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Human Hepatocyte and Discovery Core

人类肝细胞和发现核心

基本信息

批准号：
10560532
负责人：
Ype Peter De Jong
金额：
$ 40.06万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-05 至 2027-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10560532
关键词：
Address Animals Biology Blood Coagulation Factor Breeding CRISPR/Cas technology Cells Chimerism Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Cryopreservation Engraftment F8 gene Fetal Liver Gene Transfer Genes Genetic Engineering Harvest Hematopoietic stem cells Hemophilia A Hepatic Hepatocyte Hepatocyte transplantation Hepatotoxicity Human Immune Immune response Immune system Immunobiology Immunocompetent Immunologic Deficiency Syndromes In Vitro Institution Investigation Knock-out Kupffer Cells Lentivirus Vector Liver Modeling Molecular Mus Natural Killer Cells Plasma Process RNA analysis Rag1 Mouse Research Personnel Safety Services System Therapeutic Tissues Transgenic Organisms Transplantation Viral Viral Genes Viral Vector cDNA Library experience experimental study fetal gene therapy genetic manipulation humanized mouse immune activation in vivo liver injury male overexpression preservation programs response retransplantation single-cell RNA sequencing two-dimensional vector

项目摘要

PROJECT SUMMARY/ABSTRACT Gene therapy for hemophilia A holds promise to accomplish a lasting cure. Adeno-associated viral (AAV) gene transfer of clotting factor FVIII to the livers of males can achieve therapeutic correction but this is not sustained. The reasons for hepatotoxicity and the decline in FVIII expression in clinical trials are unclear. The AAV therapies currently under investigation target hepatocytes in the liver. There is, however, a limited understanding of the interactions between the vector and hepatocytes as well as effect of FVIII expression and immune responses. Core B will provide primary human hepatocyte models to address these basic and mechanistic questions related to the biology of AAV and FVIII. The central hypothesis of this proposal is that multiple interconnected features of AAV and FVIII biology limit durability of therapeutic expression and pose serious safety concerns. The objectives of Core B, the Human Hepatocyte and Discovery Core, is to provide three primary human hepatocytes models to study AAV vector and FVIII biology. The first model is primary hepatocyte cultures that retain stable hepatocyte functions in 2-dimensional format. These will be used for in vitro studies. The second model is chimeric mice whose livers are highly repopulated with primary human hepatocytes. These will be used for in vivo AAV studies. And the third model is mice doubly engrafted with hematopoietic stem cells and livers for in vivo studies of human immune cells in the liver. Core B will provide these services to investigators to address major unanswered questions in FVIII biology, gene therapy for hemophilia, liver-directed gene transfer, and molecular and immunobiology of AAV vectors.

项目概要/摘要 A 型血友病的基因疗法有望实现持久治愈。腺相关病毒（AAV）基因将凝血因子 FVIII 转移至男性肝脏可以实现治疗纠正，但这并不能持续。临床试验中肝毒性和 FVIII 表达下降的原因尚不清楚。 AAV疗法目前正在研究肝脏中的目标肝细胞。但人们对此的了解还很有限载体和肝细胞之间的相互作用以及 FVIII 表达和免疫反应的影响。核心 B 将提供初级人类肝细胞模型来解决这些相关的基本和机制问题 AAV 和 FVIII 的生物学。该提案的中心假设是多个相互关联的特征 AAV 和 FVIII 生物学特性限制了治疗表达的持久性并造成严重的安全问题。这核心 B（人类肝细胞和发现核心）的目标是提供三种原代人类肝细胞研究 AAV 载体和 FVIII 生物学的模型。第一个模型是原代肝细胞培养物，保持稳定肝细胞以二维形式发挥功能。这些将用于体外研究。第二个模型是嵌合小鼠的肝脏高度重新填充了原代人类肝细胞。这些将用于体内 AAV 研究。第三种模型是对小鼠进行造血干细胞和肝脏双重移植，以进行短期治疗。肝脏中人体免疫细胞的体内研究。核心 B 将向调查人员提供这些服务来解决 FVIII 生物学、血友病基因治疗、肝脏定向基因转移和 AAV 载体的分子和免疫生物学。