MicroRNA 142-3p attenuates spread of replicating retroviral vector in hematopoietic lineage-derived cells while maintaining an antiviral immune response.
MicroRNA 142-3p attenuates spread of replicating retroviral vector in hematopoietic lineage-derived cells while maintaining an antiviral immune response.
复制标题
MicroRNA 142-3p 减弱复制型逆转录病毒载体在造血谱系衍生细胞中的传播，同时维持抗病毒免疫反应。
DOI：
10.1089/hum.2012.216
复制
发表时间：
2014
影响因子：
4.2
通讯作者：
Jolly,DouglasJ
中科院分区：
文献类型：
--
作者：
Lin,AmyH;Timberlake,Nina;Logg,ChristopherR;Liu,Yanzheng;Kamijima,Shuichi;Diago,Oscar;Wong,Kenneth;Gammon,DawnK;Ostertag,Derek;Hacke,Katrin;Yang,EmilyC;Gruber,Harry;Kasahara,Noriyuki;Jolly,DouglasJ
我们正在开发一种编码胞嘧啶脱氨酶的逆转录病毒复制载体（RRV）作为胶质瘤的抗癌剂。尽管这种病毒对肿瘤具有天然的选择性和其他安全特性，但它可能通过有效感染健康组织而导致脱靶效应。在这里，我们研究了在RRV中掺入造血谱系特异性microRNA靶序列是否进一步限制了体外造血谱系来源的人细胞和体内小鼠淋巴组织中的复制。将与microRNA 142- 3 p的引导链完全互补的序列的一个或四个拷贝插入表达编码绿色荧光蛋白（GFP）的转基因的RRV基因组的3′非翻译区。通过qPCR、qRT-PCR和流式细胞术测量这些载体在体外和体内造血谱系细胞中的病毒传播和GFP表达。在造血谱系来源的人细胞系和原代人刺激的外周血单核细胞中，携带142- 3 pT序列的载体相对于亲本载体显示GFP表达显着降低，并且随着时间的推移没有观察到病毒传播。在同基因皮下小鼠肿瘤模型中，具有和不具有142- 3 pT序列的RRV在肿瘤细胞中同样良好地扩散;在血液、骨髓和脾脏中被强烈抑制;并产生抗病毒免疫应答。在免疫缺陷小鼠模型中，与未修饰的RRV相比，具有142- 3 pT序列的RRV在血液、骨髓和脾脏中被强烈抑制。基于组织特异性microRNA的RRV复制的选择性衰减可以维持抗病毒免疫，并且如果需要，为基因治疗应用的这种递送平台提供额外的保障。
We are developing a retroviral replicating vector (RRV) encoding cytosine deaminase as an anticancer agent for gliomas. Despite its demonstrated natural selectivity for tumors, and other safety features, such a virus could potentially cause off-target effects by productively infecting healthy tissues. Here, we investigated whether incorporation of a hematopoietic lineage-specific microRNA target sequence in RRV further restricts replication in hematopoietic lineage-derived human cellsin vitroand in murine lymphoid tissuesin vivo. One or four copies of a sequence perfectly complementary to the guide strand of microRNA 142-3p were inserted into the 3′ untranslated region of the RRV genome expressing the transgene encoding green fluorescent protein (GFP). Viral spread and GFP expression of these vectors in hematopoietic lineage cellsin vitroandin vivowere measured by qPCR, qRT-PCR, and flow cytometry. In hematopoietic lineage-derived human cell lines and primary human stimulated peripheral blood mononuclear cells, vectors carrying the 142-3pT sequence showed a remarkable decrease in GFP expression relative to the parental vector, and viral spread was not observed over time. In a syngeneic subcutaneous mouse tumor model, RRVs with and without the 142-3pT sequences spread equally well in tumor cells; were strongly repressed in blood, bone marrow, and spleen; and generated antiviral immune responses. In an immune-deficient mouse model, RRVs with 142-3pT sequences were strongly repressed in blood, bone marrow, and spleen compared with unmodified RRV. Tissue-specific microRNA-based selective attenuation of RRV replication can maintain antiviral immunity, and if needed, provide an additional safeguard to this delivery platform for gene therapy applications.
登录
查看更多内容
影响因子：
12.3
作者：
Monticelli, Silvia;Ansel, K Mark;Xiao, Changchun;Socci, Nicholas D;Krichevsky, Anna M;Thai, To-Ha;Rajewsky, Nikolaus;Marks, Debora S;Sander, Chris;Rajewsky, Klaus;Rao, Anjana;Kosik, Kenneth S
通讯作者：
Kosik, Kenneth S
影响因子：
6.7
作者：
Cawood R;Chen HH;Carroll F;Bazan-Peregrino M;van Rooijen N;Seymour LW
通讯作者：
Seymour LW
一系列鼠白血病病毒相关元件在逆转录病毒基因转移的灵长类受体中传播和表达
影响因子：
5.4
作者：
D. Purcell;Christine Broscius;E. Vanin;C. Buckler;A. Nienhuis;Malcolma . Martin
通讯作者：
Malcolma . Martin
影响因子：
12.4
作者：
Perez, Omar D.;Logg, Christopher R.;Jolly, Douglas J.
通讯作者：
Jolly, Douglas J.
造血细胞插入突变：从临床基因治疗试验中不良事件中吸取的教训
DOI：
--
发表时间：
2011
期刊：
影响因子：
--
作者：
L. Müller;M. Milsom;David A. Williams
通讯作者：
David A. Williams