Tuning tyrosine kinase signalling to improve haematopoietic stem cell expansion and bone marrow transplantation

概述。在这个项目中，我们将开发新的方法来培养脐带血来源的干细胞用于骨髓移植和再生医学。我们最近已经证明了多种新的针对造血干细胞（hsc）的治疗靶点，这些靶点可以对这些细胞的再生潜力产生重大影响。Grey博士证明了靶向受体酪氨酸激酶（RET）的有效性，以改善人类HSC的生长，Hitchcock教授开发了新的修饰细胞因子，以定制HSC中的血小板生成素信号。与我们的iCASE合作伙伴（Plasticell Ltd），我们的临床合作者（Anthony Nolan）和弗朗西斯克里克研究所（伦敦）一起，这个博士研究生将开发最有效的治疗组合，以改善HSC的生长，移植和解开这些表型背后的分子信号通路。历史上，造血和造血干细胞的研究在鉴定层次组织、命运映射和单细胞方法方面处于领先地位。事实上，造血干细胞的定义是它们在移植后改造整个造血系统的能力，骨髓移植（BMT）是第一个进入临床的再生方法。BMT最初用于治疗接受高剂量辐射并遭受骨髓衰竭的患者，并且作为一种临床方法已经发展出再生和免疫治疗的目标。已知造血干细胞存在于成人骨髓和怀孕期间的脐带血中。尽管骨髓活检是侵入性的，动员外周血需要趋化因子治疗，但收集UCB代表了一种侵入性较小的、临床重要的造血干细胞和祖细胞（HSPCs）来源，可用于治疗各种恶性和非恶性疾病。与传统供体来源相比，UCB具有较低的移植物抗宿主病发病率，供体hla匹配要求较低，因此增加了其对血液和非血液恶性肿瘤的价值。尽管UCB储备增加，祖细胞剂量有限，移植和免疫重建的延迟以及成人双UCB移植的成本，强调了提高这些细胞的扩增和效力以用于移植的必要性。目的：通过靶向酪氨酸激酶信号通路提高UCB供体造血干细胞的产生了解调节酪氨酸激酶信号通路改善HSC扩增的机制。将发现转化为临床分级系统。实验方法。在前期工作中，格蕾医生与安东尼·诺兰医院的合作伙伴组织了一大批具有不同发展潜力的UCB捐赠者。格雷博士和希区柯克教授已经开发出新的分子来调节干细胞中的关键信号通路。总之，我们将使用我们之前发表的方案，通过无数体内和体外方法评估HSC的功能和培养扩增，以及分子调节激酶信号，以改善移植的HSC扩增。Plasticell有限公司

Overview. In this project, we will develop new ways to grow umbilical cord blood derived stem cells for bone marrow transplantation and regenerative medicine. We have recently demonstrated multiple new therapeutic targets on haematopoietic (blood) stem cells (HSCs), which can have major impacts on the regenerative potential of these cells. Dr. Grey has demonstrated the efficacy of targeting the receptor tyrosine kinase, RET, to improve human HSC outgrowth and Prof. Hitchcock has developed new modified cytokines to tailor thrombopoietin signalling in HSCs. Together, with our iCASE partner (Plasticell Ltd), our clinical collaborators (Anthony Nolan) and The Francis Crick Institute (London), this PhD studentship will develop the most effective combination of treatments to improve HSC outgrowth, transplantation and unpick the molecular signalling pathways underlying these phenotypes.Background. Historically, the study of haematopoiesis and HSCs has led the way in identification of hierarchical organisation, fate mapping and single cell approaches. Indeed, HSCs are defined by their ability to reform the entire haematopoietic system upon transplantation, with bone marrow transplantation (BMT) being the first regenerative approach to enter the clinic. BMT was initially used to treat patients who had received high dose radiation and suffered bone marrow failure, and as a clinical approach has since developed both regenerative and immunotherapeutic aims.HSCs are known to reside in the bone marrow of adults and umbilical cord blood (UCB) during pregnancy. Although a bone marrow biopsy is invasive and mobilisation of peripheral blood requires chemokine treatment, collection of UCB represents a less invasive, clinically important source of HSCs and progenitors (HSPCs) for the treatment of a wide range of malignant and non-malignant disorders. UCB has a lower incidence of graft-versus-host disease, with less stringent donor HLA-matching required compared with classical donor sources, thus increasing its value for both haematologic and non-haematologic malignancies.Despite increased UCB banking, limited progenitor cell dose, delay of engraftment and immune reconstitution and the cost of double UCB transplantation in adults underscore the need to improve expansion and potency of these cells for the purposes of transplantation.Objectives.To improve the production of HSCs from UCB donors by targeting tyrosine kinase signallingUnderstand the mechanism by which tuning tyrosine kinase signalling improves HSC expansion.Translate findings to clinical grade systems.Experimental approaches.In preliminary work, Dr. Grey has curated a large cohort of UCB donors of varying expansion potential with collaborators at Anthony Nolan. Dr. Grey and Prof. Hitchcock have developed new molecules to tune key signalling pathways in stem cells. Together, we will use our previously published protocols for assessment of HSC functionality and expansion in culture through a myriad of in vivo and in vitro approaches and molecularly tune kinase signalling to improve HSC expansion for transplantation.Plasticell Ltd