Stroma penetrating and immune modulating nanoparticles for image-guided therapy of pancreatic cancer

项目摘要 纳米粒/药物的靶向递送和CAR T细胞疗法的最新进展显示出在 克服胰腺癌化疗和免疫治疗耐药性的新方法的发展。 胰腺癌CAR T治疗的一个主要障碍是存在多个屏障，以防止细胞毒 T细胞到达肿瘤并主动杀死肿瘤细胞。致密的肿瘤间质细胞和细胞外基质 物理和生物屏障使T细胞滞留在基质中并抑制T细胞功能。有一种未得到满足的需求 改善胰腺癌患者CAR T细胞转运的有效方法。肿瘤靶向，间质- 在母公司R01项目中开发的穿透透过性氧化铁纳米颗粒(IONPs)提供了一个机会 开发一种联合疗法，以改善CAR T细胞的靶向传递和在肿瘤内的分布，并 总体治疗反应。我们的结果显示，uPAR靶向和基质断裂的配体(ATFMMP14) 可克服基质细胞和细胞外基质障碍以增强纳米药物对肿瘤的输送 人胰腺PDX和转基因小鼠肿瘤模型中的T细胞。在这项补充研究项目中， 我们假设携带化疗药物的uPAR靶向、穿透基质的IONPs的结合 CAR T细胞免疫治疗剂显著增强CAR T细胞的转运和化疗 将药物转化为胰腺癌。免疫检查点PD-L1和CTLA4结合在IONPs上的封闭肽 进一步激活CAR T细胞功能。这些综合作用导致了对胰腺的强烈治疗反应 癌症和克服治疗阻力。在拟议的研究中，我们将首先确定ATFMMP14的影响- 携带化疗药物SN38的偶联IONPs对抗MUC16和/或细胞毒性的影响 人胰腺癌细胞系体外培养间充质T细胞。联合用药的临床疗效观察 带有CAR T细胞的靶向IONP/SN38将在SCID小鼠的胰腺癌PDX模型中进行评估(AIM 1)。下一步，我们将通过背包的方式开发一种新型的靶向和穿透基质的CAR T细胞传递系统 携带PD-L1和/或CTLA4封闭肽的ATFMMP14-IONP/SN38，由CXCR4抑制剂介导 (IL-8040)，用于克服胰腺癌化疗和CAR T细胞治疗的耐药性(目标2)。其效果 含ATFMMP14的背包系统靶向传递CAR T细胞及治疗效果的研究 IONP/SN38/免疫检查点抑制剂将在胰腺癌PDX模型中进行评估(AIM 2)。我们 然后将研究靶向化疗和免疫疗法IONP-CAR T细胞对 用小鼠抗MUC16 CAR T细胞构建肿瘤细胞、免疫细胞和肿瘤基质免疫微环境 在转基因小鼠胰腺癌细胞系中获得小鼠肿瘤模型(AIM 2)。最后，论证了我国铁路建设的可行性 将确定用于跟踪靶向IONP背包CAR T细胞的肿瘤内递送的MR成像 小鼠肿瘤模型(AIM 2)。这一补充项目将为博士后候选人提供多样化的 多学科研究培训，开发新的癌症免疫疗法，并完成博士论文研究。

Project Summary Recent advances in targeted delivery of nanoparticle/drugs and CAR T cell therapy have shown promises in the development of novel approaches for overcoming resistance to chemo- and immunotherapy in pancreatic cancer. A major obstacle in CAR T therapy in pancreatic cancer is the presence of multiple barriers that prevent cytotoxic T cells reaching tumors and actively killing tumor cells. Dense tumor stromal cells and extracellular matrix create physical and biological barriers to trap T cells in the stroma and inhibit T cell function. There is an unmet need of effective approaches to improve delivery of CAR T cells in pancreatic cancer. Tumor targeted, stroma- penetrating theranostic iron oxide nanoparticles (IONPs) developed in the parent R01 project offer an opportunity to develop a combination therapy to improve targeted delivery and intratumoral distribution of CAR T cells and overall therapeutic responses. Our results showed that uPAR-targeted and stroma breaking ligand (ATFmmp14) can overcome stromal cellular and extracellular matrix barriers to enhance tumor delivery of nanoparticle-drugs and T cells in human pancreatic PDX and transgenic mouse tumor models. In this supplement research project, we hypothesize that the binding of uPAR targeted, stroma-penetrating IONPs carrying chemo- and immunotherapeutic agents to CAR T cells significantly enhances delivery of CAR T cells and chemotherapy drugs into pancreatic cancer. Immune checkpoint PD-L1 and CTLA4 blocking peptides conjugated on the IONPs further activate CAR T cell function. Those combined effects lead to a strong therapeutic response in pancreatic cancer and overcome therapy resistance. In the proposed study, we will first determine the effect of ATFmmp14- conjugated IONPs carrying a chemotherapy agent, SN38, on viability and cytotoxicity of anti-MUC16 and/or Mesothelin CAR T cells in vitro in human pancreatic cancer cell lines. Therapeutic effect of co-delivery of the targeted IONP/SN38 with CAR T cells will be evaluated in a pancreatic cancer PDX model in SCID mice (Aim 1). Next, we will develop a novel targeted and stroma-penetrating CAR T cell delivery system by backpacking with ATFmmp14-IONP/SN38 carrying PD-L1 and/or CTLA4 blocking peptides, mediated by a CXCR4 inhibitor (BL-8040), for overcoming resistance to chemo- and CAR T cell therapy in pancreatic cancer (Aim 2). The effect on targeted delivery of CAR T cells and therapeutic efficacy using the backpack system containing ATFmmp14- IONP/SN38/immune checkpoint inhibitors will be evaluated in the pancreatic cancer PDX model (Aim 2). We will then investigate the therapeutic effects of the targeted chemo- and immunotherapy IONP-CAR T cells on tumor cells, immune cells and tumor stroma immune microenvironment using a mouse anti-MUC16 CAR T cells in a transgenic mouse pancreatic cancer cell line derived mouse tumor model (Aim 2). Finally, the feasibility of MR imaging for tracking intratumoral delivery of targeted IONP backpacked CAR T cells will be determined in mouse tumor models (Aim 2). This supplement project will provide a diversity pre-doctoral candidate with multidisciplinary research training to develop a novel cancer immunotherapy and to complete PhD thesis study.