Synthesis and delivery of short, noncoding RNA by B lymphocytes
Synthesis and delivery of short, noncoding RNA by B lymphocytes
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DOI：
10.1073/pnas.1311145110
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发表时间：
2013-12-10
影响因子：
11.1
通讯作者：
Zanetti, Maurizio
中科院分区：
文献类型：
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作者：
Almanza, Gonzalo;Anufreichik, Veronika;Zanetti, Maurizio
进化上保守的短(20-30个核苷酸)非编码RNA(MicroRNAs)是多种生理和病理过程中基因表达的强大调节因子。因此，有效地调节microRNA功能的手段构成了一个重要的治疗机会。在这里，我们证明了原代B淋巴细胞可以用非病毒质粒DNA进行遗传编程，以用于针对microRNA-150(miR-150)的反义序列(反microRNA)的生物发生和传递。在转染抗miR-150的18小时内，原代B淋巴细胞每个细胞分泌的抗miR-150分子的拷贝数接近3,000份。CD8T细胞在体内外交叉激发过程中，B淋巴细胞释放的抗miR-150分子被CD8 T细胞内化，导致内源性miR-150表达明显下调。然而，在没有交叉启动的情况下，没有观察到这种内化。这些结果表明，将抗miR-150分子从B淋巴细胞运送到T细胞需要通过抗原受体激活受体T细胞。最后，在B细胞中合成的抗miR-150以游离和胞外囊泡相关组分的形式分泌，但CD8T细胞仅明显地摄取与胞外囊泡相关的抗miR-150。总而言之，这些数据表明，初级B淋巴细胞是合成和传递短的非编码RNA的有效平台，为免疫基因组疗法的方法铺平了道路。
Evolutionarily conserved short (20-30 nucleotides) noncoding RNAs (microRNAs) are powerful regulators of gene expression in a variety of physiological and pathological processes. As such, means to efficiently modulate microRNA function constitute an important therapeutic opportunity. Here we demonstrate that primary B lymphocytes can be genetically programmed with nonviral plasmid DNA for the biogenesis and delivery of antisense sequences (anti-microRNA) against microRNA-150 (miR-150). Within 18 h of transfection with an anti-miR-150 construct, primary B lymphocytes secrete similar to 3,000 copies of anti-miR-150 molecules per cell. Anti-miR-150 molecules released by B lymphocytes were internalized by CD8 T lymphocytes during cross-priming in vitro and in vivo, resulting in marked down-regulation of endogenous miR-150. However, such internalization was not observed in the absence of cross-priming. These results suggest that shuttling anti-miR-150 molecules from B lymphocytes to T cells requires the activation of receiver T cells via the antigen receptor. Finally, anti-miR-150 synthesized in B cells were secreted both as free and extracellular vesicle-associated fractions, but only extracellular vesicle-associated anti-miR-150 were apparently taken up by CD8 T cells. Collectively, these data indicate that primary B lymphocytes represent an efficient platform for the synthesis and delivery of short, noncoding RNA, paving the way for an approach to immunogenomic therapies.