MUC1-Targeted Nanotherapy for Pancreatic Cancer

针对胰腺癌的 MUC1 纳米疗法

• 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
• 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

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）仍然是最具侵袭性的癌症之一，死亡率不断上升。在过去的三十年里，治疗模式没有改变，抗代谢药物化疗是治疗的主要手段。需要开发有效疗法的新方法。启动子驱动的基因疗法利用过度活性的癌症特异性启动子在癌细胞中表达所需基因。这种令人兴奋的策略已经在PDA中使用病毒载体进行了广泛的研究，但由于病毒治疗的局限性，在临床前水平上已经停滞。在先前的研究中，我们开发了一种非病毒纳米疗法，其中将可生物降解的聚合物DNA载体局部注射到肿瘤中。使用这种方法，在癌症特异性启动子的控制下递送白喉毒素A（DTA）导致卵巢癌和前列腺癌的临床前小鼠模型以及PDA的体外研究中肿瘤缩小。最近，我们证明了有效的系统靶向的DNA在前列腺癌荷瘤小鼠使用一种新的DNA树枝状聚合物共轭的靶向肽对转铁蛋白受体（TFRC）。TFRC在大多数肿瘤（包括PDA）中过表达，但在正常细胞中不表达。作为对这些研究的补充，我们使用了一种严格的方法从超过2500种假定的PDA生物标志物中鉴定出13种有希望的基因治疗启动子候选物，并测试了这些蛋白质在我们的患者样本中的表达。基于这项工作，MUC1被确定为转录驱动纳米疗法的最佳候选启动子，因为它与这些PDA样品中的侵袭性生物学密切相关，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。在这项提案中，我们详细计划开发一个MUC1启动子驱动的构建体，并使用衍生的DNA树状聚合物靶向PDA细胞系的培养和系统性的临床前PDA小鼠模型（异种移植和原位肿瘤）。我们将使用MUC1启动子驱动的荧光素酶报告基因来测试细胞系中的特异性，并使用启动子驱动的DTA构建体来 在小鼠中测试治疗功效、生物分布和非特异性毒性。我们假设MUC 1表达细胞对体外治疗非常敏感，而MUC 1非表达细胞将产生耐药性。类似地，我们预期MUC1表达异种移植物中的肿瘤生长受阻，并且MUC1表达原位肿瘤小鼠的存活率提高。如果有效，这些研究将作为快速测试PDA患者全身给药MUC1启动子驱动的纳米颗粒的基础，然后在患者中进行DTA纳米治疗的早期阶段安全性试验。