Generation of clinical grade human pluripotent stem cells (iPSCs) using safe mRNA reprogramming strategy

自2006年发现以来，诱导多能干细胞(IPSC)已经证明了它们在疾病建模、药物发现和再生医学中的重要性。利用与胚胎维持相关的基因/转录因子，“成年”体细胞被重新编程或诱导成类似胚胎的状态。这创造了具有自我更新和多潜能的细胞(这意味着它们可以用来创造任何类型的细胞)，同时缓解了通常与胚胎衍生材料相关的伦理限制。此外，IPSCs细胞可以用于自体治疗(使用患者自己的细胞)，或者，在仔细选择捐赠者的情况下，允许多个接受者使用人类白细胞抗原匹配的O阴性细胞库(使用来自不同捐赠者的细胞，这些细胞与接受干细胞治疗的患者的免疫系统相容)。到目前为止，大多数关于IPSCs的研究都是在早期的临床前阶段进行的；一个主要的挑战是使产生IPSCs的方案适合临床使用。许多现有的方案需要进行大量的质量测试，因为病毒/基因/转录因子可能会整合到基因组中，导致不良变化，如果不从细胞中清除，就会增加肿瘤的风险。信使核糖核酸重新编程避免了这些安全问题，因为信使核糖核酸不会整合到基因组中，只在细胞内停留很短一段时间。然而，合适的临床级方案尚未完全开发出来用于制造过程。该项目提出了一种新的策略，用于从成人最少的血液中提取具有与人类胚胎干细胞相关的特性的临床IPSCs。通过使用非整合的mRNA重编程方法，避免了通常与其他方法相关的肿瘤形成风险。为了使这一过程在经济上具有吸引力，制造程序将被标准化，以允许从供体材料、重编程、iPSC生成和细胞库的自动化工作流程。这将使同种异体和自体疗法更快、更安全地产生临床级IPSCs，并为产生的细胞系商业化和向生物技术部门发放许可提供空间。将RoslinCT和Reproell的专业知识结合在一起，将使临床级IPSCs的生产发生重大变化，并将这一技术和制造中心设在英国。“

"Since their discovery in 2006, induced pluripotent stem cells (iPSC) have demonstrated their importance to disease modelling, drug discovery and regenerative medicine. Using genes/transcription factors associated with embryonic maintenance, 'adult' somatic cells are reprogrammed or induced to an embryonic-like state. This creates cells with the properties of self-renewal and pluripotent potential (meaning they can be used to create any cell type) whilst alleviating the ethical constraints often associated with embryonic derived material. In addition, iPSCs cells may be used for autologous therapies (using the patient's own cells) or, with careful selection of the donor, allow HLA matched, O negative cell banks for multiple recipients (allogeneic therapies which use cells from different donors that are compatible with the immune system of the patient receiving the stem cell therapy).To date, most research in iPSCs has been conducted at an early pre-clinical stage; a major challenge is to make the protocols for generating iPSCs suitable for clinical use. Many of the existing protocols require an enormous amount of quality testing because the viruses/genes/transcription factors may integrate into the genome, causing undesirable changes and increasing the risk of tumours if they are not cleared from the cells. mRNA reprogramming avoids these safety concerns as mRNA does not integrate into the genome and remains within the cell for only a short period. However, suitable clinical-grade protocols have not yet been fully developed for the manufacturing processes.This project proposes a novel strategy for the derivation of clinical iPSCs which have the properties associated with human embryonic stem cells whilst being derived from minimal blood from adults. By using a non-integrating mRNA reprogramming method, the risk of tumour formation usually associated with other methods is avoided.To make the process economically attractive, manufacturing procedures will be standardised to allow an automated work flow from donor material, reprogramming, iPSC generation and cell-banking. This will allow quicker and safer generation of clinical grade iPSCs for both allogeneic and autologous therapies and provide scope for commercialisation and licensing of the generated cell lines to the biotechnology sector.Bringing together the expertise of RoslinCT and REPROCELL will provide a step change in the production of clinical grade iPSCs and centre this know-how and manufacturing within the UK."