Heregulin-Targeted Protein Uptake for Breast Cancer

乳腺癌的赫调节蛋白靶向蛋白质摄取

基本信息

批准号：
7119022
负责人：
LALI K MEDINA-KAUWE
金额：
$ 23.4万
依托单位：
CEDARS-SINAI MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-09-02 至 2009-08-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): HER2-positive breast cancers are highly aggressive, frequently resistant to chemotherapy, and associated with a high incidence of mortality. Therefore, therapy by alternative means, such as targeted delivery of therapeutic genes, may prove much more effective on these types of cancers than standard methods of treatment. The gene delivery system described here is novel in its utilization of recombinant adenoviral components which have been modified to achieve breast cancer cell-specific binding and delivery of conjugated DNA, while retaining the high efficiency cellular uptake features of the viral capsid. This system would require only the minimal adenoviral proteins necessary for cell surface binding and internalization, and lacks all other viral proteins and genes. Thus, this system avoids the concerns associated with using viruses such as mutation and viral recombination. The Specific Aims are to test the hypotheses that: 1. Recombinant soluble penton and fiber capsid protein translocation requires interactions of key molecular motifs found in capsid proteins with components of the cytoskeletal and intracellular trafficking machinery. This study will uncover useful information about the determinants governing breast cancer cell entry, as little is known about viral trafficking pathways in these cells. Immunofluorescence and confocal microscopy will be used here to visualize trafficking of wild-type and mutant proteins. Traffic-enhancing mutations will be incorporated into vector design and tested in Aims2&3. 2. Engineered capsid proteins mediate non-viral gene delivery to HER2+ breast cancer cells and are less toxic than whole recombinant Ad in vitro. Comparisons of cytotoxicity and gene transfer will be made between non-viral complexes and Ad. We will assess the contribution individual capsid proteins make toward the cellular response to Ad infection. 3. Engineered capsid proteins are less toxic and immunogenic than whole recombinant Ad, and can mediate targeted non-viral gene delivery to HER2+ breast cancer cells in vivo. As HER2+ cells overexpress the heregulin receptor, we examine of the capacity for heregulin to direct non-viral gene delivery to this tissue. We will assess the contribution of individual capsid proteins to the immune response to Ad, compare immunogenicity to Ad, and test a method of immune evasion.

描述（由申请人提供）： HER2 阳性乳腺癌具有高度侵袭性，经常对化疗产生耐药性，并且死亡率很高。因此，通过替代手段进行治疗，例如靶向递送治疗基因，可能比标准治疗方法更有效地治疗这些类型的癌症。这里描述的基因递送系统的新颖之处在于它利用了重组腺病毒成分，这些成分经过修饰以实现乳腺癌细胞特异性结合和缀合DNA的递送，同时保留了病毒衣壳的高效细胞摄取特征。该系统仅需要细胞表面结合和内化所需的最少腺病毒蛋白，并且缺乏所有其他病毒蛋白和基因。因此，该系统避免了与使用病毒相关的问题，例如突变和病毒重组。具体目标是测试以下假设： 1. 重组可溶性五邻体和纤维衣壳蛋白易位需要衣壳蛋白中发现的关键分子基序与细胞骨架和细胞内运输机制的组件相互作用。这项研究将揭示有关控制乳腺癌细胞进入的决定因素的有用信息，因为人们对这些细胞中的病毒运输途径知之甚少。此处将使用免疫荧光和共聚焦显微镜来可视化野生型和突变蛋白的运输。流量增强突变将被纳入载体设计并在 Aims2 和 3 中进行测试。 2. 工程衣壳蛋白介导非病毒基因向 HER2+ 乳腺癌细胞的传递，并且在体外比完整重组 Ad 的毒性更低。将在非病毒复合物和Ad之间进行细胞毒性和基因转移的比较。我们将评估各个衣壳蛋白对 Ad 感染的细胞反应的贡献。 3. 工程衣壳蛋白比完整重组Ad的毒性和免疫原性更低，并且可以介导靶向非病毒基因向体内HER2+乳腺癌细胞的递送。由于 HER2+ 细胞过度表达调蛋白受体，我们检查了调蛋白将非病毒基因递送至该组织的能力。我们将评估单个衣壳蛋白对 Ad 免疫反应的贡献，比较 Ad 的免疫原性，并测试免疫逃避方法。