In Situ Tumor Vaccination with a Nano-oligo Therapeutic to Induce Whole-body Antitumor Immune Response

意义/目标。尽管免疫检查点抑制剂（ICIs）在黑色素瘤患者中显示出令人印象深刻的结果，但大多数患者没有反应，主要是由于缺乏肿瘤浸润性T细胞。为了增强抗肿瘤T细胞库，我们开发了一种新的免疫纳米疗法，称为“增强免疫反应和抑制肿瘤抑制环境- AIRISE-02”，基于我们正在申请专利的纳米颗粒（NP）共同递送免疫激活CpG寡核苷酸和siRNA对抗STAT3。AIRISE-02依赖于“原位肿瘤疫苗接种”，它利用局部治疗的肿瘤来引发针对全身肿瘤的免疫反应。AIRISE-02的独特之处在于在瘤内注射时能够同时进入癌细胞和抗原提呈细胞（如DC、巨噬细胞），导致癌细胞死亡和肿瘤抗原释放，同时对抗原提呈细胞是安全的并能激活它们。通过siRNA敲除STAT3， AIRISE-02还能调节肿瘤的免疫抑制环境。仅对一种黑色素瘤进行免疫治疗，AIRISE-02在治疗和未治疗的远端肿瘤中均可引起肿瘤消退，显示出全身免疫治疗作用。AIRISE-02的效果与两种ICIs （PD-1 + CTLA- 4，给予i.p）的效果相同，并且在给予ICIs的8只小鼠中有5只完全治愈。小鼠至少在17个月的时间里没有肿瘤，而仅接受ICIs的小鼠存活时间不到2个月。AIRISE-02在结肠癌和乳腺癌小鼠模型中也有效，在猴子中也有良好的耐受性。的方法。该直接II期SBIR侧重于AIRISE-02的ind启用研究。在Aim 1中，我们将通过鉴定治疗作用所需的关键免疫细胞来建立AIRISE-02的作用机制，并在另一个临床相关的黑色素瘤模型中证实其疗效。在目标2中，我们将与cmo合作生产符合gmp的AIRISE-02 (siSTAT3， CpG和NP)，灭菌，填充整理和稳定性研究。在Aim 3中，我们将与Charles River毒理学合作，在小鼠和食蟹猴中进行AIRISE-02 glp依从毒理学研究。将建立最大耐受剂量和毒性动力学特征。反复给药后的潜在毒性将通过监测体重、一般健康状况、注射部位反应、血液学、血清化学/细胞因子和关键器官的组织病理学来评估。在Aim 4中，我们将与RTI International和Knight Cancer Institute的实验治疗中心合作，提交IND申请，以便在2023年初进行I期临床试验。新颖性和影响力。纳米颗粒共递送siRNA和佐剂用于原位癌症疫苗接种以前从未做过。AIRISE-02的成功将使许多可注射的肿瘤（如黑色素瘤、乳腺癌、淋巴瘤、头颈部）受益。未来的靶标发现可以利用其他sirna产生新的AIRISE管道。我们的纳米颗粒已被广泛评估和大规模合成，其中AIRISE-02限制了全身毒性，同时促进了AIRISE-02快速转化到临床。

Significance/Goal. Although immune checkpoint inhibitors (ICIs) have shown impressive outcomes in melanoma patients, the majority of patients fail to respond, mainly due to the absence of tumor-infiltrating T cells. In order to boost the anti-tumor T cell repertoire, we have developed a novel immuno-nanotherapeutic, called “Augmenting Immune Response and Inhibiting Suppressive Environment of tumors – AIRISE-02”, based on our patent-pending nanoparticle (NP) co-delivering an immune-activating CpG oligo and siRNA against STAT3. AIRISE-02 relies on “in situ tumor vaccination,” which exploits a locally treated tumor to prime immune response against tumors throughout the body. Unique to AIRISE-02 is the ability to enter both cancer and antigen presenting cells (e.g., DC, macrophage) upon intratumoral (i.t.) injection, resulting in cancer cell death and tumor antigen release, while being safe to and activating the antigen presenting cells. By knocking down STAT3 with siRNA, AIRISE-02 also modulates immunosuppressive environment of the tumor. Given i.t. to only one melanoma tumor, AIRISE-02 caused regression in both treated and untreated distant tumors, demonstrating whole-body immunotherapeutic action. AIRISE-02 was as effective as two ICIs (PD-1 + CTLA- 4, given i.p.) combined, and led to complete cures in 5 out of 8 mice when given with the ICIs. The mice have remained tumor-free for at least 17 months, while mice receiving just ICIs did not survive past 2 months. AIRISE-02 was also effective in colon and breast cancer mouse models and was well tolerated in monkeys. Approach. This direct phase II SBIR focuses on IND-enabling studies of AIRISE-02. In Aim 1, we will establish the mechanism of AIRISE-02 by identifying key immune cells required for therapeutic action and confirm efficacy in another clinically relevant melanoma model. In Aim 2, we will partner with CMOs for manufacturing GMP-compliant AIRISE-02 (siSTAT3, CpG, and NP), sterilization, fill finishes, and stability studies. In Aim 3, we will work with Charles River Toxicology to conduct GLP-compliant toxicology studies of AIRISE-02 in mice and cynomolgus monkeys. The maximum tolerated dose and toxicokinetic profile will be established. Potential toxicity after repeated doses will be assessed by monitoring body weight, general health, injection site reaction, hematology, serum chemistry/cytokines, and histopathology of key organs. In Aim 4, we will partner with RTI International and Knight Cancer Institute’s Center of Experimental Therapeutics to submit an IND application to enable Phase I clinical trial in early 2023. Novelty and Impact. Nanoparticle co-delivery of siRNA and adjuvant for in situ cancer vaccination has never been done before. Successful AIRISE-02 will benefit many injectable tumors (e.g., melanoma, breast, lymphoma, head and neck) with metastasis anywhere. Future target discovery can lead to new AIRISE pipelines utilizing other siRNAs. Our nanoparticle has been extensively evaluated and synthesized at large-scale, and i.t. AIRISE-02 limits systemic toxicity, together facilitating rapid translation of AIRISE-02 to the clinic.