Commercial scale manufacture of adult allogeneic cell therapy for regenerative medicine

细胞治疗行业的商业增长在很大程度上依赖于新平台技术的发展，这些技术提供快速、经济高效的生物处理解决方案，以生产大量高质量的细胞产品。在这项提案中，我们的目标是通过将由英国公司ReNeuron （Guildford, Surrey）首创的用于治疗中风的经监管机构批准的细胞永生方法转化为一种用于其他神经疾病的再生细胞治疗的新细胞类型来实现这一目标。患有急性神经损伤（如中风和脊髓损伤）的患者需要快速治疗，以防止功能丧失。举个例子，仅在英国，每年就有2000人因脊柱损伤而瘫痪，其中大部分是年轻人。目前尚无治疗方法。这种残疾对病人、他们的家庭、照顾者和整个医疗保健系统都有重大影响。每位患者每年的平均成本（直接医疗保健和间接社会成本）约为10万英镑。一般来说，神经组织在受损后不会再生。嗅觉系统是个例外，它的神经再生贯穿于整个生命周期，使我们在感冒后能够重新获得嗅觉。再生是可能的，这要归功于鼻子中发现的一种称为嗅鞘细胞（OECs）的独特细胞群，这种细胞通过产生引导通道来促进新的神经元连接。在过去的20年里，Raisman教授的团队已经确定并测试了oec与共定位成纤维细胞结合的能力，以实现神经再生。动物研究已经证明了这种细胞治疗方法的安全性和有效性，临床试验的适当伦理批准也已经到位。然而，有一个重大的障碍。目前的方法使用患者自己的（自体）细胞，但在急性创伤患者中，治疗需要在14天内进行。没有足够的时间进行组织活检，处理细胞，鉴定并植入可行的治疗剂量。因此，我们将逐步转向通用（同种异体）OEC细胞系，并探索平行单细胞培养和直接共培养的选择。在首席科学官John Sinden博士的建议下，我们将使用ReNeuron的方法来制造通用的OEC细胞系，这种细胞系很容易在实验室中生长，然后在移植到患者身上时可靠地沉默，使其成为一种安全的方法。oec成纤维细胞制剂将以不同的比例生产，我们将确定与共培养相关的关键特征和监测问题。我们将使用实验室开发的高通量体外筛选方法测试制剂的效力和再生潜力。然后将开始生产符合gmp要求的有效细胞系。这个项目将使那些受一系列神经损伤影响的人受益。值得注意的是，生物工艺行业将获得一个开创性的通用平台技术，该技术已经在多种细胞类型的几种适应症中进行了测试，并了解了用于治疗的细胞共培养的制造考虑因素。未来成功的基于细胞的产品可能包含两种不同的细胞类型来产生治疗效果。因此，获得适用于可扩展GMP生产的稳健共培养方法至关重要。该项目将加速神经损伤的治疗，并扩大英国在共培养领域的生物工艺知识库和技能基础，以增加英国的商业竞争优势。

Commercial growth within the cell therapy industry heavily relies on development of new platform technologies that offer rapid, cost-effective bioprocessing solutions to produce large quantities of high-quality cell product. In this proposal we aim to achieve that by translating a regulator-approved cell immortalization method pioneered by British company ReNeuron (Guildford, Surrey) for treatment of stroke to a new cell type for regenerative cell therapy for other neural disorders. Patients who suffer acute neurologic injury (such as stroke and spinal cord injury) require rapid treatment in order to prevent loss of function. As a single example, in the UK alone 2,000 mostly young people become paralyzed each year due to spinal trauma. Currently there is no cure. Such disability has major consequences for patients, their families, carers and the whole healthcare system. The average cost (direct healthcare and indirect social costs) per patient per year is in the order of £100,000. As a general rule, neural tissue does not regrow after damage. One exception is in the olfactory system where nerve regeneration happens throughout life, enabling us to regain sense of smell following a common cold. Regeneration is possible thanks to a unique cell population called olfactory ensheathing cells (OECs) found in the nose that facilitate new neuronal connections by producing guidance channels. Over the last 20 years Prof. Raisman's group have identified and tested the ability of OECs in combination with co-located fibroblasts to enable neural regeneration. Animal studies have demonstrated safety and efficacy of this cell therapy approach and appropriate ethical approvals for a clinical trial are in place. However, there is a significant hurdle. The present method uses the patient's own (autologous) cells but in acute trauma patients, treatment needs to occur within 14 days. This is not adequate time to take a tissue biopsy, process the cells, characterize and implant a viable therapeutic dose. We will therefore make a step change to universal (allogeneic) OEC cell lines and explore parallel single cell cultures and direct co-culture options. We will use the ReNeuron approach, with advice from their Chief Scientific Officer, Dr. John Sinden, to make universal OEC cell lines that are easily grown in the lab and then reliably silenced upon transplantation into the patient, making it a safe approach. OEC-fibroblast preparations will be produced at different ratios and we will identify key characterization and monitoring issues associated with co-culture. Preparations will be tested for potency and regenerative potential using robust high-throughput in vitro screening assays developed in our lab. GMP-compliant manufacture of a potent cell line will then be initiated. This project will benefit those affected by a range of neurologic injuries. Significantly, the bioprocess industry will gain a pioneering generic platform technology that has been tested in multiple cell types for several indications and gain understanding of manufacturing considerations for co-culturing of cells for therapy. Successful cell based products of the future are likely to comprise two different cell types to produce the therapeutic effect. Therefore it is crucial to derive robust co-culturing methodologies applicable to scalable GMP manufacture. This project will accelerate cures for neurologic injury and expand the UK's bioprocess knowledge pool and skills-base in the area of co-culture to increase the UK's commercial competitive advantage.