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）已经证明了它们对疾病建模，药物发现和再生医学的重要性。使用与胚胎维持相关的基因/转录因子，“成年”体细胞被重编程或诱导为胚胎样状态。这创造了具有自我更新和多能潜能特性的细胞（这意味着它们可以用于创造任何细胞类型），同时减轻了通常与胚胎衍生材料相关的伦理约束。此外，iPSC细胞可用于自体疗法，（使用患者自身的细胞），或者在仔细选择供体的情况下，允许HLA匹配的O阴性细胞库用于多个受体。（同种异体疗法，其使用与接受干细胞疗法的患者的免疫系统相容的来自不同供体的细胞）。一个主要的挑战是使产生iPSC的方案适合于临床应用。许多现有的方案需要大量的质量测试，因为病毒/基因/转录因子可能会整合到基因组中，导致不希望的变化，并增加肿瘤的风险，如果它们没有从细胞中清除。mRNA重编程避免了这些安全问题，因为mRNA不会整合到基因组中，并且仅在细胞内停留很短的时间。本项目提出了一种新的策略，用于临床iPSCs的衍生，这种iPSCs具有与人类胚胎干细胞相关的特性，同时来自成人的少量血液。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。为了使该过程在经济上具有吸引力，生产程序将标准化，以实现从供体材料，重编程，iPSC生成和细胞库的自动化工作流程。这将允许更快和更安全地产生用于同种异体和自体治疗的临床级iPSCs，并为生物技术部门提供所产生细胞系的商业化和许可范围。将RoslinCT和REPROCELL的专业知识结合在一起，将为临床级iPSCs的生产提供一个步骤性变化，并将该专业知识和制造集中在英国。"

"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."