Effects Of DPCI-23,a Glutaminyl Cyclase Inhibitor,on The Transcriptome Of PC12 Cells

Alzheimer's disease(AD),the most common form of dementia,is a progressive neurodegenerative disease.The effective and safe pharmacologic agents can be offered to prevent or cure AD in clinical are still insufficient.It has been confirmed that over-expression of glutaminyl cyclase(QC)plays a crucial role in the initiation of AD by catalyzing the generation of neurotoxic pE-A?s.In the previous research,a series of small molecule QC inhibitors,diphenyl conjugated imidazole derivatives(DPCIs),were designed,synthesized and assessed as potential anti-AD agents.To understand the mechanism of these compounds,effects of DPCI-23 on the transcriptome of PC12 cells were evaluated here:(1)QC activities and the generation of pE-A? in PC12 cells were inhibited obviously in a dose-and time-dependent manner after the treatment of DPCI-23 based on GD-QC enzyme-linked test and ELISA.(2)According to RNA-Seq analysis,8873 and 10480 genes were up-regulated and 9673 and 7925 genes were down-regulated in PC12 cells treated with DPCI-23 for 12 and 24 h respectively.Ribosome was affected notably and the levles of HSP70,HSP90 and Actin were significantly regulated after the treatment,indicating the effects of DPCI-23 on protein homeostasis.(3)The expression of RPL37,RPL38 and RPS18 in ribosome were up-regulated and the expression of RPS17 was down-regulated obviously in PC12 cells treated with DPCI-23 for 12 and 24 h.Interestingly,the phosphorylation level of RPS6 was decreased significantly after the treatment,but not for RPS6.Thus,the presence of DPCI-23 may contribute to the improvement of ribosome.(4)Normally,the expression of HSP70 and HSP90 were up-regulated and the expression of Actin was down-regulated obviously in PC12 cells treated with DPCI-23 for different time based on RT-qPCR,Western Blot,ELISA and immunofluorescence analysis.Considering the RNA-Seq analysis,DPCI-23 exhibited the influence on protein homeostasis in PC12 cells through the regulation of chaperone HSP70,HSP90 and Actin.As we mentioned above,effects of DPCI-23 on the expression and regulation of ribosome,HSP70,HSP90 and Actin in PC12 cells were demonstrated for the first time.Influence of QC inhibitors on protein homeostasis maybe important for the development of novel agents for the treatment and prevention of AD.