Optimal DNA Brain-Penetrating Nanoparticle (DNA-BPN) Formulation for Glioblastoma (GBM) Treatment

项目总结/摘要 该项目提案的重点是开发一种最佳的脑穿透DNA纳米颗粒制剂， 基因治疗在胶质母细胞瘤治疗中的用途。胶质母细胞瘤（GBM）是最常见和最具侵袭性的肿瘤。 原发性脑肿瘤，但目前可用的疗法有严重的副作用，疾病仍然 一致致命基因治疗是一种潜在的强有力的策略，在临床前研究中已显示出希望。 然而，有效的基因治疗尚未在人类中实现，这在很大程度上是由于无法实现 基因载体的广泛分布和脑中的肿瘤选择性基因转移，这是 GBM的高度侵袭性我们已经开发出了合成的携带DNA的纳米颗粒， 避免由于小颗粒尺寸（<<100 nm）和 致密PEG涂层。这些系统能够穿透大鼠大脑的整个纹状体 当通过对流增强递送（CED）施用时。我们的试验数据进一步表明，这些DNA 装载的脑穿透纳米颗粒（DNA-BPN）提供了更广泛和增加的转基因， 在CED后体内健康脑实质和脑肿瘤组织中的表达与金- 标准DNA纳米颗粒不能有效地渗透到输注部位之外。创新和 彻底测试用多种有前途的核心聚合物配制的DNA-BPN，并比较它们在 体外，离体和体内的金标准系统和领先的病毒为基础的载体。从第一代开始 DNA-BPN似乎已经能够安全地切除大部分大鼠大脑，我们计划进行研究。 其他有希望的聚合物可以提供甚至更高的转染沿着使用高肿瘤- 特异性启动子限制癌细胞的治疗性转基因表达。我们的假设是 联合方法将允许所有肿瘤细胞，包括导致肿瘤复发的高度侵袭性肿瘤细胞， 选择性转染，从而通过消除基因转移到健康的人， 细胞

Project Summary/Abstract This project proposal is focused on developing an optimal brain-penetrating DNA-nanoparticle formulation for gene therapy purposes in Glioblastoma treatment. Glioblastoma (GBM) is the most common and aggressive primary brain tumor, but currently available therapies have severe side effects and the disease remains uniformly lethal. Gene therapy is a potentially powerful strategy that has shown promise in preclinical studies. However, effective gene therapy has yet to be achieved in humans due in large part to an inability to achieve widespread distribution of gene vectors and yet tumor-selective gene transfer in the brain, which is required for the highly invasive nature of GBM. We have developed synthetic DNA-carrying nanoparticles capable of avoiding trapping within the brain parenchyma due to a combination of small particle size (<<100 nm) and dense PEG coatings. These systems are capable of penetrating throughout the entire striatum of the rat brain when administered by convection enhanced delivery (CED). Our pilot data further suggests that these DNA loaded brain-penetrating nanoparticles (DNA-BPN) provide much more widespread and increased transgene expression in healthy brain parenchyma and brain tumor tissue in vivo following CED compared to gold- standard DNA nanoparticles that do not efficiently penetrate beyond the site of infusion. We will develop and thoroughly test DNA-BPN formulated with multiple promising core polymers and compare their behavior in vitro, ex vivo and in vivo to gold-standard systems and leading virus-based vectors. Since the first-generation DNA-BPN already appear capable of safely transfecting a large part of the rat brain, we plan to investigate other promising polymers that may provide even higher transfection along with the use of a highly tumor- specific promoter to limit therapeutic transgene expression to cancerous cells. Our hypothesis is that this combined approach will allow all tumor cells, including highly invasive tumor cells that cause tumor relapse, to be selectively transfected, thereby minimizing potential side effects by eliminating gene transfer to healthy cells.