MR/V005405/1

MR/V005405/1

• 批准号: MR/V005405/1
• 负责人: Benjamin Simons
• 金额: $ 80.72万
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2020
• 资助国家: 英国
• 起止时间: 2020 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FV005405%2F1
• 关键词: MR; V005405

The transplantation of adult stem cells offers the potential to regenerate tissue. Haematopoietic stem cell transplantation has been long-established as a radical therapy for the treatment of blood cancers, while for other stem cell types, such as skin, transplantation is becoming increasing trialled in the clinic. In the context of sperm production, stem cell transplantation promises a wide range of potential applications, from the restoration of fertility of cancer patients who are affected by the sterilising effects of their treatments, to the preservation of genetic diversity of farm animals and endangered species. Yet, the low efficiency with which stem cells engraft in tissue following transplantation makes it currently unviable as a practical technology. Here, by resolving the regulatory programmes that control the fate of spermatogonial stem cells, the aim of this proposal is to harness the flexible behaviour of stem cells and to identify chemical compounds that could increase regeneration efficiency, opening new horizons in the treatment of fertility and animal conservation.Previously, through collaborative studies, we have used genetic cell labelling approaches and in vivo live-imaging to resolve the dynamics and fate behaviour of spermatogonial stem cells in mouse testes both in steady-state and during regeneration following injury. These studies have shown that stem cells are heterogeneous and dynamic both in their gene expression pattern and their renewal potential. Further, functional studies by our team have found evidence that the regulation of stable stem cell density during homeostasis involves a "quorum sensing"-like mechanism, similar to that encountered in bacterial populations or ecological settings, that allows stem cells to switch reversibly between states biased towards renewal or poised for differentiation. Yet the underlying gene regulatory programmes controlling this state choice remain in question. To address this, we propose a multi-disciplinary strategy that will exploit advances in single-cell technology to profile the heterogeneity of individual spermatogonial stem cell populations and their early differentiating progenies, both in steady-state conditions and during regeneration. By combining these approaches with genetic in vivo cell lineage tracing in the mouse testis, we will identify the key gene regulatory networks that control the balance between stem cell renewal and differentiation. Finally, to advance our understanding towards potential clinical application in humans, we will compare the activity of the key signalling pathways between mouse and human tissues by mining human single-cell datasets. We will then validate these results by performing deep-phenotypical characterisations of human spermatogonial stem cell populations using a multiplex proteomics approach.By resolving the mechanisms that regulate stem cell renewal and their ability to transition reversibly between primed states, our long-term goal is to identify chemical modulators that can increase transplantation efficiency, opening up the potential of this technology for practical use and clinical applications.

成体干细胞的移植提供了组织再生的潜力。造血干细胞移植长期以来一直被认为是治疗血癌的根本疗法，而对于皮肤等其他干细胞类型，移植在临床上的试验也越来越多。在精子生产方面，干细胞移植具有广泛的潜在应用，从恢复受治疗绝育作用影响的癌症患者的生育能力，到保护家畜和濒危物种的遗传多样性。然而，干细胞移植后植入组织的效率较低，使其目前无法成为一项实用技术。在这里，通过解决控制精原干细胞命运的调控程序，该提案的目的是利用干细胞的灵活行为，并识别可以提高再生效率的化合物，为生育治疗和动物保护开辟新视野。此前，通过合作研究，我们已经使用遗传细胞标记方法和体内实时成像来解决精原干细胞的动态和命运行为。 小鼠睾丸中处于稳态和损伤后再生过程中的精原干细胞。这些研究表明，干细胞的基因表达模式和更新潜力都具有异质性和动态性。此外，我们团队的功能研究发现证据表明，体内平衡期间稳定干细胞密度的调节涉及类似“群体感应”的机制，类似于细菌群体或生态环境中遇到的机制，该机制允许干细胞在偏向更新或准备分化的状态之间可逆地切换。然而控制这种状态选择的潜在基因调控程序仍然存在疑问。为了解决这个问题，我们提出了一种多学科策略，该策略将利用单细胞技术的进步来分析个体精原干细胞群体及其早期分化后代在稳态条件下和再生过程中的异质性。通过将这些方法与小鼠睾丸中的遗传体内细胞谱系追踪相结合，我们将确定控制干细胞更新和分化之间平衡的关键基因调控网络。最后，为了加深我们对人类潜在临床应用的理解，我们将通过挖掘人类单细胞数据集来比较小鼠和人体组织之间关键信号通路的活性。然后，我们将通过使用多重蛋白质组学方法对人类精原干细胞群进行深层表型表征来验证这些结果。通过解决调节干细胞更新的机制及其在启动状态之间可逆转换的能力，我们的长期目标是确定可以提高移植效率的化学调节剂，从而开启该技术在实际使用和临床应用中的潜力。

项目成果

Transient suppression of transplanted spermatogonial stem cell differentiation restores fertility in mice.

短暂抑制移植的精原干细胞分化可恢复小鼠的生育能力。

• DOI: 10.17863/cam.66203
• 发表时间: 2021
• 作者: Nakamura Y