Discovery of therapeutic agents targeting PKLR for NAFLD using drug repositioning
Cheng Zhang,
Mengnan Shi,
Woonghee Kim,
Muhammad Arif,
Martina Klevstig,
Xiangyu Li,
Hong Yang,
Cemil Bayram,
Ismail Bolat,
Özlem Özdemir Tozlu,
Ahmet Hacımuftuoglu,
Serkan Yıldırım,
Jihad Sebhaoui,
Shazia Iqbal,
Yongjun Wei,
Xiaojing Shi,
Jens Nielsen,
Hasan Turkez,
Mathias Uhlen,
Jan Boren,
Adil Mardinoglu
Affiliations
Cheng Zhang
Science for Life Laboratory, KTH - Royal Institute of Technology, Stockholm, Sweden; School of Pharmaceutical Sciences & Key Laboratory of Advanced Drug Preparation Technologies, Ministry of Education, Zhengzhou University, Zhengzhou, Henan Province, 450001, PR China
Mengnan Shi
Science for Life Laboratory, KTH - Royal Institute of Technology, Stockholm, Sweden
Woonghee Kim
Science for Life Laboratory, KTH - Royal Institute of Technology, Stockholm, Sweden
Muhammad Arif
Science for Life Laboratory, KTH - Royal Institute of Technology, Stockholm, Sweden
Martina Klevstig
Department of Molecular and Clinical Medicine, University of Gothenburg, Sahlgrenska University Hospital, Gothenburg, Sweden
Xiangyu Li
Science for Life Laboratory, KTH - Royal Institute of Technology, Stockholm, Sweden
Hong Yang
Science for Life Laboratory, KTH - Royal Institute of Technology, Stockholm, Sweden
Cemil Bayram
Department of Medical Pharmacology, Faculty of Medicine, Atatürk University, 25240, Erzurum, Turkey
Ismail Bolat
Department of Pathology, Veterinary Faculty, Ataturk University, Erzurum, 25240, Turkey
Özlem Özdemir Tozlu
Department of Molecular Biology and Genetics, Faculty of Science, Erzurum Technical University, 25200 Erzurum, Turkey
Ahmet Hacımuftuoglu
Department of Medical Pharmacology, Faculty of Medicine, Atatürk University, 25240, Erzurum, Turkey
Serkan Yıldırım
Department of Pathology, Veterinary Faculty, Ataturk University, Erzurum, 25240, Turkey
Jihad Sebhaoui
Trustlife Laboratories, Drug Research & Development Center, Istanbul, Turkey
Shazia Iqbal
Trustlife Laboratories, Drug Research & Development Center, Istanbul, Turkey
Yongjun Wei
School of Pharmaceutical Sciences & Key Laboratory of Advanced Drug Preparation Technologies, Ministry of Education, Zhengzhou University, Zhengzhou, Henan Province, 450001, PR China
Xiaojing Shi
School of Pharmaceutical Sciences & Key Laboratory of Advanced Drug Preparation Technologies, Ministry of Education, Zhengzhou University, Zhengzhou, Henan Province, 450001, PR China
Jens Nielsen
Department of Biology and Biological Engineering, Chalmers University of Technology, Gothenburg, Sweden
Hasan Turkez
Department of Medical Biology, Faculty of Medicine, Atatürk University, Erzurum, Turkey
Mathias Uhlen
Science for Life Laboratory, KTH - Royal Institute of Technology, Stockholm, Sweden
Jan Boren
Department of Molecular and Clinical Medicine, University of Gothenburg, Sahlgrenska University Hospital, Gothenburg, Sweden; Corresponding authors at: Science for Life Laboratory, Tomtebodavägen 23A, Solna, SE-17165, Sweden.
Adil Mardinoglu
Science for Life Laboratory, KTH - Royal Institute of Technology, Stockholm, Sweden; Centre for Host-Microbiome Interactions, Faculty of Dentistry, Oral & Craniofacial Sciences, King's College London, London, SE1 9RT, United Kingdom; Corresponding authors at: Science for Life Laboratory, Tomtebodavägen 23A, Solna, SE-17165, Sweden.
Summary: Background: Non-alcoholic fatty liver disease (NAFLD) encompasses a wide spectrum of liver pathologies. However, no medical treatment has been approved for the treatment of NAFLD. In our previous study, we found that PKLR could be a potential target for treatment of NALFD. Here, we investigated the effect of PKLR in in vivo model and performed drug repositioning to identify a drug candidate for treatment of NAFLD. Methods: Tissue samples from liver, muscle, white adipose and heart were obtained from control and PKLR knockout mice fed with chow and high sucrose diets. Lipidomics as well as transcriptomics 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 both tested in in vitro and in vivo models to evaluate their toxicity and efficacy. Findings: The Pklr KO reversed the increased hepatic triglyceride level in mice fed with high sucrose diet and partly recovered the transcriptomic changes in the liver as well as in 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 drug candidates were identified as potential inhibitor of PKLR using our 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 we found that these drugs attenuate the hepatic steatosis without side effect on other tissues. Interpretation: In 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 studies. Funding: ScandiEdge Therapeutics and Knut and Alice Wallenberg Foundation.
