Development of CIDEA reporter mouse model and its application for screening thermogenic drugs

Yeonho Son,Yoonjoe Joh,Cheol Song,Ju Seung Son,Hyeonyeong Im,Yoon Keun Cho,Cheoljun Choi,Sungug Joo,Young Jae Lee,Yun-Hee Lee,Je Kyung Seong

doi:10.1038/s41598-021-97959-0

Yeonho Son, Yoonjoe Joh + Show 9 more

Open Access

https://doi.org/10.1038/s41598-021-97959-0

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Journal: Scientific Reports	Publication Date: Sep 16, 2021
Citations: 7	License type: open-access

Affiliation: Seoul National University, Gachon University

Abstract

Cell death-inducing DNA fragmentation factor-like effector A (CIDEA) is a lipid droplet-associated protein and is a known marker of the thermogenic capacity of brown/beige adipocytes. To monitor the expression of CIDEA in live mice in a non-invasive manner, we generated CIDEA reporter mice expressing multicistronic mRNAs encoding CIDEA, luciferase 2, and tdTomato proteins under the control of the Cidea promoter. The expression level of endogenous CIDEA protein in adipose tissue was not affected by the expression of polycistronic reporters. The two CIDEA reporters, luciferase 2 and tdTomato, correctly reflected CIDEA protein levels. Importantly, luciferase activity was induced by cold exposure and the treatment with β3-adrenergic receptor agonist CL316,243 in interscapular and inguinal adipose tissue, which was detectable by in vivo bioluminescence imaging. We further evaluated the effects of candidate brown adipogenic agents using this CIDEA reporter system and demonstrated a positive correlation between drug-induced luciferase activity and thermogenic gene expression levels both in vitro and in vivo. Collectively, we established a dual CIDEA reporter mouse model in which fluorescence and luminescence signals correctly reflect CIDEA expression, and therefore, suggested that this reporter system can be used to evaluate the thermogenic efficacy of candidate molecules.

Highlights

Cell death-inducing DNA fragmentation factor-like effector A (CIDEA) is a lipid droplet-associated protein and is a known marker of the thermogenic capacity of brown/beige adipocytes
The Cidea-P2A-luciferase 2 (Luc2)-T2A-tandem-dimer Tomato (tdT) (CideaLuc2-tdT) multicistronic transcript was produced under the control of the Cidea promoter, and the translated protein was split into CIDEA, Luc[2], and tdT proteins through the 2A self-cleaving peptides Porcine teschovirus-1 2A (P2A) and Thosea asigna virus 2A (T2A)[22,23]
To examine the tissue-specific expression of CIDEA in wild type (WT) mice, we analyzed Cidea gene expression levels in various tissues, including adipose tissue, liver, and heart, which play an important role in lipid metabolism

Summary

Introduction

Cell death-inducing DNA fragmentation factor-like effector A (CIDEA) is a lipid droplet-associated protein and is a known marker of the thermogenic capacity of brown/beige adipocytes. We further evaluated the effects of candidate brown adipogenic agents using this CIDEA reporter system and demonstrated a positive correlation between drug-induced luciferase activity and thermogenic gene expression levels both in vitro and in vivo. Brown and white adipocytes are known to originate from distinct mesenchymal stem cell lineages, white adipocytes can convert into brown-like adipocytes (beige adipocytes) under thermogenic stimuli, such as cold temperature[3] This process, known as WAT browning, is characterized by the induction of uncoupling protein 1 (UCP1) expression and an increase in the mitochondrial mass and oxidative metabolism[4]. Pharmacological activation using small molecules that mimic thermogenic stimuli, such as treatment with sustained β-adrenergic receptor agonist, induces mitochondrial biogenesis and increases oxidative metabolism and thermogenesis via UCP1-dependent and -independent mechanisms[2,12]

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