High-throughput Optimization of Polymeric Nanoparticles for Small RNA Delivery to Treat Glioblastoma

用于治疗胶质母细胞瘤的小 RNA 递送的聚合物纳米颗粒的高通量优化

• 批准号: 10314020
• 负责人: Yuan Rui
• 金额: --
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-06 至 2021-07-07
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10314020
• 关键词: 3-Dimensional; Aftercare; Biocompatible Materials; Biological Assay; Blood - brain barrier anatomy; Bypass; Cell Communication; Cell model; Cells; Cerebrospinal Fluid; Chemotherapy and/or radiation; Clinical Trials; Data; Diagnosis; Diffuse; Diffusion; Drug Delivery Systems; Engineering; Environment; Epigenetic Process; Esters; Excision; Formulation; Generations; Genetic; Glioblastoma; Group Structure; Heterogeneity; Human; In Vitro; Infiltration; Knowledge; Libraries; Malignant Neoplasms; Malignant neoplasm of brain; Mediating; Methods; MicroRNAs; Mus; Neuraxis; Neurons; Operative Surgical Procedures; Pathway interactions; Patients; Penetration; Performance; Permeability; Phenotype; Polymer Chemistry; Polymers; Property; Pump; RNA delivery; Recurrence; Reporter Genes; Research; Signal Transduction; Small Interfering RNA; Small RNA; Specificity; Structure; Structure-Activity Relationship; Surface; System; Techniques; Technology; Therapeutic; Tumor Burden; Tumor Tissue; United States; Untranslated RNA; Validation; Work; Xenograft procedure; aggressive therapy; base; biodegradable polymer; brain tissue; cancer cell; cancer diagnosis; cell type; clinical application; clinical translation; clinically translatable; colloidal nanoparticle; combinatorial; design; experimental study; gene therapy; high throughput screening; human model; improved; in vitro Assay; in vivo; innovation; insight; knock-down; migration; nanomedicine; nanoparticle; nanoparticle delivery; neoplastic cell; next generation; novel; novel therapeutic intervention; screening; self-renewal; surface coating; targeted treatment; therapeutic nanoparticles; tumor; tumor growth; tumorigenesis; uptake

Nearly 50,000 new cases of glioblastoma (GBM) are diagnosed in the United States each year, with a dismal median survival of 14.6 months. Currently available therapeutics are largely ineffective due to the genetic, epigenetic, and signaling heterogeneity within the GBM tumor. Non-coding small RNAs such as short interfering RNA and micro-RNA are emerging as potent epigenetic regulators of cell fate and oncogenesis, and represent a promising tailored therapeutic strategy to counter tumor cell heterogeneity. However, clinical translation of small RNAs has been limited by significant knowledge gaps regarding their safe and effective delivery to GBM cells. The overall objective of this study is to use high-throughput screening approaches to optimize poly(beta-amino ester) (PBAE) polymeric nanoparticles for therapeutic small RNA delivery to treat GBM. Our preliminary data have shown that 1st generation PBAE materials enabled small RNA delivery to inhibit GBM proliferative phenotype in vitro and significantly slowed GBM tumor growth in GBM xenografts in vivo. However, these nanoparticles need to be optimized in delivery efficiency, biomaterial-mediated tumor targeting, long-term nanoparticle colloidal stability, and permeation throughout the tumor bulk to further their clinical translatability. To develop optimized 2nd generation PBAE nanoparticle formulations, the proposed work will utilize novel high-throughput approaches to generate polymer structural diversity and screen hundreds of unique polymer structures in parallel to identify delivery materials of improved potency and cancer targeting. In Aim 1, innovative in vitro assays examining nanoparticle performance in overcoming critical intracellular delivery barriers such as nanoparticle uptake and endosomal escape will be performed in primary patient- derived GBM cell models to better predict nanoparticle performance in vivo. Furthermore, these assays will yield important structure-functional relationships on how biomaterial structures can be altered to control their interactions with cells in a cancer-selective manner. In Aim 2, nanoparticle surface engineering techniques will be employed to enhance nanoparticle stability and tumor penetration capabilities. Orthotopic GBM tumor bearing mice will be treated with optimized nanoparticle formulations to characterize nanoparticle diffusion throughout the tumor bulk. This is critical in achieving uniform nanoparticle delivery as well as in reaching infiltrative GBM cells at the tumor periphery, which are primarily responsible for tumor recurrence after treatment. Finally, in Aim 3, nanoparticles carrying two GBM-inhibiting micro-RNAs will be evaluated for their ability to reduce GBM proliferation and self-renewal. State of the art primary human GBM cell models will be used to assess nanoparticle-induced phenotypic changes in vitro, and nanoparticles will also be infused into tumor-bearing mice to assess therapeutic delivery in the 3D tumor environment in vivo. These findings will have substantial positive impact on developing a scalable, bio-degradable small RNA delivery system to treat brain cancer.

在美国，每年诊断出近50，000例胶质母细胞瘤（GBM）新病例， 中位生存期为14.6个月。目前可用的治疗剂由于遗传原因在很大程度上是无效的， GBM肿瘤内的表观遗传和信号异质性。非编码小RNA，如短 干扰RNA和微小RNA正在成为细胞命运和肿瘤发生的有效表观遗传调节因子， 代表了一种有前景的定制治疗策略，以对抗肿瘤细胞异质性。但临床 小RNA的翻译受到关于其安全性和有效性的重大知识缺口的限制， 递送至GBM细胞。本研究的总体目标是使用高通量筛选方法， 优化用于治疗性小RNA递送的聚（β-氨基酯）（PBAE）聚合物纳米颗粒， GBM。我们的初步数据表明，第一代PBAE材料能够将小RNA递送到 抑制体外GBM增殖表型并显著减缓GBM异种移植物中GBM肿瘤生长， vivo.然而，这些纳米颗粒需要在递送效率、生物材料介导的肿瘤 靶向、长期纳米颗粒胶体稳定性和贯穿肿瘤块的渗透，以进一步提高其 临床可译性为了开发优化的第二代PBAE纳米颗粒制剂，所提出的工作 将利用新的高通量方法来产生聚合物结构多样性，并筛选数百种 独特的聚合物结构，以确定提高效力和癌症靶向的递送材料。在 目的1，创新的体外试验，检查纳米颗粒在克服关键细胞内 将在原发性患者中进行递送屏障，例如纳米颗粒摄取和内体逃逸， 衍生的GBM细胞模型，以更好地预测体内纳米颗粒的性能。此外，这些分析将 产生重要的结构-功能关系，关于如何改变生物材料的结构以控制其 以癌症选择性的方式与细胞相互作用。在目标2中，纳米颗粒表面工程技术将 用于增强纳米颗粒稳定性和肿瘤穿透能力。原位GBM肿瘤 将用优化的纳米颗粒制剂处理荷瘤小鼠以表征纳米颗粒扩散 在整个肿瘤块中。这对于实现均匀的纳米颗粒递送以及实现纳米颗粒的纳米化是至关重要的。 在肿瘤周围的浸润性GBM细胞，其主要负责肿瘤复发后， 治疗最后，在目标3中，将评估携带两种GBM抑制性微RNA的纳米颗粒的生物学活性。 减少GBM增殖和自我更新的能力。现有技术水平的原代人GBM细胞模型将是 用于评估纳米颗粒诱导的体外表型变化，纳米颗粒也将被注入 荷瘤小鼠体内3D肿瘤环境中评估治疗递送。这些发现将 对开发一种可扩展的、可生物降解的小RNA递送系统以治疗 脑癌