Effects of extracellular vesicles on miRNA activity in the skin

项目摘要/摘要 由于每年新诊断的病例超过100万例，皮肤癌占全世界所有癌症的近一半 美国鳞状细胞癌(SCC)是皮肤癌的一种主要形式，具有共同的遗传特征，但 没有明确的致癌机制。为鳞状细胞癌开发靶向治疗一直具有挑战性，因为 这种疾病的分子病因学描绘了一幅不同的图景。一个新兴的概念是，协同效应 癌细胞和肿瘤微环境(TME)之间的串扰对于肿瘤的发生和发展至关重要。 进步。这种细胞-间质串扰的关键是膜衍生的纳米级细胞外的作用 由肿瘤细胞分泌的囊泡(EV)释放到血液和其他携带其功能的体液中 调节本地和远程TME的分子大分子货物。除了有丝分裂信号蛋白 和mRNAs，EV携带miRNAs，即抑制对细胞至关重要的互补mRNAs的非编码短RNA 细胞周期调控与细胞增殖分化。我们研究的长期目标是了解 在分子和细胞水平上，发育期间的正常信号事件是如何被颠覆的 鳞癌进展过程中的致病信号。在恶变过程中，异常表达或 观察桥粒钙粘附素-桥粒-桥粒蛋白-2(Dsg2)的定位。Dsg2在正常组织中起着关键作用 细胞生长和发育，强调这一点，因为基因消融会导致胚胎细胞 小鼠的生长缺陷和致死性。在人类中，Dsg2基因突变与心肌病有关， 呼吸道和尿路感染，以及阿尔茨海默病。Dsg2的过度表达也发生在 包括前列腺、结肠和皮肤在内的各种其他恶性肿瘤，表明Dsg2在 致癌作用。Dsg2促进细胞增殖、迁移和过度增殖的机制 人们对疾病的认识还不够充分。这项提案的目的是证明Dsg2促进肿瘤发生 通过促进携带能够启动角质形成细胞的癌细胞状态特异性货物的EVS的释放 和成纤维细胞。我们的假设是，细胞表面Dsg2的呈现调制了EV相关的输出 可作为鳞状细胞癌进展和/或侵袭的生物标志物和关键调控因子的miRNAs 为他们提供了新的治疗靶点。目标1将确定Dsg2调节 电动汽车的生物合成。目的2将研究EV相关miR146a和miR155的细胞效应。目标3将 评价血清EV miRNA水平作为鳞癌预后危险因素的价值。在这里获得的结果将是 阐明Dsg2的新功能，并为Dsg2如何影响细胞生长和迁移提供新的线索： 对于适当的组织形态发生和肿瘤发生至关重要的生物学步骤。在成功完成 在这项工作中，我们将定义控制蜂窝电动汽车出口的机制；电动汽车如何调节 并揭示了可作为诊断和预后生物标志物的新的miRNAs 恶性鳞状细胞癌和其他上皮源性癌症。

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.