Modeling Primary Tumor Organoids for lmmunotherapy

摘要 癌症是第二大死因，在美国每 4 人死亡中就有近 1 人死于癌症。当前 癌症免疫治疗，尤其是检查点阻断，很大程度上受肿瘤中PD-L1表达的影响； PD-L1 阳性表达有限的患者通常对免疫疗法的反应较差。这些 研究结果表明，需要新技术来治疗 PD-L1 耐药肿瘤。循环 [G(3',5')pA(3',5')p] (cGAMP) 最近出现了一种令人兴奋的新型疫苗佐剂，它可以依次激活先天免疫 反应并协调适应性免疫。肿瘤相关巨噬细胞（TAM），一种主要的免疫细胞类型 肿瘤微环境中的细胞是辅助治疗的有效靶点。颗粒系统是理想的载体 用于靶向肿瘤内的 TAM 群体。此外，联合疗法对肿瘤治疗具有巨大的前景 预防和治疗。最重要的是，癌症微生物组的关键作用正在引起越来越多的兴趣 用于癌症治疗。我们假设靶向 TAM 的颗粒 cGAMP 佐剂可以增强抗肿瘤作用 免疫反应并逆转 PD-L1 反应性和耐药性中的促肿瘤微环境 癌症，并与检查点封锁和肿瘤切除相结合提高抗肿瘤效率。 我们已经并将继续使用 B16F10 黑色素瘤（皮下和肺）来测试该假设 转移模型； PD-L1 反应）和 C3(1)Tag 基底样乳腺癌（原位和自发性 GEM 模型； PD-L1 耐药），并测试以下具体目标： 目标 1. STING 激动剂的颗粒递送 抗癌免疫疗法：（F99期）。我们优化了脂质体 NP 和 Ace-DEX MP 递送系统 用于cGAMP评估细胞摄取和M2->M1偏斜能力，并检测主要组织相容性 (MHC) 基因和共刺激基因表达以及细胞因子的体外产生。我们还研究了组织 颗粒cGAMPs在上述肿瘤模型中的分布及其抗肿瘤功效。目标2。 STING 激动剂颗粒递送发挥抗癌免疫治疗作用的机制 联合疗法：（F99期）。我们将研究巨噬细胞、CD4 和 CD8 T 细胞以及 NK 的作用 颗粒 cGAMP 的抗肿瘤反应中的细胞。我们还将监测长期生存和肿瘤复发 通过颗粒 cGAMP 与抗 PD-L1 抗体的联合治疗以及上述肿瘤切除术 提到了肿瘤模型。目标3.博士后研究方向（K00阶段）。我会追求我的兴趣 了解肠道微生物群在宿主对癌症发展的反应中的机制和功能作用 与此同时，治疗特别关注研究口腔微生物群对头颈反应的作用 癌症治疗和恢复。

Abstract Cancer ranks as the second leading cause of death, contributing to nearly 1 in every 4 death in the US. Current cancer immunotherapies, especially checkpoint blockade, is largely influenced by PD-L1 expression in tumors; and patients with limited PD-L1 positive expression are normally less responsive to the immunotherapy. These findings suggest new technologies are needed to for PD-L1 resistant tumors. Cyclic [G(3',5')pA(3',5')p] (cGAMP) has recently emerged as an exciting new class of vaccine adjuvants, which sequentially activate innate immune responses and orchestrate adaptive immunity. Tumor associated macrophages (TAMs), a major type of immune cells in tumor microenvironment, are a potent target for adjuvant therapy. Particulate systems are ideal vehicles for targeting TAM population within tumor. Additionally, combination therapy holds great promise for tumor prevention and treatment. Most important, the critical role of cancer microbiome is gaining increased interests for cancer therapy. We hypothesize that Particulate cGAMP adjuvants targeting TAMs can enhance anti-tumor immune response and reverse the pro-tumorigenic microenvironment in both PD-L1 responsive and resistant cancers, and improve the anti-tumor efficiency in combination with checkpoint blockade and tumor resection. We have tested and will continue to test the hypothesis using B16F10 melanoma (subcutaneous and lung metastasis models; PD-L1 responsive) and C3(1)Tag basal-like breast cancer (orthotopic and spontaneous GEM models; PD-L1 resistant), and test the following specific aims: Aim 1. Particulate delivery of STING agonist as anti-cancer immuotherapeutics: (F99 phase). We optimized liposomal NP and Ace-DEX MP delivery systems for cGAMP to evaluate the cellular uptake and M2->M1 skewing capacity, and detect major histocompatibility (MHC) gene and costimulatory gene expression and cytokine production in vitro. We also investigated the tissue distribution of particulate cGAMPs and their anti-tumor efficacy in above-mentioned tumor models. Aim 2. Mechanism by which particulate delivery of STING agonist acts an as anti-cancer immuotherapeutic and combination therapies: (F99 phase). We will study the role of macrophage, CD4+ and CD8+ T cells, and NK cells in anti-tumor response of particulate cGAMPs. We will also monitor long term survival and tumor recurrence by combination therapies of particulate cGAMPs with anti-PD-L1 antibodies and tumor resection in above- mentioned tumor models. Aim 3. The Postdoctoral Research Direction (K00 phase). I will pursue my interest in understanding the mechanism and functional role of gut microbiota in host response to cancer development and therapy, meanwhile, with a particular interest in studying the role of oral microbiota in response to head and neck cancer therapeutics and restoration.