Overcoming metastatic spread of osteosarcoma with RNA loaded nanoparticles

项目摘要 免疫疗法已显示出对成人患者的深远好处，但尚未完全用于儿童固体 肿瘤，如骨肉瘤(OSA)，相当大比例的儿童/青少年死于 肺转移的存在。像许多肿瘤免疫原性差的肿瘤一样，阻塞性睡眠呼吸暂停综合征的定义是缺乏肿瘤。 特定的靶点和调节的肿瘤微环境(TME)。针对劣质产品推出免疫疗法 免疫原性癌症需要新的技术来激活TME，同时同时参与两者 免疫系统的先天和适应性手臂，以产生持续的细胞免疫。 我们已经开发出一种新型的(FDA批准的)RNA纳米颗粒(RNA-NP)疫苗，它可以同时 穿透TME/对TME重新编程，同时诱导OSA特异性T细胞反应。这种疫苗使用了一种新的 将肿瘤来源的信使核糖核酸层叠到脂质纳米颗粒(NP)“洋葱状”包装中的工程设计。我们有 表明全身注射rna-nps可以安全地模拟病毒血症，激活静止的免疫系统。 在几个免疫原性差的小鼠肿瘤中仅用几个小时就能诱导出强大的抗肿瘤效果 对免疫检查点抑制剂具有抵抗力。这些RNA-NPs激活树突状细胞(DC)，取代调节性 瘤内髓系细胞诱导抗原召回反应对小鼠的长期存活期益处 肺转移性阻塞性睡眠呼吸暂停模型。我们已经确定了RNA-NPs在急性/慢性小鼠毒性中的安全性 研究，并启动了一项大型动物犬OSA试验，证明了RNA-NP给药是 可行、安全且具有免疫活性。虽然RNA-NPs介导了大量的抗肿瘤活性，但一些动物 遭受肿瘤生长，这需要探索我们的非幸存者的耐药机制。我们已经展示了 RNA-NPs可以富含肿瘤特异性抗原或配置有针对相关靶点的siRNAs 调节轴(即PD-L1)，这可以在我们的小鼠/犬OSA模型中进行研究。科学的前提是 这项工作是骨肉瘤被一种调节的髓系微环境所包围，这种微环境积极地颠覆 适应性免疫。我们假设转移性阻塞性睡眠呼吸暂停综合征的髓系重新编程将导致安全根除。 疾病的威胁。我们的具体目标是： 1.建立可被适应性RNA-NPs克服的OSA治疗抗性机制。 2.在比较肿瘤学犬OSA模型中确定疫苗反应和逃逸的相关因素。 3.开展多机构I/II阶段研究，评估最有希望的 复发性阻塞性睡眠呼吸暂停患者的RNA-NP制剂。 这项研究的成功完成将导致一种新的阻塞性睡眠呼吸暂停疗法，并从机制上理解其 作为生物反应的治疗效果将在人体临床试验中相互关联。 。

Project Summary Immunotherapy has shown profound benefit for adult patients but has yet to be fully unlocked for pediatric solid tumors such as osteosarcoma (OSA) where a significant percentage of children/adolescents succumb due to the presence of lung metastasis. OSA, like many poorly tumor immunogenic tumors, is defined by a lack of tumor specific targets and a regulatory tumor microenvironment (TME). Unleashing immunotherapy against poorly immunogenic cancers requires new technologies that activate the TME, while concomitantly engaging both innate and adaptive arms of the immune system to generate sustained cellular immunity. We have developed a novel (FDA approved) RNA-nanoparticle (RNA-NP) vaccine that simultaneously penetrates/reprograms the TME while inducing an OSA specific T cell response. This vaccine utilizes a novel engineering design that layers tumor derived mRNA into a lipid-nanoparticle (NP) “onion-like” package. We have shown that systemic administration of RNA-NPs safely mimics viremia, activating the quiescent immune system in only a few hours for induction of potent anti-tumor efficacy in several poorly immunogenic murine tumors resistant to immune checkpoint inhibitors. These RNA-NPs activate dendritic cells (DCs) that supplant regulatory intratumoral myeloid populations inducing antigen-recall response with long-term survivor benefits in murine metastatic pulmonary OSA models. We have established safety of RNA-NPs in acute/chronic murine toxicity studies, and launched a large animal canine OSA trial which demonstrated that RNA-NP administration is feasible, safe and immunologically active. While RNA-NPs mediate substantial anti-tumor activity, some animals suffer tumor outgrowth that warrant exploration of resistance mechanisms in our non-survivors. We have shown that RNA-NPs can be enriched for tumor specific antigens or configured with siRNAs to target pertinent regulatory axes (i.e. PD-L1), which can be studied in our murine/canine OSA models. The scientific premise for this work is that osteosarcoma is encased by a regulatory myeloid microenvironment that actively subverts adaptive immunity. We hypothesize that myeloid reprogramming of metastatic OSA will lead to safe eradication of disease. Our SPECIFIC AIMS are: 1. Establish mechanisms of OSA treatment resistance that can be overcome with adaptable RNA-NPs. 2. Identify correlates for vaccine response and escape in a comparative oncology canine OSA model. 3. Conduct a multi-institutional phase I/II study evaluating the safety and activity of the most promising RNA-NP formulation in recurrent OSA patients. Successful completion of this study will lead to a novel OSA therapy and a mechanistic understanding of its therapeutic effects that will be co-opted as biologic response correlates in a human clinical trial. .