Yunfei Bai,Zhenghao Yang,Xiaochan Xu,Wanqiu Ding,Juntian Qi,Feng Liu,Xiaoxiao Wang,Bin Zhou,Wenpeng Zhang,Xiaomei Zhuang,Guanglu Li,Yang Zhao^✉

Abstract

Background and Aims: Cell fate can be directly reprogrammed from acces-sible cell types (e.g., fibroblasts) into functional cell types by exposure to small molecule stimuli. However, no chemical reprogramming method has been re-ported to date that successfully generates functional hepatocyte-like cells that can repopulate liver tissue, casting doubt over the feasibility of chemical repro-gramming approaches to obtain desirable cell types for therapeutic applications.

Approach and Results: Here, through chemical induction of phenotypic plas-ticity,  we  provide  a  proof-of-concept  demonstration  of  the  direct  chemical  re-programming  of  mouse  fibroblasts  into  functional  hepatocyte-like  cells  using  exposure to small molecule cocktails in culture medium to successively stimulate endogenous expression of master transcription factors associated with hepato-cyte development, such as hepatocyte nuclear factor 4a, nuclear receptor sub-family 1, group I, member 2, and nuclear receptor subfamily 1, group H, member 4. RNA sequencing analysis, metabolic assays, and in vivo physiological experi-ments show that chemically induced hepatocytes (CiHeps) exhibit comparable activity and function to primary hepatocytes, especially in liver repopulation to rescue liver failure in fumarylacetoacetate hydrolase^−/−recombination activatinggene 2^−/−interleukin 2 receptor, gamma chain^−/− mice in vivo. Single-cell RNA-seq  further  revealed  that  gastrointestinal-like  and  keratinocyte-like  cells  were  induced along with CiHeps, resembling the activation of an intestinal program within hepatic reprogramming as described in transgenic approaches.

Conclusions: Our  findings  show  that  direct  chemical  reprogramming  can  generate hepatocyte-like cells with high-quality physiological properties, pro-viding a paradigm for establishing hepatocyte identity in fibroblasts and dem-onstrating  the  potential  for  chemical  reprogramming  in  organ/tissue  repair  and regeneration therapies.

https://aasldpubs.onlinelibrary.wiley.com/doi/abs/10.1002/hep.32686