Tumor Stroma Breaking System for Efficient Delivery of Therapeutic Agents

过继T细胞转移肿瘤免疫疗法在血液病治疗中的应用前景看好 恶性肿瘤。尽管付出了巨大的努力，但使用肿瘤浸润性淋巴细胞或 人类实体肿瘤中的工程化CAR-T细胞没有产生预期的治疗反应。实心 对于肿瘤，有多种屏障阻止细胞毒性T细胞到达肿瘤细胞。第一，异常血液 血管限制T细胞进入实体肿瘤，尤其是肿瘤中心。致密的肿瘤基质细胞的存在 和细胞外基质形成物理屏障，将T细胞困在免疫抑制基质中 微环境。因此，开发新的方法以提高交付效率是一个尚未得到满足的需求 实体瘤中的细胞毒性T细胞。拟议的第一阶段研究的翻译目标是开发 并在人体固体中验证用于免疫治疗的打破基质的治疗性T细胞递送系统 肿瘤，重点是胰腺癌和结肠癌。我们已经开发出一种靶向受体和基质 以新颖的设计打破药物输送平台以克服肿瘤间质细胞和细胞外基质 障碍。我们已经证明了它有能力运送纳米药物穿过肿瘤血管，通过 肿瘤间质，穿透基底膜到达肿瘤细胞。这种重组的靶向配体 由尿激酶型纤溶酶原激活剂受体(UPAR)结合域的uPA与催化 基质金属蛋白酶-14的结构域(ATFMMP14)。我们的初步结果显示，T细胞的联合传递 使用ATFMMP14配体导致肿瘤中T细胞的数量显著增加 静脉给药在小鼠结肠癌和胰腺癌模型以及人结肠癌患者中的应用 衍生异种移植(PDX)模型。在这个第一阶段的项目中，我们的目标是确定ATFMMP14-T细胞的作用 以MHC非限制性细胞毒性ɣδT细胞为靶细胞的肿瘤内传递和分布传递系统 人结肠癌和胰腺癌pdx模型中的T细胞模型系统。第一，我们要发展最好的 健康供者ɣδT细胞构建ATFMMP14结合T细胞传递平台的研究 人胰腺和结肠PDX模型(Aim1)。接下来，我们将确定ATFmmp14配体是否可以 改善ɣδT细胞对胰腺和结肠肿瘤的选择性输送，并通过 间质和基底膜阻隔到达肿瘤细胞(目标2)。大鼠的生物分布和全身毒性 还将对正常器官和组织进行评估(目标2)。最后，靶向给药的治疗效果 将在胰腺和结肠中检测不使用或联合使用化疗药物的ɣδT细胞 PDX模型(目标3)。上述拟议研究的结果应能为我们奠定坚实的基础 在第二阶段研究项目中进行进一步的翻译研究，以将这种方法带入临床试验。 这项研究的结果也将导致这一独特方法的未来开发和商业化 适应性T细胞转移疗法，如CAR-T、肿瘤特异性T细胞和肿瘤浸润性T细胞。

Cancer immunotherapy using adoptive T cell transfer has shown its promises in the treatment of hematological malignancies. Despite tremendous efforts, results of clinical trials using tumor infiltrating lymphocytes or engineered CAR-T cells in human solid tumors have not produced desired therapeutic responses. In solid tumors, there are multiple barriers that prevent cytotoxic T cells reaching tumor cells. First, abnormal blood vessels limit T cell delivery into solid tumors, especially tumor center. The presence of dense tumor stromal cells and extracellular matrix create a physical barrier that traps T cells in an immunosuppressive stroma microenvironment. Therefore, there is an unmet need to develop novel approaches to improve delivery efficiency of cytotoxic T cells in solid tumors. The translational goal of the proposed phase I research is to develop and validate a stroma-breaking therapeutic T cell delivery system for immunotherapy in human solid tumors with a focus on pancreatic and colon cancers. We have developed a receptor targeted and stroma breaking drug delivery platform with novel designs to overcome tumor stromal cellular and extracellular matrix barriers. We have demonstrated its ability to deliver nanoparticle-drug crossing tumor vessels, migrating through tumor stroma, and penetrating the basement membrane to reach tumor cells. This recombinant targeting ligand consists of urokinase plasminogen activator receptor (uPAR) binding domain of uPA fused with the catalytic domain of matrix metalloproteinase-14 (ATFmmp14). Our preliminary results showed that co-delivery of T cells with ATFmmp14 ligands led to significant increases in the numbers of T cells delivered in tumors following intravenous delivery in mouse colon and pancreatic tumor models as well as a human colon cancer patient derived xenograft (PDX) model. In this phase I project, we aim to determine the effect of ATFmmp14-T cell delivery system on intratumoral delivery and distribution using the MHC-non-restricted cytotoxic ɣδ T cells as a T cell model system in human colon and pancreatic cancer PDX models. First, we will develop the best formulation of ATFmmp14 conjugated T cell delivery platform using ɣδ T cells obtained from healthy donors in human pancreatic and colon PDX models (Aim1). Next, we will determine whether ATFmmp14 ligand can improve selective delivery of ɣδ T cells into pancreatic and colon PDX tumors and promote cell migration through stroma and basement membrane barriers to reach tumor cells (Aim 2). Biodistribution and systemic toxicity in normal organs and tissues will also be evaluated (Aim 2). Finally, the therapeutic efficacy of targeted delivery of ɣδ T cells without or with the combination of a chemotherapy drug will be determined in pancreatic and colon PDX models (Aim 3). Results of the above proposed studies should allow us building a strong foundation for conducting further translational studies in the Phase II research project to bring this approach into a clinical trial. Result of this study will also lead to future development and commercialization of this unique approach for other adaptive T transfer therapies, such as CAR-T, tumor specific T cells, and tumor infiltrating T cells.