Next generation gamma Peptide Nucleic Acids (yPNAs) for the treatment of ischemic stroke

摘要： Micro RNAs(MiRNAs)是一类短的非编码RNA，已被发现具有潜在的功能 用于研究和治疗许多疾病的工具，包括缺血性中风。由于它们的保守性 序列，使用合成的抗microRNA(anti-miR)试剂靶向特定的miRNAs使它们具有吸引力 药物开发的目标。我们最近证实了中风和影响中风反应的因素， 如社会隔离，可以调节miRNAs，特别是miR-141-3p。因此，miR-141-3p代表一个 有望成为中风治疗的新分子靶点。因此，通过开发量身定做的疗法 拮抗miR-141-3p，我们希望能为卒中提供一种有效的治疗方法。在这里，我们提出了一种多维的 应用现代技术推进一种有前途的中风疗法的学科项目，该疗法针对miR-141- 3P使用基于新一代反miR的新工具。在过去，我们展示了纳米技术- 递送肽核酸(PNA)为基础的miRNA抑制剂可以靶向miR-155用于淋巴瘤治疗。不像 大多数核酸，PNA是人工合成的DNA模拟物，其中磷酸二酯主链被替换为 一种中性的N-(2-氨基乙基)甘氨酸骨架。PNAs可以结合单链靶标，具有高度的特异性和 PNA具有亲和力，并且不受蛋白酶的影响，使其成为靶向miRNAs的理想分子。为了提高 进一步，我们将开发一类新的PNA类似物，命名为Gamma PNA(PNA)，它们是构象预先组织的，因此具有有利的结合和溶解能力 应增加其作为抗miR试剂的有效性的特性。在送货方面，我们将采用聚乳酸- 以聚乙醇酸(PLGA)为基础的纳米制剂。作为我们稳定的下一代PNA的原则证明- 基于抗miR-141-3p作为卒中治疗药物，我们建议测试其在卒中基础上的传递和疗效 患病的小鼠模型。在这里，我们将追求两个独立的具体目标：1)合成和质量控制 一组基于核酸的抗miR-141-3p在细胞培养检测中的分析；2)体内疗效测试 在患病小鼠模型中研究基于核酸的抗miR-141-3p变异体。我们已经组建了一个跨学科的 凭借在核酸化学、药物配方和疾病生物学方面的专业知识，我们的团队将实现我们的目标。

Summary: Micro RNAs (miRNAs) are a class of short non-coding RNAs that have been identified as potentially powerful tools for the study and treatment of many diseases, including ischemic stroke. Due to their conserved sequence, targeting specific miRNAs using synthetic anti-microRNA (anti-miR) reagents makes them attractive targets for drug development. We recently established that stroke and factors modifying stroke responses, such as social isolation, can modulate miRNAs, especially miR-141-3p. Hence, miR-141-3p represents a promising new molecular target for stroke therapy. Therefore, by developing therapeutics tailored to antagonize miR-141-3p, we hope to generate an effective therapy for stroke. Herein, we propose a multi- disciplinary project that applys modern technology to advance a promising stroke therapy that targets miR-141- 3p using novel next-generation anti-miR-based tools. In the past, we demonstrated that nanotechnology- delivered peptide nucleic acid (PNA)-based miRNA inhibitors can target miR-155 for lymphoma therapy. Unlike most nucleic acids, PNAs are synthetic DNA mimics in which the phosphodiester backbone is substituted with a neutral N-(2-aminoethyl) glycine backbone. PNAs can bind single-stranded targets with high specificity and affinity and are not susceptible to proteases, making PNAs ideal molecules for targeting miRNAs. To improve the effectiveness of anti-miR PNAs further, we will exploit a new class of PNA analogs designated gamma PNAs (PNAs), which are conformationally pre-organized and so have advantageous binding and solubility properties that should increase their effectiveness as anti-miR agents. For delivery, we will employ poly (lactic- co-glycolic acid) (PLGA)-based nanoformulations. As proof of principle for our stable next generation PNA- based anti-miR-141-3p as a stroke therapeutic, we propose to test delivery and efficacy in stroke-based diseased mouse models. Here, we will pursue two independent specific aims: 1) Synthesis and quality control analysis of an array of PNA-based anti-miR-141-3p in cell culture-based assays; and 2) Test in vivo efficacy of PNA-based anti-miR-141-3p variants in a diseased mouse model. We have assembled an interdisciplinary team to achieve our goals, with expertise in nucleic acid chemistry, drug formulation, and disease biology.