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

免疫抑制或免疫排斥肿瘤微环境(TME)在限制肿瘤生长的过程中起着关键作用。 多种肿瘤类型对免疫治疗的反应。实现淋巴细胞增加的一个有吸引力的策略 肿瘤的侵袭是细胞因子的使用，它可以直接影响多条免疫途径和重新编程。 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， 由于毒性方面的考虑，许多细胞因子在临床上受到限制。合理的药物输送策略，可以 挽救细胞因子的治疗潜力可以作为我们仔细操纵的能力的重要一步 TME中的抗肿瘤免疫反应，为更有效的免疫治疗打开了大门。 纳米颗粒(NPs)是一种很有前途的解救有毒细胞因子的载体。虽然许多研究都使用了 NPS要提高疗效和毒性，围绕NP的作用仍有很大的认识差距 增强递送的生物物理特性。关于纳米粒子如何 以及这些差异如何影响治疗结果。我们处于独特的地位，可以调查 鉴于我们在NP设计方面的丰富经验，NP的生物物理性质在细胞因子传递中的作用 靶向肿瘤细胞递送和聚电解质逐层(LBL)组装。LBL-NP系统可以是 旨在调节多种药物从核心和周围层的释放，通常是随着时间的推移 依赖的阶段性释放；然而，操纵外层以拥有某些表面化学和 靶向部分可以显著影响颗粒在解剖和细胞水平上的运输。vbl.使用 这一系统将允许系统地研究这些独特的NP属性对有效细胞因子的作用 送货。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、本地化和释放动力学 粒子和有效载荷。 我们的工作将集中在白介素12(IL-12)，一种最有效和毒性最大的促炎细胞因子 我们最近已经证明了通过使用LBL-NPs来提高疗效和降低全身毒性 结合在卵巢癌细胞表面的膜上。我们的研究将在先进的 浆液性卵巢癌(OC)，对现有免疫疗法反应有限，非小细胞 肺癌(NCSLC)，这是高度敏感的，但只适用于特定的患者亚群。IL-12负载的NPs 对于具有一系列表面化学和靶向部分的外层，将检查细胞 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， 纳米粒子系统将在体内检查单独和联合抗病毒药物传递细胞因子的情况。 同种异体原位动物模型中PD1的处理。 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