Systemic Gene Silencing in Primary T Lymphocytes Using Targeted Lipid Nanoparticles
Systemic Gene Silencing in Primary T Lymphocytes Using Targeted Lipid Nanoparticles
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DOI：
10.1021/acsnano.5b02796
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发表时间：
2015-07-01
期刊：
影响因子：
17.1
通讯作者：
Peer, Dan
中科院分区：
文献类型：
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作者：
Ramishetti, Srinivas;Kedmi, Ranit;Peer, Dan
通过使用RNA干扰（RNAi）下调特定基因来调节T细胞功能，在推进许多免疫相关疾病（包括癌症、炎症、自身免疫和病毒感染）的靶向治疗方面具有巨大潜力。一般来说，造血细胞，特别是初级T淋巴细胞，众所周知很难用小干扰RNA（siRNA）进行干扰。在此，我们描述了一种新的策略，使用靶向脂质纳米颗粒（tLNP）特异性地将siRNA递送至小鼠CD4（+）T细胞。为了增加siRNA递送的功效，这些tLNP已经与设计用于改善siRNA递送的稳定性和功效的几种脂质一起配制。用抗CD4单克隆抗体对tLNP进行表面功能化，以允许siRNA特异性地递送至CD4（+）T淋巴细胞。离体，tLNP通过仅靶向原代CD4（+）T淋巴细胞而不靶向其他细胞类型来证明特异性。这些颗粒的全身静脉内给药导致在几个解剖部位（包括脾脏、腹股沟淋巴结、血液和骨髓）中有效结合和吸收到CD4（+）T淋巴细胞中。tLNP的沉默发生在循环和静息CD 4（+）T淋巴细胞的亚群中。有趣的是，我们表明tLNP内化而不是内体逃逸是早在全身给药后1小时发生的基本事件，并决定了tLNP的功效。总之，这些结果表明，tLNPs可能为操纵T细胞功能开辟新的途径，并可能有助于建立RNAi作为白细胞相关疾病的治疗方式。
Modulating T cell function by down-regulating specific genes using RNA interference (RNAi) holds tremendous potential in advancing targeted therapies in many immune-related disorders including cancer, inflammation, autoimmunity, and viral infections. Hematopoietic cells, in general, and primary T lymphocytes, in particular, are notoriously hard to transfect with small interfering RNAs (siRNAs). Herein, we describe a novel strategy to specifically deliver siRNAs to murine CD4(+) T cells using targeted lipid nanoparticles (tLNPs). To increase the efficacy of siRNA delivery, these tLNPs have been formulated with several lipids designed to improve the stability and efficacy of siRNA delivery. The tLNPs were surface-functionalized with anti-CD4 monoclonal antibody to permit delivery of the siRNAs specifically to CD4(+) T lymphocytes. Ex vivo, tLNPs demonstrated specificity by targeting only primary CD4(+) T lymphocytes and no other cell types. Systemic intravenous administration of these particles led to efficient binding and uptake into CD4(+) T lymphocytes in several anatomical sites including the spleen, inguinal lymph nodes, blood, and the bone marrow. Silencing by tLNPs occurs in a subset of circulating and resting CD4(+) T lymphocytes. Interestingly, we show that tLNP internalization and not endosome escape is a fundamental event that takes place as early as 1 h after systemic administration and determines tLNPs' efficacy. Taken together, these results suggest that tLNPs may open new avenues for the manipulation of T cell functionality and may help to establish RNAi as a therapeutic modality in leukocyte-associated diseases.