Addressing Chemoresistance in Pancreatic and Ovarian Cancers: Photodynamic Priming and Repurposing of Tetracyclines using Targeted Photo-Activable Multi-Inhibitor Liposome

摘要 晚期卵巢癌或胰腺癌患者的预后仍然很差， 几十年反应率低的部分原因是对补救化疗的抵抗，包括 拓扑异构酶I（Topl）抑制剂，如伊立替康和拓扑替康。Top1抑制剂的全部潜力是 主要受到两种机制的阻碍：（1）ATP结合盒（ABC）转运蛋白（即，p-糖蛋白和 ABCG 2），积极泵出癌细胞的药物，和（2）上调的DNA修复酶，酪氨酰- DNA磷酸二酯酶1，它分解拓扑异构酶I-DNA可切割复合物， 再连接和细胞增殖。越来越清楚的是，没有一种单一的治疗方法可能克服 这个复杂的问题，以及新兴模式的组合治疗可能提供最有希望的。 在这里，我们介绍了一种互补的、双管齐下的方法来解决化疗耐药性：（i）采用 光动力引发（PDP）破坏ABC转运蛋白，改善Top1抑制剂的递送， 正常组织毒性，和（ii）重新使用四环素抗生素来抑制DNA损伤修复 酪氨酰-DNA磷酸二酯酶1. PDP是一种临床相关的、基于光化学的模式， 涉及光敏剂的光激活以调节附近的组织或生物分子而不杀死细胞。 该提案利用图像引导策略和纳米级工程来开发有针对性的照片， 可激活的多抑制剂脂质体（TPMIL），可在体内共同递送PDP、Top1抑制剂和四环素类抗生素， 考虑到它们的机械相互作用，确定适当的顺序。除了共同包装Top1 抑制剂和抗生素，TPMIL是表面修饰的抗体-光敏剂缀合物，以靶向 表皮生长因子受体，其在胰腺癌和卵巢癌中频繁扩增。使用一种新 高光谱荧光显微内窥镜成像系统，我们将纵向监测光敏剂 递送和ABC转运蛋白表达的变化，以改善体内PDP和化学增敏作用（目的1）。 将在体内研究Top1抑制剂和四环素类抗生素之间的相互作用机制， 有和没有PDP（目标2）。还将鉴定预测化疗增敏的生物标志物。TPMIL将 定制以靶向卵巢癌和胰腺癌细胞，同时共同递送光敏剂，Top1 抑制剂和四环素类抗生素（Aim 3）。将确定TPMIL的安全性和治疗效果 在PDX小鼠模型中（目标4）。我们已经证明了PDP在局部肺结核患者中的临床可行性。 晚期胰腺癌对于转移性卵巢癌，我们设想了一种简单可行的修改， 标准临床框架。TPMIL将在卵巢切除术后腹腔内递送。 肿瘤，然后光激活以触发PDP并释放化疗和抗生素。的 所获得的知识可能在开发改进的治疗方案方面发挥变革性作用， 针对个体患者中晚期胰腺癌和卵巢癌的分子特征进行定制。

ABSTRACT The prognosis for patients with advanced stage ovarian or pancreatic cancer has remained dismal for decades. The poor response rates result, in part, from resistance to salvage chemotherapies, including topoisomerase I (Top1) inhibitors such as irinotecan and topotecan. The full potential of Top1 inhibitors is hindered mainly by two mechanisms: (1) ATP-binding cassettes (ABC) transporters (i.e., P-glycoprotein and ABCG2) that actively pump drugs out of cancer cells, and (2) Upregulation of the DNA repair enzyme, tyrosyl- DNA phosphodiesterase 1, which resolves the topoisomerase I-DNA cleavable complexes to allow DNA religation and cell proliferation. It is becoming increasingly clear that no single treatment is likely to overcome this complex problem, and combination treatments of newly emerging modalities may offer the most promise. Here, we introduce a complementary, two-pronged approach to address chemoresistance: (i) Employing photodynamic priming (PDP) to damage ABC transporters, improve the delivery of Top1 inhibitors, and reduce the normal tissue toxicity, and (ii) Repurposing tetracycline antibiotics to inhibit the DNA damage repair enzyme tyrosyl-DNA phosphodiesterase 1. PDP is a clinically relevant, photochemistry-based modality that involves light activation of photosensitizers to modulate nearby tissues or biomolecules without killing the cells. This proposal leverages image-guided strategies and nanoscale engineering to develop Targeted Photo- Activable Multi-Inhibitor Liposome (TPMIL) that co-delivers PDP, Top1 inhibitors, and tetracycline antibiotics in the appropriate sequence with consideration of their mechanistic interactions. In addition to co-packaging Top1 inhibitors and antibiotics, TPMIL is surface modified with antibody-photosensitizer conjugates to target epidermal growth factor receptor, which is frequently amplified in pancreatic and ovarian cancer. Using a novel hyperspectral fluorescence microendoscope imaging system, we will longitudinally monitor photosensitizer delivery and changes in ABC transporter expression to improve PDP and chemosensitization in vivo (Aim 1). The mechanistic interactions between Top1 inhibitors and tetracycline antibiotics will be investigated in vivo, with and without PDP (Aim 2). Biomarkers predictive of chemosensitization will also be identified. TPMILs will be customized to target ovarian and pancreatic cancer cells while co-delivering photosensitizers, Top1 inhibitors, and tetracycline antibiotics (Aim 3). The safety and therapeutic efficacy of TPMIL will be determined in PDX mouse models (Aim 4). We have demonstrated the clinical feasibility of PDP in patients with locally advanced pancreatic cancer. For metastatic ovarian cancer, we envision a simple and feasible modification to the standard clinical framework. TPMILs will be delivered intraperitoneally after surgical debulking of ovarian tumors, and then light activated to trigger PDP and the release of chemotherapy and antibiotics. The knowledge gained may play a transformative role in the development of improved therapeutic regimens that are tailored to the molecular profile of advanced pancreatic and ovarian cancer in individual patients.