Development of drugs targeting liver fibrosis by modulation of protein/lipid glycosylation

2019 Fiscal Year Final Research Report

• Project/Area Number: 18K19530
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 53:Organ-based internal medicine and related fields
• Research Institution: Hokkaido University
• Principal Investigator: Sakamoto Naoya 北海道大学, 医学研究院, 教授 (10334418)
• Co-Investigator(Kenkyū-buntansha): 須田 剛生 北海道大学, 大学病院, 特任助教 (20447460) ; 古川 潤一 北海道大学, 医学研究院, 特任准教授 (30374193)
• Project Period (FY): 2018-06-29 – 2020-03-31
• Keywords: 肝線維化

Outline of Final Research Achievements

In order to search for novel therapeutic targets targeting liver fibrosis, which was previously thought to be an irreversible pathological condition, the complex sugar chain structure of the liver tissue and serum of liver cirrhosis cases, and the surface protein of cultured hepatic stellate cells was investigated. The purpose of this study was to analyze the changes along with the dissease stage and their related mechanisms. (1) Using cultured hepatic stellate cell-derived RI-T cell line and primary human hepatic stellate cells, set culture conditions for constructing activated stellate cells by LPS stimulation, TGF-β addition culture, etc. under Kuppfer cell co-culture. (2) We found that 4 secretory micro RNAs was overexpressed in relation to the activation of cultured hepatic stellate cells. Gene expression of alpha-SMA, type I collagen, and chemokine genes associated with fibrosis was observed when each micro RNA was introduced into cultured stellate cells.

Free Research Field

消化器内科

Academic Significance and Societal Importance of the Research Achievements

肝線維化に関連する患者血清中の複合糖鎖解析とその機能解析から抗体薬等の開発の可能性を見いだした。また、血清分泌型microRNAの網羅的探索、MSC培養液から肝線維化抑制効果を担う蛋白、小分子の探索など、多くの治療標的を同定し、創薬につながる基盤を形成することができた。
今後、同定された候補miRNAの制御遺伝子のpashway解析、さらにanti-miRNAによる発現抑制が星細胞活性化抑制、肝線維化抑制効果をもたらすか否かを検証する。さらに、SILAC法による活性化星細胞のプロテオーム解析および候補蛋白の抽出をおこなう。