Selective Lentiviral Gene Delivery to CD133-Expressing Human Glioblastoma Stem Cells
Selective Lentiviral Gene Delivery to CD133-Expressing Human Glioblastoma Stem Cells
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DOI：
10.1371/journal.pone.0116114
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发表时间：
2014-12-26
期刊：
影响因子：
3.7
通讯作者：
Placantonakis, Dimitris G.
中科院分区：
文献类型：
--
作者：
Bayin, N. Sumru;Modrek, Aram S.;Placantonakis, Dimitris G.
多形性胶质母细胞瘤（GBM）是一种致命的原发性脑恶性肿瘤。胶质母细胞瘤干细胞（GSC）具有自我更新和分化成肿瘤谱系的能力，由于其对当前治疗的抵抗，被认为会导致肿瘤复发。GSCs的一个子集以CD133的细胞表面表达为标志，CD133是一种糖基化的五跨膜蛋白。表达cd133的GSCs的研究一直受到特异性靶向它们的遗传工具相对缺乏的限制。在这里，我们提出了CD133- lv，这是一种慢病毒载体，在其包膜上呈现针对CD133的单链抗体，作为表达CD133的GSCs的选择性转导载体。我们发现CD133- lv以剂量依赖的方式选择性地转导CD133+人GSCs，并且转导的细胞保持其干细胞样特性。CD133- lv的转导效率被一种识别与病毒包膜上CD133相同表位的抗体和shrna介导的CD133的敲低所降低。相反，在原代人GBM培养物中，CD133- lv的转导率通过CD133的过表达而增强。CD133-LV在NOD患者颅内GBM异种移植物中选择性地转导cd133表达细胞。SCID小鼠，但保留正常小鼠脑组织，来源于人胚胎干细胞和原代人星形胶质细胞的神经元。我们的研究结果表明，CD133-LV代表了一种对表达cd133的GSCs进行选择性遗传操作的新工具，可以用来回答有关这些细胞如何促进肿瘤生物学和治疗耐药性的重要问题。
Glioblastoma multiforme (GBM) is a deadly primary brain malignancy. Glioblastoma stem cells (GSC), which have the ability to self-renew and differentiate into tumor lineages, are believed to cause tumor recurrence due to their resistance to current therapies. A subset of GSCs is marked by cell surface expression of CD133, a glycosylated pentaspan transmembrane protein. The study of CD133-expressing GSCs has been limited by the relative paucity of genetic tools that specifically target them. Here, we present CD133-LV, a lentiviral vector presenting a single chain antibody against CD133 on its envelope, as a vehicle for the selective transduction of CD133-expressing GSCs. We show that CD133-LV selectively transduces CD133+ human GSCs in dose-dependent manner and that transduced cells maintain their stem-like properties. The transduction efficiency of CD133-LV is reduced by an antibody that recognizes the same epitope on CD133 as the viral envelope and by shRNA-mediated knockdown of CD133. Conversely, the rate of transduction by CD133-LV is augmented by overexpression of CD133 in primary human GBM cultures. CD133-LV selectively transduces CD133-expressing cells in intracranial human GBM xenografts in NOD. SCID mice, but spares normal mouse brain tissue, neurons derived from human embryonic stem cells and primary human astrocytes. Our findings indicate that CD133-LV represents a novel tool for the selective genetic manipulation of CD133-expressing GSCs, and can be used to answer important questions about how these cells contribute to tumor biology and therapy resistance.