机制; EV如何调节细胞的运动。 基质环境;并揭示可作为诊断和预后的生物标志物的新型miRNA 恶性SCC和其他上皮来源的癌症。

PROJECT SUMMARY/ABSTRACT With more than one million new cases diagnosed yearly, skin cancer accounts for nearly half of all cancers in the U.S. Squamous cell carcinoma (SCC) is a major form of skin cancer with common genetic signatures, but without a clear mechanism of oncogenesis. Developing targeted therapies for SCCs has been challenging as the molecular etiology of the disease paints a heterogeneous picture. An emerging concept is that the synergy or crosstalk between cancer cells and the tumor microenvironment (TME) is critical for tumor initiation and progression. Essential for this cell-stroma crosstalk is the role of membrane-derived, nano-sized extracellular vesicles (EVs) secreted by tumor cells and released into the blood and other bodily fluids carrying their molecular macromolecular cargo to modulate local and distant TME. In addition to mitogenic signaling proteins and mRNAs, EVs carry miRNAs, short non-coding RNAs that repress complementary mRNAs critical for cell cycle regulation and cell proliferation and differentiation. The long-term goal of our research is to understand at the molecular and cellular level, how normal signaling events during development are subverted for pathogenic signaling during SCC progression. During malignant transformation, aberrant expression or localization of the desmosomal cadherin desmoglein 2 (Dsg2) is observed. Dsg2 plays a critical role in normal cell growth and development, which is emphasized by the fact that genetic ablation results in embryonic cell growth defects and lethality in mice. In human, mutations in Dsg2 are associated with cardiomyopathy, respiratory and urinary tract infections, and Alzheimer's disease. Overexpression of Dsg2 also occurs in various other malignancies including prostate, colon and skin, suggesting a more general role for Dsg2 in oncogenesis. The mechanism by which Dsg2 promotes cell proliferation, migration, and hyperproliferative disorders is not fully understood. The goal of this proposal is demonstrate that Dsg2 promotes oncogenesis by enhancing the release of EVs carrying cancer cell-state specific cargos capable of priming keratinocytes and fibroblasts. It is our hypothesis that cell surface presentation of Dsg2 modulates export of EV-associated miRNAs that can serve as biomarkers and key regulators of SCC cancer progression and/or invasion rendering them novel therapeutic targets. Aim 1 will determine the mechanisms by which Dsg2 modulates the biosynthesis of EVs. Aim 2 will study the cellular effects of EV-associated miR146a and miR155. Aim 3 will evaluate the utility of serum EV miRNA levels as prognostic risk factors in SCC. The results obtained here will elucidate novel functions of Dsg2 and shed new light on how Dsg2 can impact cell growth and migration: biological steps critical for proper tissue morphogenesis and tumorigenesis. Upon the successful completion of this work, we will define the mechanisms by which cellular EV export are controlled; how EVs can modulate the stromal environment; and reveal novel miRNAs that can serve as biomarkers for the diagnosis and prognosis of malignant SCCs and other epithelial-derived cancers.