CRISPR/Cas9-Based Screening of FDA-Approved Drugs for NRF2 Activation: A Novel Approach to Discover Therapeutics for Non-Alcoholic Fatty Liver Disease.
CRISPR/Cas9-Based Screening of FDA-Approved Drugs for NRF2 Activation: A Novel Approach to Discover Therapeutics for Non-Alcoholic Fatty Liver Disease.
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DOI：
10.3390/antiox12071363
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发表时间：
2023-06-29
期刊：
影响因子：
7
通讯作者：
Perez-Leal, Oscar
中科院分区：
文献类型：
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作者：
Li, James;Arest, Sandra;Olszowy, Bartlomiej;Gordon, John;Barrero, Carlos A.;Perez-Leal, Oscar
随着肥胖患病率的上升，非酒精性脂肪性肝病（NAFLD）现在影响着全球20-25%的人口。NAFLD是一种与氧化应激相关的进行性疾病，可分别导致10%和3%的NAFLD患者发生肝硬化和肝癌。目前治疗选择有限，强调需要新的治疗方法。在这项研究中，我们研究了激活转录因子NRF 2的潜力，NRF 2是对抗氧化应激的关键因素，是治疗NAFLD的创新方法。利用CRISPR/Cas9工程改造的人HEK 293 T细胞系，我们能够使用Nanoluc荧光素酶标签监测血红素加氧酶-1（HMOX 1）（NRF 2靶标）的表达。我们的模型使用已知的NRF 2激活剂进行了验证，之后我们筛选了1200种FDA批准的药物，发现了六种化合物（双硫仑，硫链丝菌酮，金诺芬，噻吗替林，卤泛群和伏立诺他），这些化合物增强了NRF 2活性和抗氧化反应。这些化合物在体外对肝癌HUH-7细胞表现出对过氧化氢诱导的氧化应激和脂滴积累的保护作用。我们的研究强调了用Nanoluc荧光素酶标记CRISPR/Cas9在识别潜在的NRF 2激活剂方面的实用性，为潜在的NAFLD治疗铺平了道路。
With the rising prevalence of obesity, non-alcoholic fatty liver disease (NAFLD) now affects 20–25% of the global population. NAFLD, a progressive condition associated with oxidative stress, can result in cirrhosis and liver cancer in 10% and 3% of patients suffering NAFLD, respectively. Therapeutic options are currently limited, emphasizing the need for novel treatments. In this study, we examined the potential of activating the transcription factor NRF2, a crucial player in combating oxidative stress, as an innovative approach to treating NAFLD. Utilizing a CRISPR/Cas9-engineered human HEK293T cell line, we were able to monitor the expression of heme oxygenase-1 (HMOX1), an NRF2 target, using a Nanoluc luciferase tag. Our model was validated using a known NRF2 activator, after which we screened 1200 FDA-approved drugs, unearthing six compounds (Disulfiram, Thiostrepton, Auranofin, Thimerosal, Halofantrine, and Vorinostat) that enhanced NRF2 activity and antioxidant response. These compounds demonstrated protective effects against oxidative stress induced by hydrogen peroxide and lipid droplets accumulation in vitro with hepatoma HUH-7 cells. Our study underscores the utility of CRISPR/Cas9 tagging with Nanoluc luciferase in identifying potential NRF2 activators, paving the way for potential NAFLD therapeutics.
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DOI：
10.3390/antiox9101025
发表时间：
2020-10-21
期刊：
Antioxidants (Basel, Switzerland)
影响因子：
--
作者：
Ibrahim L;Mesgarzadeh J;Xu I;Powers ET;Wiseman RL;Bollong MJ
通讯作者：
Bollong MJ
影响因子：
82.9
作者：
Friedman SL;Neuschwander-Tetri BA;Rinella M;Sanyal AJ
通讯作者：
Sanyal AJ
影响因子：
16.6
作者：
Lei, Yuanyuan;Tang, Li;Chen, Qiao;Wu, Lingyi;He, Wei;Tu, Dianji;Wang, Sumin;Chen, Yuyang;Liu, Shuang;Xie, Zhuo;Wei, Hong;Yang, Shiming;Tang, Bo
通讯作者：
Tang, Bo
影响因子：
--
作者：
Kumagai Y;Kanda H;Shinkai Y;Toyama T
通讯作者：
Toyama T
影响因子：
9.8
作者：
Buzzetti, Elena;Pinzani, Massimo;Tsochatzis, Emmanuel A.
通讯作者：
Tsochatzis, Emmanuel A.