The Study Of Combination Of Metformin And 2-deoxyglucose To Inhibit The Progress Of Polycystic Kidney Disease In Mini-pig Model

Objective: Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disease and there is no specific prevention or treatment. Studies have reported that PKD cells, similar to tumour cells, mainly depends on glycolysis to produce ATP.Abnormal glucose metabolism is maybe involved in hyper-proliferation of renal cyst epithelial cells. Mini-pigs are more similar to humans than rodents in metabolism and physiology, and therefore, are an ideal large animal model. In this study, for the first time,we systematically investigated the changes in glucose metabolism and cell proliferation signaling pathways in the kidney tissues of chronic progressive PKD mini-pig models created by knock-outing PKD1 gene and observed the therapeutic effect of metformin and 2-Deoxy-D-glucose.Methods: (1) The ADPKD mini-pig model was established using the novel zinc finger nuclease (ZFN) genome editing technology to induce PKD1 gene deletion mutation.Wild-type male wild type Mini-pigs were obtained from the State Key Laboratory for Agrobiotechnology. Both groups were raised to 20 months of age and were sacrificed after anesthesia. Blood, urine, and kidney tissue specimens were collected., we observed blood biochemistry, urine protein, renal pathology, cell cycle-related protein expression and,changes in metabolism and proliferation. (2) The ADPKD mini-pigs were divided into four groups(control group, 2DG intenrvention group, metformin intenrvention group,combination of 2DG and metformin group). In order to examine the effects of metformin and 2-Deoxy-D-glucose on polycystic kidney disease, some of these pigs were raised to 10 months of age and were sacrificed after anesthesia. Blood, urine, and kidney tissue specimens were collected.To observe the long-term therapeutic effect, we take some kidney tisssues from the 20 months of age mini-pigs by local operation.Results: (1) The results showed that in the kidneys of PKD mini-pigs, the glycolysis is increased and the mTOR and ERK signaling pathway were significantly activated. (2)Some of the animials were sacrificed after anesthesia after 10 months treatment. The results of kidney appearance and cyst index detected by CT showed that the drug intervention groups have significant differences compared with control group, but the level of blood and urine biochemical results have no obvious change. The results of biochemistry showed that the serum creatinine and urinary protein/creatinine in drug intervention groups reduced significantly compared with control group after 20 months intervention.To observe the long-term effect of the two drugs, we used local surgery technique to take some kidney tissue from the 20 months intervention mini-pigs rather than killed them. The results showed that the expression of molecules in mTOR and ERK signaling pathway were decreased for different levels in drug intervention groups compared with control group, and the most obvious is the combination group.Conclusion: This study showed that in the kidneys of PKD mini-pigs, the level of glycolysis significantly increased, and cell proliferation signaling pathways significantly activated. Combination of metformin and 2DG can delay the progress of polycystic kidney disease. Our data provide proof of principle support for the combination use of 2DG and metformin as a therapeutic strategy in ADPKD.