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A NOVEL TARGETED THERAPEUTIC USING VIRAL CAPSID PROTEIN

使用病毒衣壳蛋白的新型靶向治疗

基本信息

批准号：
7394404
负责人：
LALI K MEDINA-KAUWE
金额：
$ 19.08万
依托单位：
CEDARS-SINAI MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-04-10 至 2010-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7394404
关键词：
A Mouse Address Adenoviruses Affect Alkanesulfonates Alternative Therapies Animal Model Animals Apoptosis Applications Grants Biodistribution Biological Biological Assay Blood Cells Blood Circulation Breast Cancer Cell Breast Cancer Treatment Capsid Capsid Proteins Carrier Proteins Categories Cell Death Cell Line Cell Nucleus Cell Surface Receptors Cell Survival Cells Condition Cytochrome P450 Cytosol Cytotoxic agent DNA Development Drug Delivery Systems Drug Kinetics ERBB2 gene Elements Enzymes Epidermal Growth Factor Receptor Figs - dietary Flow Cytometry Fluorescence Fluorescence Microscopy Funding Future Gene Delivery Gene Expression Gene Transduction Agent Genes Hair Cells Harvest Health Heregulin Human Immune In Vitro Injection of therapeutic agent Investigation Ligands Measures Mediating Membrane Metabolic Methods Modeling Mus Neoplasm Metastasis Non-Viral Vector Normal Cell Penetration Pharmaceutical Preparations Pilot Projects Preparation Process Protein Overexpression Proteins Public Health Publishing Purpose Recombinant Proteins Research Research Project Grants Risk Serum Specificity Staging Standards of Weights and Measures System Temperature Testing Therapeutic Therapeutic Uses Time Tissue Banks Toxic effect Treatment Efficacy Vesicle Viral Work amphiphilicity base cancer therapy cancer type concept corrole cytokine cytotoxicity design dosage experience falls gastrointestinal epithelium gene therapy gene transfer vector immunogenicity improved in vivo malignant breast neoplasm mixed cell culture mortality neutralizing antibody new technology non-viral gene therapy novel novel strategies novel therapeutics outcome forecast particle pathogen penton base prevent receptor receptor mediated endocytosis response therapeutic target trafficking tumor tumor xenograft uptake vector viral vector development

项目摘要

DESCRIPTION (provided by applicant): This proposal will test the hypothesis that the cell entry and cell penetration features of a novel recombinant protein derived from the adenovirus (Ad) capsid mediates non-viral delivery of a novel toxic corrole compound specifically to breast cancer cells for targeted breast cancer therapy. We will test this new therapeutic for targeting HER2+ breast cancer, which is characterized by aggressive tumor formation, metastasis, chemoresistance, a poor prognosis, high mortality, and amplification of the human epidermal growth factor receptor HER2 subunit. About 30% of breast cancers are HER2+, comprising a significant subset of breast cancers requiring novel alternatives to standard treatment. The targeting aspect of this conjugate will direct therapy specifically to HER2+ cells, thus avoiding normal cells and other dividing cells such as blood cells, hair cells, and gastrointestinal epithelium. The Specific Aims of this project are to test the hypotheses that: 1) HerPBK10 targets corroles to HER2+ cells in vitro. Standard metabolic assays for measuring cell survival and proliferation will be used to determine the cytotoxicity of conjugates on a panel of HER2+ and HER2- cells in vitro, and to assess the effect of serum on conjugate activity, and determine optimal dosage. Spectrophotometric absorbance assays will be used to assess the assembly parameters of the conjugates under different storage times and temperatures. Flow cytometry, fluorescence microscopy, and cytotoxicity assays will be used to assess targeting specificity by competitive inhibition with a free ligand, delivery to HER2+ cells in a mixed cell culture, and delivery to isogenic cell lines expressing different levels of receptor subunits. Finally, apoptosis as a mechanism of cell death will be examined. 2) HerPBK10 targets corroles to HER2+ cells in vivo. A mouse xenograft tumor model of HER2+ breast cancer will be used to assess the therapeutic efficacy of conjugate delivery by both intratumoral injection and systemic delivery, and the presence of circulating heregulin will be assessed. The biodistribution and pharmacokinetics of conjugates will also be examined by harvesting and immunohistological processing of tissues, and collection of serum, after in vivo circulation. We will use ELISA-based assays to examine cytokine induction in both tumor model and immune competent mice and to examine induction of neutralizing antibodies in immune competent mice. Apoptosis as a mechanism of cell death in vivo will also be examined. This research is relevant to public health because the proposed project will develop a novel therapeutic that can specifically target HER2+ breast cancer. This targeted therapy should be improved over conventional treatment methods because normal cells should not be affected. As HER2+ breast cancer does not respond well to conventional therapies, this alternative therapy could provide a significant contribution to breast cancer treatment.

描述（由申请人提供）：本提案将检验以下假设：源自腺病毒（Ad）衣壳的新型重组蛋白的细胞进入和细胞穿透特征介导新型毒性咔咯化合物特异性非病毒递送至乳腺癌细胞，用于靶向乳腺癌治疗。我们将测试这种针对HER 2+乳腺癌的新疗法，其特征是侵袭性肿瘤形成，转移，化疗耐药性，预后不良，死亡率高，以及人表皮生长因子受体HER 2亚基的扩增。大约30%的乳腺癌是HER 2+，包括需要新的替代标准治疗的乳腺癌的重要子集。该缀合物的靶向方面将特异性地将治疗引导至HER 2+细胞，从而避免正常细胞和其他分裂细胞如血细胞、毛细胞和胃肠上皮。本项目的具体目标是检验以下假设： 1)HerPBK 10靶点在体外与HER 2+细胞结合。将使用用于测量细胞存活和增殖的标准代谢试验来确定缀合物对一组HER 2+和HER 2-细胞的体外细胞毒性，并评估血清对缀合物活性的影响，并确定最佳剂量。将使用分光光度吸光度测定法评估不同储存时间和温度下结合物的组装参数。将使用流式细胞术、荧光显微镜和细胞毒性试验，通过与游离配体的竞争性抑制、递送至混合细胞培养物中的HER 2+细胞以及递送至表达不同水平受体亚基的等基因细胞系来评估靶向特异性。最后，将检查作为细胞死亡机制的凋亡。 2)HerPBK 10靶点在体内与HER 2+细胞结合。将使用HER 2+乳腺癌的小鼠异种移植肿瘤模型来评估通过肿瘤内注射和全身递送两者的缀合物递送的治疗功效，并且将评估循环调蛋白的存在。在体内循环后，还将通过组织的收获和免疫组织学处理以及血清的收集来检查缀合物的生物分布和药代动力学。我们将使用基于ELISA的测定来检查肿瘤模型和免疫活性小鼠中的细胞因子诱导，并检查免疫活性小鼠中中和抗体的诱导。还将检查作为体内细胞死亡机制的细胞凋亡。这项研究与公共卫生有关，因为拟议的项目将开发一种新的治疗方法，可以特异性靶向HER 2+乳腺癌。这种靶向治疗应该比传统的治疗方法有所改进，因为正常细胞不应该受到影响。由于HER 2+乳腺癌对常规化疗反应不佳，这种替代疗法可以为乳腺癌治疗做出重大贡献。