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

• 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
• 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

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 末期肝病与发病率和死亡率有关，通常需要肝移植。功能障碍 肝脏正弦内皮细胞（LSEC）的毛细管化可能有助于肝修复和 肝硬化。因此，发现LSEC获得其专业功能以支持的机制 肝修复可以使疗法开发用于无疤痕的肝脏修复。我们已经表明 LSEC中ID1和CXCR7的激活诱导血管分泌因子，从而指导肝脏再生而无需 纤维化。但是，LSEC获得其亲肝功能的机制尚不清楚。我们显示 尽管转录因子（TFS）FLI1和ERG决定了LSEC的血管命运和稳态（Gomez-- Salinero JM等，自然心血管研究，2022年），TF C-MAF的诱导指定表型和 LSEC的再生功能（Gomez-Salinero JM等，细胞干细胞，2022年）。诱导C-MAF 通用的人内皮细胞（EC）开关肝特异性LSEC特征和血管分泌因子 支持肝细胞功能。在成人EC中删除C-MAF的小鼠中，从CCL4结果恢复 在纤维化和LSEC在动脉细胞命运中的回归。因此，我们假设LSEC血管的维持 细胞命运需要组成型FLI1或ERG表达，以维持LSEC稳态功能。诱导c- MAF执行专业的促进功能以及LSEC与肝细胞，Kupffer和 星状细胞可防止应激诱导的LSEC动脉化，从而促进肝修复而无需纤维化。对此 最后，我们已将人类通用EC重新编程为适应性的肾变生成状态。这些重置 - 血管 内皮细胞（R-VECS）自组装成3D血管网络，运输人血和树木 肝细胞（Palikuqi B等人自然，2020年）。具有星状和库普弗的血管化肝骨料 可扩展和灌注的微流体设备内的细胞建立了人类肝内血管台平台，即 在人肝细胞和LSEC之间对生理适应性跨语的研究。这 假设将通过执行以下目标来检验：目标1：定义层次fli1的机制 通过C-MAF诱导表达ERG表达在抗肝期间的LSEC稳定状态下的专业化 再生和CCL4诱导肝损伤。目标2：揭示C-MAF在 通过强制执行和维持LSEC血管属性来调节肝稳态的细胞 肝脏再生和CCL4诱导肝损伤。目的3：使用人类肝肝平台到 发现在人肝EC中诱导C-MAF形成专门LSEC的机制。 确定人C-MAF诱导的LSEC（ILSEC）是否可以恢复肝后再生。具体来说， 在不引起的乙酰氨基氨基酚（APAP）损伤的情况下，注入人ILSEC在恢复肝修复中的作用 将评估纤维化。这些研究将发现人类LSEC的分子决定因素，并允许 采用ILSEC来维持其促进性和抗纤维化属性的策略用于肝脏维修。