poster session iii abstracts 1195 1637

HEPATOLOGY, VOLUME 64, NUMBER (SUPPL) AASLD ABSTRACTS 601A 1194 Clinical study of liver regenerative therapy of cirrhosis using autologous adipose tissue-derived stromal cells and the characterization of the obtained stromal cells 1195 ♦ Therapeutic targeting of Hsp90 alleviates liver macrophage-specific NLRP3 inflammasome activation in alcoholic liver injury Yoshio Sakai1, Akihiro Seki1, Hajime Sunagozaka1, Takeshi Terashima1, Kazunori Kawaguchi1, Hatsune Mochida2, Alessandro Nasti2, Geraldine B Buffa1, Masatoshi Yamato2, Kosuke Ishida4, Masaaki Takamura2, Soichiro Usui2, Takashi Wada3, Masao Honda1, Shuichi Kaneko4,1; 1Department of Gastroenterology, Kanazawa University, Kanazawa, Japan; 2Disease Control and Homeostasis, Kanazawa University, Kanazawa, Japan; 3Department of Nephrology, Kanazawa University, Kanazawa, Japan; 4System Biology, Kanazawa University, Kanazawa, Japan Daniel Bullock, Asmita Choudhury, Pranoti Mandrekar; Medicine, University of Massachusetts Medical Center, Worcester, MA Adipose tissue is enriched with mesenchymal stromal/stem cells We conducted the clinical study for liver regenerative therapy of cirrhosis by intrahepatic arterial administration of freshly isolated autologous adipose tissue derived stromal/stem (regenerative) cells (ADRCs) (UMIN000009122, NCT01062750) We also characterized the obtained fresh cells (ADRCs) as well as the cultured expanded stromal cells (ADSCs) [Methods] The objectives were cirrhosis patients who provided the written informed consent The patients underwent liposuction of their subcutaneous adipose tissues in their abdomen or buttock The obtained adipose tissues were immediately processed using the adipose-tissue dissociation equipment (Celution®, Cytori Therapeutics Inc.) to obtain autologous ADRCs The designated number of cells (3.3x10^5/Kg (n=2), 6.6x10^5/Kg (n=2)) were infused through the catheter positioned at the hepatic artery Safety evaluation was assessed month after the treatment Surface antigen of ADRCs and cultured cells (ADSCs) were assessed by FACS Serum cytokine/chemokine concentration was measured by Bio-Plex® We also analyzed gene expression of the freshly isolated ADRCs compared to the cultured ADSCs by DNA microarray [Result] Four liver cirrhosis patients were enrolled, (HI-01(type C), HI-03 (type C), HI-04 (NASH), HI-05(type B)) The number of infused ADRCs were 2.2x10^7 ~ 4.4x10^7 Among ADRCs and ADSCs, 10.3 ~45.8 % and 80.998.9% of cells, respectively, expressed the mesenchymal stem cell surface marker CD44 No severe adverse events occurred Three among treated patients improved serum albumin concentration during 36 months after treatment Serum HGF, M-CSF, MIF, IL18, and IL-6 were elevated in all patients one day after treatment The surplus ADRCs after treatment were successfully expanded and the spindle-like shape of mesenchymal stem cell was observed Gene expression profile analysis using clusters analysis showed the two distinct clusters discerning ADRCs from ADSCs, completely, regardless of the patient’s etiologies of cirrhosis The freshly isolated ADRCs were shown to involve inflammatory features, suggesting that they were related to immunomodulatory biological effects [Conclusion] Intrahepatic arterial administration of autologous freshly isolated ADRCs with immunomodulatory biological effects were confirmed to be safely conducted without serious adverse events Disclosures: Background/Aims: Inflammasomes are multimeric protein complexes that respond to PAMPs or DAMPs that serve as a scaffold to promote cleavage of pro-caspase-1 and subsequent active IL-1β and IL-18 production Previous studies show activation of NLRP3 inflammasome in alcoholic liver disease (ALD) Further the role of stress-mediated protein heat shock protein 90 (hsp90) as a chaperone for NLRP3 inflammasome scaffold was identified Studies in our laboratory have demonstrated that chronic alcohol induces hsp90 and its inhibition alleviates alcoholic liver injury Here we hypothesize that hsp90 is required for alcohol mediated NLRP3 inflammasome activity in the liver, specifically in macrophages/Kupffer cells and its therapeutic targeting reduces active IL-1β in the alcoholic liver Methods: C57/BL/6J mice were subjected to weeks and weeks of chronic-single binge or chronic-multiple binge alcohol feeding respectively Specific hsp90 inhibitor, 17-DMAG was injected intraperitoneally either every alternate day (5mg/ kg) or at the end of the alcohol feeding (50mg/kg) Whole livers were harvested and subjected to inflammasome qPCR and IL-1β ELISA Hepatocytes and liver macrophages were isolated at the end of the feeding and NLRP3 and IL-1β analyzed For mechanistic studies, bone marrow derived macrophages (BMDM) were differentiated in vitro and stimulated with LPS ± ATP and DMAG (0.5mM-2.0mM) or directly heat shocked and stimulated with ATP, and analyzed for inflammasome mRNA and protein Results: Chronic alcohol mediated induction of NLRP3 expression (p