Synergistic effects of ECM and heterotypic crosstalk on cellular responses in non-alcoholic fatty liver disease

ECM 和异型串扰对非酒精性脂肪肝细胞反应的协同作用

基本信息

批准号：
10744973
负责人：
Salman R Khetani
金额：
$ 59.33万
依托单位：
UNIVERSITY OF ILLINOIS AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-02-08 至 2027-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10744973
关键词：
3-Dimensional ATAC-seq Affect Animal Model Antibodies Biochemical Cell Communication Cell physiology Cells Cirrhosis Clinical Coculture Techniques Cues Data Data Set Development Disease Disease Outcome Disease Pathway Drug Screening Endothelial Cells Engineering Epidemic Epigenetic Process Exposure to Extracellular Matrix Extracellular Matrix Proteins FDA approved Fatty Acids Fibrosis Fructose Gene Expression Profile Genes Glucose Goals Hepatic Stellate Cell Hepatocyte Human Hydrogels In Vitro Individual Inflammation Insulin Investigation Kupffer Cells Liver Liver Dysfunction Liver Fibrosis Liver diseases Mechanics Mediator Microfluidics Modification Myofibroblast Normal Range Pathway interactions Patients Pharmaceutical Preparations Pharmacotherapy Phenotype Physiological Primary carcinoma of the liver cells Publishing Regulation Regulator Genes Role Series Signal Transduction Single Nucleotide Polymorphism Stimulus System Testing Therapeutic Tissue Engineering Tissues Work adverse outcome cell type cellular targeting combinatorial drug metabolism extracellular human model improved innovation insight liver biopsy metabolic-associated fatty liver disease non-alcoholic fatty liver disease nonalcoholic steatohepatitis novel novel therapeutics quantitative imaging response screening tool transcription factor transcriptome sequencing

项目摘要

ABSTRACT / PROJECT SUMMARY Non-alcoholic steatohepatitis (NASH) represents a rapidly growing epidemic of liver disease that can predispose affected individuals to advanced fibrosis and hepatocellular carcinoma. Strategies for targeting cellular mediators of liver fibrosis, such as activated hepatic stellate cells (HSCs), remain limited due to an incomplete understanding of the mechanisms governing myofibroblastic differentiation and the signaling between liver parenchymal and non-parenchymal cell (NPC) types. The importance of microenvironmental signal crosstalk, including interactions between extracellular matrix (ECM) protein composition and mechanical stiffness in cell phenotypic alterations, is increasingly appreciated. Although animal models have provided insights into NASH, significant differences across species in drug metabolism and disease pathways exist. Thus, there is a need for human-relevant in vitro approaches that enable the investigation of hepatocellular phenotypes within physiological and NASH-like microenvironments and could facilitate the high-throughput discovery of novel therapeutics. The goal of this project is to implement engineered culture platforms for selectively modulating microenvironmental signals and utilize these systems to reveal key phenotypic programming pathways and interaction mechanisms within the context of a NASH-like microenvironment. Our approach will enable hypothesis-driven studies incorporating controlled perturbations of extracellular signals. In Aim 1, we will investigate the microenvironmental regulation of myofibroblastic phenotype. Utilizing defined ECM compositions and mechanical stiffness regimes in engineered cultures, we will examine the role of epigenetic gene regulatory mechanisms and test the hypothesis that combinatorial microenvironmental cues regulate epigenetic changes that are critical for the myofibroblastic programming of human hepatic stellate cells within the context of normal and NASH-like soluble triggers. In Aim 2, we will examine the influence of ECM composition and stiffness on Kupffer cell (KC) and primary human hepatocyte (PHH) functions including reciprocal intercellular interactions and cooperative effects of NASH-like soluble stimuli. Our approach will facilitate modular control and deconvolution of the effects of ECM composition, substrate stiffness, and soluble signals. In Aim 3, we will develop and implement multicellular 3D liver microtissues for the systematic analysis of multicellular phenotype regulation in the context of disease-like ECM alterations. We will investigate heterotypic cellular crosstalk mechanisms between non-parenchymal cell types (HSC, KC, and liver sinusoidal endothelial cells), and their collective influence on primary human hepatocyte functions. We will further establish capabilities for assessing NASH-relevant therapeutics within the multicellular liver platform. The proposed studies will provide a) fundamental insights into the microenvironmental cues and signaling mechanisms underlying cellular phenotypic alterations in NASH, which could aid the development of novel drug therapies, and b) novel engineered liver culture platforms that can serve as enabling tools for a broad range of physiological and disease outcomes.

摘要 /项目摘要非酒精性脂肪性肝炎（NASH）代表肝病快速增长的流行病影响个体的纤维化和肝细胞癌。针对蜂窝介质的策略由于不完整而导致的肝纤维化（例如活化的肝星状细胞（HSC））仍然有限了解控制肌纤维细胞分化的机制和肝之间的信号实质和非核细胞（NPC）类型。微环境信号串扰的重要性，包括细胞外基质（ECM）蛋白组成与细胞中的机械刚度之间的相互作用表型变化受到越来越多的理解。尽管动物模型为纳什提供了见解，但存在药物代谢和疾病途径的物种之间的显着差异。因此，需要与人相关的体外方法，可以研究肝细胞表型生理和类似纳什的微环境，可以促进新颖的高通量发现疗法。该项目的目的是实施用于选择性调节的工程文化平台微环境信号并利用这些系统揭示关键的表型编程途径和在纳什样微环境的背景下的相互作用机制。我们的方法将使假设驱动的研究结合了细胞外信号的受控扰动。在AIM 1中，我们将研究肌纤维细胞表型的微环境调节。利用定义的ECM组成和机械刚度在工程培养物中，我们将检查表观遗传基因调节的作用机制和检验组合微环境线索调节表观遗传变化的假设对于在正常情况下，人类肝星状细胞的肌纤维细胞编程至关重要和纳什般的可溶性触发器。在AIM 2中，我们将检查ECM组成和刚度对 Kupffer细胞（KC）和原代人肝细胞（PHH）功能，包括相互互相互作用纳什样刺激的合作效应。我们的方法将促进模块化控制和 ECM组成，底物刚度和可溶性信号的效果的反卷积。在AIM 3中，我们将开发和实施多细胞3D肝微作用，以进行多细胞表型的系统分析在类似疾病的ECM改变的背景下进行调节。我们将研究异型细胞串扰非副质细胞类型（HSC，KC和肝窦内皮细胞）之间的机制及其集体影响对原代人肝细胞功能。我们将进一步建立评估的功能多细胞肝平台内的NASH相关治疗。拟议的研究将提供a）对微环境提示和信号传导机制的基本见解 NASH的改变，可以帮助开发新的药物疗法，b）新型肝脏的新型肝脏可以作为广泛生理和疾病结果的促成工具的文化平台。

项目成果

期刊论文数量（10）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Combinatorial Microgels for 3D ECM Screening and Heterogeneous Microenvironmental Culture of Primary Human Hepatic Stellate Cells.

用于原代人肝星状细胞的 3D ECM 筛选和异质微环境培养的组合微凝胶。

DOI：
10.1101/2023.05.05.539608
发表时间：
2023
期刊：
bioRxiv : the preprint server for biology
影响因子：
0
作者：
Ryoo,Hyeon;Underhill,GregoryH
通讯作者：
Underhill,GregoryH

High throughput interrogation of human liver stellate cells reveals microenvironmental regulation of phenotype.

DOI：
10.1016/j.actbio.2021.11.015
发表时间：
2022-01-15
期刊：
Acta biomaterialia
影响因子：
9.7
作者：
Brougham-Cook A;Jain I;Kukla DA;Masood F;Kimmel H;Ryoo H;Khetani SR;Underhill GH
通讯作者：
Underhill GH

Effect of distinct ECM microenvironments on the genome-wide chromatin accessibility and gene expression responses of hepatic stellate cells.

不同 ECM 微环境对肝星状细胞全基因组染色质可及性和基因表达反应的影响。

DOI：
10.1016/j.actbio.2023.06.018
发表时间：
2023
期刊：
Acta biomaterialia
影响因子：
9.7
作者：
Jain,Ishita;Brougham-Cook,Aidan;Underhill,GregoryH
通讯作者：
Underhill,GregoryH

Regulatory role of cholesterol in modulating actin dynamics and cell adhesive interactions in the trabecular meshwork.

胆固醇在调节小梁网中肌动蛋白动力学和细胞粘附相互作用中的调节作用。

DOI：
10.1101/2024.02.02.578717
发表时间：
2024
期刊：
bioRxiv : the preprint server for biology
影响因子：
0
作者：
Wang,Ting;Kimmel,HannahRC;Park,Charles;Ryoo,Hyeon;Liu,Jing;Underhill,GregoryH;Pattabiraman,PadmanabhanP
通讯作者：
Pattabiraman,PadmanabhanP

Spatially Defined Cell-Secreted Protein Detection Using Granular Hydrogels: μGeLISA.

使用颗粒水凝胶进行空间定义的细胞分泌蛋白检测：μGeLISA。