Unravelling the molecular mechanisms that drive mitotic defects in human Pluripotent Stem Cells and their potential impact in tumourigenic potential

未来的再生医学将依赖于能够产生各种各样的“设计”组织（神经元、心脏细胞、肝细胞等），这些组织可以用于临床的组织替代。由于十年前获得诺贝尔奖的人类诱导多能干细胞（human induced Pluripotent Stem Cells, hiPSCs）的发现，这一梦想得以实现，这是一项革命性的技术，可以将我们体内的任何细胞转化为多能干细胞，通过体外分化，几乎可以从任何想要的目标组织中获得。然而，在干细胞治疗的前景成为现实之前，还需要克服一些关键的挑战，特别是hipsc衍生的分化组织通常由于尚不清楚的原因而保留增殖和致瘤潜能。最近，Rafael Carazo Salas（英国布里斯托尔大学）实验室发现，人类多能干细胞（hPSCs，即人类胚胎干细胞（hESCs）和hiPSCs）显示出多种有丝分裂缺陷，表明基因组不稳定，这表明其中一些缺陷可能与hPSC衍生细胞和组织未来的肿瘤发生潜力有关。因此，本项目的目的是确定驱动造血干细胞有丝分裂缺陷的分子机制，以及这些有丝分裂缺陷在干细胞致瘤潜能中的作用。具体来说，我们的目标是：1。鉴定在人乳头状细胞（特别是hipsc）中观察到的有丝分裂缺陷负责/参与的基因。2. 研究它们的失调是否在分化后持续存在，并可能导致hPSC衍生的分化细胞/组织的致瘤潜能。3. 询问他们的放松管制是否可以在药理学上恢复，以及是否有可能确定可能减轻这些缺陷的潜在小分子化合物。寻求定义预测特性和特征（“精确诊断”预测因子），这些特性和特征可能告诉细胞、群体或细胞系它们的有丝分裂受损程度，哪些分子机制功能失调，并允许预测可能的致瘤潜力。特别是，研究这些精确诊断是否可以在没有外源性基因编码报告的情况下以无标签的方式获得。5. 在时间允许的情况下，建立/测试这些“精确诊断”的预测能力是否可以转化为更简单的组织学分析和读数，更接近于在更多医疗环境中使用的方法。

The Regenerative Medicine of the future will rely on being able to produce a wide variety of "designer" tissues of choice (neurons, heart cells, liver cells, etc) that can be used for tissue replacement in the clinic. This dream has become achievable thanks to the Nobel Prize winning discovery a decade ago of human induced Pluripotent Stem Cells (hiPSCs), a revolutionary technology allowing any cells in our body to be converted into pluripotent stem cells from which almost any desired target tissue of choice could be derived by differentiation in vitro. However, key challenges have to be overcome before the promise of stem cell therapeutics becomes a reality, particularly the fact hiPSC-derived differentiated tissues often retain proliferative and tumourigenic potential for reasons that are not well understood. Recently, the lab of Rafael Carazo Salas (University of Bristol, UK) discovered that human Pluripotent Stem Cells (hPSCs, that is human Embryonic Stem Cells (hESCs) and hiPSCs) display multiple mitotic defects indicative of genome instability, suggesting that some of those defects might be involved in underpinning future tumourigenic potential of hPSC derived cells and tissues. Therefore, the aim of this project is to identify the molecular mechanisms that drive mitotic defects in hPSCs and the role of these mitotic defects in the tumourigenic potential of stem cells. Specifically, we aim to:1. Identify genes that are responsible/involved in the mitotic defects observed in hPSCs, particularly hiPSCs. 2. Investigate whether their deregulation persists beyond differentiation and could be responsible for tumourigenic potential in hPSC derived differentiated cells/tissues. 3. Ask whether their deregulation could be pharmacologically reverted, and whether it is possible to identify potential small molecule compounds that may alleviate those defects.4. Seek to define predictive properties and features ("Precision Diagnostics" predictors) that might tell for cells, populations or cell lines how mitotically compromised they are, what molecular machineries are dysfunctional, and allow to predict possible tumourigenic potential. In particular, investigate if those Precision Diagnostics can be obtained without exogenous genetically encoded reporters in a label free manner. 5. Time permitting, establish/test whether the predictive power of those "Precision Diagnostics" can be translated to simpler histological assays and readouts, closer to what could be used in more medical settings.