A Stem Cell Based Exosomal Vaccine for the Prevention of Cancer

项目摘要/摘要 肺癌是一种流行的疾病，每年夺去许多人的生命。疾病复发、侵袭和 转移是导致死亡的主要原因。最近的发现提供了令人信服的证据，至少有一些 不同类型的癌症是由一小群称为致癌细胞的恶性细胞引发和维持的 干细胞(CIC)。复发、侵袭和转移的原因是CICs有不同的 在生物学上比所有其他肿瘤细胞都要强，而且重要的是，它对化疗和辐射都有抵抗力。肺 CICS已被证明约占所有肿瘤细胞的1%-15%，并可以在注射低剂量的情况下形成肿瘤 作为100个细胞。已发表的研究证据表明，人类和啮齿动物的癌症 包含表达胚胎干细胞(ES)抗原的细胞群。含有这些蛋白质的细胞 也表达了用于识别肺CICs的标志物；因此，我们假设ES细胞和CICs共享 几个常见的分子特征。为了验证这一假设，我们给小鼠接种了从ES中提取的外切体。 细胞表达GM-CSF，并研究是否诱导了抗肿瘤免疫反应。我们 发现基于ES细胞衍生的外切体的疫苗策略(ES-exo疫苗)在 防止植入性和致癌物诱导的肺腺癌的发展 可检测到的毒性或自身免疫的迹象。我们实验室最近公布的结果显示， ES细胞免疫的小鼠脾细胞对肺CICs具有较强的细胞毒作用。建议进行的实验 这项申请试图扩展这些新的发现，以令人信服地证明ES-exo疫苗 针对肺癌相关CICs免疫以及抗CIC免疫负责预防肺 腺癌的发展。ES-exo疫苗的抗肿瘤活性及其免疫刺激作用 将在体外和体内研究肺癌小鼠模型的特性。本报告中提出的实验 研究将解决我们的新型ES-exo疫苗作为无细胞预防性疫苗的翻译潜力 可移植、转基因和异种移植的小鼠肺癌模型的形态。这次会议的主要目标之一是 应用于识别对ES-exo疫苗抗肺癌效果重要的肿瘤抗原。 基于蛋白质组学的筛选方法。为实现既定目标，提出了以下目标：1) 调查肺癌起始细胞是否为ES细胞衍生外切体(ES-exo)疫苗的靶点- 诱导抗肿瘤免疫；2)评估ES细胞来源的外切体(ES-exo)的翻译潜能 作为一种新型的肺癌无细胞疫苗。我们建议的研究将为以下方面提供重要的见解 开发一种安全的预防肺癌发病和/或复发的疫苗。

PROJECT SUMMARY/ABSTRACT Lung cancer is a prevalent disease and consume many lives every year. Disease relapse, invasion and metastases are the main causes of death. Recent discoveries provide compelling evidence that at least some types of cancer are initiated and maintained by a small population of malignant cells called cancer-initiating stem cells (CICs). Relapse, invasion, and metastases are explained by the fact that CICs have a different biology than all the other tumor cells and, importantly, are resistant to chemotherapies and radiation. Lung CICs have been shown to represent about 1–15% of all tumor cells and can form tumors with injections as low as 100 cells. Evidence from published studies have demonstrated that human, as well as rodent, cancers contain populations of cells that express embryonic stem (ES) cell antigens. Cells containing these proteins also express markers used to identify lung CICs; therefore, we hypothesized that ES cells and CICs share several common molecular traits. To test this hypothesis, we vaccinated mice with exosomes derived from ES cells expressing GM-CSF and investigated whether an anti-tumor immune response was elicited. We discovered that ES cell-derived exosome-based vaccination strategy (ES-exo vaccine) is very effective in preventing both implantable and carcinogen-induced lung adenocarcinoma development without any detectable toxicity or signs of autoimmunity. Recently published results from our laboratory reveal that splenocytes from ES cell-immunized mice are preferentially cytotoxic to lung CICs. Experiments proposed in this application seek to expand these novel findings to convincingly demonstrate that ES-exo vaccine immunize against lung cancer-associated CICs and that anti-CIC immunity is responsible for preventing lung adenocarcinoma development. The anti-tumor activities of ES-exo vaccine as well as their immunostimulatory properties will be investigated in in vitro and in vivo lung cancer mouse models. Experiments proposed in this study will address the translational potential of our novel ES-exo vaccine as a cell-free prophylactic vaccine modality in implantable, transgenic and xenograft mouse models of lung cancer. One of the major goals of this application is to identify tumor antigens important for anti-lung cancer efficacy of ES-exo vaccine using a proteomics-based screening methodology. To fulfill the stated objectives, the following aims are proposed: 1) Investigate whether lung cancer-initiating cells are targets of ES cell-derived exosomes (ES-exo) vaccination- induced anti-tumor immunity, and 2) Evaluate the translational potential of ES cell-derived exosomes (ES-exo) as a novel cell-free vaccine for lung cancer. Our proposed study will provide important insights towards developing a safe prophylactic vaccine for lung cancer onset and/or recurrence.