12F). Collectively, targeting AR in BM-MSCs with either AR-siRNA or ASC-J9® may represent a new potential therapeutic approach to battle liver cirrhosis. Androgen/AR signaling, as a critical determining factor in sexual phenotypes and maturation, is well documented. It is generally agreed that androgen/AR signaling stimulates differentiation

of prostate, bone, and muscle.35-37 During the embryonic stem cell differentiation process, AR amount gradually increases.38 In natural development, androgen/AR signaling seems to enhance the differentiation process, except for adipocyte differentiation. This observation triggered us to think about whether this fact exists in disease treatment when BM-MSCs transplantation therapy is performed. Indeed, it has check details been suggested that gender difference does exist using BM-MSCs transplantation therapy in heart diseases.13 Although gender differences have been suggested in BM-MSC transplantation Rucaparib therapy with the observation that female BM-MSCs

have better therapeutic potential than male BM-MSCs,13 effects of AR, a key factor in male sexual phenotype, in BM-MSC therapeutic application remain unclear. In this study, we used a genetic deletion approach to prove that deletion of AR in BM-MSCs increases their self-renewal potential (consistent results were observed in female ARKO mice; data not shown), migration capacity, anti-inflammatory actions, and anti-fibrotic effects to better treat liver cirrhosis. Because loss of BM-MSCs after transplantation cannot be prevented as a result of microischemia, our finding that knockout of AR in BM-MSCs however increases functions and numbers of BM-MSCs after transplantation can provide the future basis for improving BM-MSC transplantation in liver cirrhosis

as well as other diseases that have currently adopted BM-MSC transplantation therapy in clinical trials. In therapeutic approaches, we also clearly demonstrated that targeting AR using either AR-siRNA or ASC-J9® in BM-MSCs both led to better therapeutic outcomes in treating liver cirrhosis, suggesting that targeting BM-MSC AR could exert better BM-MSC transplantation therapeutic efficacy in treating cirrhotic liver. The advantage of targeting AR with ASC-J9® is that we can handle AR degradation in in vitro cultured BM-MSCs before transplanting into liver, and several early in vivo mice studies all proved that ASC-J9® has little side effects and mice have normal sexual activity with healthy serum testosterone level. Our mechanism studies pointed out that targeting AR in BM-MSCs leads to better therapeutic efficiency to treat liver cirrhosis through four major pathways (as illustrated in Fig. 7). Targeting AR in BM-MSCs could (1) increase EGFR to enhance their self-renewal potential (Fig. 3A), (2) elevate the MMP-9 to stimulate their migration (Fig. 3B), and (3) promote IL1Ra production to inhibit macrophage infiltration and HSCs proliferation (Fig. 4A,B).