Control of hepatic T cell responses in biliary atresia

胆道闭锁中肝 T 细胞反应的控制

基本信息

批准号：
10378600
负责人：
Alexander Miethke
金额：
$ 41.29万
依托单位：
CINCINNATI CHILDRENS HOSP MED CTR
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-15 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10378600
关键词：
ABCB1 gene Acute Adjuvant Therapy Agonist Anti-Inflammatory Agents Archives Attenuated Bile Acids Bile Duct Epithelium Bile fluid Biliary Biliary Atresia Bilirubin Biological Markers Bone Marrow Transplantation CD4 Positive T Lymphocytes CD8-Positive T-Lymphocytes CD8B1 gene Candidate Disease Gene Cells Childhood Cholestasis Chronic Clinical Clinical Data Cluster Analysis DNA Methylation Data Dendritic Cells Diagnosis Drainage procedure Duct (organ) structure Epigenetic Process Epithelial Etiology FOXP3 gene Fibrosis Funding Future GPBAR1 gene Gene Expression Gene Expression Profile Genes Grant Hepatic Hepatobiliary Hepatology Homeostasis Icterus Image Analysis Immune Immune response Immunofluorescence Immunologic In Vitro Infant Inflammatory Injury Intestines Intrahepatic bile duct Knockout Mice Kupffer Cells Leukocytes Lipids Liver Liver Fibrosis Liver diseases Lymphocyte Lymphocyte Activation Mediating Molecular Molecular Target Mononuclear Mus National Institute of Diabetes and Digestive and Kidney Diseases Natural Killer Cells Neonatal Obstruction Operative Surgical Procedures Outcome Parents Pathway interactions Patients Pharmacology Phenotype Population Pre-Clinical Model Process Production Publishing Regulation Regulatory T-Lymphocyte Reporting Research Rotavirus STAT3 gene Sclerosing Cholangitis Serum Shapes Surveys T cell response T-Lymphocyte Testing Time Tissues Validation Work base bile duct biliary tract clinical phenotype cohort cytokine efflux pump end stage liver disease epigenetic silencing epithelial injury experimental analysis experimental study human disease improved individualized medicine intrahepatic liver biopsy liver injury mRNA Expression macrophage mouse model neonate new therapeutic target novel osteopontin patient subsets polarized cell prospective receptor recruit response restoration reuptake segregation surgery outcome targeted treatment therapeutic target transcription factor transcriptome transcriptome sequencing uptake

项目摘要

Project Summary/ Abstract Biliary atresia (BA) refers to idiopathic obliteration of the biliary tree in neonates. Unfortunately, up to 50% of infants who undergo attempts at surgical restoration of bile flow by hepatoportoenterostomy (HPE) remain cholestatic and rapidly progress to end-stage liver disease. Potential therapeutic targets include hepatic NK, CD8 and Th17-lymphocytes which have been identified as drivers of inflammatory ductal obstruction and progressive injury of the intrahepatic biliary epithelium. Our group has reported that regulatory T cells (Tregs) are able to restrain hepatic lymphocyte responses protecting from bile duct injury and initiation of liver fibrosis. Therefore, we explored the mechanisms controlling Treg/Th17 homeostasis under cholestatic conditions recapitulating progressive BA. Our preliminary findings demonstrate that conjugated bile acids (CBA) repress expression of the Treg- transcription factor Foxp3 and inhibit Treg-function in vitro. In the Mdr2-/- mouse model of “toxic bile” induced progressive cholestasis, inhibition of intestinal bile acid reuptake resulted in dramatic reduction in serum bile acid concentration which shifted cytokine production by Kupffer cells accompanied by rapid hepatic expansion of Tregs. In rotavirus induced murine BA, treatment with Fxr agonists induced expression of Il-10 in hepatic mononuclear cells, expanded Tregs, and attenuated the BA phenotype. We hypothesize that bile acids are epigenetic modifiers that control Treg homeostasis and determine the inflammatory microenvironment in the liver through activation of the bile acid receptors Tgr5 and Fxr. We will test this hypothesis by investigating the molecular mechanisms mediating bile acid induced silencing of Foxp3 in vitro and by deleting candidate molecular targets for CBA in CD4 lymphocytes in mice with acute or chronic cholestasis (Aim 1). In Aim 2, the effects of activation of Tgr5 and Fxr on regulation of macrophage derived cytokine production and Treg homeostasis, and on the fibrosing cholangiopathy phenotype will be studied in bone marrow transplant experiments and in conditional bile acid receptor knockout mice. For validation in human disease, we will investigate whether hepatic T lymphocyte responses are associated with distinct clinical phenotypes by analyzing liver RNAseq and clinical data and performing immunofluorescence and image analysis on archived liver biopsies from an inception cohort of 203 patients with BA followed by the multicenter Childhood Liver Disease Research Network (Aim 3). In preliminary studies we generated a gene list of Th17 associated genes from in vitro polarized neonatal Th17 lymphocytes. Hierarchical cluster analysis of these experimentally derived Th17 candidate genes in liver RNAseq data from 137 patients with BA revealed clusters of patients with similar gene expression profile. Serum conjugated bilirubin levels differed between these clusters, supporting our hypothesis. Future studies will interrogate segregation of clusters defined by integrated RNAseq- and immunofluorescence - based enumeration of liver infiltrating T cells with BA phenotype and HPE outcome.

项目概要/摘要胆道闭锁 (BA) 是指新生儿胆管树的特发性闭塞，不幸的是，高达 50% 的新生儿发生胆道闭锁。尝试通过肝门肠造口术 (HPE) 恢复胆汁流量的婴儿仍保留胆汁淤积并迅速进展为终末期肝病的潜在治疗靶点包括肝脏 NK、 CD8 和 Th17 淋巴细胞已被确定为炎症性导管阻塞的驱动因素我们的研究小组报道了调节性 T 细胞 (Treg) 的进行性损伤。能够抑制肝淋巴细胞反应，防止胆管损伤和肝纤维化的发生。因此，我们探索了胆汁淤积条件下控制Treg/Th17稳态的机制我们的初步研究结果表明，共轭胆汁酸 (CBA) 会抑制。在 Mdr2-/- 小鼠模型中表达 Treg 转录因子 Foxp3 并抑制 Treg 功能。 “有毒胆汁”诱导进行性胆汁淤积，抑制肠道胆汁酸再摄取导致显着的血清胆汁酸浓度降低，从而改变库普弗细胞产生细胞因子的能力，并伴有在轮状病毒诱导的小鼠 BA 中，用 Fxr 激动剂治疗可诱导 Tregs 的快速肝扩张。肝单核细胞中 IL-10 的表达、Treg 细胞扩增以及 BA 表型减弱。研究表明，胆汁酸是表观遗传修饰剂，可控制 Treg 稳态并决定我们将测试通过激活胆汁酸受体 Tgr5 和 Fxr 来调节肝脏炎症微环境。通过研究介导胆汁酸诱导 Foxp3 沉默的分子机制，提出了这一假设体外并通过删除急性或慢性小鼠 CD4 淋巴细胞中 CBA 的候选分子靶标胆汁淤积（目标 1）。在目标 2 中，Tgr5 和 Fxr 的激活对巨噬细胞衍生的调节的影响。细胞因子的产生和 Treg 稳态，以及纤维化胆管病表型的研究将在骨髓移植实验和条件性胆汁酸受体敲除小鼠用于人体验证。疾病，我们将研究肝 T 淋巴细胞反应是否与不同的临床相关通过分析肝脏 RNAseq 和临床数据并进行免疫荧光和图像分析来确定表型对 203 名 BA 患者的初始队列中存档的肝活检进行了研究，随后进行了多中心儿童研究肝脏疾病研究网络（目标 3）。在初步研究中，我们生成了 Th17 相关基因列表。来自体外极化新生儿 Th17 淋巴细胞的基因对这些实验进行分层聚类分析。从 137 名 BA 患者的肝脏 RNAseq 数据中衍生的 Th17 候选基因揭示了一组患有相似的基因表达谱在这些簇之间存在差异，支持了这一点。我们的假设是，未来的研究将探讨由整合的RNAseq-和定义的簇的分离。基于免疫荧光的具有 BA 表型和 HPE 结果的肝脏浸润 T 细胞计数。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Farnesoid X receptor antagonizes macrophage-dependent licensing of effector T lymphocytes and progression of sclerosing cholangitis.

DOI：
10.1126/scitranslmed.abi4354
发表时间：
2022-12-14
期刊：
SCIENCE TRANSLATIONAL MEDICINE
影响因子：
17.1
作者：
Shi, Tiffany;Malik, Astha;vom Hofe, Annika Yang;Matuschek, Louis;Mullen, Mary;Lages, Celine S.;Kudira, Ramesh;Singh, Ruchi;Zhang, Wujuan;Setchell, Kenneth D. R.;Hildeman, David;Pasare, Chandrashekhar;Wagner, Brandee;Miethke, Alexander G.
通讯作者：
Miethke, Alexander G.

Serum Matrix Metalloproteinase 7 Is a Diagnostic Biomarker of Biliary Injury and Fibrosis in Pediatric Autoimmune Liver Disease.

DOI：
10.1002/hep4.1589
发表时间：
2020-11
期刊：
Hepatology communications
影响因子：
5.1
作者：
Lam S;Singh R;Dillman JR;Trout AT;Serai SD;Sharma D;Sheridan R;Su W;Fei L;Karns R;Haramija MM;Ridgway G;Goldfinger M;Squires JE;Denson LA;Hyams JS;Miethke AG
通讯作者：
Miethke AG

Regulatory T cells control the CD8 adaptive immune response at the time of ductal obstruction in experimental biliary atresia.

DOI：
10.1002/hep.25662
发表时间：
2012-07
期刊：
HEPATOLOGY
影响因子：
13.5
作者：
Lages, Celine S.;Simmons, Julia;Chougnet, Claire A.;Miethke, Alexander G.
通讯作者：
Miethke, Alexander G.

The dendritic cell-T helper 17-macrophage axis controls cholangiocyte injury and disease progression in murine and human biliary atresia.

DOI：
10.1002/hep.28851
发表时间：
2017-01
期刊：
HEPATOLOGY
影响因子：
13.5
作者：
Lages, Celine S.;Simmons, Julia;Maddox, Avery;Jones, Keaton;Karns, Rebekah;Sheridan, Rachel;Shanmukhappa, Shiva Kumar;Mohanty, Sujit;Kofron, Matthew;Russo, Pierre;Wang, Yui-Hsi;Chougnet, Claire;Miethke, Alexander G.
通讯作者：
Miethke, Alexander G.

Fishing for biliary atresia susceptibility genes.