Noninvasive optical imaging of cysteine protease activity using fluorescently quenched activity-based probes
Noninvasive optical imaging of cysteine protease activity using fluorescently quenched activity-based probes
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DOI：
10.1038/nchembio.2007.26
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发表时间：
2007-10-01
影响因子：
14.8
通讯作者：
Bogyo, Matthew
中科院分区：
文献类型：
--
作者：
Blum, Galia;von Degenfeld, Georges;Bogyo, Matthew
我们已经产生了一系列基于淬灭的近红外荧光活性的探针（qNIRF-ABP），其共价靶向先前显示在肿瘤发生的多个阶段中重要的木瓜蛋白酶家族半胱氨酸蛋白酶.这些“智能”探针仅在共价修饰特定蛋白酶靶标后才发射荧光信号。将NIRF-ABPs静脉注射到移植瘤小鼠体内后，非侵入性全身成像可以直接监测组织蛋白酶活性。重要的是，探针的永久性还允许第二次离体生物化学分析以鉴定特定蛋白酶并将其活性与全身图像相关联。最后，我们证明，这些探针可以用于监测小分子抑制蛋白酶的目标生化和直接成像方法。因此，NIRF-ABP是（i）用于疾病诊断的潜在有价值的新成像剂和（ii）用于体内小分子治疗剂的临床前和临床测试的有力工具。
We have generated a series of quenched near- infrared fluorescent activity- based probes ( qNIRF- ABPs) that covalently target the papain- family cysteine proteases shown previously to be important in multiple stages of tumorigenesis. These ` smart' probes emit a fluorescent signal only after covalently modifying a specific protease target. After intravenous injection of NIRF- ABPs into mice bearing grafted tumors, noninvasive, whole- body imaging allowed direct monitoring of cathepsin activity. Importantly, the permanent nature of the probes also allowed secondary, ex vivo biochemical profiling to identify specific proteases and to correlate their activity with whole- body images. Finally, we demonstrate that these probes can be used to monitor small- molecule inhibition of protease targets both biochemically and by direct imaging methods. Thus, NIRF- ABPs are ( i) potentially valuable new imaging agents for disease diagnosis and ( ii) powerful tools for preclinical and clinical testing of small- molecule therapeutic agents in vivo.