A novel optical imaging probe for targeted visualization of NLRP3 inflammasomes in a mouse model of age-related macular degeneration.
A novel optical imaging probe for targeted visualization of NLRP3 inflammasomes in a mouse model of age-related macular degeneration.
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DOI：
10.3389/fmed.2022.1047791
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发表时间：
2022
影响因子：
3.9
通讯作者：
中科院分区：
文献类型：
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作者：
湿性年龄相关性黄斑变性（湿性AMD）是一种进行性血管疾病，主要影响老年人，并导致严重和不可逆的视力丧失。湿性AMD的关键并发症是脉络膜新生血管形成（CNV），其可能部分由与巨噬细胞迁移至CNV病变相关的NLRP 3炎性体驱动。由于活化的NLRP 3与CNV相关，因此可视化NLRP 3炎性小体及其相关巨噬细胞对于监测湿性AMD进展和开发针对其的有效疗法具有极大的意义。因此，在这项研究中，我们开发了InflammaProbe-1，一种用于CNV病变中NLRP 3炎性小体靶向可视化的光学成像探针。通过将临床相关的荧光团俄勒冈州绿色® 488缀合至选择性NLRP 3抑制剂CY-09来合成InflammaProbe-1。用酶联免疫吸附测定法通过比较InflammaProbe-1抑制LPS致敏和尼日利亚菌素刺激的BMDM中NLRP 3介导的IL-1β分泌的能力与CY-09的能力来评估InflammaProbe-1靶向NLRP 3的能力。在已经用LPS诱导表达NLRP 3的炎症探针-1染色的BMDM上进行NLRP 3的体外共聚焦成像。在LCNV后第4天腹膜内注射10 mg/kg的InflammaProbe-1后6小时，在小鼠激光诱导的脉络膜新生血管形成（LCNV）（AMD模型）上进行NLRP 3的体内成像。InflammaProbe-1在抑制IL-1β分泌方面与CY-09一样有效（相对于对照，InflammaProbe-1和CY-09组在10 μM时p < 0.01）。已被诱导表达NLRP 3的经InflammaProbe-1染色的BMDM显示出比未处理的细胞显著更亮的荧光（对于LPS处理组p < 0.0001，对于LPS和尼日利亚菌素处理组p < 0.001）。此外，在小鼠LCNV中实现了NLRP 3的体内分子成像。我们认为InflammaProbe-1可能是一种有用的分子成像探针，用于监测AMD和其他NLRP 3介导的疾病的发病、进展和治疗反应。
Wet form of age-related macular degeneration (wet AMD) is a progressive vascular disease that mainly affects older adults and causes severe and irreversible vision loss. A key complication of wet AMD is choroidal neovascularization (CNV), which may be driven in part by NLRP3 inflammasomes that are associated with macrophages migration to CNV lesions. Since activated NLRP3 is correlated with CNV, visualizing NLRP3 inflammasomes and their associated macrophages is of great interest to monitor wet AMD progression and develop effective therapies against it. However, to the best of our knowledge, current ophthalmic imaging systems do not permit such targeted imaging. Therefore, in this study, we developed InflammaProbe-1, an optical imaging probe for targeted visualization of NLRP3 inflammasomes in CNV lesions. InflammaProbe-1 was synthesized by conjugating a clinically relevant fluorophore, Oregon Green® 488, to the selective NLRP3 inhibitor, CY-09. The ability of InflammaProbe-1 to target NLRP3 was assessed with an enzyme-linked immunosorbent assay by comparing its ability to inhibit NLRP3-mediated secretion of IL-1β to that of CY-09 in LPS-primed and nigericin-stimulated BMDMs. In vitro confocal imaging of NLRP3 was performed on InflammaProbe-1-stained BMDMs that had been induced to express NLRP3 with LPS. In vivo imaging of NLRP3 was conducted on mouse laser induced choroidal neovascularization (LCNV), a model of AMD, 6 h after an intraperitoneal injection of InflammaProbe-1 at 10 mg/kg on day 4 post-LCNV. InflammaProbe-1 was just as effective as CY-09 at inhibiting IL-1β secretion (p < 0.01 at 10 μM for both the InflammaProbe-1 and CY-09 groups relative to the control). InflammaProbe-1-stained BMDMs that had been induced to express NLRP3 showed significantly brighter fluorescence than untreated cells (p < 0.0001 for LPS treatment group and p < 0.001 for LPS and nigericin treatment group). Furthermore, in vivo molecular imaging of NLRP3 was achieved in mouse LCNV. We propose that InflammaProbe-1 may be a useful molecular imaging probe to monitor the onset, progression, and therapeutic response of AMD and other NLRP3-mediated diseases.
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DOI：
10.1006/bbrc.1996.1112
发表时间：
1996-07-25
影响因子：
3.1
作者：
Imai, Y;Ibata, I;Kohsaka, S
通讯作者：
Kohsaka, S
影响因子：
6
作者：
Tobe, T;Ortega, S;Campochiaro, PA
通讯作者：
Campochiaro, PA
影响因子：
7.7
作者：
Malsy J;Alvarado AC;Lamontagne JO;Strittmatter K;Marneros AG
通讯作者：
Marneros AG
影响因子：
20.3
作者：
Fantin, Alessandro;Vieira, Joaquim M.;Ruhrberg, Christiana
通讯作者：
Ruhrberg, Christiana
影响因子：
4.6
作者：
Tisi, A.;Parete, G.;Maccarone, R.
通讯作者：
Maccarone, R.