Delivery of cytokines for cancer immunotherapy using nanolayer-controlled trafficking of liposomal nanoparticles

免疫抑制性或免疫排斥性肿瘤微环境（TME）在限制肿瘤微环境中起着关键作用。 许多肿瘤类型对免疫疗法的反应。一个有吸引力的策略，以实现增加淋巴细胞 肿瘤中的浸润是细胞因子的使用，其可以直接影响多种免疫途径和重编程 TME能够对癌细胞产生强大的免疫反应。不幸的是，尽管有这种明显的潜力， 由于毒性问题，许多细胞因子在临床上受到限制。合理的药物输送策略， 拯救细胞因子的治疗潜力可以作为我们能够仔细操纵的重要一步， TME中的抗肿瘤免疫反应，并为更有效的免疫疗法打开大门。 纳米颗粒（NPs）是用于拯救毒性细胞因子的有前途的载体。虽然许多研究使用 NP改善疗效和毒性，但围绕NP的作用仍存在很大的知识差距 生物物理特性对增强递送的影响。关于纳米颗粒如何作用，还有很多尚不清楚的地方 交通以及这些差异如何影响治疗结果。我们有独特的优势来调查 考虑到我们在NP设计和细胞因子递送方面的丰富经验， 靶向肿瘤细胞递送和在体外逐层（LbL）组装。LbL-NP系统可以是 其设计用于调节多种药物从核心和周围层的释放，通常随着时间的推移 依赖性阶段释放;然而，操纵外层以具有某些表面化学性质， 靶向部分可以在解剖学和细胞水平上显著影响颗粒的运输。使用 该系统将允许系统地研究这些独特的NP性质对有效细胞因子的作用 交付.这项工作的目标是了解和控制细胞因子对实体瘤的传递 使用LbL-NPs作为工具，重点是贩运的影响， 粒子和有效载荷 我们的工作将集中在白细胞介素-12（IL-12），一种最有效和毒性的促炎细胞因子， 我们最近通过使用LbL-NP证明了其改善的功效和降低的全身毒性， 与卵巢癌细胞的表面膜结合。我们的研究将在先进的背景下进行 浆液性卵巢癌（OC）对现有免疫疗法的反应有限，而非小细胞卵巢癌（OC） 肺癌（NCSLC），这是高度响应，但仅适用于一个定义的子集的患者。负载IL-12的NP 具有一系列表面化学性质和靶向部分的外层将被检查用于细胞 以及亚细胞摄取和免疫细胞刺激。将检查和优化细胞因子释放动力学， 和纳米颗粒系统将在体内检查单独的细胞因子和与抗肿瘤药物组合的细胞因子递送。 原位同基因动物模型中的PD 1治疗。 1

An immunosuppressive or immune excluded tumor microenvironment (TME) plays a key role in limiting the response of many tumor types to immunotherapy. One attractive strategy to accomplish increased lymphocyte infiltration in tumors is the use of cytokines, which can directly impact multiple immune pathways and reprogram the TME to enable a robust immune response against cancer cells. Unfortunately, despite this obvious potential, many cytokines have been limited clinically due to toxicity concerns. Rational drug delivery strategies that can rescue the therapeutic potential of cytokines could act as an important step in our ability to carefully manipulate the anti-tumor immune response in the TME and open the door for more effective immunotherapies. Nanoparticles (NPs) are a promising vehicle for the rescue of toxic cytokines. While many studies have used NPs to improve efficacy and toxicity, there remains a substantial knowledge gap surrounding the role of NP biophysical properties on enhanced delivery. There is much that is not yet understood about how nanoparticles traffic and how these differences can affect therapeutic outcomes. We are uniquely positioned to investigate the role of NP biophysical properties on cytokine delivery given our extensive experience in both NP design for targeted tumor cell delivery and in polyelectrolyte layer-by-layer (LbL) assembly. LbL-NP systems can be designed to modulate the release of multiple drugs from the core and from surrounding layers, often with time dependent staged release; whereas, manipulating the outer layer to possess certain surface chemistries and targeting moieties can significantly impact trafficking of particles on both the anatomical and cellular level. Using this system will allow for a systematic investigation of the role of these unique NP properties on effective cytokine delivery. The goal of this work is to understand and control the delivery of cytokines against solid tumors using LbL-NPs as a tool, with a focus on the impact of trafficking, localization and release kinetics of the particle and payload. Our work will focus on interleukin-12 (IL-12), one of the most potent and toxic proinflammatory cytokines for which we have recently demonstrated improved efficacy and lowered systemic toxicity by using LbL-NPs that bind to the surface membrane of ovarian cancer cells. Our studies will take place within the context of advanced serous ovarian cancer (OC), which has shown limited response to existing immunotherapies, and non-small cell lung cancers (NCSLC), which is highly responsive, but only for a defined subset of patients. IL-12 loaded NPs with external layers possessing a range of surface chemistries and targeting moieties will be examined for cellular and subcellular uptake and immune cell stimulation. Cytokine release kinetics will be examined and optimized, and nanoparticle systems will be examined in vivo for delivery of cytokines alone and in combination with anti- PD1 treatments in orthotopic syngeneic animal models. 1