Developing Blood Cell Production Platforms for Therapeutic Applications

离体或体外血细胞的产生和修饰在基于细胞的治疗和再生医学中具有巨大的潜力。实现这一潜力的一个重要瓶颈是难以在体外有效地生产大量血液制品。解决这个问题的一种方法是找到新的方法来有效地操纵血液前体和祖细胞，以创建可以扩增的细胞系，同时仍然保留其发育潜力。这些细胞可以潜在地代表治疗相关的细胞产物的无限来源，例如修饰的自然杀伤细胞、嵌合抗原受体T细胞、修饰的红细胞、血小板或造血干细胞。转录因子指导的重编程、转分化或诱导增殖已经通过异位基因表达实现。新的非遗传方法，如蛋白质转导或使用化学化合物，已被证明是产生类似结果的遗传修饰的潜在替代方案，为基于基因的细胞调节策略的新发现建立了临床路径。本博士项目旨在通过建立新的血细胞系，开发新的方法来生产大量的造血细胞和分化的造血祖细胞。该项目还旨在建立用于先进细胞疗法的改良血细胞平台。诱导增殖的理想靶点是造血内皮细胞（HE），其是胚胎发育期间产生的罕见瞬时细胞群，并且是包括造血干细胞（HSC）在内的所有成体血细胞的前体。HE细胞系将是生产任何成人血液谱系（包括HSC）的强大平台，并为研究驱动和指导血液形成的分子途径提供工具。该项目旨在通过异位基因表达诱导HE细胞的广泛增殖，同时保持这些细胞从内皮谱系细胞向HSC及其后代过渡的能力。将探索修改这些HE和血细胞系以生产定制血细胞产品的潜力，最终目标是开发用于治疗癌症和自身免疫性疾病的新的潜在细胞免疫疗法。

Generation and modification of blood cells ex vivo or in vitro harbours a huge potential in cell based therapies and regenerative medicine. An important bottleneck in realising this potential is the difficulty in efficiently producing large amounts of blood products in vitro. One way to address this problem is to find new ways to effectively manipulate blood precursors and progenitors to create cell lines that can be amplified while still retaining their developmental potential. These cells could potentially represent an unlimited source of therapeutically relevant cell products such as modified natural killer cells, chimeric antigen receptor T-Cells, modified red blood cells, platelets or haematopoietic stem cells.The seminal demonstration that pluripotency can be induced by the ectopic expression of specific transcription factors has instigated a lot of interest in developing new strategies to manipulate the developmental potential of cells. Transcription factor directed reprogramming, trans-differentiation, or induced proliferation have been achieved through ectopic gene expression. Novel non-genetic methods, such as protein transduction or use of chemical compounds, have been demonstrated as potential alternatives to genetic modification producing similar results, establishing a clinical path for new discoveries in gene based cellular modulation strategies. This PhD project aims to develop new methods to produce large quantities of haematopoietic cells and differentiated haematopoietic progenitor cells through the establishment of novel blood cell lines. The project also aims to establish modified blood cell platforms for applications in advanced cellular therapies. An ideal target to induce proliferation of is the haemogenic endothelium (HE), which is a rare transient cell population generated during embryonic development, and the precursor to all adult blood cells including haematopoietic stem cells (HSCs). A HE cell line would be a powerful platform for the production of any adult blood lineage including HSCs and provide a tool for investigating the molecular pathways that drive and direct blood formation. The project aims to induce extensive proliferation of HE cells though ectopic gene expression, while maintaining the ability for these cells to transition from endothelial lineage cells to HSCs and their progeny. The potential of modifying these HE and blood cell lines to produce customized blood cell products will be explored with the ultimate goal of developing new potential cellular immunotherapies for the treatment of cancer and autoimmune diseases.