Microenvironmental Control of Liver Progenitor Cell Differentiation and Spatial Patterning

肝祖细胞分化和空间模式的微环境控制

• 批准号: 10598474
• 负责人: Gregory H Underhill
• 金额: $ 47.33万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-15 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10598474
• 关键词: 3-Dimensional; Address; Adult; Anatomy; Animal Model; Biliary; Biochemical; Biological; Biological Models; Biomechanics; Cell Communication; Cell Culture System; Cell Culture Techniques; Cell Differentiation process; Cells; Chronic; Communication; Cues; Degenerative Disorder; Development; Developmental Process; Disease; Duct (organ) structure; Embryonic Development; Engineering; Epithelial Cells; Exhibits; Functional disorder; Geometry; Goals; Hepatocyte; Human; Individual; Injury; Intrahepatic bile duct; Investigation; Ligands; Link; Liver; Liver Regeneration; Liver diseases; Malignant Neoplasms; Malignant neoplasm of liver; Maps; Mechanical Stress; Mechanics; Mesenchyme; Modeling; Morphogenesis; Natural regeneration; Nature; Notch Signaling Pathway; Pattern; Play; Portal vein structure; Process; Proliferating; Reaction; Regulation; Research; Research Personnel; Role; Signal Pathway; Signal Transduction; Specific qualifier value; Structure; Technology; Testing; Tissue Engineering; Tissues; Work; bile duct; bile formation; cell fate specification; cholangiocyte; combinatorial; design; differentiation protocol; disease mechanisms study; drug testing; experience; experimental study; in vitro Model; induced pluripotent stem cell; insight; jagged1 protein; liver development; mechanical signal; notch protein; novel; novel strategies; novel therapeutics; progenitor; response; single-cell RNA sequencing; stem cell differentiation; stem cell function; stem cells; therapeutic development; tissue culture; tool; two-dimensional

Project Summary The bipotential differentiation of liver progenitor cells to hepatocytes and biliary epithelial cells (cholangiocytes) is integral to liver development, regeneration, and diseases including bile duct paucity and liver cancer. In particular, the most common congenital liver diseases are associated with bile duct dysfunction. In addition to the developmental processes during embryogenesis, duct morphogenesis also occurs in the adult liver in response to severe and chronic injury. These so-called ductular reactions exhibit highly variable differentiation patterns, and although these reactions significantly contribute to the proliferative responses in the liver, they remain poorly characterized. Notably, ductular proliferations in the adult liver are concentrated near the portal vein region, similar to the formation of bile ducts during development. Despite substantial research efforts, the structural complexity and dynamic nature of liver development and regeneration has limited the comprehensive understanding of disease mechanisms as well as the advancement of new therapeutic options. The long-term goal of this project is to develop complementary two-dimensional and three-dimensional engineered tissue platforms that can be applied towards the investigation of liver progenitor cell differentiation mechanisms that are presently inaccessible with current cell culture systems and animal models. Towards this end, we will pursue the following research objectives, which are specifically targeted towards deconstructing the combined influence of biochemical and biomechanical signals in liver progenitor cell fate specification. In Aim 1, we will utilize a cell microarray platform to investigate the influence of spatial gradients of Notch signaling and cell mechanical stresses in progenitor cell differentiation. Our approach will enable the independent control of cell- cell interactions, defined by multicellular geometry, and specific exogenous microenvironmental signals presented within the array platform. In Aim 2, we will develop and utilize a three-dimensional microtissue culture platform to systematically investigate the effects of three-dimensional geometry and determine how distinct multicellular geometries regulate differentiation patterns. These research efforts will establish microscale tissue engineering tools that enable the controlled presentation and systematic perturbation of a range of microenvironmental signals. In Aim 3, we will extend our studies towards the direct analysis of human liver differentiation mechanisms through the integration of human induced pluripotent stem (iPS) cell-derived liver progenitor cells. Collectively, our approach will allow for novel studies into the mechanisms of liver progenitor cell differentiation, including the unique examination of the combinatorial influence of cell mechanical stress gradients and Notch signaling. Further, we envision that these platforms are generalizable, and could be implemented as enabling technologies for fundamental biological investigation and therapeutic development efforts for a broad range of cell and tissue contexts.

项目摘要 肝脏祖细胞对肝细胞和胆道上皮细胞的双性分化（胆管细胞） 是肝脏发育，再生和疾病包括胆管衰弱和肝癌的组成部分。在 特别是，最常见的先天性肝脏疾病与胆管功能障碍有关。此外 胚胎发生过程中的发育过程，导管形态发生也发生在成年肝脏中 对严重和慢性损伤的反应。这些所谓的导管反应表现出高度可变的分化 模式，尽管这些反应显着有助于肝脏的增殖反应 保持不佳的特征。值得注意的是，成年肝脏中的延展性浓缩在门户附近 静脉区域，类似于发育过程中胆管的形成。尽管进行了大量研究，但 肝发育和再生的结构复杂性和动态性质限制了综合性 了解疾病机制以及新的治疗选择的发展。长期 该项目的目标是开发互补的二维和三维工程组织 可以应用于研究肝脏祖细胞分化机制的平台 目前与当前的细胞培养系统和动物模型无法访问。为此，我们将 追求以下研究目标，这些目标是针对解构合并的目标 生化和生物力学信号在肝祖细胞命运规范中的影响。在AIM 1中，我们将 利用细胞微阵列平台来研究Notch信号和细胞的空间梯度的影响 祖细胞分化的机械应力。我们的方法将使细胞独立控制 - 细胞相互作用，由多细胞几何形状和特定的外源微环境信号定义 在数组平台内呈现。在AIM 2中，我们将开发和利用三维微动物 培养平台系统地研究三维几何形状的影响，并确定如何 不同的多细胞几何形状调节分化模式。这些研究工作将确定 显微镜组织工程工具，可实现受控的演示和系统扰动 微环境信号范围。在AIM 3中，我们将把研究扩展到对人类的直接分析 通过整合人类诱导的多能茎（IPS）细胞来源的肝分化机制 肝祖细胞。总的来说，我们的方法将允许对肝脏机制进行新颖的研究 祖细胞分化，包括对细胞组合影响的独特检查 机械应力梯度和缺口信号传导。此外，我们设想这些平台是可推广的， 并可以作为基本生物学研究和治疗的能力实施 针对各种细胞和组织环境的开发工作。

项目成果

Engineered Platforms for Maturing Pluripotent Stem Cell-Derived Liver Cells for Disease Modeling.

• DOI: 10.1016/j.jcmgh.2023.01.013
• 发表时间: 2023
• 期刊: CELLULAR AND MOLECULAR GASTROENTEROLOGY AND HEPATOLOGY
• 影响因子: 7.2
• 作者: Yuan, Yang;Cotton, Kristen;Samarasekera, Dinithi;Khetani, Salman R.
• 通讯作者: Khetani, Salman R.

Role of Extracellular Matrix Biomolecules on Endometrial Epithelial Cell Attachment and Cytokeratin 18 Expression on Gelatin Hydrogels.

• DOI: 10.1021/acsbiomaterials.2c00247
• 发表时间: 2022-09-12
• 期刊: ACS BIOMATERIALS SCIENCE & ENGINEERING
• 影响因子: 5.8
• 作者: Zambuto, Samantha G.;Jain, Ishita;Clancy, Kathryn B. H.;Underhill, Gregory H.;Harley, Brendan A. C.
• 通讯作者: Harley, Brendan A. C.

Delineating cooperative effects of Notch and biomechanical signals on patterned liver differentiation.

• DOI: 10.1038/s42003-022-03840-9
• 发表时间: 2022-10-07
• 期刊: Communications biology
• 影响因子: 5.9