Super Natural Killer Cells That Target Metastases in the Tumor-Draining Lymph Nodes

项目总结/摘要 肿瘤引流淋巴结（LN）是大多数类型癌症的第一个转移部位。转移的程度 在LN中，常用于分期癌症进展。值得注意的是，在最近的工作中，申请人描述了新颖的 纳米级TRAIL包被的脂质体，当与人自然杀伤（NK）细胞缀合时， 在体外和体内杀死癌细胞的内源性治疗潜力。在这项概念验证研究中， 申请人将通过将这些脂质体与NK细胞缀合而将它们靶向LN，并将研究它们的 防止小鼠结肠癌肿瘤淋巴扩散的能力。将显示靶向NK细胞 与TRAIL脂质体一起使用可以增加脂质体在区域淋巴结内的保留时间，以诱导细胞凋亡。 癌细胞如果成功的话，这种方法可以用来杀死肿瘤引流淋巴结中的癌细胞 来防止癌症的淋巴扩散拟议的工作分为三个具体目标。具体 目的1：探讨TRAIL/Anti-NK 1.1脂质体治疗肿瘤的机制，并检测其抗肿瘤药物的疗效。 皮下LN转移模型中的耐药结肠癌。次级目标1.1：审查不同组织的作用 自然杀伤细胞受体对超级NK细胞毒性的影响。子目标1.2：测试TRAIL/抗NK1.1的功效 脂质体治疗奥沙利铂耐药结肠癌。奥沙利铂是临床上重要的铂类药物 然而，奥沙利铂的长期治疗已被证明会导致患者获得耐药性。 大肠癌细胞具体目标2：表征以下物质的生物分布、药代动力学和毒性： 腹腔内引入TRAIL/抗NK1.1脂质体。脂质体注射的腹膜内途径将是 检查以使得能够在目标3的原位结肠癌模型中进行功效研究。次级目标2.1：审查 TRAIL/Anti-NK 1.1脂质体的全身生物分布和LN药代动力学，特别关注 肠系膜淋巴结子目标2.2：评估重复腹腔内给药后的毒性反应 注射TRAIL/抗NK1.1脂质体。具体目的3：评估TRAIL/抗NK1.1脂质体在肿瘤中的功效。 结肠癌转移至肠系膜淋巴结和脾至肝转移的原位模型。次 目的3.1：表征TRAIL/抗NK1.1脂质体治疗原位结肠癌转移至 肠系膜淋巴结子目标3.2：治疗从脾到肝的继发性转移， 静脉内TRAIL/抗NK1.1脂质体。结肠癌细胞将被注射到脾脏，一个淋巴管 器官，以检查静脉内TRAIL/抗NK1.1脂质体治疗是否也可以预防或减少 从脾到肝的继发性转移。影响：这种创新的基于TRAIL脂质体的干预 将证明NK细胞可用于消除肿瘤引流LN中的致瘤细胞， 防止LN转移的形成，这是目前未满足的需求。该项目的成功将建立一个新的 基于细胞的受体-配体治疗剂递送的平台技术， 癌症和其他疾病。

Project Summary/Abstract Tumor-draining lymph nodes (LN) are the first site of metastasis in most types of cancer. The extent of metastasis in the LN is often used in staging cancer progression. Notably, in recent work the applicants described novel nanoscale TRAIL-coated liposomes that when conjugated to human natural killer (NK) cells enhance their endogenous therapeutic potential in killing cancer cells both in vitro and in vivo. In this proof-of-concept study, the applicants will target these liposomes to the LN by conjugating them to NK cells, and will investigate their ability to prevent the lymphatic spread of colon cancer tumors in mice. It will be shown that targeting NK cells with TRAIL liposomes can enhance liposome retention time within regional lymph nodes to induce apoptosis in cancer cells. If successful, the proposed approach could be used to kill cancer cells within the tumor draining LN to prevent the lymphatic spread of cancer. The proposed work is organized into three Specific Aims. Specific Aim 1: To examine the mechanism of TRAIL/Anti-NK1.1 liposome therapy and test their efficacy against drug- resistant colon carcinoma in a subcutaneous LN metastasis model. Sub-aim 1.1: Examine the roles of different natural killer cell receptors on super NK cytotoxicity. Sub-aim 1.2: To test the efficacy of TRAIL/Anti-NK1.1 liposomes to treat oxaliplatin-resistant colon cancer. Oxaliplatin is a clinically important platinum-based drug however long-term treatments with oxaliplatin have been shown to lead to the acquisition of drug resistance in colorectal cancer cells. Specific Aim 2: To characterize the biodistribution, pharmacokinetics and toxicity of TRAIL/Anti-NK1.1 liposomes introduced intraperitoneally. Intraperitoneal route of liposome injection will be examined to enable efficacy studies in the orthotopic colon cancer model of Aim 3. Sub-aim 2.1: To examine the whole body biodistribution and LN pharmacokinetics of TRAIL/Anti-NK1.1 liposomes, with special focus on the mesenteric lymph nodes. Sub-aim 2.2: To assess for toxicity in response to repeated intraperitoneal injections of TRAIL/Anti-NK1.1 liposomes. Specific Aim 3: To evaluate TRAIL/Anti-NK1.1 liposome efficacy in an orthotopic model of colon cancer metastasis to the mesenteric lymph nodes and spleen-to-liver metastasis. Sub- aim 3.1: Characterize the efficacy of TRAIL/Anti-NK1.1 liposomes to treat orthotopic colon cancer metastasis to the mesenteric lymph nodes. Sub-aim 3.2:Treatment of secondary metastasis from the spleen to the liver with intravenous TRAIL/Anti-NK1.1 liposomes. Colon carcinoma cells will be injected into the spleen, a lymphatic organ, to examine whether intravenous TRAIL/Anti-NK1.1 liposome treatment can also prevent or reduce secondary metastasis from the spleen to the liver. IMPACT: This innovative TRAIL-liposome based intervention will demonstrate that NK cells can be used to eliminate tumorigenic cells in the tumor-draining LN, and thus prevent the formation of LN metastases, a currently unmet need. The success of this project will establish a new platform technology for the cellular-based delivery of receptor-ligand therapeutics for the treatment of various cancers and other diseases.