Matrix in pre-cirrhotic HCC

项目摘要/摘要 非酒精性脂肪性肝炎(NASH)是肝细胞癌的主要病因。临床上认识到 NASH中的肝细胞癌通常发生在肝硬变前阶段，然而，NASH中肝细胞癌的病理机制 非肝硬变的肝脏还没有被很好地了解。由于血糖控制不佳的T2 DM是一种 肝细胞癌的独立危险因素有可能存在不同的途径，从而产生有利于- 非肝硬化性T2 DM/NASH的致癌生态位。我们的研究表明， 晚期糖基化终产物(AGEs)在T2 DM/NASH患者和动物模型中的表达 是坏死性炎症和氧化性肝损伤的关键。年龄限制的下调 受体AGER1促进了年龄的沉积，体内校正AGER1改善了NASH。 为了研究高龄环境如何为转化细胞创造允许的条件，我们 在流体动力注射HMET/突变型β-连环蛋白之前调整饮食/年龄含量：1)高 年龄背景导致更早和更具侵袭性的肝癌，2)年龄积累与 矩阵动力学的显著变化--能量耗散或损失增加以及 对变形的响应更快的应力松弛-在年龄和年龄的受体中 (RAGE)依赖方式，3)抑制AGE产生逆转基质变化 体内的粘弹性，降低了肿瘤负担。我们将检验假设，在非 肝硬变T2 DM/NASH积聚的AGEs有助于增加基质粘弹性 而母细胞的变化创造了一个有利于入侵的环境。目标1：我们建议 研究饮食/年龄含量、基质粘弹性、性别和肝癌表型之间的关系 以及一种新的肝细胞癌模型的结果。我们将特别关注年龄是如何调节的 特定氨基酸上的胶原交联物可以改变分子间的识别和相互作用 从而影响基质动力学。目标2：我们将剖析 RAGE和AGER1信号和基质粘弹性参数的年龄累积与肝细胞癌 表型。在目标3中，我们建议开发一种具有可调粘弹性的三维水凝胶体系 并将研究AGEs修饰的基质对细胞行为的影响。基于RNAseq 数据我们将集中于细胞如何感知粘弹性变化的病理机制，以及 关键的母细胞信号，赋予侵袭性和迁移性。 这些研究将证实年龄对非肝硬化性NASH患者肝基质的影响。 并概述了支持侵入性环境的病理机制。定义关键的矩阵单元 驱动入侵的信号将使我们能够在未来进行翻译研究。

Project Summary/Abstract Non-alcoholic steatohepatitis (NASH) is a major cause of HCC. It is clinically recognized that HCC in NASH often arises at a pre-cirrhotic stage, however the pathomechanism of HCC in non-cirrhotic livers is not well understood. As T2DM with poor glycemic control is an independent risk factor for HCC it is plausible that there are distinct pathways that create a pro- carcinogenic niche in non-cirrhotic T2DM/NASH. We showed that the accumulation of advanced glycation end products (AGEs) in patients with T2DM/NASH and in an animal model are key to necroinflammation and oxidative liver injury. Downregulation of the AGE clearance receptor AGER1 accelerated AGE deposition, and correcting AGER1 in vivo improved NASH. To study how high AGE environment creates permissive conditions for transformed cells, we modulated diet/AGE content prior to hydrodynamic injection of hMET/mutant β-catenin: 1) High AGE background induced an earlier and more invasive HCC, 2) AGE accumulation was linked to significant changes in matrix dynamics-with an increase in energy dissipation or loss and faster stress relaxation in response to a deformation - in an AGE and receptor for AGEs (RAGE)-dependent manner, 3) Inhibiting AGE production reversed changes in matrix viscoelasticity in vivo and lowered tumor burden. We will test the hypothesis that in non- cirrhotic T2DM/NASH accumulation of AGEs contribute to an increase in matrix viscoelasticity and matricellular changes creating a pro-invasive environment. Aim 1: We propose to investigate the link between diet/AGE content, matrix viscoelasticity, sex and HCC phenotypes and outcomes in a novel HCC model. In particular we will focus on how AGE-mediated collagen crosslinks on specific amino acids can alter intermolecular recognition and interaction with proteoglycans thereby affecting matrix dynamics. Aim 2: We will dissect the effects of RAGE and AGER1 signals and AGE accumulation on matrix viscoelastic parameters and HCC phenotypes. In Aim 3 we propose to develop a 3D hydrogel system with tunable viscoelasticity and the impact of AGEs-modified matrix on cell behavior will be studied. Based on the RNAseq data we will focus on the pathomechanism of how viscoelastic changes are sensed by cells, and the key matricellular signals that confer invasive and migratory properties. These studies will demonstrate the impact of AGEs on the liver matrix in non-cirrhotic NASH and outline the pathomechanism for a pro-invasive environment. Defining the key matricellular signals that drive invasion will enable us to pursue translational studied in the future.