Generation of functionally mature pancreatic organoids as a replacement strategy for animal-models of pancreatitis and pancreatic cancer

生成功能成熟的胰腺类器官作为胰腺炎和胰腺癌动物模型的替代策略

• 批准号: NC/V002260/1
• 负责人: Jean-Francois Darrigrand
• 金额: $ 16.72万
• 依托单位: King's College London
• 依托单位国家: 英国
• 项目类别: Fellowship
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=NC%2FV002260%2F1
• 关键词: Generation; functionally; mature; pancreatic; organoids

The pancreas consists of two organs in one. The endocrine part houses insulin-secreting cells controlling blood glucose levels and the exocrine part produces enzymes controlling the digestion. Diseases affecting the exocrine pancreas, such as pancreatitis and pancreatic cancer, present dramatic mortality rates, as highlighted by the 95% mortality of patients diagnosed with pancreatic cancer. To alleviate the societal burden caused by these diseases, research teams all over the world are trying to better understand pancreatic diseases and establish early diagnosis and innovative therapeutic strategies. To develop and test the efficacy of these strategies, genetically engineered mouse models replicating most human pathophysiological features are routinely used in research labs. Although necessary for research purposes, the use of these models leads to the culling of thousands of animals worldwide every year.Organoids are emerging 3D in vitro systems derived from stem cells, which represent a very promising alternative to animal models. Organoids are simplified versions of organs, of which they should recapitulate the physiology and microanatomy. Unfortunately, the pancreatic organoids generated so far do not faithfully recapitulate the pancreas complexity. This strongly limits the adoption of pancreas organoids as valuable replacement models in the labs working on pancreatic diseases. The goal of this project is thus to develop physiologically relevant organoid models for studying pancreatitis and pancreatic cancer in vitro. The formation of the pancreas in the embryo is known to be tightly regulated by its local microenvironment, whose precise composition is still poorly known. The first goal of the project is to define the pancreas microenvironment observed in embryos and to reproduce it artificially in vitro for the maturation of organoids. To that end, I will use cutting-edge sequencing technologies on mouse embryonic pancreas and precisely define the biochemical cues that compose its microenvironment. I will then use the identified cues to supplement artificial microcavities in which organoids will be cultured. Cultured organoids will be monitored for the acquisition of mature morphological and physiologic pancreatic markers. This approach will allow me to identify the best microcavities composition to mature pancreatic organoids. To model pancreatitis, organoids will be treated with caerulein, a drug used to induce pancreatitis in animal models. To model pancreatic cancer, I will generate organoids using genetically modified stem-cells that over-activate a variant of a gene known to induce pancreatic cancer in mice and humans. I will assess whether the obtained organoids are valuable models to study pancreatitis and pancreatic cancer by comparing their pathophysiological features with those observed in animal models. The completion of this project will surely enable a significant reduction of animal procedures and provide research labs with new tools to screen the effect of next-generation drugs for pancreatic diseases treatment.

胰腺由两个器官合二为一组成。内分泌部分容纳控制血糖水平的胰岛素分泌细胞，外分泌部分产生控制消化的酶。影响胰腺外分泌的疾病，如胰腺炎和胰腺癌，具有显著的死亡率，如诊断为胰腺癌的患者的95%死亡率所强调的。为了减轻这些疾病造成的社会负担，世界各地的研究团队正在努力更好地了解胰腺疾病，并建立早期诊断和创新的治疗策略。为了开发和测试这些策略的功效，复制大多数人类病理生理学特征的基因工程小鼠模型通常用于研究实验室。虽然这些模型对于研究目的是必要的，但每年全世界都有成千上万的动物被淘汰。类器官是来自干细胞的新兴3D体外系统，它代表了一种非常有前途的动物模型替代品。类器官是器官的简化版本，它们应该概括生理和显微解剖。不幸的是，到目前为止产生的胰腺类器官并不能忠实地概括胰腺的复杂性。这严重限制了胰腺类器官作为胰腺疾病实验室中有价值的替代模型的采用。因此，本项目的目标是开发用于体外研究胰腺炎和胰腺癌的生理学相关的类器官模型。已知胚胎中胰腺的形成受到其局部微环境的严格调控，其精确组成仍然知之甚少。该项目的第一个目标是定义在胚胎中观察到的胰腺微环境，并在体外人工复制它以使类器官成熟。为此，我将在小鼠胚胎胰腺上使用尖端的测序技术，并精确定义组成其微环境的生化线索。然后，我将使用识别的线索来补充人工微腔，在其中培养类器官。将监测培养的类器官是否获得成熟的形态和生理胰腺标志物。这种方法将使我能够确定成熟胰腺类器官的最佳微腔组成。为了模拟胰腺炎，将用雨蛙肽（一种用于在动物模型中诱导胰腺炎的药物）处理类器官。为了模拟胰腺癌，我将使用基因修饰的干细胞来产生类器官，这些干细胞过度激活已知在小鼠和人类中诱导胰腺癌的基因变体。我将通过比较它们的病理生理学特征与在动物模型中观察到的那些来评估所获得的类器官是否是研究胰腺炎和胰腺癌的有价值的模型。该项目的完成必将大大减少动物手术，并为研究实验室提供新的工具来筛选下一代胰腺疾病治疗药物的效果。