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

项目摘要/摘要 该项目的重点是开发一种最佳的脑穿透DNA纳米颗粒配方，用于 基因治疗在胶质母细胞瘤治疗中的作用。胶质母细胞瘤是最常见和最具侵袭性的肿瘤。 原发脑瘤，但目前可用的治疗方法有严重的副作用，这种疾病仍然存在 一律致命。基因治疗是一种潜在的强大策略，已在临床前研究中显示出前景。 然而，有效的基因治疗尚未在人类身上实现，这在很大程度上是由于无法实现 基因载体在大脑中的广泛分布和肿瘤选择性基因转移，这是 基底膜的高度侵袭性。我们已经开发出合成的携带DNA的纳米颗粒，能够 避免由于小颗粒(&lt；&lt；100 nm)和 致密的聚乙二醇涂层。这些系统能够穿透整个大鼠大脑的纹状体。 当通过对流强化输送(CED)给药时。我们的试验数据进一步表明，这些DNA 载药脑穿透纳米粒(DNA-BPN)提供更广泛和更多的转基因 CED后健康脑实质和脑肿瘤组织中表达的比较 标准的DNA纳米颗粒不能有效地穿透输液部位。我们将发展和 彻底测试由多种有前景的核心聚合物组成的DNA-BPN，并比较它们在 从体外、体外和体内到黄金标准系统和领先的基于病毒的载体。从第一代开始 DNA-BPN似乎已经能够安全地转染大鼠大脑的一大部分，我们计划调查 其他有希望的聚合物，可能提供更高的转染率，以及使用高度肿瘤- 特定启动子将治疗性转基因表达限制在癌细胞中。我们的假设是，这是 联合疗法将允许所有肿瘤细胞，包括导致肿瘤复发的高侵袭性肿瘤细胞， 被选择性地转基因，从而通过消除对健康的基因转移而将潜在的副作用降至最低 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.