Construction of Escherichia coli Whole-Cell Biosensors for Statin Efficacy and Production Test.

Huanjie Li,Yunshan Wang,Qingda Wang,Rui Zhao,Huaiwei Liu,Luying Xun

doi:10.3389/fcell.2020.00404

Abstract

Statins are widely used cholesterol-lowering drugs. Their potential application in anti-cancer treatment is also under investigation. The individual variance in statin response has been observed, which may be caused by the variation in human HMG-CoA reductase (hHMGR)—the inhibition target of statin drugs. Herein, we reported the design and construction of two Escherichia coli whole-cell biosensors. The first one is statin-efficacy testing sensor, which is composed of two separate modules: a hybrid mevalonate (MVA) pathway and a HMG-CoA sensing system. A truncated hHMGR was used as the key enzyme of the MVA pathway and a promiscuous transcription factor (TF) BsFapR was used as the HMG-CoA sensor. When hHMGR was inhibited by statins, HMG-CoA accumulated intracellularly and was sensed by BsFapR, which subsequently turned on its cognate promoter. This biosensor has the potential to be used as a “precision medicine” tool—selecting potent statin drugs for individual patients. The second one is a statin-production testing sensor, which is based on another promiscuous TF AraCM that can sense statins. This biosensor can be used in optimization of statin-producing strains. The prototypes of these two biosensors were successfully constructed and their further optimization is highly expected.

Highlights

Statins are a class of cholesterol-lowering drugs that are widely used in clinical practice (Rizzo et al, 2012; Garattini and Padula, 2017; Hennekens et al, 2017)
Human HMG-CoA reductase (HMGR) has 1677 single nucleotide variants (NCBI), and diverse statin drugs are in the market as well; it is beneficial to construct a biosensor for pre-testing the inhibition effect of different statin drugs upon an individual hHMGR clinical treatment
The whole design consists of two separate modules: the hHMGR-based MVA pathway module and the MVA pathway monitoring module (Figures 1A,B)

Summary

Introduction

Statins are a class of cholesterol-lowering drugs that are widely used in clinical practice (Rizzo et al, 2012; Garattini and Padula, 2017; Hennekens et al, 2017). They are competitive inhibitors of the HMG-CoA reductase (HMGR) that catalyzes the conversion of HMG-CoA to mevalonate (Endo, 1992; Istvan, 2003). Despite the generally good treatment effect of statins, notable individual variation in response has been reported (Armitage, 2007; Wysockakapcinska et al, 2009; Sopková et al, 2017) This could be caused by the HMGR variance in different individuals. Human HMGR (hHMGR) has 1677 single nucleotide variants (NCBI), and diverse statin drugs are in the market as well; it is beneficial to construct a biosensor for pre-testing the inhibition effect of different statin drugs upon an individual hHMGR clinical treatment

Methods

Results

Conclusion