Innovative Research for Cancer Nanotechnology (IRCN) for Enhancing Melanoma-specific Immune Responses by the Rational Design of Spherical Nucleic Acids

项目摘要/摘要 这项研究将利用球形核酸(SNA)纳米结构来开发有效的、结构信息丰富的 晚期黑色素瘤疫苗。传统的治疗方法(即化疗或放射治疗)不太成功 黑色素瘤是因为难以辨别黑色素瘤细胞的表型。免疫疗法必须确保 这种黑色素瘤具有很高的突变负担，不能轻易逃脱免疫系统。SNA可以发挥作用 通过精确控制多发性黑色素瘤相关的表现，作为强大的癌症疫苗 以免疫细胞为靶标，从而降低其逃避免疫的可能性。SNA是由纳米颗粒组成的 具有致密的放射状核酸外壳的核心。当使用免疫刺激“佐剂”合成时 寡核苷酸，SNAs诱导免疫反应。事实上，这种仅有佐剂的结构展示了 与线性佐剂相比，从3D结构产生的增强的响应形成了 正在进行的1b/2期临床试验。我们利用了化学合成的简便性和模块化架构 SNAs，并合成它们以包括佐剂和单一的肿瘤相关多肽(“抗原”)。这些 在模型系统中，结构增强了抗肿瘤反应并提供了长期保护性免疫，并且在 特别是，疫苗结构和疗效之间有很强的关系。在我们提议的工作中，我们的目标是 通过精确整合和呈现多种基因来开发抗黑色素瘤的SNA疫苗 免疫刺激提示免疫系统。将用多种临床相关的方法合成RNA 黑色素瘤抗原(MHC-I和-II限制性的、肿瘤相关的新抗原)，在结构上如何变化 给出了佐剂和抗原。对结构的控制，结合体外和体内评估 免疫刺激，将阐明结构-活性关系，这将为癌症疫苗的未来提供信息 设计。使用结构来控制多个免疫系统提示的呈现具有提升 对黑色素瘤的免疫反应和改善临床结果。在目标1中，我们将合成包含 多种抗原和不同的稳定性，以增强抗原特异性T细胞反应。我们将分析他们的吸收情况 通过免疫细胞、SNA组分的亚细胞运输和多条途径的激活动力学 (例如抗原提呈、共刺激标记物表达)。在目标2中，我们将比较不同的管理方式 途径和分析体内的生物分布和摄取动力学以及抗原特异性免疫反应 由SNAs在具有免疫能力的小鼠体内产生。我们将评估在交付包含以下内容的SNA后提出的反应 人源化小鼠和病人标本的人抗原。在目标3中，我们将评估SNA在 体内单独和与免疫检查点封锁相结合，并确定SNA作为进一步的候选 临床前研究和临床翻译。值得注意的是，这种方法将生成基于结构的 了解SNA作为疫苗的性能，并通过产生广泛的T细胞来改进免疫治疗 效果出众的反应。

PROJECT SUMMARY/ABSTRACT This research will utilize spherical nucleic acid (SNA) nanostructures to develop effective, structure-informed vaccines for advanced melanoma. Traditional treatments (i.e. chemotherapy or radiation) are less successful for melanoma because of difficulties in discerning melanoma cells phenotypically. Immunotherapeutics must ensure that melanoma—with a high mutational burden—cannot easily evade the immune system. SNAs can function as robust cancer vaccines through the precise control over the presentation of multiple melanoma-associated targets to immune cells which lowers its potential for immune evasion. SNAs are composed of a nanoparticle core with a dense radial shell of nucleic acids. When synthesized using immunostimulatory “adjuvant” oligonucleotides, SNAs induce immune responses. Indeed, this adjuvant only structure demonstrates the enhanced responses generated from a 3D structure compared to linear adjuvant and forms the basis of an ongoing Phase 1b/2 clinical trial. We have exploited the ease of chemical synthesis and modular architecture of SNAs, and synthesized them to include both adjuvant and a single tumor-associated peptide (“antigen”). These structures enhance antitumor responses and provide long-term protective immunity in model systems, and in particular, there is a strong relationship between vaccine structure and efficacy. In our proposed work, we aim to develop SNA vaccines against melanoma by precisely incorporating and presenting multiple immunostimulatory cues to the immune system. SNAs will be synthesized with multiple clinically-relevant melanoma antigens (MHC-I and -II restricted, tumor-associated, neoantigens), with structural variations in how the adjuvant and antigen are presented. Control over structure, combined with in vitro and in vivo evaluations of immunostimulation, will elucidate structure-activity relationships that will inform the future of cancer vaccine design. Using structure to control the presentation of multiple immune system cues has the power to elevate immune responses to melanoma and improve clinical outcomes. In Aim 1, we will synthesize SNAs containing multiple antigens and varied stabilities to enhance antigen-specific T cell responses. We will analyze their uptake by immune cells, subcellular trafficking of the SNA components, and kinetics of activation of multiple pathways (e.g. antigen presentation, co-stimulatory marker expression). In Aim 2, we will compare different administration routes and analyze in vivo biodistribution and uptake kinetics, as well as the antigen-specific immune responses raised by SNAs in immunocompetent mice. We will assess raised responses after delivery of SNAs containing human antigens to humanized mice and patient specimens. In Aim 3, we will evaluate SNA antitumor efficacy in vivo alone and in combination with immune checkpoint blockade, and identify SNAs as candidates for further preclinical studies and clinical translation. Significantly, this approach will generate a structure-based understanding of SNA performance as vaccines, and improve immunotherapy by generating a breadth of T cell responses with superior efficacies.