Screening of FDA-approved drugs identifies sutent as a modulator of UCP1 expression in brown adipose tissue

Abstract

BackgroundThe pharmacological activation of thermogenesis in brown adipose tissue has long been considered promising strategies to treat obesity. However, identification of safe and effective agents remains a challenge. In this study, we addressed this challenge by developing a cellular system with a fluorescence readout, and applied in a high-throughput manner to screen for FDA-approved drugs that may activate endogenous UCP1 expression in adipocytes.MethodsWe have generated a Ucp1-2A-GFP reporter mouse, in which GFP intensity serves as a surrogate of the endogenous expression level of UCP1 protein; and immortalized brown adipocytes were derived from this mouse model and applied in drug screening. Candidate drugs were further tested in mouse models either fed with normal chow or high fat diet to induce obesity.FindingsBy using the cellular screening platform, we identified a group of FDA-approved drugs that can upregulate UCP1 expression in brown adipocyte, including previously known UCP1 activators and new candidate drugs. Further studies focusing on a previously unreported drug—sutent, revealed that sutent treatment could increase the energy expenditure and inhibit lipid synthesis in mouse adipose and liver tissues, resulting in improved metabolism and resistance to obesity.InterpretationThis study offered an easy-to-use cellular screening system for UCP1 activators, and provided a candidate list of FDA-approved drugs that can potentially treat obesity. Further study of these candidates may shed new light on the drug discovery towards obesity.FundNational Key Research and Development Program and the Strategic Priority Research Program of the Chinese Academy of Sciences, etc. (250 words).

Indirubin, a small molecular deriving from connectivity map (CMAP) screening, ameliorates obesity-induced metabolic dysfunction by enhancing brown adipose thermogenesis and white adipose browning

“…Since the recent discovery of functional BAT or BATlike tissue in adult humans, the development and function of brown or beige adipocyte has been widely and deeply researched for its potential as a target of anti-obesity therapeutics [63]. Although many screening approaches (in vitro and in vivo) have been adopted for the discovery of chemicals to activate UCP1 expression [64][65][66][67], to identify safe and effective drugs for upregulating the activity of brown and (or) beige adipocytes is still a challenge, especially given the high-cost and time-consuming traditional methods for screening UCP1 activators. In order to address this challenge, we took a multi-pronged approach to screen UCP1 activators (through drug repurposing), including in silico predictions, in vitro assays, as well as in vivo experiments.…”

Glyburide Regulates UCP1 Expression in Adipocytes Independent of KATP Channel Blockade

“…We have previously developed a cellular screening platform with the Ucp1-2A-GFP reporter cell lines, which allows high-throughput screenings using a fluorescence signal as readout ( Qiu et al., 2018 ). Using this reporter line, we have identified a group of drugs that can upregulate UCP1 expression in brown adipocytes and reported sutent as an effective modulator of UCP1 expression in brown adipose tissue ( Qiu et al., 2018 ). Sutent belongs to the tyrosine kinase (RTK) inhibitor family and has been used clinically to treat certain kidney, pancreatic, and gastrointestinal cancers.…”

Identification of a rhodanine derivative BML-260 as a potent stimulator of UCP1 expression

“…Despite these findings, more efficacious reagents and comprehensive understanding of the UCP1 regulation in different adipocytes are still lacking. Several cellular screening systems for UCP1 activators have been previously established that allow for high-throughput and unbiased identification of effective compounds 19-21. For example, we have generated an immortalized brown adipocyte cell line that harbors an Ucp1 -2A-GFP reporter system.…”

“…For example, we have generated an immortalized brown adipocyte cell line that harbors an Ucp1 -2A-GFP reporter system. In this system, the 2A-GFP cassette was knocked-in immediately before the stop codon of the endogenous Ucp1 gene, the GFP intensity thus serves as a surrogate of the endogenous expression level of the UCP1 protein 21. Using this reporter system, we have screened ~1,000 FDA-approved compounds and identified 42 drugs that may activate UCP1 expression in brown adipocytes 21.…”

“…In this system, the 2A-GFP cassette was knocked-in immediately before the stop codon of the endogenous Ucp1 gene, the GFP intensity thus serves as a surrogate of the endogenous expression level of the UCP1 protein 21. Using this reporter system, we have screened ~1,000 FDA-approved compounds and identified 42 drugs that may activate UCP1 expression in brown adipocytes 21. In this study, with an initial purpose to discover possible phosphatases that may involve in UCP1 regulation, we applied the same screening platform to a phosphatase inhibitor library.…”