For in vivo studies C57BL/6 male GPCR Compound Library in vitro mice at 12 weeks of age were subjected

to bile duct ligation or sham surgery, or injected with lipopolysaccharide (LPS, 2.0 mg/kg) or saline vehicle by i.p. injection. In vivo ChIP assays done with liver nuclei obtained from mice after 3 days of CBDL or 5 hours post LPS injection showed a markedly increased recruitment of NF-kB p65 to the Bsep and Fxr promoters in both BDL and LPS-treated mice compared to controls. There was also increased recruitment of the SMRT and of HDAC2 and 3 to the Bsep locus as part of a corepressor complex. The compensatory transporter Osta-Ostp is up-regulated in cholestasis. In contrast to the inhibitory effect of p65 on expression of the BSEP and FXR promoters, we found that p65 expression dose-dependently activated a Osta-lucifer-ase construct in Huh7 cells. Expression of the NF-kB p50 sub-unit, alone, which lacks a C-terminal activation domain, had no effect. Combined expression of NF-kB p65 and p50 subunits further enhanced Osta promoter ERK inhibitor activity. The overexpression of inhibitors IkBa and IkBSR blocked induction by p65 and p50 in Huh7 cells. Taken together, these studies provide mechanistic insight into how NF-kB impairs normal pathways for bile acid excretion and participates in an adaptive response in cholestasis.

Disclosures: The following people have nothing to disclose: Natarajan Balasubramaniyan, Meena Ananthanarayanan, Frederick J. Suchy Background: Liver repopulation of FRG mice (immune-deficient mouse model of tyrosinemia) with transplanted hepatocytes is possible with donor hepatocytes from diverse species.

Use of donor mouse or rat cells in this model produces robust repop-ulation of the liver, with the repopulated liver remaining the same size as the un-repopulated liver, and a normal sized bile acid pool. In contrast, when donor cells are human hepato-cytes, the resulting repopulated liver is roughly 3 times larger than the un-repopulated liver, and the bile acid pool is significantly expanded. Aims 1. 上海皓元医药股份有限公司 Create a genetic model to correct aberrant bile acid signaling in the human hepatocyte repopu-lated mouse liver 2. Determine if aberrant bile acid signaling in the human hepatocyte repopulated mouse liver is responsible for the enlarged liver size in these animals Methods: A 150 kb BAC containing the human FGF19 gene in the middle of the sequence was introduced into FRG mice. FGF19+ transgene mice (and their FGF19- littermates) underwent intra-splenic transplantation of human or mouse hepatocytes. After full repopulation, livers were examined for size and repopu-lation by histology. Bile acid pools, and intestinal and liver bile acid signaling were quantified. Deep RNA sequencing of repopulated livers was performed. Results: Mouse hepatocyte repopulated livers averaged 5.3% and 5.7% of body weight (p =0.43) in FGF19- and FGF19+ recipients. In contrast, human hepatocyte repopulated livers averaged 12.8% and 7.7% of body weight (p<0.