Intravesical Delivery of Small Activating RNA Formulated into Lipid Nanoparticles Inhibits Orthotopic Bladder Tumor Growth
Intravesical Delivery of Small Activating RNA Formulated into Lipid Nanoparticles Inhibits Orthotopic Bladder Tumor Growth
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DOI：
10.1158/0008-5472.can-12-1871
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发表时间：
2012-10-01
期刊：
影响因子：
11.2
通讯作者：
Li, Long-Cheng
中科院分区：
文献类型：
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作者：
Kang, Moo Rim;Yang, Glen;Li, Long-Cheng
提高体内蛋白质生产的实用方法仍然是一个挑战。RNA激活（RNAa）作为一种潜在的解决方案正在兴起，它利用双链RNA（dsRNA）来增加内源基因表达。这种方法虽然与RNA干扰（RNAi）相关，但引发的反应与基因沉默相反。双链dsP21 - 322及其化学修饰变体是基于RNAa的药物的例子，它们通过诱导肿瘤抑制因子p21（WAF1/CIP1）（p21）的表达来抑制癌细胞生长。在这项研究中，我们通过将一种2'-氟修饰衍生物（dsP21 - 322 - 2'F）制成脂质纳米颗粒（LNP）用于膀胱内给药，来研究dsP21 - 322在膀胱癌原位模型中的治疗潜力。LNP的组成基于在基于RNAi的疗法中使用的临床相关配方，由用脂质DLin - KC2 - DMA构建的聚乙二醇稳定的单层脂质体组成。我们证实了用LNP配制的dsP21 - 322 - 2'F（LNP - dsP21 - 322 - 2'F）或其一种未配制的变体处理后，在体外可诱导p21、引起细胞周期停滞和细胞凋亡。2'-氟修饰和LNP配方也提高了双链在尿液中的稳定性。将LNP - dsP21 - 322 - 2'F膀胱内递送到小鼠膀胱中会导致尿路上皮摄取，并延长患有原位人膀胱癌小鼠的生存期。LNP - dsP21 - 322 - 2'F治疗还促进体内p21激活，导致40%的治疗小鼠肿瘤消退/消失。我们的结果为通过膀胱内给予LNP配制的RNA双链体治疗膀胱癌的新方法提供了临床前概念验证。《癌症研究》；72(19)；5069 - 79。（C）2012美国癌症研究协会。
Practical methods for enhancing protein production in vivo remain a challenge. RNA activation (RNAa) is emerging as one potential solution by using double-stranded RNA (dsRNA) to increase endogenous gene expression. This approach, although related to RNA interference (RNAi), facilitates a response opposite to gene silencing. Duplex dsP21-322 and its chemically modified variants are examples of RNAa-based drugs that inhibit cancer cell growth by inducing expression of tumor suppressor p21(WAF1/CIP1) (p21). In this study, we investigate the therapeutic potential of dsP21-322 in an orthotopic model of bladder cancer by formulating a 2'-fluoro-modified derivative (dsP21-322-2'F) into lipid nanoparticles (LNP) for intravesical delivery. LNP composition is based upon clinically relevant formulations used in RNAi-based therapies consisting of PEG-stabilized unilamellar liposomes built with lipid DLin-KC2-DMA. We confirm p21 induction, cell-cycle arrest, and apoptosis in vitro following treatment with LNP-formulated dsP21-322-2'F (LNP-dsP21-322-2'F) or one of its nonformulated variants. Both 2'-fluoro modification and LNP formulation also improve duplex stability in urine. Intravesical delivery of LNP-dsP21-322-2'F into mouse bladder results in urothelium uptake and extends survival of mice with established orthotopic human bladder cancer. LNP-dsP21-322-2'F treatment also facilitates p21 activation in vivo leading to regression/disappearance of tumors in 40% of the treated mice. Our results provide preclinical proof-of-concept for a novel method to treat bladder cancer by intravesical administration of LNP-formulated RNA duplexes. Cancer Res; 72(19); 5069-79. (C)2012 AACR.