MUC1-Targeted Nanotherapy for Pancreatic Cancer

针对胰腺癌的 MUC1 纳米疗法

基本信息

批准号：
8683135
负责人：
Jordan M Winter
金额：
$ 16.71万
依托单位：
THOMAS JEFFERSON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-07-01 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8683135
关键词：
Address Adenocarcinoma Cell Adjuvant Antimetabolite Chemotherapy Biodistribution Biological Markers Biology Cancer Death Rates Candidate Disease Gene Cell Line Cells Cessation of life Clinical Code Collaborations Complex DNA DNA Sequence DNA analysis DNA delivery Data Death Rate Dendrimers Development Diphtheria Toxin Dose Drug resistance Enzymes Event Excision Female Foundations Genes Genetic Enhancer Element Gleevec Histology In Vitro Injection of therapeutic agent KRAS2 gene Lead Ligand Binding Ligands Luc Gene Luciferases MUC1 gene Malignant Neoplasms Malignant neoplasm of ovary Malignant neoplasm of pancreas Malignant neoplasm of prostate Metabolic Molecular Molecular Target Mucin-1 Staining Method Mus Normal Cell Nude Mice Oncogenes Organ Pancreatic Ductal Adenocarcinoma Pathway interactions Patients Peptides Pharmaceutical Preparations Phase Polymers Prostate Proteins Reporter Resistance Safety Sampling Scientist Signal Transduction Specificity Stromal Neoplasm Suicide Gene Therapy System Testing Therapeutic Toxic effect Toxin Transfection Transferrin Receptor Translating Transplantation Treatment Efficacy Tumor Biology Viral Viral Vector Work Xenograft procedure base biodegradable polymer cancer cell cell killing chemotherapy effective therapy gain of function mutation gene therapy improved innovation mesothelin mouse model nanoparticle nanotherapy neoplastic cell novel novel strategies optical imaging overexpression pancreatic cancer cells pancreatic neoplasm pancreatic tumorigenesis pre-clinical preclinical study promoter protein expression public health relevance research study response screening skills success treatment response treatment strategy tumor tumor growth vector

项目摘要

DESCRIPTION (provided by applicant): Pancreatic ductal adenocarcinoma (PDA) remains one of the most aggressive cancers with a rising death rate. The treatment paradigm has not changed over the past three decades, with antimetabolite chemotherapy as the mainstay of treatment. Novel approaches to develop effective therapies are needed. Promoter-driven gene therapy exploits an overactive cancer- specific promoter to express a desired gene in a cancer cell. This exciting strategy has been studied extensively in PDA using viral vectors, but has stalled at the pre-clinical level due to limitations of viral therapy. In prior studies, we developd a non-viral nanotherapy in which a biodegradable polymer DNA vector is injected locally into tumors. Using this approach, delivery of diphtheria toxin A (DTA) placed under the control of a cancer-specific promoter resulted in tumor shrinkage in pre-clinical mouse models of ovarian and prostate cancer, as well as in vitro studies of PDA. We recently demonstrated effective systemic targeting of DNA in prostate cancer-bearing mice using a novel DNA dendrimer conjugated to a targeting peptide against the transferrin receptor (TFRC). TFRC is overexpressed in most tumors (including PDA), but not in normal cells. Complementary to these studies, we used a rigorous approach to identify 13 promising promoter candidates for gene therapy from over 2500 putative PDA biomarkers, and tested these proteins for expression in our patient samples. Based on this work, MUC1 was identified as the best candidate promoter for transcriptionally-driven nanotherapy, since it was tightly correlated with aggressive biology i these PDA samples, and previous gene therapy studies (all viral) established efficacy of the MUC1 promoter. In this proposal, we detail plans to develop a MUC1 promoter-driven construct and use a derivatized DNA dendrimer to target PDA cell lines in culture and systemically in pre-clinical PDA mouse models (xenografts and orthotopic tumors). We will use a MUC1 promoter-driven luciferase reporter to test specificity in cell lines and a promoter-driven DTA construct to test treatment efficacy, biodistribution, and non-specific toxicity in mice. We hypothesize that MUC1-expressing cells will be exquisitely sensitive to treatment in vitro, and MUC1 non-expressing cells will be resistant. Similarly, we expect stunted tumor growth in MUC1-expressing xenografts and improved survival in mice with MUC1-expressing orthotopic tumors. If effective, these studies will serve as a foundation for rapid testing of systemically administerd MUC1 promoter-driven nanoparticles in patients with PDA, followed by early phase-safety trials of DTA nanotherapy in patients.

描述（由申请人提供）：胰腺导管腺癌（PDA）仍然是最具侵略性的癌症之一，死亡率上升。在过去的三十年中，治疗范式没有变化，抗代谢物化学疗法是治疗的主要。需要开发有效疗法的新颖方法。启动子驱动的基因治疗利用过度活跃的癌症启动子在癌细胞中表达所需的基因。这种令人兴奋的策略已在PDA中使用病毒载体进行了广泛的研究，但由于病毒疗法的局限性，在临床前的水平停滞了。在先前的研究中，我们开发了一种非病毒纳米疗法，其中可生物降解的聚合物DNA载体在局部注入肿瘤。使用这种方法，将放置在癌症特异性启动子控制下的白喉毒素A（DTA）的递送导致卵巢前小鼠和前列腺癌的临床小鼠模型以及PDA的体外研究导致肿瘤收缩。最近，我们使用一种与靶向肽针对转铁蛋白受体（TFRC）相结合的新型DNA树枝状聚合物（TFRC）的新型DNA树枝状聚合物对前列腺癌的小鼠进行了有效的DNA有效靶向。 TFRC在大多数肿瘤（包括PDA）中过表达，但在正常细胞中却没有表达。与这些研究相辅相成，我们使用了一种严格的方法来鉴定来自2500多个推定PDA生物标志物的13个有希望的启动子候选者，用于基因治疗，并测试了这些蛋白质在我们的患者样品中的表达。基于这项工作，MUC1被确定为转录驱动纳米疗法的最佳候选促进剂，因为它与这些PDA样本的侵略性生物学I紧密相关，并且先前的基因治疗研究（所有病毒）确定了MUC1启动子的功效。在此提案中，我们详细介绍了开发MUC1启动子驱动的构建体，并使用衍生化的DNA树状聚合物来靶向培养物中的PDA细胞系，并在临床前PDA小鼠模型（异种移植物和原位肿瘤）中系统地靶向PDA细胞系。我们将使用MUC1启动子驱动的荧光素酶报告基因测试细胞系中的特异性，并使用启动子驱动的DTA构建体到测试治疗功效，生物分布和小鼠的非特异性毒性。我们假设表达MUC1的细胞将对体外治疗非常敏感，而MUC1非表达细胞将具有抗性。同样，我们预计在表达MUC1的异种移植物中肿瘤的发育迟缓，并改善了表达MUC1的原位肿瘤的小鼠的生存率。如果有效，这些研究将是对PDA患者的系统施用MUC1启动子驱动的纳米颗粒的快速测试的基础，然后是患者DTA纳米疗法的早期阶段安全试验。