Noninvasive de novo imaging of human embryonic stem cell-derived teratoma formation.
Noninvasive de novo imaging of human embryonic stem cell-derived teratoma formation.
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DOI：
10.1158/0008-5472.can-08-4122
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发表时间：
2009-04-01
期刊：
影响因子：
11.2
通讯作者：
Wu JC
中科院分区：
文献类型：
--
作者：
Cao F;Li Z;Lee A;Liu Z;Chen K;Wang H;Cai W;Chen X;Wu JC
畸胎瘤的形成可能是人类胚胎干细胞(HES)治疗性移植后的一个严重缺陷。因此，畸胎瘤的非侵入性成像可能是监测接受HES细胞治疗的患者的有价值的工具。在这里，我们研究了人胚胎干细胞来源的畸胎瘤内血管生成的过程，现在报道了第一个使用64Cu标记的RGD四聚体(64CuDOTA-RGD4)通过靶向αvβ3整合素进行畸胎瘤形成的正电子发射断层扫描成像的例子。将稳定表达萤火虫荧光素酶和增强型绿色荧光蛋白(Fluc-EGFP)的H9 HES细胞(2×106)注射到12只成年裸鼠皮下。移植8周后，纵向生物发光成像证实，这些HES细胞移植物演变成畸胎瘤。在微正电子发射断层扫描下，2-脱氧-2-[18F]氟-D-葡萄糖和3‘-脱氧-3’-[18F]-氟胸苷均未能检出HES细胞来源的畸胎瘤(分别为0.8±0.5和1.1±0.4%ID/g；与背景信号相比，P=不显著)。与之相比，64Cu-DOTA-RGD4在血管性畸胎瘤中有明显的特异性摄取，而在低整合表达的对照肿瘤(人卵巢癌2008细胞系)中摄取显著降低(10.1±3.4比1.4±1.2%ID/g；P<；0.01)。CD31和β3整合素的免疫荧光染色也支持我们的体内成像结果(P<；0.05)。此外，我们还发现，从畸胎瘤分离的细胞比2008年的细胞表现出更高的αvβ3整合素表达。综上所述，通过靶向αvβ3整合素，我们首次成功地展示了64Cu-DOTA-RGD4无创性地显示体内畸胎瘤形成的能力。
Teratoma formation can be a serious drawback after the therapeutic transplantation of human embryonic stem (hES) cells. Therefore, noninvasive imaging of teratomas could be a valuable tool for monitoring patients undergoing hES cell treatment. Here, we investigated the angiogenic process within teratomas derived from hES cells and now report the first example of using 64Cu-labeled RGD tetramer (64Cu-DOTA-RGD4) for positron emission tomography imaging of teratoma formation by targeting αvβ3 integrin. H9 hES cells (2 × 106), stably expressing firefly luciferase, and enhanced green fluorescence protein (Fluc-eGFP) were injected into adult nude mice (n = 12) s.c. Eight weeks after transplantation, these hES cell grafts evolved into teratomas as confirmed by longitudinal bioluminescence imaging. Under micropositron emission tomography imaging, 2-deoxy-2-[18F]fluoro-D-glucose and 3′-deoxy-3′-[18F]-fluorothymidine both failed to detect hES cell–derived teratomas (0.8 ± 0.5 versus 1.1 ± 0.4 %ID/g, respectively; P = not significant versus background signals). By contrast, 64Cu-DOTA-RGD4 revealed specific and prominent uptake in vascularized teratoma and significantly lower uptake in control tumors (human ovarian carcinoma 2008 cell line), which had low intergrin expression (10.1 ± 3.4 versus 1.4 ± 1.2 %ID/g; P < 0.01). Immunofluorescence staining of CD31 and β3 integrin also supported our in vivo imaging results (P < 0.05). Moreover, we found that the cells dissociated from teratomas showed higher αvβ3 integrin expression than the 2008 cells. In conclusion, by targeting αvβ3 integrin, we successfully showed the ability of 64Cu-DOTA-RGD4 to noninvasively visualize teratoma formation in vivo for the first time.