Tumor Targeted Corroles for Detection and Intervention

摘要 这一提议将检验非共价Corole组件同时调节两者的假设 在单个自组装复合体中进行肿瘤靶向检测和干预。 磺化腐蚀剂是一种可溶于水的大环化合物，可被金属化，并可释放出 强烈的荧光。我们已经发现，腐蚀剂会自发地与载体蛋白组装，载体蛋白是 需要促进细胞进入，一旦进入细胞，必须释放到细胞质中以诱导 细胞毒性，但仍被排除在核外，因此暗示胞液因子是靶点 科罗尔介导的毒性。我们的靶向细胞穿透蛋白HerPBK10使Corole能够摄取到 HER2+癌细胞的体外和体内实验。HerPBK10由细胞靶向和内化配体组成 来源于HERG蛋白，以及来源于腺病毒(Ad)的膜穿透结构域 卡西德·潘顿基地。Corole荧光使体外和体内肿瘤细胞靶向的可视化成为可能， 体内肿瘤靶向导致以近300倍的剂量进行肿瘤生长干预 与直接瘤内注射化疗药物阿霉素进行比较。 HER2+癌症已经成为我们实验室测试新的靶向治疗的模型系统。作为 HER2(或ErbB2)亚单位的过表达增强了受体的亲和力，HER2+细胞类型是理想的 测试配基导向疗法的模型。更重要的是，随着HER2在乳腺癌中的过度表达 与侵袭性的化疗耐药肿瘤相关，并预测预后不良，替代治疗 标准方案可能被证明对这类乳腺癌更有效，虽然不包括 大多数病例都是最致命的乳腺癌之一。尽管如此，我们已经确定了其他 HERG导向疗法的潜在靶点，包括卵巢、胶质瘤和前列腺癌细胞 高水平表达不同的HER亚基。因此，此处提供的针对HER的系统可能具有 除了HER2+乳腺癌外，更广泛的应用于几种不同的肿瘤类型。 此建议结合了多个合作者的专业知识，以进一步开发Corole程序集 可成像的肿瘤靶向剂。我们将评估靶细胞和免疫与携带者的相互作用 在引入修饰方面引导努力的蛋白质，可提高治疗效果和安全性。一 我们将探索的激动人心的方向是应用定向进化来选择载体蛋白结构域进行改进 靶细胞相互作用和免疫逃避。我们将在体外测试这些改良剂对科罗尔的释放。 和体内，并利用腐蚀剂独特的光电子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.