Liposome-based mRNA cancer immunotherapy targeting ion channels

项目总结 免疫检查点抑制物，如抗程序性细胞死亡1(PD1)的单抗 彻底改变了治疗实体恶性肿瘤的方式。然而，尽管一些患者反应剧烈， 大多数人要么没有反应，要么产生了抵抗力。我们的研究重点是头部和颈部。 鳞状细胞癌(HNSCC)是一种高死亡率和高耐药性的侵袭性癌症 检查点抑制药。我们的目标是开发新的治疗方案，改善免疫监控并减少 HNSCC患者对PD1治疗的耐药性。 免疫疗法在癌症中的疗效取决于该疗法增加肿瘤迁移的能力- 特异性T细胞进入肿瘤，并在免疫抑制肿瘤的情况下维持其肿瘤杀伤能力 微环境(TME)。提高T细胞对肿瘤的侵袭能力和在敌对状态下的功能 TME仍然是免疫治疗的最大挑战。T淋巴细胞的这些功能依赖于Kv1.3和 KCa3.1钾通道，控制膜电位，促进钙离子内流 细胞因子的产生、细胞毒性和趋化性。我们发现，这些渠道有助于 HNSCC免疫监测失败及免疫治疗抵抗。KCa3.1通道被抑制 通过腺苷，一种存在于TME中的免疫抑制分子，这种机制限制了T细胞的渗透 进入肿瘤。此外，Kv1.3通道在肿瘤浸润性T细胞中减少，从而导致低效 细胞毒性。最后，我们观察到HNSCC患者对PD1阻断有反应的细胞毒性T细胞 表现为Kv1.3的特征性增加和对腺苷的反应丧失。因此，一种能够增强 Kv1.3和选择性地赋予T细胞对腺苷的耐药性将对HNSCC大有裨益。 我们将开发脂质纳米粒(LNPs)，用于定向向T细胞递送编码信使RNA(MRNA) Kv1.3和一种通过脂质体纳米粒对腺苷产生抵抗力的多肽。腺苷- 阻断肽可阻断腺苷受体下游的信号通路。在这里，将测试 一种诱导Kv1.3过表达的细胞靶向脂质体mRNA制剂的假说 T淋巴细胞通道和腺苷敏感性丧失可降低对抗PD1的抵抗力 HNSCC患者的治疗。在目标1中，我们将生产携带编码Kv1.3的mRNA和多肽的LNPs 这将阻断腺苷的作用，并用抗体装饰，以定向传递给T淋巴细胞。在……里面 目的2，我们将确定这些LNPs对HNSCC患者衍生的有机化合物和肿瘤的影响 人性化的老鼠。这些研究将确定一种新配方的可行性，这种配方可以改善T细胞 TME的细胞毒性和耐药性及其对单药或联合癌症的有效性 免疫疗法。

PROJECT SUMMARY Immune checkpoint inhibitors like monoclonal blocking antibodies against programmed cell death 1 (PD1) have revolutionized the way solid malignancies are treated. However, despite a dramatic response in some patients, the majority either have no response or develop resistance. The focus of our research is on head and neck squamous cell carcinoma (HNSCC), an aggressive cancer with high mortality rates and high resistance to checkpoint inhibitors. Our goal is to develop new treatment options that improve immunosurveillance and reduce the resistance to PD1 therapy in HNSCC patients. The efficacy of immunotherapy in cancer relies on the therapy's capability to increase their migration of tumor- specific T cells into the tumor, and sustain their cancer killing capacity despite the immunosuppressive tumor microenvironment (TME). Improving the ability of the T cells to infiltrate the tumor and to function in the hostile TME remains the greatest challenge of immunotherapy. These functions of T lymphocytes rely on Kv1.3 and KCa3.1 potassium channels that control the membrane potential and facilitate the Ca2+ influx necessary for cytokine production, cytotoxicity and chemotaxis. We have discovered that these channels contribute to the failure of immune surveillance and the resistance to immunotherapy in HNSCC. KCa3.1 channels are inhibited by adenosine, an immunosuppressive molecule present in the TME, and this mechanism limits T cell infiltration into the tumors. In addition, Kv1.3 channels are reduced in tumor infiltrating T cells and contribute to inefficient cytotoxicity. Lastly, we observed that cytotoxic T cells of HNSCC patients that respond to PD1 blockade display a characteristic increase in Kv1.3 and loss of response to adenosine. Therefore, a therapy that enhances Kv1.3 and confer resistance to adenosine selectively in T cells would be highly beneficial in HNSCC. We will develop lipid nanoparticles (LNPs) for targeted delivery to T cells of a messenger RNA (mRNA) encoding Kv1.3 and a peptide that confers resistance to adenosine through liposomal nanoparticles. The adenosine- blocking peptide blocks the signaling pathway downstream to the adenosine receptor. Herein, will test the hypothesis that a cell-targeted liposomal mRNA formulation that induces overexpression of Kv1.3 channels and loss of adenosine-sensitivity in T lymphocytes can reduce the resistance to anti-PD1 therapy in HNSCC patients. In Aim 1, we will produce LNPs that carry an mRNA encoding Kv1.3 and a peptide that blocks the effect of adenosine and are decorated with antibodies for targeted delivery to T lymphocytes. In Aim 2, we will determine the impact of these LNPs on tumors utilizing HNSCC patient-derived organoids and humanized mice. These studies will establish the feasibility of a new formulation that can improve the T cell cytotoxicity and confer resistance to the TME, and its validity for single-agent or combinatorial cancer immunotherapy.