Perfusion decellularization and recellularization for the creation of a bioengineered transplantable liver

灌注脱细胞和再细胞化用于创建生物工程可移植肝脏

• 批准号: 10018893
• 负责人: Jeffrey James Ross
• 金额: $ 86.79万
• 依托单位: MIROMATRIX MEDICAL, INC.
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-17 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10018893
• 关键词: Activities of Daily Living; Acute; Acute Liver Failure; Address; Affect; Albumins; Allogenic; Ammonia; Animal Model; Animals; Anticoagulant therapy; Bile Acids; Bile Duct Epithelium; Bile fluid; Biological Assay; Biomedical Engineering; Blood; Blood Vessels; Blood flow; Cause of Death; Cells; Chronic; Clinical Research; Coculture Techniques; Conditioned Culture Media; Data; Development; Donor person; Endothelial Cells; Endothelium; Engraftment; Excision; Family suidae; Goals; Hepatocyte; Hour; Human; Implant; In Vitro; Legal patent; Liver; Liver Failure; Liver diseases; Measures; Medical; Modeling; Organ; Organ Procurements; Perfusion; Phase; Production; Recovery; Recovery of Function; Reporting; Seeds; Serum; Small Business Innovation Research Grant; Structure; Survival Rate; Technology; Testing; Therapeutic; Transplantation; Umbilical vein; United States; Urea; Vascular Patency; Very low density lipoprotein; base; bile duct; cell type; cholangiocyte; clinically translatable; graft function; in vivo; liver function; liver transplantation; meetings; novel therapeutics; reconstitution; scaffold; screening; transplant model

ABSTRACT Liver transplantation currently represents the only treatment for end stage liver disease (ESLD), the 8th most frequent cause of death in the United States (US). Efforts to overcome the chronic shortage of transplantable human livers include attempts at alternative organ procurement and replacement strategies. The most promising approach to date that provides the appropriate matrix composition and supporting vascular structures has been the development of perfusion decellularization, which enables the removal of cellular material for a native organ while maintaining the native matrix, structure and vascular of the liver. Utilizing perfusion decellularization and recellularization technology, several groups have demonstrated the ability to seed a variety of liver-specific cell types into decellularized liver constructs, however reconstitution of the endothelial cell lining of the vascular networks in these scaffolds has remained a significant challenge to the development of a therapeutic bioengineered liver (BEL). Miromatrix Medical Inc. has recently reported revascularizing a clinically translatable porcine-derived liver scaffold and demonstrated in vivo graft patency in a large animal model without sustained administration of anticoagulant therapies. Solving the critical vascular constraint allows this Direct to Phase II SBIR project to focus on production and testing of a fully functional BEL to address the chronic shortage of transplantable livers and develop new therapeutic options to those with ESLD utilizing Miromatrix’ broad proprietary perfusion decellularization and recellularization technology. In Specific Aim #1, we will demonstrate recellularized liver graft function in an acute liver failure model. The optimal media and seeding conditions for hepatocyte functionality will be selected (1.1). Equivalency of human hepatocytes to porcine hepatocytes will be demonstrated (1.2), and vascular patency of endothelial and hepatocyte recellularized livers will be confirmed in acute blood loops (1.3). Forty-eight hour functionality of this liver graft will be demonstrated in pigs (1.4). In Specific Aim #2, we will optimize the seeding of cholangiocytes into liver grafts recellularized with endothelial cells and hepatocytes and identify assays that will allow assessment of cholangiocyte function after seeding to verify successful engraftment (2.1). We will then create tri-culture grafts using only human cells and test for equivalency to the porcine+human grafts (2.2) and determine whether addition of cholangiocytes to endothelial+hepatocyte liver grafts adversely affects graft patency (2.3). Finally, in Specific Aim #3, we will demonstrate the BEL functionality in an orthotopic chronic liver failure model by developing a transplant model and evaluating baseline data (3.1) and by demonstrating liver functionality for fourteen days in a recovery model in pigs (3.2). Data generated will be used in a Pre-IND submission meeting to define the testing required for complete human recellularization and functional testing of the BEL grafts to initiate clinical studies.

摘要 肝移植目前是终末期肝病（ESLD）的唯一治疗方法，在世界上排名第8位。 在美国常见的死亡原因（US）。努力克服可移植器官的长期短缺 人类肝脏包括替代器官获取和替代策略的尝试。最有前途的 迄今为止，提供适当基质组成和支持血管结构的方法已经 灌注去细胞化的发展，这使得能够去除天然器官的细胞物质 同时保持肝脏的天然基质、结构和血管。利用灌注脱细胞和 利用再细胞化技术，几个研究小组已经证明了接种各种肝脏特异性细胞的能力 类型的脱细胞肝结构，然而，血管内皮细胞衬里的重建， 这些支架中的网络仍然是开发治疗药物的重大挑战。 生物工程肝（BEL）。Miromatrix Medical Inc.最近报道了一种临床上可以翻译为 猪源性肝脏支架，并在大型动物模型中证明了体内移植物通畅性， 给予抗凝治疗。解决关键的血管约束允许直接进入阶段II SBIR项目的重点是生产和测试功能齐全的BEL，以解决长期短缺的问题。 可移植的肝脏，并开发新的治疗选择，以那些与ESLD利用Miromatrix的广泛 专有的灌注脱细胞和再细胞化技术。 在具体目标#1中，我们将在急性肝功能衰竭模型中证明再细胞化肝移植功能。的 将选择用于肝细胞功能的最佳培养基和接种条件（1.1）。人体等效性 将证明肝细胞到猪肝细胞（1.2），以及内皮和 肝细胞再细胞化肝脏将在急性血液循环中得到证实（1.3）。48小时的功能， 肝脏移植将在猪中得到证实（1.4）。在具体目标#2中，我们将优化胆管细胞的接种 用内皮细胞和肝细胞再细胞化的肝移植物中，并确定允许 评估接种后的胆管细胞功能以验证成功的植入（2.1）。我们将创造 仅使用人细胞进行三代培养移植物，并测试与猪+人移植物的等效性（2.2），并确定 向内皮细胞+肝细胞肝移植物中添加胆管细胞是否会对移植物通畅性产生不利影响（2.3）。 最后，在具体目标#3中，我们将在原位慢性肝功能衰竭模型中证明BEL的功能 通过开发移植模型和评估基线数据（3.1），并通过证明肝脏功能， 在猪恢复模型中持续14天（3.2）。生成的数据将用于Pre-IND提交会议 定义完成人体再细胞化所需的试验和BEL移植物的功能试验， 启动临床研究。