TARGETED DELIVERY OF siRNA TO CELL DEATH PROTEINS IN SEPSIS
TARGETED DELIVERY OF siRNA TO CELL DEATH PROTEINS IN SEPSIS
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DOI：
10.1097/shk.0b013e318194bcee
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发表时间：
2009-08-01
期刊：
影响因子：
3.1
通讯作者：
Hotchkiss, Richard S.
中科院分区：
文献类型：
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作者：
Brahmamdam, Pavan;Watanabe, Eizo;Hotchkiss, Richard S.
免疫抑制是脓毒症患者发病和死亡的主要原因。免疫效应细胞如CD4 T和B细胞的凋亡损失是脓毒症中免疫能力损失的关键组成部分。淋巴细胞凋亡的抑制导致败血症动物模型的存活率提高。使用定量实时聚合酶链反应分离的脾脏CD4 T和B细胞，我们确定，Bim和CD4A，两个关键的细胞死亡蛋白，在脓毒症过程中显着上调。众所周知，淋巴细胞很难用小干扰RNA（siRNA）进行干扰。因此，一种新的，环糊精聚合物为基础的，转铁蛋白受体靶向的，传递媒介物被用来共同管理siRNA的Bim和BiNA的小鼠盲肠结扎和穿刺后立即。基于抗凋亡siRNA的治疗显著降低淋巴细胞凋亡，并防止脾脏CD4 T和B细胞的丢失。流式细胞术证实了siRNA向CD4 T和B细胞的体内递送，并且还证实了细胞内Bim和CD4A蛋白的减少。总之，Bim和CD4A是脓毒症中免疫细胞死亡的两个关键介质。使用一种新的基于环糊精聚合物的转铁蛋白受体靶向siRNA递送载体，能够有效地将抗凋亡siRNA施用至淋巴细胞，并逆转免疫细胞耗竭，这是这种高度致命性疾病的标志。
Immune suppression is a major cause of morbidity and mortality in the patients with sepsis. Apoptotic loss of immune effector cells such as CD4 T and B cells is a key component in the loss of immune competence in sepsis. Inhibition of lymphocyte apoptosis has led to improved survival in animal models of sepsis. Using quantitative real-time polymerase chain reaction of isolated splenic CD4 T and B cells, we determined that Bim and PUMA, two key cell death proteins, are markedly upregulated during sepsis. Lymphocytes have been notoriously difficult to transfect with small interfering RNA (siRNA). Consequently a novel, cyclodextrin polymer-based, transferrin receptor-targeted, delivery vehicle was used to coadminister siRNA to Bim and PUMA to mice immediately after cecal ligation and puncture. Antiapoptotic siRNA-based therapy markedly decreased lymphocyte apoptosis and prevented the loss of splenic CD4 T and B cells. Flow cytometry confirmed in vivo delivery of siRNA to CD4 T and B cells and also demonstrated decreases in intracellular Bim and PUMA protein. In conclusion, Bim and PUMA are two critical mediators of immune cell death in sepsis. Use of a novel cyclodextrin polymer-based, transferrin receptor-targeted siRNA delivery vehicle enables effective administration of antiapoptotic siRNAs to lymphocytes and reverses the immune cell depletion that is a hallmark of this highly lethal disorder.