The availability of an autologous transplantable auxiliary liver would dramatically affect the treatment of liver disease. Assembly and function in vivo of a bioengineered human liver derived from induced pluripotent stem cells (iPSCs) has not been previously described. By improving methods for liver decellularization, recellularization, and differentiation of different liver cellular lineages of human iPSCs in an organ-like environment, we generated functional engineered human mini livers and performed transplantation in a rat model. Whereas previous studies recellularized liver scaffolds largely with rodent hepatocytes, we repopulated not only the parenchyma with human iPSC-hepatocytes but also the vascular system with human iPS-endothelial cells, and the bile duct network with human iPSC-biliary epithelial cells. The regenerated human iPSC-derived mini liver containing multiple cell types was tested in vivo and remained functional for 4 days after auxiliary liver transplantation in immunocompromised, engineered (IL2rg À/À ) rats.

Objectives T‐cell immunoglobulin domain and mucin domain‐4 (TIM‐4) is selectively expressed on antigen‐presenting cells (APCs) and modulates various immune responses. However, the role of TIM‐4 expressed by Kupffer cells (KCs) in liver fibrosis remains unclear. The present study aimed to explore whether and how TIM‐4 expressed by KCs is involved in liver fibrosis. Materials and Methods Mice chronic liver fibrosis models were established and divided into the olive‐induced control group, CCL4‐induced control group, olive‐induced TIM‐4 interference group and CCL4‐induced TIM‐4 interference group. Different techniques were used to monitor the fibrotic effects of TIM‐4, including histopathological assays, Western blotting, ELISA and transmission electron microscopy. Additionally, mice liver transplant models were established to determine the fibrotic effects of TIM‐4 on fibrosis after liver transplantation (LT). Results We found that the induction of liver fibrosis by CCL4 was associated with TIM‐4 expression in KCs. TIM‐4 interference essentially contributed to liver fibrosis resolution. KCs from the TIM‐4 interference group had decreased levels of pro‐fibrotic markers, reduced TGF‐β1 secretion and inhibited hepatic stellate cell (HSC) differentiation into myofibroblast‐like cells. In addition, we used GdCl3 to verify that KCs are the primary source of TGF‐β1 during fibrosis progression. Moreover, KCs from CCL4‐induced mice showed increased ROS production, mitophagy activation and TGF‐β1 secretion. However, TIM‐4 interference in the KCs inhibited Akt1‐mediated ROS production, resulting in the suppression of PINK1, Parkin and LC3‐II/I activation and the reduction of TGF‐β1 secretion during liver fibrosis. Additionally, TIM‐4 interference potentially attenuated development of fibrosis after LT. Conclusions Our findings revealed the underlying mechanisms of TIM‐4 interference in KCs to mitigate liver fibrosis.

Methodology for accurate quantification of intra-hepatic cccDNA has long been a technical challenge, yet it is highly desired in the clinic. Here, we developed a sensitive method for quantification of intrahepatic cccDNA in liver biopsies from patients, which allowed to predict patient’s response to interferon therapy at baseline. Twenty-five patients with HBeAg+ CHB were recruited and liver biopsies were obtained at baseline and 1-year after interferon treatment, respectively. Both intrahepatic cccDNA and HBV DNA were absolutely quantified by a droplet digital PCR amplification system. Patients were categorized as either responder or non-responder group based on their HBeAg status 1-year after interferon therapy. Levels of both intrahepatic HBV DNA and HBV cccDNA were significantly reduced after interferon treatment among the responders, but not the non-responders, in comparison with their levels at baseline. Baseline values of intrahepatic HBV DNA over cccDNA significantly correlated with patient’s response to PEG-IFN therapy (P = 0.000). In addition, HBeAg seroconversion also correlates with a significant reduction in intrahepatic pgRNA production among the responders after interferon therapy (P = 0.030). In conclusion, our results suggest that baseline value of intrahepatic HBV DNA over cccDNA may be a preferable indicator for selecting appropriate patients for IFN-based therapy in the clinic.

Bladder cancer is a highly heterogeneous and aggressive malignancy with a poor prognosis. EGF/EGFR activation causes the detachment of SHC-binding protein 1 (SHCBP1) from SHC adapter protein 1 (SHC1), which subsequently translocates into the nucleus and promotes cancer development via multiple signaling pathways. However, the role of the EGF-SHCBP1 axis in bladder cancer progression remains unexplored. Herein, we report that SHCBP1 is upregulated in bladder cancer tissues and cells, with cytoplasmic or nuclear localization. Released SHCBP1 responds to EGF stimulation by translocating into the nucleus following Ser273 phosphorylation. Depletion of SHCBP1 reduces EGF-induced cell migration and invasiveness of bladder cancer cells. Mechanistically, SHCBP1 binds to RACGAP1 via its N-terminal domain of amino acids 1 ~ 428, and this interaction is enhanced following EGF treatment. Furthermore, SHCBP1 facilitates cell migration by inhibiting RACGAP-mediated GTP-RAC1 inactivation, whose activity is indispensable for cell movement. Collectively, we demonstrate that the EGF-SHCBP1-RACGAP1-RAC1 axis acts as a novel regulatory mechanism of bladder cancer progression, which offers a new clinical therapeutic strategy to combat bladder cancer.