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生产各种可用于临床组织替代的“设计者”组织（神经元、心脏细胞、肝细胞等）。这个梦想已经成为可以实现的，这要归功于十年前获得诺贝尔奖的人类诱导多能干细胞（hiPSC）的发现，这是一项革命性的技术，允许我们体内的任何细胞转化为多能干细胞，几乎任何所需的目标组织都可以通过体外分化获得。然而，在干细胞疗法的前景成为现实之前，必须克服关键挑战，特别是hiPSC衍生的分化组织由于尚不清楚的原因而通常保留增殖和致瘤潜力的事实。最近，Rafael Carazo Salas（英国布里斯托大学）的实验室发现，人类多能干细胞（hPSC，即人类胚胎干细胞（hESC）和hiPSC）显示出指示基因组不稳定性的多种有丝分裂缺陷，这表明这些缺陷中的一些可能涉及支持hPSC衍生细胞和组织的未来致瘤潜力。因此，该项目的目的是确定驱动hPSC中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力中的作用。具体而言，我们的目标是：1。鉴定负责/参与hPSC，特别是hiPSC中观察到的有丝分裂缺陷的基因。2.研究它们的失调是否在分化后持续存在，并且可能是hPSC衍生的分化细胞/组织中的致瘤潜力的原因。3.询问他们的放松管制是否可以逆转，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。试图定义预测性质和特征（“精确诊断”预测因子），这些预测性质和特征可以告诉细胞、群体或细胞系它们在有丝分裂方面受到了多大的损害，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，研究是否可以在无外源基因编码报告基因的情况下以无标签方式获得这些Precision Diagnostics。5.时间允许的话，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测定和读数，更接近于可以在更多医疗环境中使用的内容。

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.