Defining the regenerative capacity of ductular cells from non-transplantable human liver

定义不可移植的人肝脏导管细胞的再生能力

• 批准号: MR/P016839/1
• 负责人: Stuart Forbes
• 金额: $ 237.26万
• 依托单位: University of Edinburgh
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2017
• 资助国家: 英国
• 起止时间: 2017 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FP016839%2F1
• 关键词: Defining; regenerative; capacity; ductular; cells

Liver disease killed 16,087 people in the UK in 2008, and causes 1 in 50 deaths in Scotland. Although a liver transplant is curative for these patients, a global shortage of donor organs means that many patients die waiting for suitable liver. Development of renewable therapeutic alternatives to whole organ transplants is imperative to alleviate this clinical shortage. Because of this clinical need a cellular treatment strategy would be highly attractive. Hepatocyte transplantation has successfully been used to: help keep patients alive who are waiting for whole liver transplantation; support pediatric patients in acute liver failure and correct metabolic liver disease. However, hepatocytes are not readily obtained and are usually sourced from discarded livers not suitable for transplantation. These livers are either significantly fatty or have undergone prolonged ischaemia (time without blood flow). These liver's cells are damaged and therefore high quality hepatocytes are difficult to obtain from such discarded livers. Furthermore it is not possible to expand the numbers of hepatocytes in the laboratory. An alternative source of transplantable cells for either biliary or hepatocyte regeneration would be a significant healthcare advance.We have previously shown in the mouse that cells in liver called ductal cells or hepatic progenitor cells (HPCs) can act like stem cells and regenerate bile ducts and hepatocytes when the liver is severely damaged. Ductal cells/HPCs can be readily frozen and thawed without damage and then grown in the laboratory increasing the numbers of cells. We now aim to develop human ductal cells/HPCs for cell therapy with the eventual aim of developing a new type of cell therapy for liver patients. We aim to isolate these cells from livers that are too damaged to transplant whole.Potential donor organs are increasingly undergoing normothermic perfusion prior to potential whole organ transplantation. For those livers deemed unsuitable for transplantation or where for logistical reasons the transplantation cannot proceed we will move seemlessly to efficient cell isolation of biliary ductal cells. Putative bipotential HPCs will be purified using the clinical cell sorter. These cells can be expanded and frozen/defrosted prior to use as a clinical cell therapy. Over the course of the grant (48 months) we will: 1. Optimise the isolation and cell culture of ductular cells/HPCs from non-transplantable human livers.2. Test the ability of HPCs to regenerate bile ducts +/- hepatocytes in mouse models. We need to know if the human cells are bipotential (regenerate bile ducts and hepatocytes) like the mouse ductular/HPC cells we described or are limited to bile duct regeneration alone. This result would guide future potential clinical conditions that the cells could be used for.3. Develop protocols to isolate, purify and expand ductular/HPCs in a GMP compatible (suitable for use in man) manner as a potential clinical cell product.4. Assess long term stability and safety of the transplanted ductular/HPCs. 5. If the above components are successful we will apply for MHRA (Medicines and Healthcare products Regulatory Agency) approval. This would pave the way for a fist in human clinical trial of ductular/HPC cell therapy. The above results would guide the clinical target to conditions requiring biliary or hepatocyte regeneration.

2008年，肝病在英国造成16，087人死亡，在苏格兰造成50人死亡。虽然肝移植对这些患者是有疗效的，但全球供体器官短缺意味着许多患者在等待合适的肝脏时死亡。开发可再生的治疗替代整个器官移植是缓解这种临床短缺的当务之急。由于这种临床需要，细胞治疗策略将是非常有吸引力的。肝细胞移植已成功用于：帮助等待全肝移植的患者存活;支持急性肝功能衰竭的儿科患者和纠正代谢性肝病。然而，肝细胞不容易获得，并且通常来源于不适合移植的废弃肝脏。这些肝脏要么是明显的脂肪，要么经历了长时间的缺血（没有血流的时间）。这些肝细胞受损，因此难以从这些丢弃的肝脏中获得高质量的肝细胞。此外，不可能在实验室中扩大肝细胞的数量。我们之前已经在小鼠中证明，肝脏中被称为导管细胞或肝祖细胞（HPC）的细胞可以像干细胞一样发挥作用，在肝脏严重受损时再生胆管和肝细胞。导管细胞/HPC可以很容易地冷冻和解冻而不会损坏，然后在实验室中生长，增加细胞数量。我们现在的目标是开发用于细胞治疗的人导管细胞/HPC，最终目标是为肝脏患者开发一种新型的细胞治疗。我们的目标是从受损严重无法进行整体移植的肝脏中分离出这些细胞。在潜在的整体器官移植之前，越来越多的潜在供体器官正在接受常温灌注。对于那些被认为不适合移植的肝脏或由于后勤原因无法进行移植的肝脏，我们将毫不犹豫地进行胆管细胞的有效细胞分离。将使用临床细胞分选仪纯化推定的双能HPC。这些细胞可以在用作临床细胞疗法之前进行扩增和冷冻/解冻。在补助金的过程中（48个月），我们将：1。优化非移植性人肝脏中胆管细胞/HPC的分离和细胞培养.在小鼠模型中测试HPC再生胆管+/-肝细胞的能力。我们需要知道人类细胞是否像我们描述的小鼠导管/HPC细胞那样是双能的（再生胆管和肝细胞），或者仅限于胆管再生。这一结果将指导未来潜在的临床条件，细胞可用于。开发方案以GMP兼容（适用于人类）的方式分离、纯化和扩增导管/HPC作为潜在的临床细胞产品。评估移植的导管/HPC的长期稳定性和安全性。5.如果上述组成部分成功，我们将申请MHRA（药品和保健品管理局）批准。这将为导管/HPC细胞疗法的人类临床试验铺平道路。上述结果将指导临床靶向需要胆汁或肝细胞再生的疾病。

项目成果

Paracrine cellular senescence exacerbates biliary injury and impairs regeneration.

• DOI: 10.1038/s41467-018-03299-5
• 发表时间: 2018-03-09
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Ferreira-Gonzalez S;Lu WY;Raven A;Dwyer B;Man TY;O'Duibhir E;Lewis PJS;Campana L;Kendall TJ;Bird TG;Tarrats N;Acosta JC;Boulter L;Forbes SJ
• 通讯作者: Forbes SJ

Inhibition of nuclear factor (erythroid-derived 2)-like 2 promotes hepatic progenitor cell activation and differentiation.

• DOI: 10.1038/s41536-021-00137-z
• 发表时间: 2021-05-26
• 期刊: NPJ Regenerative medicine
• 影响因子: 7.2
• 作者: Bellanti F;di Bello G;Iannelli G;Pannone G;Pedicillo MC;Boulter L;Lu WY;Tamborra R;Villani R;Vendemiale G;Forbes SJ;Serviddio G
• 通讯作者: Serviddio G