Matrilysin-Sensing Gene Delivery Vectors for Colorectal Cancer Therapy

用于结直肠癌治疗的基质溶素感应基因递送载体

基本信息

批准号：
8749004
负责人：
Junghae Suh
金额：
$ 21.43万
依托单位：
RICE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-10 至 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8749004
关键词：
Achievement Adverse effects Animals Area Behavior Binding Biodistribution Biological Assay Biological Markers Cancer Model Cancer Patient Cancer Survivor Capsid Cell Surface Receptors Cell surface Cells Colorectal Colorectal Cancer Colorectal Neoplasms Data Dependovirus Detection Digestion Disease Dose Environment Europe Exposure to Gene Delivery Generations Genes Genome Heterogeneity High temperature of physical object Histology Human Immune response In Vitro Knowledge Lead Libraries Malignant Neoplasms Matrilysin Measures Mediating Metric Modality Modeling Molecular Models Mutagenesis Neoplasm Metastasis Nucleic Acids Organ Outcome Patients Peptide Hydrolases Performance Positioning Attribute Pre-Clinical Model Process Property Proteolysis RNA Interference Research Personnel Reverse Transcriptase Polymerase Chain Reaction Serum Site Solutions Specificity Staging Survival Rate Technology Testing Therapeutic Tissues Transgenes Treatment Efficacy Viral Vector Virus Work adeno-associated viral vector base bioluminescence imaging cancer cell cancer site cancer therapy cellular transduction combat combinatorial design extracellular gene therapy in vivo metastatic colorectal molecular modeling neoplastic cell overexpression programs prototype public health relevance receptor receptor binding screening skills targeted delivery therapeutic gene transduction efficiency tumor tumor microenvironment uptake vector

项目摘要

PROJECT SUMMARY Once colorectal cancer metastasizes, it becomes a lethal disease with a 5-year survival rate of approximately 10%. Effective therapeutics that can specifically target metastatic colorectal tumor cells are sorely needed. Delivery of nucleic-acids (e.g. genes or RNAi) to combat cancer is a highly promising therapeutic approach; unfortunately, targeted delivery of gene vectors to tumor cells has been largely difficult to achieve. Most vector targeting approaches to date have relied on cell surface receptors overexpressed on some subpopulation of target cancer cells. Unfortunately, there is no unique cell surface biomarker that specifically identifies all cells in a tumor. To overcome this limitation, we propose to develop protease- activatable viruses (PAVs) that use extracellular proteases overexpressed in metastatic colorectal tumor microenvironments as the biomarkers to achieve targeted delivery. Specifically, matrilysin (also known as matrix metalloproteinase 7, MMP7) has been shown to be overexpressed in colorectal cancer. High levels of MMP7 in the tumor microenvironment will activate the PAVs in a localized manner and enable the vectors to bind cellular receptors that are broadly expressed, including on colorectal cancer cells, and mediate efficient gene delivery. Our PAV technology is based on the clinically promising adeno-associated virus (AAV), which has recently been approved as the first human gene therapy product in Europe. We have key pilot data demonstrating we have created MMP7-sensing PAVs that dramatically increase their gene delivery efficiency once exposed to the protease. Moreover, in an orthotopic cancer model, a PAV prototype is able to significantly increase transgene delivery and expression in tumors. In aim 1, we will synthesize and characterize a panel of MMP7-sensing PAVs. Our design process will harness both rational and combinatorial approaches in order to expedite achievement of the design solution. In aim 2, we will test the gene delivery performance of PAVs in vitro on colorectal cancer cells, and mechanistic studies will be done to probe the interaction of PAVs with the cells. Finally, we will test the PAVs in an orthotopic model of metastatic colorectal cancer in order to determine their in vivo specificity and therapeutic efficacy. If successful, this project will generate protease-responsive AAV vectors that may become viable therapeutic options for metastatic colorectal cancer.

项目摘要一旦结直肠癌转移，它就会成为一种致命疾病，生存率为5年约10％。可以专门针对转移性结直肠肿瘤细胞的有效治疗剂是非常需要。将核酸（例如基因或RNAi）递送到打击癌症是一种高度有希望的治疗方法；不幸的是，针对性的基因载体向肿瘤细胞递送一直很困难实现。迄今为止，大多数矢量靶向方法都依赖于过表达的细胞表面受体目标癌细胞的一些亚群。不幸的是，没有独特的细胞表面生物标志物明确识别肿瘤中的所有细胞。为了克服这一限制，我们建议开发蛋白酶 - 可活化病毒（PAVS），使用过表达的细胞外蛋白酶在转移性结直肠肿瘤中过表达微环境作为实现目标递送的生物标志物。具体而言，天生莲（也称为基质金属蛋白酶7，MMP7）已显示在结直肠癌中过表达。高水平肿瘤微环境中的MMP7将以局部方式激活PAV，并使向量能够到达媒介结合广泛表达的细胞受体，包括在结直肠癌细胞上，并介导有效基因输送。我们的PAV技术基于临床上有希望的腺相关病毒（AAV），该病毒（AAV）最近已被批准为欧洲的第一个人类基因治疗产品。我们有关键的飞行员数据证明我们已经创建了MMP7感应PAV，从而显着提高了基因递送效率一旦暴露于蛋白酶。此外，在原位癌模型中，PAV原型能够显着增加转基因的递送和肿瘤的表达。在AIM 1中，我们将合成并表征一组MMP7感应PAVS。我们的设计过程将利用理性和组合方法是为了加快设计解决方案的实现。在AIM 2中，我们将测试基因输送 PAV在结直肠癌细胞上的表现，并将进行机理研究以探测 PAV与细胞的相互作用。最后，我们将在转移性结直肠的原位模型中测试PAVS 为了确定其体内特异性和治疗功效。如果成功，这个项目将产生蛋白酶反应的AAV载体，可能成为转移性的可行治疗选择结直肠癌。