Photonanomedicine against EpCAM and Ki-67 positive head and neck tumors

针对 EpCAM 和 Ki-67 阳性头颈肿瘤的光子纳米医学

• 批准号: 361889716
• 负责人: Privatdozent Dr. Ramtin Rahmanzadeh
• 金额: --
• 依托单位: Institut für Biomedizinische Optik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2017
• 资助国家: 德国
• 起止时间: 2016-12-31 至 2021-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/361889716?language=en
• 关键词: Photonanomedicine; against; EpCAM; Ki; 67

Head and neck tumor is a malignancy with rising incidence and to date, surgical resection is the primary treatment option, which is associated with facial disfigurement and a high risk for recurrence. Therefore, the development of new therapies for this tumor type is of high clinical value. We plan to use photonanomedicine as a basis to develop such an approach and to evaluate it in animal experiments. In a previous project, we successfully developed the toolbox for a targeting concept against Ki-67 positive cells, which is based on nano-constructs and light irradiation. Antibody conjugates of TuBB-9 (an anti-Ki-67 antibody) with different photosensitizers suitable for in vivo applications were prepared and tested. Coupling to different cell and nucleus penetrating peptides or encapsulation into liposomes ensures cellular und nuclear delivery. Intracellular release of the antibodies from endososomes by photochemical internalization (PCI) drastically increased efficiency. We were able to demonstrate the following points, which provide strong evidence for the effectiveness of the new method:1. Reliable distribution of the antibody-dye conjugates in tissues and uptake in cells by liposomes and photochemical internalization (PCI).2. A pronounced tumor selectivity by targeting of the tumor specific membrane protein EpCAM and by local light application.3. An effective mechanism that leads to cell death. This is ensured by the light inactivation of Ki-67, which is feasible with antibody concentration in the nanomolar range.In the proposed project, the antibody-assisted photomedical therapy shall be simplified and tested in a mouse model for head and neck squamous cell carcinoma. To increase the tumor selectivity, liposomes directed against the EpCAM protein will be used. EpCAM is not only used as a target structure for the transport of liposomes, but it will also be investigated as a direct target for light inactivation. For the tumor mouse model, GFP transfected cells will be used, which enable fluorescence assisted identification of tumor cells and mechanistic studies in living mice by intra-vital multiphoton microscopy. For photochemical internalization, the photosensitizer Amphinex, which is already used in clinical studies, will be applied. With the help of cell-penetrating immunoconjugates, proliferating cells will be effectively damaged after irradiation. Short-term and long-term treatment successes in the animal model will be evaluated and cellular effects of Ki-67 inactivation will be investigated with the help of multiphoton microscopy.A successful elimination of the tumor in the animal model would be an important first step towards the application of photonanotherapy for the treatment of head and neck tumors in humans, providing new treatment options for this crippling disease.

头颈部肿瘤是一种发病率不断上升的恶性肿瘤，手术切除是目前的主要治疗方法，但手术切除后患者面部畸形严重，复发风险高。因此，针对这种肿瘤类型开发新的治疗方法具有很高的临床价值。我们计划使用光纳米医学作为基础来开发这种方法，并在动物实验中对其进行评估。在之前的一个项目中，我们成功地开发了针对Ki-67阳性细胞的靶向概念工具箱，该工具箱基于纳米结构和光照射。制备并测试TuBB-9（抗Ki-67抗体）与适合于体内应用的不同光敏剂的抗体缀合物。偶联到不同的细胞和核穿透肽或封装到脂质体中确保细胞和核递送。通过光化学内化（PCI）从内体细胞内释放抗体显著提高了效率。我们能够证明以下几点，这为新方法的有效性提供了强有力的证据：1。抗体-染料缀合物在组织中的可靠分布以及通过脂质体和光化学内化（PCI）在细胞中的吸收。2.通过靶向肿瘤特异性膜蛋白EpCAM和通过局部光应用，具有显著的肿瘤选择性。导致细胞死亡的有效机制。这是通过Ki-67的光灭活来确保的，这在纳摩尔范围内的抗体浓度是可行的。在拟议的项目中，抗体辅助的光医学治疗将被简化，并在头颈部鳞状细胞癌的小鼠模型中进行测试。为了增加肿瘤选择性，将使用针对EpCAM蛋白的脂质体。EpCAM不仅用作脂质体转运的靶结构，而且还将作为光灭活的直接靶点进行研究。对于肿瘤小鼠模型，将使用GFP转染的细胞，其能够通过活体多光子显微镜在活小鼠中荧光辅助鉴定肿瘤细胞和机制研究。对于光化学内化，将应用已用于临床研究的光敏剂Amphinex。在细胞穿透免疫偶联物的帮助下，增殖细胞将在辐射后被有效地破坏。将评估动物模型的短期和长期治疗成功率，并将借助多光子显微镜研究Ki-67灭活的细胞效应。在动物模型中成功消除肿瘤将是将光鼻疗法应用于治疗人类头颈部肿瘤的重要第一步，为这种致残性疾病提供新的治疗选择。