Discovery of therapeutic agents targeting PKLR for NAFLD using drug repositioning

Zhang, C.; Shi, M.; Kim, W.; Arif, M.; Klevstig, M.; Li, X.; Yang, H.; Bayram, C.; Bolar, I.; Tozlu, O. O.; Hacimuftuoglu, A.; Yildirim, S.; Wei, Y.; Shi, X.; Nielsen, J.; Turkez, H.; Uhlen, M.; Boren, J.; Mardinoglu, A.

doi:10.1101/2022.02.15.480557

bioRxiv · 10.1101/2022.02.15.480557

Discovery of therapeutic agents targeting PKLR for NAFLD using drug repositioning

Zhang, C.·Shi, M.·Kim, W.·Arif, M.·Klevstig, M.·Li, X.·Yang, H.·Bayram, C.·Bolar, I.·Tozlu, O. O.·Hacimuftuoglu, A.·Yildirim, S.·Wei, Y.·Shi, X.·Nielsen, J.·Turkez, H.·Uhlen, M.·Boren, J.·Mardinoglu, A.

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Abstract

Background & AimsNon-alcoholic fatty liver disease (NAFLD) encompasses a wide spectrum of liver pathologies. However, not medical treatment has been approved for the treatment of the disease. In our previous study, we found PKLR could be a potential target for treatment of NALFD. Here, the aim is to investigate the effect of PKLR in in vivo model and perform drug repositioning to identify a drug candidate for treatment of NAFLD. MethodsBiopsies from liver, muscle, white adipose tissue and heart were obtained from control and PKLR knockout mice fed with chow and high sucrose diets. Lipidomics as well as transcriptmics analyses were conducted using these tissue samples. In addition, a computational drug repositioning analysis was performed and drug candidates were identified. The drug candidates were finally tested in both in vitro and in vivo models to evaluated their toxicity and efficacy. ResultsThe Pklr KO reversed the increased hepatic triglyceride level in mice fed with high sucrose diet and partly recovered the transcriptomic changes in liver as well as other three tissues. Both liver and white adipose tissues exhibited dysregulated circadian transcriptomic profiles, and these dysregulations were reversed by hepatic knockout of Pklr. In addition, 10 small molecule drugs were identified as potential inhibitor of PKLR by the drug repositioning pipeline, and two of them significantly inhibited both the PKLR expression and triglyceride level in in vitro model. Finally, the two selected small molecule drugs were evaluated in in vivo rat models and it was demonstrated that these drugs attenuated hepatic steatosis without side effect on other tissues. ConclusionIn conclusion, our study provided biological insights about the critical role of PKLR in NAFLD progression and proposed a treatment strategy for NAFLD patients, which has been validated in preclinical experiment.

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Zhang, C., Shi, M., Kim, W., Arif, M., Klevstig, M., Li, X., Yang, H., Bayram, C., Bolar, I., Tozlu, O. O., Hacimuftuoglu, A., Yildirim, S., Wei, Y., Shi, X., Nielsen, J., Turkez, H., Uhlen, M., Boren, J., Mardinoglu, A.. 2022-02-17. Discovery of therapeutic agents targeting PKLR for NAFLD using drug repositioning. https://doi.org/10.1101/2022.02.15.480557

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