In vivo imaging of protease activity in arthritis -: A novel approach for monitoring treatment response
In vivo imaging of protease activity in arthritis -: A novel approach for monitoring treatment response
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DOI：
10.1002/art.20379
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发表时间：
2004-08-01
影响因子：
--
通讯作者：
Mahmood, U
中科院分区：
文献类型：
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作者：
Wunder, A;Tung, CH;Mahmood, U
目标。监测类风湿关节炎(RA)治疗反应的灵敏非侵入性策略将有助于促进适当的治疗和剂量，评估临床结果，并开发更有效的药物。由于不同的蛋白水解酶在RA中高度上调，并对关节破坏有显著贡献，在本研究中，我们研究了这些酶是否适合体内成像生物标记物，用于早期评估RA小鼠模型的治疗反应。使用蛋白酶激活的近红外荧光(NIRF)成像“SMART”探针，我们通过非侵入性荧光成像和组织学两种方法检测了胶原性关节炎小鼠关节中荧光的存在和分布。以Lys-Lys裂解位点为靶点的蛋白酶，包括组织蛋白酶B，激活探针荧光。获得甲氨蝶呤(MTX)治疗后的治疗监测数据。静脉注射蛋白水解酶传感器24小时后，关节炎小鼠患病的脚趾和脚掌的荧光强度明显高于健康小鼠的脚趾和脚掌。蛋白水解酶探针的荧光和组织蛋白酶B抗体的组织学染色定位于炎症滑膜的绝大多数细胞。注射MTX前48小时，注射MTX(35 mg/kg)的关节炎动物与未治疗的关节炎动物相比，荧光信号明显减弱(爪子发炎50%，脚趾发炎70%)。蛋白酶激活的NIRF探针是一种灵敏的手段，可以成像关节中靶酶的存在，并可用于早期监测抗风湿药物(如MTX)的治疗反应。
Objective. Sensitive noninvasive strategies for monitoring treatment response in rheumatoid arthritis (RA) would be valuable for facilitating appropriate therapy and dosing, evaluating clinical outcome, and developing more effective drugs. Because different proteases are highly up-regulated in RA and contribute significantly to joint destruction, in the present study we investigated whether such enzymes are suitable in vivo imaging biomarkers for early evaluation of treatment response in a murine model of RA.Methods. Using a protease-activated near-infrared fluorescence (NIRF) imaging "smart" probe, we examined the presence and distribution of fluorescence in arthritic joints of mice with collagen-induced arthritis by both noninvasive fluorescence imaging and histology. Proteases that target the Lys-Lys cleavage site, including cathepsin B, activate probe fluorescence. Treatment monitoring data were obtained following methotrexate (MTX) therapy.Results. Twenty-four hours after intravenous injection of the protease sensor, affected toes and paws of arthritic mice showed significantly higher fluorescence intensity than did toes and paws of healthy mice. Fluorescence from the protease probe and cathepsin B antibody histologic staining were localized in the vast majority of cells in the inflamed synovium. In arthritic animals treated with MTX (35 mg of MTX/kg 48 hours prior to probe injection), a significantly lower fluorescent signal (inflamed paws 50%, inflamed toes 70%) was observed as compared with untreated arthritic animals.Conclusion. Protease-activated NIRF probes are sensitive means of imaging the presence of target enzymes in arthritic joints and can be used for early monitoring of treatment response to antirheumatic drugs such as MTX.