Development of Complex Liver Organoids Using Cell-Specific Patterned Biomaterials

使用细胞特异性图案化生物材料开发复杂的肝脏类器官

基本信息

批准号：
10654156
负责人：
Muhammad Rizwan
金额：
$ 44.34万
依托单位：
MICHIGAN TECHNOLOGICAL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-01 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10654156
关键词：
3-Dimensional Adult Affect Alagille Syndrome Area Bile Duct Diseases Bile fluid Biocompatible Materials Biological Assay Biomechanics Biomedical Research Cells Childhood Coculture Techniques Complex Cues Culture Techniques Data Development Differentiation Antigens Disease model Ductal Epithelial Cell Engineering Foundations Generations Genes Growth Hepatocyte Human Ink Ligands Liver Liver Cirrhosis Liver diseases Modeling Monitor Morphogenesis Mus Organoids Pattern Pharmaceutical Preparations Printing Public Health Relaxation Reporter Research Research Project Grants Signal Transduction Stress Structure System Technology Therapeutic Time Tissue Engineering Tissue Model Training Transplantation Tubular formation Work bile duct bioink bioprinting cancer cell cell type cholangiocyte density drug testing experience improved liver development liver function liver transplantation mechanotransduction new technology notch protein photoactivation response spatiotemporal stem cells three dimensional cell culture undergraduate student viscoelasticity

3-Dimensional Adult Affect Alagille Syndrome Area Bile Duct Diseases Bile fluid Biocompatible Materials Biological Assay Biomechanics Biomedical Research Cells Childhood Coculture Techniques Complex Cues Culture Techniques Data Development Differentiation Antigens Disease model Ductal Epithelial Cell Engineering Foundations Generations Genes Growth Hepatocyte Human Ink Ligands Liver Liver Cirrhosis Liver diseases Modeling Monitor Morphogenesis Mus Organoids Pattern Pharmaceutical Preparations Printing Public Health Relaxation Reporter Research Research Project Grants Signal Transduction Stress Structure System Technology Therapeutic Time Tissue Engineering Tissue Model Training Transplantation Tubular formation Work bile duct bioink bioprinting cancer cell cell type cholangiocyte density drug testing experience improved liver development liver function liver transplantation mechanotransduction new technology notch protein photoactivation response spatiotemporal stem cells three dimensional cell culture undergraduate student viscoelasticity

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项目摘要

Liver bile-duct diseases are one of the main causes of liver transplantation, and often result in liver cirrhosis, affecting millions of US citizens. Current human liver organoids lack integrated bile ducts, which makes it difficult to accurately model liver diseases due to a lack of bile-transport system. Liver organoids containing bile ducts have been difficult to create because the hepatocytes and bile-duct cells in the liver have vastly different needs for both Notch signaling and biomechanical cues, which the current 3D-culture techniques are not capable of providing simultaneously. Thus, there is a critical need for a technology that can simultaneously deliver targeted Notch signaling and biomechanical cues to both types of liver cells in co-culture, in order to maintain their maturation. Exciting preliminary studies indicate that the dual-bioink-based bioprinted constructs can provide cell-type-specific signals within a co-culture matrix. Moreover, building on their previous work, the PI has developed new engineered matrix to precisely tune the bio-mechanical cues (stiffness and viscoelasticity) in a cell-specific manner, which supports the growth of 3D human bile-duct network. Accordingly, the objective of this proposal is to evaluate and optimize the effect of patterned Notch signaling and bio-mechanical cues in co-cultured liver cells to develop liver organoids with integrated bile-flow system. The rationale is that liver organoids with integrated bile-flow system will mimic liver function, thus will improve disease modelling and drug testing. The proposed research will pursue two specific aims: (1) Determine the effect of spatio-temporal Notch activation on liver organoid functions, and (2) Optimize the maturation of liver organoid via cell-type-specific biomechanical cues. In the first aim, bioprinted constructs will be developed using two distinct, cell-laden bioinks with and without photo- activatable Notch ligands, to achieve targeted Notch activation in co-culture. The organoids will be analyzed using Notch target genes and a range of liver functional assays. In the second aim, an 18-condition matrix screen will be developed to systematically evaluate and optimize the effect of patterned biomechanical cues on liver organoids in a bioprinted co-culture construct. Finally, the mechano-sensing mechanism will be evaluated in co-culture construct. The proposed research is expected to be significant because it will leverage targeted Notch signaling and biomechanical cues to inform the development of liver organoids with integrated bile ducts for therapeutic applications, and will train a diverse group of undergraduate students in the area of bioprinting, biomaterials, and liver tissue engineering.

肝胆管疾病是肝移植的主要原因之一，通常导致肝硬化，影响数百万美国公民。当前的人肝脏类器官缺乏综合的胆管，这使得由于缺乏胆汁传输系统，难以准确地对肝病进行建模。含有胆管的肝脏类器官很难产生，因为肝脏中的肝细胞和胆管细胞对Notch信号传导和生物力学提示都有很大不同的需求，而当前的3D文化技术无法同时提供。因此，对于可以同时向两种类型的肝细胞传递有针对性的缺口信号传导和生物力学提示的技术，以维持其成熟。令人兴奋的初步研究表明，基于双生物联体生物的生物打印构建体可以在共培养基质中提供细胞类型的特异性信号。此外，在他们先前的工作的基础上，PI开发了新的工程矩阵，以特定于细胞的方式准确调整生物力学提示（刚度和粘弹性），从而支持3D人类胆管网络的生长。因此，该提案的目的是评估和优化共培养肝细胞中图案化的Notch信号传导和生物力学提示的效果，以开发具有整合的胆汁流系统的肝癌。理由是，具有整合胆液系统的肝脏类器官将模仿肝功能，从而改善疾病建模和药物测试。拟议的研究将追求两个具体的目的：（1）确定时空缺口激活对肝脏器官功能的影响，以及（2）通过细胞型特异性生物力学提示优化肝脏器官的成熟。在第一个目标中，将使用有和没有光活化的凹口配体的两个不同的，含有细胞的生物学来开发生物打印构建体，以实现共培养中的靶向缺陷激活。将使用Notch靶基因和一系列肝功能测定法分析器官。在第二个目标中，将开发一个18条件矩阵屏幕，以系统地评估和优化图案化的生物力学提示对生物培养共同培养构建体中肝脏类器官的影响。最后，将在共培养构建体中评估机械感应机制。拟议的研究预计将是重要的，因为它将利用有针对性的Notch信号传导和生物力学提示来告知肝脏器官的开发，该肝脏类器官具有用于治疗应用的集成胆管，并将在生物植物，生物材料和肝组织工程学领域培训各种各样的本科生。