Choi Ki Young, Correa Santiago, Min Jouha, Li Jiahe, Roy Sweta, Laccetti Kristiana H, Dreaden Erik, Kong Stephanie, Heo Roun, Roh Young Hoon, Lawson Edward C, Palmer Peter A, Hammond Paula T
Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
Adv Funct Mater. 2019 May 16;29(20). doi: 10.1002/adfm.201900018. Epub 2019 Mar 27.
Using siRNA therapeutics to treat hematologic malignancies has been unsuccessful because blood cancer cells exhibit remarkable resistance to standard transfection methods. Herein we report the successful delivery of siRNA therapeutics with a dual-targeted, layer-by-layer nanoparticle (LbL-NP). The LbL-NP protects siRNA from nucleases in the bloodstream by embedding it within polyelectrolyte layers that coat a polymeric core. The outermost layer consists of hyaluronic acid (a CD44-ligand) covalently conjugated to CD20 antibodies. The CD20/CD44 dual-targeting outer layer provides precise binding to blood cancer cells, followed by receptor-mediated endocytosis of the LbL-NP. We use this siRNA delivery platform to silence B-cell lymphoma 2 (BCL-2), a pro-survival protein, and . The dual-targeting approach significantly enhanced internalization of BCL-2 siRNA in lymphoma and leukemia cells, which led to significant downregulation of BCL-2 expression. Systemic administration of the dual-targeted, siRNA-loaded nanoparticle induced apoptosis and hampered proliferation of blood cancer cells both in cell culture and in orthotopic non-Hodgkin's lymphoma animal models. These results provide the basis for approaches to targeting blood-borne cancers and other diseases, and suggest that LbL nanoassemblies are a promising approach for delivering therapeutic siRNA to hematopoetic cell types that are known to evade transfection by other means.
使用小干扰RNA（siRNA）疗法治疗血液系统恶性肿瘤一直未获成功，因为血癌细胞对标准转染方法表现出显著抗性。在此，我们报告了利用一种双靶向逐层纳米颗粒（LbL-NP）成功递送siRNA疗法。LbL-NP通过将siRNA包埋于覆盖聚合物核心的聚电解质层内，保护其免受血流中核酸酶的降解。最外层由共价连接至CD20抗体的透明质酸（一种CD44配体）组成。CD20/CD44双靶向外层可与血癌细胞精确结合，随后LbL-NP通过受体介导的内吞作用进入细胞。我们利用这个siRNA递送平台使促生存蛋白B细胞淋巴瘤-2（BCL-2）沉默。这种双靶向方法显著增强了淋巴瘤和白血病细胞中BCL-2 siRNA的内化，导致BCL-2表达显著下调。在细胞培养以及原位非霍奇金淋巴瘤动物模型中，全身给予负载siRNA的双靶向纳米颗粒均可诱导血癌细胞凋亡并抑制其增殖。这些结果为靶向血源性癌症和其他疾病的方法提供了依据，并表明逐层纳米组装体是一种很有前景的方法，可将治疗性siRNA递送至已知会通过其他方式逃避转染的造血细胞类型。
