RIG-I Activating Nanoparticles for Immunopotentiation

项目总结 可以对先天免疫系统进行药理学编程，以达到预期的免疫效果。 视黄酸诱导基因i(rig-i)是一种模式识别受体，是一种很有前途的天然受体。 免疫增强的免疫靶点。RIG-I在识别5‘-三磷酸化，双- 胞浆中的链状RNA(3pRNA)，刺激抗病毒样炎症程序，可 用来治疗或预防各种疾病的。 然而，3pRNA的效力和功效一直是 受限于主要的药物传递障碍，包括核酸酶降解、细胞摄取效率低下和胞浆 快递，快速通关。为了应对这些挑战，我们开发了RIG-I激活纳米颗粒 (RANS)。RAN是一种聚合物纳米颗粒，旨在促进合成的， 我们团队最近开发的分子定义的高亲和力茎环RNA(SLR)RIG-I激动剂。 此R01应用程序的目标是优化和推进RAN，使其成为一种通用平台 RIG-I的药理活性。我们将通过以下具体目标实现这一目标。首先，我们将 设计具有改进性能的下一代RAN，通过优化 SLR和聚合物的电荷和疏水性。这种方法将利用组合化学多样性来 访问3pRNA交付的新设计空间，我们预计这将产生具有更高性能的下一代RAN SLR负载效率，降低细胞毒性，防止核酸酶降解，提高稳定性，以及 增强免疫刺激活性。其次，我们将在RAN属性之间建立关系， 免疫激活，药代动力学，聚合物和SLR生物分布，以及毒性。这些研究是 在新的免疫治疗模式的临床前开发中至关重要，并将产生新的见解 研究如何设计纳米载体以安全有效地激活RIG-I。我们预计这些研究将 产生下一代RAN，这些RAN针对SLR疗法的系统管理进行了优化。第三，虽然 RANS具有广泛的临床应用潜力，我们将评估其作为一种全身用药的疗效。 免疫原性差的黑色素瘤小鼠模型的癌症免疫治疗作为临床重要的测试案例。 我们期望证明系统地给药先导候选核糖核酸将激活肿瘤中的RIG-I。 微环境，导致肿瘤部位的免疫重新编程，抑制肿瘤生长和 与免疫检查点抑制剂协同作用。总的来说，这些研究将把RAN定位为一种使能 免疫增强平台，有可能满足对新的癌症免疫疗法的重大需求， 抗病毒药物和疫苗佐剂。

PROJECT SUMMARY The innate immune system can be pharmacologically programed to elicit desired immunological outcomes. Retinoic acid-inducible gene I (RIG-I) is a pattern recognition receptor that has emerged as a promising innate immune target for immunopotentiation. RIG-I is activated upon recognizing 5’-triphosphorylated, double- stranded RNA (3pRNA) in the cytosol, which stimulates an antiviral-like inflammatory program that can be harnessed to treat or prevent a diversity of diseases. However, the potency and efficacy of 3pRNA has been limited by major drug delivery barriers, including nuclease degradation, inefficient cellular uptake and cytosolic delivery, and rapid clearance. To address these challenges, we have developed RIG-I activating nanoparticles (RANs). RANs are polymer nanoparticles that are engineered to promote the cytosolic delivery of synthetic, molecularly-defined, and high-affinity stem-loop RNA (SLR) RIG-I agonists recently developed by our team. The objective of this R01 application is to optimize and advance RANs as a versatile platform for pharmacological activation of RIG-I. We will accomplish this through the following Specific Aims. First, we will engineer next-generation RANs with improved properties for systemic administration through optimization of SLR and polymer charge and hydrophobicity. This approach will leverage combinatorial chemical diversity to access a new design space for 3pRNA delivery, which we expect will yield next-generation RANs with higher SLR loading efficiency, reduced cytotoxicity, protection from nuclease degradation, improved stability, and enhanced immunostimulatory activity. Second, we will establish relationships between RAN properties, innate immune activation, pharmacokinetics, polymer and SLR biodistribution, and toxicity. These studies are essential in the preclinical development of new immunotherapeutic modalities and will also yield new insight into how nanocarriers can be engineered for safe and effective activation of RIG-I. We expect these studies to yield next-generation RANs that are optimized for systemic administration of SLR therapeutics. Third, while RANs have broad potential clinical applications, we will evaluate their efficacy as a systemically administered cancer immunotherapy in poorly immunogenic mouse models of melanoma as a clinically important test case. We expect to demonstrate that systemic administration of lead-candidate RANs will activate RIG-I in the tumor microenvironment, resulting in an immunological reprograming of tumor sites that inhibits tumor growth and synergizes with immune checkpoint inhibitors. Collectively, these studies will position RANs as an enabling platform for immunopotentiation with potential to address the significant need for new cancer immunotherapies, antiviral agents, and vaccine adjuvants.