A self-quenched galactosamine-serum albumin - RhodamineX conjugate: A"Smart" fluorescent molecular Imaging probe synthesized with clinically applicable material for detecting peritoneal ovarian cancer Metastases
A self-quenched galactosamine-serum albumin - RhodamineX conjugate: A"Smart" fluorescent molecular Imaging probe synthesized with clinically applicable material for detecting peritoneal ovarian cancer Metastases
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DOI：
10.1158/1078-0432.ccr-07-1004
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发表时间：
2007-11-01
影响因子：
11.5
通讯作者：
Kobayashi, Hisataka
中科院分区：
文献类型：
--
作者：
Hama, Yukihiro;Urano, Yasuteru;Kobayashi, Hisataka
目的：在细胞内化靶向荧光标记缀合物后激活荧光团是一种有效的分子成像策略，可以增加目标与背景的比率。卵巢癌细胞上的d -半乳糖受体已被用于靶向自猝灭的亲和素罗丹明x偶联物，其中亲和素成分与d -半乳糖受体结合，罗丹明只有在细胞内化后才能通过脱猝而被光学激活。半乳糖胺偶联血清白蛋白（GmSA）作为一种非免疫原性的亲和蛋白替代品，以更高的结合亲和力靶向d -半乳糖受体，并且比亲和蛋白具有更多的rhodamineX偶联位点。实验设计：将GmSA与20个罗丹明x分子（GmSA- 20rox）偶联形成自猝灭复合物，并与GmSA与单个罗丹明x偶联物（GmSA- 1rox）在体外化学活化特性、细胞内活化及体内分子成像检测SHIN3卵巢癌腹膜微转移的效果进行比较。结果：与SHIN3卵巢癌细胞孵育3小时，GmSA-20ROX的亮度是GmSA-1ROX的5倍。由于肿瘤与背景比高，体内亚毫米SHIM卵巢癌植入物在腹腔内的光谱荧光成像清晰可见。GmSA-20ROX检测种植体的敏感性和特异性是通过将罗丹明x发射物与SHIN3肿瘤种植体中组成性表达的红色荧光蛋白共定位来确定的。在336个病变中，GmSA-20ROX的敏感性和特异性分别为99%/99%，而GmSA-1ROX的结果仅为24%/100% (n = 388)，对于直径类似于0.8 mm或更大的病变。结论：自淬灭的GmSA-20ROX比以往的d -半乳糖靶向荧光偶联物效率更高。
Purpose: Fluorophore activation after cellular internalization of a targeted fluorescently labeled conjugate is an effective molecular imaging strategy to increase target-to-background ratios. The D-galactose receptor on ovarian cancer cells has been used to target self-quenched avidinrhodamineX conjugates in which the avidin component binds to D-galactose receptor and the rhodamines are optically activated by dequenching only after cellular internalization. As a non-immunogenic alternative of avidin, galactosamine-conjugated serum albumin (GmSA) targets the D-galactose receptor with higher binding affinity and has more conjugation sites available for rhodamineX than avidin.Experimental Design: GmSA was conjugated with 20 rhodamineX molecules (GmSA-20ROX) to create a self-quenching complex, which was compared with a conjugate consisting of GmSA and a single rhodamineX (GmSA-1ROX) in ex vivo chemical activation characteristics, intracellular activation, and in vivo molecular imaging for detecting peritoneal micrometastases of SHIN3 ovarian cancer.Results: GmSA-20ROX was five times brighter than GmSA-1ROX when incubated with SHIN3 ovarian cancer cells for 3 h. Submillimeter SHIM ovarian cancer implants in the peritoneal cavity were clearly visualized in vivo with spectral fluorescence imaging due to the high tumor-to-background ratio. The sensitivity and specificity of GmSA-20ROX for implant detection were determined by colocalization of the rhodamineX emission with red fluorescent protein expressed constitutively in the SHIN3 tumor implants. Among 336 lesions, sensitivity and specificity were 99%/99%, respectively, for GmSA-20ROX, whereas the results for GmSA-1ROX were only 24%/100% (n = 388), respectively, for lesions similar to 0.8 mm or greater in diameter.Conclusion: Self-quenched GmSA-20ROX is more efficient than previous D-galactose-targeted fluorescent conjugates.