Tumor Targeted Corroles for Detection and Intervention

摘要 该提议将检验非共价的咔咯组装体同时介导 在单个自组装复合物中进行肿瘤靶向检测和干预。 磺化的可腐蚀物是水溶性的大环化合物，其可以被金属化并且可以释放出 强烈的荧光我们已经发现，corroles自发地组装载体蛋白，这是 需要促进细胞进入，并且一旦进入胞浆，必须释放到细胞质中以引起 细胞毒性，同时保持排除从核，从而牵连胞质因子的目标， 腐蚀介导的毒性我们的靶向细胞渗透蛋白HerPBK10使corrole摄取进入 体外和体内HER2+癌细胞。HerPBK10由细胞靶向和内化配体组成 衍生自调蛋白的膜穿透结构域和衍生自腺病毒（Ad）的膜穿透结构域 衣壳五邻体碱基。Corrole荧光能够在体外和体内可视化肿瘤细胞靶向， 并且体内肿瘤靶向导致肿瘤生长干预的剂量减少近300倍， 与化疗剂阿霉素的直接肿瘤内递送相比。 HER2+癌症已成为我们实验室测试新靶向疗法的模型系统。为 HER2（或ErbB2）亚基的过表达增强受体亲和力，HER2+细胞类型是理想的 用于测试配体导向疗法的模型。更重要的是，随着乳腺癌中HER2的过度表达， 与侵袭性化疗耐药肿瘤相关，并预测预后不良， 标准方案可能证明对这类乳腺癌更有效，虽然不包括 大多数情况下，是最致命的乳腺癌。然而，我们发现了更多的 我们的调蛋白导向治疗的潜在靶点，包括卵巢癌、神经胶质瘤和前列腺癌细胞 表达高水平的不同HER亚基。因此，这里提出的HER靶向系统可能具有 除HER2+乳腺癌外，还可广泛应用于几种不同的肿瘤类型。 该提案结合了多个合作者的专业知识，进一步将corrole程序集开发为 可成像的肿瘤靶向剂。我们将评估靶细胞和免疫与载体的相互作用 蛋白质，以指导在引入修饰，可以提高治疗效果和安全性的努力。一 我们将探索的一个令人兴奋的方向是应用定向进化来选择载体蛋白结构域， 靶细胞相互作用和免疫逃避。我们将在体外测试这些修饰的可罗传递 并利用Corroles独特的光电发射特性来检测体内肿瘤靶向。

ABSTRACT This proposal will test the hypothesis that noncovalent corrole assemblies simultaneously mediate both tumor targeted detection and intervention in a single self-assembled complex. Sulfonated corroles are water soluble, macrocyclic compounds that may be metallated and can emit an intense fluorescence. We have found that corroles spontaneously assemble with carrier proteins, which are required to facilitate cel entry, and once entering cels, must be released into the cytoplasm to elicit cytotoxicity while remaining excluded from the nucleus, thus implicating cytosolic factors as the targets of corrole-mediated toxicity. Our targeted cell penetration protein, HerPBK10, enables corrole uptake into HER2+ cancer cells in vitro and in vivo. HerPBK10 is comprised of a cell-targeting and internalizing ligand derived from the heregulin protein, and membrane penetration domain derived from the adenovirus (Ad) capsid penton base. Corrole fluorescence enables visualization of tumor cell targeting in vitro and in vivo, and tumor targeting in vivo results in tumor growth intervention at nearly 300x less dosage in comparison to direct intratumoral delivery of the chemotherapy agent, doxorubicin. HER2+ cancer has served as a model system for testing new targeted therapeutics in our lab. As the overexpression of the HER2 (or ErbB2) subunit enhances receptor affinity, the HER2+ cell type is an ideal model for testing ligand-directed therapies. More importantly, as HER2 overexpression in breast cancer correlates with aggressive chemoresistant tumors and predicts a poor prognosis, alternative treatments to standard regimens may prove more effective on this subset of breast cancers that, while not comprising a majority of cases, are among the most deadly of breast cancers. Nevertheless, we have identified additional potential targets of our heregulin-directed therapeutics, including ovarian, glioma, and prostate cancer cells that express high levels of different HER subunits. Thus, the HER-targeted system presented here may have a broader application to several different tumor types in addition to HER2+ breast cancer. This proposal combines the expertise of multiple collaborators to further develop corrole assemblies into image-able tumor targeting agents. We will assess target cell and immune interactions with the carrier protein to direct efforts in introducing modifications that may enhance therapeutic efficacy and safety. One exciting direction we will explore is to apply directed evolution to select carrier protein domains to improve target cell interactions and immune evasion. We will test these modifications for corrole delivery in vitro and in vivo, and utilize the unique photoemission properties of corroles to detect in vivo tumor targeting.