沉默CIDEB基因治疗非酒精性脂肪肝病的siRNA筛选与评价OA

This study aimed to develop small interfering RNA(siRNA)therapeutics targeting the cell death-indu-cing DNA fragmentation factor-like effector B(CIDEB),a key player in nonalcoholic steatohepatitis(NASH),to achieve long-term and safe protein inhibition.Different humanized siRNAs sequences were designed to target CI-DEB,and cell models were constructed by introducing humanized psiCHECK-CIDEB double luciferase reporter genes at the cellular level for preliminary in vitro activity screening.The selected siRNAs were chemically modified and further screened and optimized.Subsequently,the GalNAc-siRNA conjugate was chemically synthesized for the optimized siRNA,and a CIDEB transgenic animal model was constructed for in vivo activity evaluation to identify candidate sequences with a significant silencing effect on CIDEB messenger RNA(mRNA).Three dose gradients(10,30,and 50 mg/kg)were used for the early in vivo safety evaluation of the optimal sequence.In vivo activity experiments showed that the optimized siRNA could significantly and long-term inhibit CIDEB expression in trans-genic mice(with a relative inhibition rate>80%for 4 weeks).A preliminary safety evaluation was conducted based on serum levels of glutamic oxalic aminotransferase,glutamic pyruvic aminotransferase,organ coefficients and HE staining,which showed that high-dose administration did not cause significant liver toxicity.Therefore,tar-geting the CIDEB gene with siRNA therapy provides a safer and more durable treatment option for patients with chronic liver disease who require long-term medication by precisely regulating a key targets in NASH progression.