Beasock Damian, Chandler Morgan, Radwan Yasmine, Liao Zhen, Ma Fuying, Webb Ken, Panigaj Martin, Lee Jeoung Soo, Afonin Kirill A
UNC Charlotte, NC, USA.
Drug Design Development and Delivery (4D) Laboratory, Department of Bioengineering, Clemson University, SC 29631, USA.
RNA Nanomed. 2024;1(1):79-90. doi: 10.59566/isrnn.2024.0101079.
Primary injury of the spinal cord is caused by a mechanical traumatic event which is rapidly followed by a secondary injury cascade that may evolve for several months leading to biological and functional changes. During the secondary injury, many pathophysiological pathways and process are activated including inflammation, oxidative stress, demyelination, excitotoxicity, axon degeneration, and cell death. The RhoA/Rho kinase pathway significantly contributes to spinal degeneration and regeneration and therefore represents a potential therapeutic target. Nucleic acid nanoparticles (NANPs) offer easy rational and programable design with the potential to carry on multiple synergistic therapeutic nucleic acid functional motifs. In this context we designed, synthesized, and assembled several representative NANPs decorated with multiple copies of siRNAs targeting RhoA. Subsequently we assessed NANPs' physicochemical properties, toxicity, and immunorecognition upon delivery with the nanocarrier PLGA-g-PEI (PgP), developed with the aim to select for the most immunoquiescent type of formulations. In addition, we observed that L1 neural cell adhesion molecule conjugated PgP (L1-PgP) efficiently delivered NANP-siRhoA in cultured neuroblastoma (B35) cells. RhoA mRNA expression was significantly reduced by all L1-PgP/ NANP-siRhoA relative to the untreated control, while no significant differences were observed between the different NANP-siRhoAs.
脊髓原发性损伤由机械性创伤事件引起，随后迅速出现继发性损伤级联反应，这种反应可能持续数月，导致生物学和功能变化。在继发性损伤期间，许多病理生理途径和过程被激活，包括炎症、氧化应激、脱髓鞘、兴奋毒性、轴突退变和细胞死亡。RhoA/Rho激酶途径对脊髓退变和再生有显著影响，因此是一个潜在的治疗靶点。核酸纳米颗粒（NANPs）具有易于合理设计和可编程的特点，有潜力携带多种协同治疗性核酸功能基序。在此背景下，我们设计、合成并组装了几种用靶向RhoA的多个小干扰RNA（siRNAs）拷贝修饰的代表性NANPs。随后，我们评估了NANPs的物理化学性质、毒性以及与纳米载体聚乳酸-羟基乙酸共聚物-聚乙二醇化聚乙烯亚胺（PLGA-g-PEI，PgP）一起递送时的免疫识别情况，开发PgP的目的是选择免疫反应最小的制剂类型。此外，我们观察到L1神经细胞黏附分子偶联的PgP（L1-PgP）能有效地将NANP-siRhoA递送至培养的神经母细胞瘤（B35）细胞中。相对于未处理的对照，所有L1-PgP/NANP-siRhoA组的RhoA mRNA表达均显著降低，而不同的NANP-siRhoA之间未观察到显著差异。
