Optimization of the engineered 3D hepatic microenvironment enhances pluripotent stem cell derived hepatocyte

项目总结/摘要 肝病在美国和全世界是一个日益严重的临床问题，影响着3000万美国人 导致每年750，000人住院治疗和36，000人死亡。不幸的是，与其他常见疾病不同 如心血管疾病，肝病及其相关并发症的发病率 增加。鉴于肝病患者的稳步上升，对肝移植的需求 继续增加，而器官供应保持不变，迫切需要解决 器官短缺的问题器官移植的替代方案是细胞移植，但人类 肝细胞同样难以获得。因此，目前没有替代方案来获得稳健的 功能肝细胞，这是显着阻碍科学研究和发展更有效的 临床治疗;我们的工作解决了这一重大的研究和临床差距。多能干细胞源 肝细胞样细胞经常被引用为器官移植的潜在替代细胞来源， 制药和科学研究的平台。然而，目前无法获得健全和充分成熟的 肝细胞限制了目前生产的细胞的临床和科学应用。多能干细胞的命运和 肝细胞的成熟是由复杂的内部信号和来自肝细胞的外部信号决定的。 微环境不幸的是，我们目前的分化平台不能模拟细胞外， 基质（ECM）和体内肝脏微环境的3D复杂性，因此ECM细胞的作用 并且在分化过程中的细胞-细胞相互作用在很大程度上仍然未知。我们的核心假设是， 3D微环境和随后的细胞相互作用（例如细胞-ECM和细胞-细胞相互作用）决定了 细胞命运的决定，正是通过这些因素，胎儿细胞类型成熟为终末分化 细胞类型。在最近的工作中，我们的小组已经开发出强大的工具来系统和有效地操纵 3D微环境这项建议的中心目标是确定和检查重要的诱导细胞 体外定向多能干细胞分化的相互作用， 和功能成熟的肝细胞。我们将系统地研究和确定细胞和细胞的作用， ECM相互作用在调节胎儿到成人肝细胞的转变中起作用，并将这些发现纳入其中。 我们目前的分化和组织工程构建。这些优化的3D多细胞球体 这些平台将使科学研究和开发更有效的临床疗法成为可能。

Project Summary/Abstract Liver disease is a growing clinical problem in the United States and worldwide affecting 30 million Americans resulting in 750,000 hospitalizations and 36,000 deaths yearly. Unfortunately, unlike other common diseases such as cardiovascular diseases, the incidence of liver disease and its associated complications are increasing. Given the steady rise of patients with liver disease, the demand for liver transplantation has continued to increase while the supply of organs has remained unchanged creating a pressing need to address this organ scarcity. Proposed alternatives to organ transplantation are the transplantation of cells but human hepatocytes are similarly difficult to source. As a result, there are currently no alternatives for obtaining robust functional hepatocytes which is significantly hampering scientific study and the development of more effective clinical therapies; our work addresses this significant research and clinical gap. Pluripotent stem cell derived hepatocyte-like cells are often cited as a potential alternative cell source for organ transplants and as a platform for pharmaceutical and scientific study. However, the current inability to derive robust and fully mature hepatocytes limits the clinical and scientific utility of currently produced cells. Pluripotent stem cell fate and hepatocyte maturation is determined by the complex array of internal signals and external cues from the microenvironment. Unfortunately our current differentiation platforms do not mimic the cellular, extracellular matrix (ECM), and 3D complexities of the in vivo hepatic microenvironment and therefore the role of ECM-cell and cell-cell interactions in the differentiation process remains largely unknown. Our central hypothesis is that the 3D microenvironment and subsequent cellular interactions (e.g. cell-ECM and cell-cell interactions) dictates cell fate decisions and it is through these factors by which fetal cell types mature into terminally differentiated cell types. In recent work our group has developed robust tools to systematically and efficiently manipulate the 3D microenvironment. The central goal of this proposal is to identify and examine important inductive cellular interactions for directed pluripotent stem cell differentiation in vitro to enable the generation of phenotypically and functionally mature hepatocytes. We will systematically examine and identify the role cell-cell and cell- ECM interactions play in regulating the fetal to adult hepatocyte transition and will incorporate these findings into our current differentiation and tissue engineered constructs. These optimized 3D multicellular spheroids platforms will enable scientific study and the development of more effective clinical therapies.