荧光成像指导的双靶向透明质酸纳米颗粒/维替泊芬复合物用于葡萄膜黑色素瘤治疗研究

Uveal melanoma (UM) is the most common primary ocular malignancy in adults and severely threaten patients’ life. Currently no effective systemic treatment can prolong survival of UM patients with metastases. Previous studies have shown the efficacy of one promising medicine Verteporfin (VP) in UM; either via its photosensitizer property involved in photodynamic therapy (PDT), or through its intrinsic characteristic of inhibiting the oncogene YAP without light activation. However, some disadvantages hamper the effectiveness of Verteporfin，such as low water solubility and poor targeting. Therefore, it is urgent to optimize the treatment strategy or modify the current chemical compound. Our previous studies have found that hyaluronic acid nanoparticle (HANP) can encapsulate photosensitizers or chemotherapy drugs, and target tumors more efficiently and precisely compared to traditional treatment. We have validated the low cytotoxicity, excellent biocompatibility and good tumor targetability of HANP. Furthermore, by conjugating Indocyanine green (ICG) to HANP, we successfully monitored targeted accumulation of the nanocomplex using the fluorescent imaging system in vivo. Here, we propose to encapsulate VP with ICG-modified HANP, and therefore form a new nanomedicine ICG-HANP/VP (IHV). We will test the characteristic of the new nanocomplex with TEM and HPLC, and examine its anti-cancer effectiveness and safety of targeted PDT/chemotherapy integrative treatment in vitro and in vivo. In addition, we plan to study several factors involved in VP-related signaling pathways，and to investigate the mechanisms using RT-PCR, Western blot, etc. This study will preliminarily explore novel therapeutic options targeting UM and provide support for anti-cancer nanodrugs development and have a great translational potency.

葡萄膜黑色素瘤（UM）是成人最常见的眼内恶性肿瘤，目前尚无有效药物方案可提高转移性UM患者的生存期。维替泊芬（VP）的光动力疗效及非光敏化疗作用为UM治疗带来新希望。然而VP因水溶性差、靶向性不够好而影响肿瘤反应率并造成一定程度的视力损伤。我们研究证实透明质酸纳米颗粒（HANP）可携载光敏剂或化疗药物对恶性肿瘤发挥更具优势的靶向疗效，用吲哚青绿（ICG）修饰HANP可实现活体荧光成像实时监测肿瘤内的纳米药物聚集。本项目拟用ICG修饰的HANP装载VP自组装成新型纳米药物ICG-HANP/VP（IHV），透射电镜及高效液相色谱等检测纳米药物IHV的表征及性能，通过体内外实验验证IHV较传统VP给药发挥更安全有效的靶向光动力/化疗疗效，RT-PCR、Western blot等检测VP相关通路蛋白的表达，探索IHV的抗癌机制。本课题的完成将为UM的靶向纳米药物开发提供临临床前参考。

葡萄膜黑色素瘤（UM）是成人最常见的眼内恶性肿瘤，具有侵袭性高、恶性程度强、转移发生早、预后差等特点。目前尚无有效的治疗方案来改善转移性UM患者的生存期。在临床上，光敏剂维替泊芬（VP）具有既能作为光敏剂介导光动力治疗又能作为化疗药发挥抗肿瘤作用的特点，为UM的治疗带来了希望。但由于VP生物相容性不理想、靶向性差以及全身毒性等问题，使其临床应用受限。因此，优化现有UM治疗方案意义重大。透明质酸纳米颗粒（HANP）因其所具有的提高药物稳定性、改善药物靶向性等优势，成为理想的药物载体。本课题组通过物理包埋的方式将VP装载入HANP中，制备出新型纳米药物HANP/VP复合物。因VP具有自发荧光，可实现肿瘤部位纳米颗粒聚集程度的可视化。同时通过实验证明HANP/VP具有良好的稳定性和水溶性。我们分别在体外细胞实验和体内小鼠肿瘤模型中验证了HANP/VP具有更强的肿瘤靶向性和药物递送能力。通过荷瘤小鼠体内抗肿瘤实验证明HANP/VP在激光照射条件下可显著降低UM细胞活力、抑制细胞增殖和肿瘤生长。综上，本研究设计构建了双靶向透明质酸纳米药物HANP/VP，显著提高了临床药物VP的靶向肿瘤递送效果，减少了全身毒副作用，提高了其抗肿瘤疗效，具有潜在的临床应用价值。本项目的实施也为临床优化UM治疗的新型靶向纳米药物的设计、合成提供新思路和有力的实验参考。除上述研究工作外，我们同时开展了其他方向研究，拓展了本课题组的研究方向，为后续课题的开展打下了坚实基础，例如本课题组成功构建了GPC-3靶向FANP复合物用于荧光/ MRI/PA成像引导肿瘤光热治疗；揭示了圆锥角膜发生机制中可能涉及的调控网络; 研究了人类和小鼠视网膜母细胞瘤相关的microRNA-mRNA调控网络和通路；分析了细菌性角膜炎与真菌性角膜炎差异表达的基因，以便快速的区分识别感染性角膜炎的致病因素。