Strategy for dual-analyte luciferin imaging: in vivo bioluminescence detection of hydrogen peroxide and caspase activity in a murine model of acute inflammation.
Strategy for dual-analyte luciferin imaging: in vivo bioluminescence detection of hydrogen peroxide and caspase activity in a murine model of acute inflammation.
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DOI：
10.1021/ja309078t
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发表时间：
2013-02-06
影响因子：
15
通讯作者：
Chang, Christopher J.
中科院分区：
文献类型：
--
作者：
Van de Bittner, Genevieve C.;Bertozzi, Carolyn R.;Chang, Christopher J.
体内分子成像有望了解健康，损伤，衰老和疾病的潜在机制，因为它可以检测不同的生化过程，如酶活性，反应性小分子通量或翻译后修饰。目前的成像技术往往只检测一个单一的生化过程，但在整个生物体，多种类型的生化事件有助于生理和病理表型。在这份报告中，我们提出了一个通用的策略，用于双分析物检测活体动物，采用原位形成的萤火虫荧光蛋白从两个互补的笼前体，可以通过不同的生化过程被揭露。为了建立这种方法，我们已经开发了过氧笼状荧光素-2（PCL-2），一种H2 O2响应性硼酸探针，在与这种活性氧（ROS）反应时释放6-羟基-2-氰基苯并噻唑（HCBT），以及一种基于肽的探针，Ile-Glu-Thr-Asp-D-Cys（IETDC），其在活性半胱天冬酶8的存在下释放D-半胱氨酸。一旦释放，HCBT和D-半胱氨酸在原位形成萤火虫荧光，当且仅当两种化学触发剂都进行时，才会产生生物发光信号。因此，该系统构成了一个AND型分子逻辑门，报告同时存在的H2 O2和半胱天冬酶8活性。使用这些探针，在体外和体内进行H2 O2或caspase 8活性的化学选择性成像。此外，PCL-2和IETDC在体内的同时使用建立了在活小鼠的急性炎症期间H2 O2和半胱天冬酶8活性的同时增加。总之，这种方法提供了一种潜在的强大的新的化学工具，用于研究在损伤，衰老和疾病期间活动物中同时发生的氧化应激和炎症过程，以及一种通用的方法，用于同时监测多种分析物，使用基于生物发光成像技术。
In vivo molecular imaging holds promise for understanding the underlying mechanisms of health, injury, aging, and disease, as it can detect distinct biochemical processes such as enzymatic activity, reactive small-molecule fluxes, or post-translational modifications. Current imaging techniques often detect only a single biochemical process, but, within whole organisms, multiple types of biochemical events contribute to physiological and pathological phenotypes. In this report, we present a general strategy for dual-analyte detection in living animals that employs in situ formation of firefly luciferin from two complementary caged precursors that can be unmasked by different biochemical processes. To establish this approach, we have developed Peroxy Caged Luciferin-2 (PCL-2), a H2O2-responsive boronic acid probe that releases 6-hydroxy-2-cyanobenzothiazole (HCBT) upon reacting with this reactive oxygen species (ROS), as well as a peptide-based probe, Ile-Glu-Thr-Asp-D-Cys (IETDC) which releases D-cysteine in the presence of active caspase 8. Once released, HCBT and D-cysteine form firefly luciferin in situ, giving rise to a bioluminescent signal if and only if both chemical triggers proceed. This system thus constitutes an AND-type molecular logic gate that reports on the simultaneous presence of H2O2 and caspase 8 activity. Using these probes, chemoselective imaging of either H2O2 or caspase 8 activity was performed in vitro and in vivo. Moreover, concomitant use of PCL-2 and IETDC in vivo establishes a concurrent increase in both H2O2 and caspase 8 activity during acute inflammation in living mice. Taken together, this method offers a potentially powerful new chemical tool for studying simultaneous oxidative stress and inflammation processes in living animals during injury, aging, and disease, as well as a versatile approach for concurrent monitoring of multiple analytes using luciferin-based bioluminescence imaging technologies.
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影响因子：
15
作者：
Dickinson BC;Huynh C;Chang CJ
通讯作者：
Chang CJ
影响因子：
29
作者：
Cochemé HM;Quin C;McQuaker SJ;Cabreiro F;Logan A;Prime TA;Abakumova I;Patel JV;Fearnley IM;James AM;Porteous CM;Smith RA;Saeed S;Carré JE;Singer M;Gems D;Hartley RC;Partridge L;Murphy MP
通讯作者：
Murphy MP
影响因子：
4.7
作者：
Choi SH;Aid S;Kim HW;Jackson SH;Bosetti F
通讯作者：
Bosetti F
影响因子：
16.6
作者：
Conley, Nicholas R.;Dragulescu-Andrasi, Anca;Rao, Jianghong;Moerner, W. E.
通讯作者：
Moerner, W. E.
DOI：
10.1073/pnas.2637010100
发表时间：
2004-01-06
影响因子：
11.1
作者：
Cao, YA;Wagers, AJ;Contag, CH
通讯作者：
Contag, CH