Multi-omics approach to investigate the immune-cholangiocyte spatial relationship to stratify liver diseases

多组学方法研究免疫-胆管细胞空间关系以对肝脏疾病进行分层

• 批准号: 2887376
• 金额: --
• 依托单位: University of Edinburgh
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2887376
• 关键词: Multi; omics; approach; investigate; immune

The involvement of proinflammatory cytokine IL17 (Interleukin-17) has been linked to multiple chronic inflammatory diseases, impairing wound healing and promoting fibrosis progression. The increase in IL17 level has been identified in a wide range of chronic liver diseases such as Non-Alcoholic Fatty Liver Disease (NAFLD), Primary Biliary Cholangitis (PBC), and Primary Sclerosing Cholangitis (PSC). Chronic liver disease is estimated to affect 800 million people worldwide with no effective anti-fibrotic treatments. It is well known that IL17 promotes fibrogenesis through the activation of fibroblasts. However, the role of IL17 on epithelial cell fate and regeneration is less clear, especially with recent work suggesting that IL17 signalling primes epithelial proliferation in response to injury, fitting with preliminary data in our lab showing that IL17 signalling primes liver stem cell activation in response to liver injury. We have shown previously that cholangiocytes in the liver become regenerative liver stem cells in response to injury but the exact mechanisms which control this remains to be investigated. Furthermore, the emergence of the regenerative cholangiocytes tends to correlate with scar deposition around the regenerative niche which has a mixture of proliferating epithelial cells, immune cells, fibroblasts, and scar. It is unclear whether the emergence of IL17 cells and signalling in the liver is a protective mechanism or an indicator of poor prognosis in human diseases. Indeed, treatments involving pan-blockade of IL17 such as Secukinumab are on Phase 1/2 clinical trials for psoriasis, however, with the wide spectrum of chronic liver diseases and pathology, we wonder whether "one treatment fits all"?Here, we aim to analyse published, publicly available single cell RNA (scRNA) sequencing datasets across a spectrum of liver diseases with variable aetiology such as cirrhosis, NAFLD, PSC and analyse whether IL17 has implications on regenerative cholangiocytes. Furthermore, we aim to spatially analyse the implications each disease has on IL17-mediated immune infiltrates in humans. This will stratify the role of IL17 and cholangiocytes across a spectrum of liver diseases and characterise the cholangiocytes and immune cells in different diseases transcriptionally and spatially, answering whether IL17 makes regenerative cholangiocytes pathogenic? To complement the findings from scRNA sequencing datasets from clinical samples, we will use a mouse model where IL17 receptor is conditionally deleted in the cholangiocytes (Krt19CreERT2tdTomatoloxSTOPloxIL17Raflox) to inhibit IL17 signalling on cholangiocytes. Models of biliary or hepatocyte damage will be used on the transgenic mice in the presence or absence of cholangiocyte IL17 signalling. scRNA sequencing will be performed to investigate whether the absence of IL17 signalling on cholangiocytes alters the immune and regenerative landscape in the liver. Furthermore, this dataset will be used to match with the human datasets available publicly to identify matching cholangiocyte populations in human diseases. Qualitative & quantitative proteomics analysis will be performed on isolated murine cholangiocytes to identify pathogenic vs regenerative protein signatures. The cholangiocyte specific IL17 inhibition strategy will show whether IL17 signalling differs in response to different injury mechanisms (i.e biliary vs hepatic), stratifying IL17 targeting treatment based on disease aetiology and complementing the pan-IL17 blockade strategies.Dissecting the cell specific effect of IL17 on tissue regeneration and fibrosis, we aim to work towards the overarching aim to understand the role of IL17 signalling in chronic liver diseases - a friend or foe? More importantly, leading to potential targeted treatments for chronic liver disease based on aetiology.

促炎症细胞因子IL-17(IL-17)参与多种慢性炎症性疾病，损害伤口愈合，促进纤维化进展。IL17水平在多种慢性肝病如非酒精性脂肪性肝病(NAFLD)、原发性胆管炎(PBC)和原发性硬化性胆管炎(PSC)中被发现升高。据估计，在没有有效的抗纤维化治疗的情况下，全球有8亿人受到慢性肝病的影响。众所周知，IL17通过激活成纤维细胞促进纤维化形成。然而，IL17在上皮细胞命运和再生中的作用还不是很清楚，特别是最近的研究表明，IL17信号转导损伤后的上皮细胞增殖，与我们实验室的初步数据相吻合，表明IL17信号转导肝脏干细胞激活以响应肝损伤。我们以前已经证明，肝脏中的胆管细胞在损伤后会变成再生的肝干细胞，但控制这一过程的确切机制仍有待研究。此外，再生胆管细胞的出现往往与再生壁龛周围的瘢痕沉积有关，再生壁龛由增殖的上皮细胞、免疫细胞、成纤维细胞和疤痕组成。目前尚不清楚肝脏中IL17细胞和信号的出现是一种保护机制，还是预示着人类疾病预后不良的指标。事实上，涉及对IL17的泛阻断的治疗，如Secukinumab，正在进行银屑病的1/2期临床试验，然而，由于慢性肝病和病理的广泛范围，我们想知道“一种治疗方法是否适用于所有人”？在这里，我们旨在分析已发表的、公开可用的单细胞RNA(ScRNA)测序数据，这些数据涵盖了不同病因的肝脏疾病，如肝硬变、NAFLD、PSC，并分析IL17是否对再生胆管细胞有影响。此外，我们的目标是在空间上分析每种疾病对IL17介导的人类免疫渗透的影响。这将对IL17和胆管细胞在一系列肝脏疾病中的作用进行分层，并在转录和空间上表征不同疾病中的胆管细胞和免疫细胞，回答IL17是否使再生胆管细胞致病？为了补充来自临床样本的scRNA测序数据集的发现，我们将使用一个小鼠模型，其中IL17受体在胆管细胞(Krt19CreERT2tdTomatoloxSTOPloxIL17Raflox)中有条件地删除，以抑制胆管细胞上的IL17信号。在胆管细胞IL17信号存在或不存在的情况下，将在转基因小鼠上使用胆汁或肝细胞损伤模型。将进行单链RNA测序，以调查胆管细胞上IL17信号的缺失是否改变了肝脏的免疫和再生格局。此外，该数据集将用于与公开可用的人类数据集进行匹配，以识别人类疾病中匹配的胆管细胞群体。将对分离的小鼠胆管细胞进行定性和定量的蛋白质组学分析，以确定致病和再生蛋白的特征。胆管细胞特异性IL17抑制策略将显示IL17信号在不同损伤机制(即胆道和肝脏)中的反应是否不同，根据疾病病因对IL17靶向治疗进行分层，并补充PAN-IL17阻断策略。剖析IL17对组织再生和纤维化的细胞特异性作用，我们旨在朝着总体目标努力，以了解IL17信号在慢性肝病中的作用--是朋友还是敌人？更重要的是，这将导致基于病因学的慢性肝病的潜在靶向治疗。