Macrophage Cell Subset Specific Biomarkers for Disease Prognosis in Biliary Atresia

项目概要： 胆道闭锁（BA）是一种婴儿期胆汁淤积性肝病，是儿童肝脏的主要原因 移植虽然有证据支持异常免疫应答在BA发病机制中的作用，但 疾病的确切机制仍然未知。科学知识的这一差距限制了免疫系统的识别。 诊断时的生物标志物以分层患者预后。虽然巨噬细胞（M β）以前曾被牵连 在人类和小鼠BA中，M β在性质上是异质的， 尚未发现肝损伤。克服科学知识的这一差距将确定BA特异性 致病性M β亚群的研究，并导致新的细胞亚群特异性生物标志物预测患者预后。 我们是第一个在儿童胆汁淤积性肝病中发现3种新的肝M β亚群的人， 通过利用单细胞RNA测序的能力， （scRNA-seq）。我们已经将这些M β亚群定义为脂质相关M β（参与脂质代谢的基因的高表达）。 脂质代谢）、单核细胞样M（单核细胞中也发现了基因表达）和适应性M （富集参与淋巴细胞活化的基因）。此外，单核细胞样和自适应M-子集 与Luo等人建立的诊断时的不良预后特征正相关，表明这些 亚群可能驱动与疾病结果有关的纤维化和CD 8阳性T细胞活化。我们从而 假设血清细胞因子网络驱动M β向致病性单核细胞样极化， 适应性M β亚群;诊断时组织学检查显示这些M β亚群的数量增加， 与不良结局相关。 为了研究这一假设，我们将：1）确定脂质相关的单核细胞样的， 人BA的适应性M受体诊断与3个月时黄疸清除定义的结局相关 Kasai门肠吻合术（KPE）后，以及2）定义血清中与 预后和组织学发现。这项创新的研究计划应用了之前scRNA-seq 对来自ChiLDReN的人类样本进行分析，以鉴定M β亚群特异性组织学和血清学 在诊断时用于预后的生物标志物。这项研究的结果将促进对患有 BA，并允许在KPE时改善预后咨询。

PROJECT SUMMARY: Biliary atresia (BA) is a cholestatic liver disease of infancy that is the leading cause of pediatric liver transplantation. While evidence supports a role for an aberrant immune response in the pathogenesis of BA, the exact mechanism of disease remains unknown. This gap in scientific knowledge limits identification of immune biomarkers at diagnosis to stratify patient prognosis. While macrophages (M) have previously been implicated in both human and murine BA, M are heterogeneous by nature and the pathogenic subsets required for ongoing hepatic injury have not been identified. Overcoming this gap in scientific knowledge will identify the BA-specific pathogenic M subsets and lead to novel cell-subset specific biomarkers predictive of patient prognosis. We have been the first to identify 3 novel hepatic M subsets in pediatric cholestatic liver disease at the time of liver transplant that are distinct from non-diseased M by leveraging the ability of single cell RNA-sequencing (scRNA-seq). We have defined these M subsets as lipid-associated M (high expression of genes involved in lipid metabolism), monocyte-like M (expression of genes also identified in monocytes), and adaptive M (enrichment for genes involved in lymphocyte activation). Furthermore, monocyte-like and adaptive M subsets positively correlate with the poor prognostic signature at diagnosis established by Luo et al suggesting these subsets may drive fibrosis and activation of CD8-positive T cells implicated in disease outcome. We thereby hypothesize that serum cytokine networks drive M polarization towards pathogenic monocyte-like and adaptive M subsets; increased numbers of these M subsets by histology at diagnosis will be associated with poor outcome. To investigative this hypothesis we will: 1) determine if the numbers of lipid-associated, monocyte-like, and adaptive M at diagnosis in human BA correlate with outcome as defined by clearance of jaundice by 3 months post-Kasai Portoenterostomy (KPE), and 2) define M-specific cytokine networks in serum that correlate with prognosis and histologic findings. This innovative research plan applies findings from previous scRNA-seq analysis to human samples available from ChiLDReN to identify M-subset specific histologic and serum biomarkers for prognosis at the time of diagnosis. Findings from the study will advance the care of patients with BA and allow for improved prognostic counseling at the time of KPE.