Molecular Determinants of liver sinusoidal endothelial cells for hepatic regeneration

肝窦内皮细胞肝再生的分子决定因素

基本信息

批准号：
10682071
负责人：
Shahin Rafii
金额：
$ 46.9万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
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：
2023-05-01 至 2028-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10682071
关键词：
3-Dimensional Acetaminophen Adult Angiopoietin-2 Anticoagulation Biochemical Biophysics Blood Blood Vessels Capillarity Cardiovascular system Cell Line Cell physiology Cells Central Vein Cicatrix Cirrhosis Development EGF gene Endothelial Cells FGF2 gene FLI1 Transcription Factor Fibrosis Functional disorder GATA4 gene Hepatectomy Hepatic Hepatic artery Hepatocyte Homeostasis Human Immune Impairment Inflammation Mediators Inflammatory Injury Kupffer Cells Liver Liver Dysfunction Liver Regeneration Liver diseases Maintenance Microfluidic Microchips Molecular Morbidity - disease rate Mus Natural regeneration Nature Organ Outcome Study Partial Hepatectomy Pathogenesis Phenotype Physiological Portal vein structure Procedures Recovery Research Role Signal Transduction Specific qualifier value Stress Testing Tissues VEGFA gene Vascular Endothelial Cell Vascularization Vasomotor WNT2 gene constitutive expression design efficacious treatment end stage liver disease factor C fetal healing liver function liver injury liver repair liver transplantation mortality organ repair prevent regeneration function regenerative repaired self assembly stellate cell stem cells therapy development transcription factor

项目摘要

ABSTRACT End-stage liver disease is associated with morbidity and mortality often requiring a liver transplant. Dysfunction and capillarization of liver sinusoidal endothelial cells (LSECs) could contribute to impaired hepatic repair and cirrhosis. Thus, uncovering the mechanisms by which LSECs acquire their specialized functions to support hepatic repair could enable the development of therapies for scar-free liver repair. We have shown that activation of Id1 and Cxcr7 in LSECs induce angiocrine factors, that guide hepatic regeneration without fibrosis. However, the mechanism by which LSECs acquire their pro-hepatic functions is unknown. We show that while transcription factors (TFs) Fli1 and Erg dictate the vascular fate and homeostasis of LSECs (Gomez- Salinero JM, et al, Nature Cardiovascular Research, 2022), induction of TF c-Maf specifies the phenotype and regenerative functions of LSECs (Gomez-Salinero JM, et al, Cell Stem Cell, 2022). Induction of c-Maf in generic human endothelial cells (ECs) switches on liver-specific LSEC signatures and angiocrine factors supporting hepatocyte functionality. In mice in which c-Maf is deleted in adult ECs, recovery from CCl4 results in fibrosis and LSECs regression to arterial cell fate. Thus, we hypothesize that maintenance of LSEC vascular cell fate requires constitutive Fli1 or Erg expression, sustaining LSECs homeostatic functions. Induction of c- Maf enforces specialized pro-regenerative functions and interactions of LSECs with hepatocytes, Kupffer and stellate cells that prevents stress-induced LSEC arterialization promoting hepatic repair without fibrosis. To this end, we have reprogrammed human generic ECs to an adaptable tubulogenic state. These Reset-Vascular Endothelial Cells (R-VECs) self-assemble into a 3D vascular network, transporting human blood and arborizing hepatocytes (Palikuqi B et al. Nature, 2020). The vascularized hepatic aggregates with stellate and Kupffer cells within scalable and perfusable microfluidic devices establish a human Hepatic-on-VascularNet platform, enabling study of physiologically adaptive cross-talk between human hepatocytes and LSECs. This hypothesis will be tested by performing these Aims: Aim 1: Define the mechanism by which hierarchical Fli1 and Erg expression through c-Maf induction sustains specialization of LSECs at steady state, during hepatic regeneration and after CCl4 induced liver injury. AIM 2: Uncover the contribution of c-Maf expressed in the Kupffer cells that by enforcing and sustaining LSEC vascular attributes regulate hepatic homeostasis during liver regeneration and CCl4 induced liver injury. AIM 3: Employ the human Hepatic-On-VascularNet platform to uncover the mechanism by which c-Maf is induced functionally in human liver ECs to form specialized LSECs. Determine whether human c-Maf induced LSECs (iLSECs) can restore hepatic regeneration. Specifically, the role of infusing human iLSECs in restoring hepatic repair post-acetaminophen (APAP) injury without provoking fibrosis will be assessed. These studies will uncover the molecular determinants of human LSECs and allow strategies to employ iLSECs to sustain its pro-regenerative and anti-fibrotic attributes for liver repair.

抽象的终末期肝病与通常需要肝移植的发病率和死亡率相关。功能障碍肝窦内皮细胞（LSEC）的毛细血管化可能导致肝脏修复受损肝硬化。因此，揭示 LSEC 获得其专门功能以支持的机制肝脏修复可以促进无疤痕肝脏修复疗法的开发。我们已经证明 LSEC 中 Id1 和 Cxcr7 的激活可诱导血管分泌因子，从而指导肝再生，而无需纤维化。然而，LSEC 获得亲肝功能的机制尚不清楚。我们展示转录因子 (TF) Fli1 和 Erg 决定 LSEC 的血管命运和稳态 (Gomez- Salinero JM 等人，《自然心血管研究》，2022），TF c-Maf 的诱导指定了表型和 LSEC 的再生功能（Gomez-Salinero JM 等人，Cell Stem Cell，2022）。 c-Maf 的感应通用人内皮细胞 (EC) 开启肝脏特异性 LSEC 特征和血管分泌因子支持肝细胞功能。在成年 EC 中删除了 c-Maf 的小鼠中，CCl4 结果恢复纤维化和 LSEC 回归动脉细胞命运。因此，我们假设 LSEC 血管的维持细胞命运需要 Fli1 或 Erg 的组成型表达，维持 LSEC 的稳态功能。感应 c- Maf 强制执行特殊的促再生功能以及 LSEC 与肝细胞、Kupffer 和肝细胞的相互作用星状细胞可防止应激诱导的 LSEC 动脉化，促进肝脏修复而不发生纤维化。对此最后，我们将人类通用 EC 重新编程为适应性强的肾小管发生状态。这些重置血管内皮细胞 (R-VEC) 自组装成 3D 血管网络，输送人体血液并形成树枝肝细胞（Palikuqi B 等人，Nature，2020）。具有星状和库普弗的血管化肝聚集体可扩展和可灌注的微流体装置中的细胞建立了人类肝血管网络平台，能够研究人肝细胞和 LSEC 之间的生理适应性串扰。这将通过执行以下目标来检验假设：目标 1：定义分层 Fli1 的机制肝脏过程中，通过 c-Maf 诱导表达和 Erg 维持 LSEC 在稳态下的特化再生和 CCl4 诱导的肝损伤后。目标 2：揭示 c-Maf 在库普弗细胞通过执行和维持 LSEC 血管属性来调节肝脏稳态肝再生和CCl4诱导的肝损伤。目标 3：利用人类 Hepatic-On-VasularNet 平台揭示 c-Maf 在人肝脏 EC 中功能性诱导形成专门的 LSEC 的机制。确定人 c-Maf 诱导的 LSEC (iLSEC) 是否可以恢复肝再生。具体来说，输注人 iLSEC 在不引起对乙酰氨基酚 (APAP) 损伤后肝脏修复中的作用将评估纤维化情况。这些研究将揭示人类 LSEC 的分子决定因素，并允许利用 iLSEC 维持其促再生和抗纤维化属性以进行肝脏修复的策略。