Department of Medicine, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, AB, Canada.
Department of Chemical and Materials Engineering, Faculty of Engineering, University of Alberta, Edmonton, AB, Canada.
PLoS One. 2021 Jun 22;16(6):e0251719. doi: 10.1371/journal.pone.0251719. eCollection 2021.
Overexpression and persistent activation of STAT5 play an important role in the development and progression of acute lymphoblastic leukemia (ALL), the most common pediatric cancer. Small interfering RNA (siRNA)-mediated downregulation of STAT5 represents a promising therapeutic approach for ALL to overcome the limitations of current treatment modalities such as high relapse rates and poor prognosis. However, to effectively transport siRNA molecules to target cells, development of potent carriers is of utmost importance to surpass hurdles of delivery. In this study, we investigated the use of lipopolymers as non-viral delivery systems derived from low molecular weight polyethylenimines (PEI) substituted with lauric acid (Lau), linoleic acid (LA) and stearic acid (StA) to deliver siRNA molecules to ALL cell lines and primary samples. Among the lipid-substituted polymers explored, Lau- and LA-substituted PEI displayed excellent siRNA delivery to SUP-B15 and RS4;11 cells. STAT5A gene expression was downregulated (36-92%) in SUP-B15 and (32%) in RS4;11 cells using the polymeric delivery systems, which consequently reduced cell growth and inhibited the formation of colonies in ALL cells. With regard to ALL primary cells, siRNA-mediated STAT5A gene silencing was observed in four of eight patient cells using our leading polymeric delivery system, 1.2PEI-Lau8, accompanied by the significant reduction in colony formation in three of eight patients. In both BCR-ABL positive and negative groups, three of five patients demonstrated marked cell growth inhibition in both MTT and trypan blue exclusion assays using 1.2PEI-Lau8/siRNA complexes in comparison with their control siRNA groups. Three patient samples did not show any positive results with our delivery systems. Differential therapeutic responses to siRNA therapy observed in different patients could result from variable genetic profiles and patient-to-patient variability in delivery. This study supports the potential of siRNA therapy and the designed lipopolymers as a delivery system in ALL therapy.
STAT5 的过度表达和持续激活在急性淋巴细胞白血病（ALL）的发展和进展中起着重要作用，ALL 是最常见的儿科癌症。小干扰 RNA（siRNA）介导的 STAT5 下调代表了 ALL 的一种有前途的治疗方法，可以克服当前治疗方法的局限性，例如高复发率和预后不良。然而，为了有效地将 siRNA 分子递送到靶细胞，开发有效的载体对于克服递送来的障碍至关重要。在这项研究中，我们研究了使用脂质聚合物作为源自低分子量聚乙烯亚胺（PEI）的非病毒递送系统，该系统用月桂酸（Lau）、亚油酸（LA）和硬脂酸（StA）取代，以将 siRNA 分子递送到 ALL 细胞系和原代样本。在所探索的脂质取代聚合物中，Lau 和 LA 取代的 PEI 显示出对 SUP-B15 和 RS4;11 细胞的出色 siRNA 递送。使用聚合物递送系统，SUP-B15 和 RS4;11 细胞中的 STAT5A 基因表达下调（36-92%），这相应地降低了 ALL 细胞的生长并抑制了集落的形成。关于 ALL 原代细胞，使用我们的领先聚合物递送系统 1.2PEI-Lau8，在八位患者细胞中的四位观察到 STAT5A 基因沉默，同时在八位患者中的三位患者中显著减少了集落形成。在 BCR-ABL 阳性和阴性组中，与对照 siRNA 组相比，使用 1.2PEI-Lau8/siRNA 复合物在五名患者中的三名患者中，MTT 和台盼蓝排除测定法均观察到明显的细胞生长抑制。我们的递送系统对三位患者样本没有显示出任何阳性结果。不同患者对 siRNA 治疗的不同治疗反应可能是由于遗传谱的差异和患者间的递送变异性所致。这项研究支持 siRNA 治疗和设计的脂质体作为 ALL 治疗中的递送系统的潜力。
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