The role of mechanosensing in the selective advantage of genetically variant human pluripotent stem cells

机械传感在遗传变异人类多能干细胞的选择优势中的作用

• 批准号: MR/X000028/1
• 负责人: Ivana Barbaric
• 金额: $ 68.86万
• 依托单位: University of Sheffield
• 依托单位国家: 英国
• 项目类别: Research Grant
• 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=MR%2FX000028%2F1
• 关键词: role; mechanosensing; selective; advantage; genetically

Cell replacement therapies aim to heal diseased or injured tissues and organs by replacing the missing or defective cells in the body. Human pluripotent stem cells (hPSC) represent an essential pillar of such efforts due to their dual ability to proliferate extensively in culture and to differentiate to any cell type. Several clinical trials using hPSC-derived differentiated cells for treatment of diseases and injuries, such as macular degeneration, Parkinson's disease and spinal cord injury are currently ongoing, with many more therapies also progressing towards the clinical trials. The generation of large numbers of differentiated cells required for cell replacement therapies typically entails expanding undifferentiated hPSC and keeping them in culture for long periods of time. However, a key issue with this process is the appearance of non-random genetic changes (such as gains of chromosomes 1, 12, 17, 20 or X) in hPSC upon prolonged culture. Genetically variant hPSC harbouring recurrent genetic changes outcompete the euploid, wild-type cells, thereby achieving clonal dominance in cultures. Such recurrent variants show signs of neoplastic transformation and defects in differentiation, raising concerns that they may pose significant dangers for their use in regenerative medicine. These changes may also affect the use of hPSC in other applications including drug discovery, toxicology and disease modelling. Thus, resolving the mechanisms that confer fitness advantage to variant hPSC is pivotal for developing strategies aimed at minimising their appearance during hPSC scale-up for use in research and regenerative medicine.Our recent work established that mechanosensing, i.e. the ability of cells to sense mechanical cues from their environment, is implicated in the selective advantage of variant hPSC. Here, our overarching aim is to investigate the perturbed mechanosensing of variant hPSC in order to design culture conditions that suppress the variant overtake and allow the scale up of genetically normal hPSCs for research and regenerative medicine. First, we will characterise the key mechanosensing machinery (i.e. cell-cell and cell-ECM adhesions) in wild-type and variant cells and investigate the role of mechanosensing in the variant cells' selective advantage. Next, we will study how mechanosensing in hPSC is coupled with signalling pathways that instruct hPSC fates. Finally, using an informed iterative approach, we will engineer cellular environments that reduce the selective advantage of variant hPSC.We expect that the outputs of our research will encompass a mechanistic understanding of variant hPSC's selective advantage and improved culture conditions that minimise the appearance of variants.

细胞替代疗法旨在通过替换体内缺失或有缺陷的细胞来治愈患病或受伤的组织和器官。人类多能干细胞（hPSC）由于其在培养中广泛增殖和分化为任何细胞类型的双重能力，代表了这些努力的重要支柱。目前正在进行一些使用hpsc衍生的分化细胞治疗黄斑变性、帕金森病和脊髓损伤等疾病和损伤的临床试验，还有更多的治疗方法也在进行临床试验。细胞替代疗法所需的大量分化细胞的产生通常需要扩增未分化的hPSC并将其长时间培养。然而，该过程的一个关键问题是hPSC在长时间培养后出现非随机遗传变化（如染色体1、12、17、20或X的增加）。遗传变异的hPSC具有复发性遗传变化，与整倍体、野生型细胞竞争，从而在培养中获得克隆优势。这种复发性变异表现出肿瘤转化和分化缺陷的迹象，引起人们对它们在再生医学中的应用可能构成重大危险的关注。这些变化也可能影响hPSC在其他应用中的使用，包括药物发现、毒理学和疾病建模。因此，解决赋予变异hPSC适应度优势的机制对于在研究和再生医学中使用hPSC扩大规模期间制定旨在最大限度地减少其外观的策略至关重要。我们最近的研究表明，机械感应，即细胞从环境中感知机械信号的能力，与变异hPSC的选择优势有关。在这里，我们的首要目标是研究变异hPSC的受干扰的机械传感，以便设计抑制变异超量的培养条件，并允许扩大遗传正常的hPSC用于研究和再生医学。首先，我们将描述野生型和变异细胞中的关键机械传感机制（即细胞-细胞和细胞- ecm粘附），并研究机械传感在变异细胞的选择优势中的作用。接下来，我们将研究hPSC中的机械传感如何与指示hPSC命运的信号通路相结合。最后，使用一种知情的迭代方法，我们将设计减少变异hPSC的选择优势的细胞环境。我们期望我们的研究成果将包括对变异hPSC的选择优势和改进的培养条件的机制理解，从而最大限度地减少变异的出现。

项目成果

Modeling the selective growth advantage of genetically variant human pluripotent stem cells to identify opportunities for manufacturing process control

• DOI: 10.1016/j.jcyt.2024.01.010
• 发表时间: 2024-04-03
• 期刊: CYTOTHERAPY
• 影响因子: 4.5
• 作者: Beltran-Rendon，Catherine;Price，Christopher J.;Thomas，Robert J.
• 通讯作者: Thomas，Robert J.