A Thermoreversible 3D Manufacturing Platform for Scalable Production of Dopaminergic Neurons for Parkinson's Disease Therapy

用于可扩展生产用于帕金森病治疗的多巴胺能神经元的热可逆 3D 制造平台

• 批准号: 10459577
• 负责人: Riya Muckom
• 金额: $ 26.37万
• 依托单位: AXENT BIOSCIENCES INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10459577
• 关键词: 3-Dimensional; Adult; Affect; Animal Disease Models; Animals; Antibodies; Area; Benchmarking; Biological Sciences; Bioreactors; Brain; Cell Count; Cell Culture Techniques; Cell Survival; Cell Therapy; Cells; Chemistry; Clinic; Clinical; Contralateral; Corpus striatum structure; Cyclic GMP; Development; Dimensions; Disease; Dopamine; Dopaminergic Cell; Dose; Excipients; Formulation; Goals; Harvest; Histologic; Human; Hydrogels; Implant; In Vitro; Industrialization; Industry Standard; Injectable; Investigational Drugs; Investigational New Drug Application; Methods; Midbrain structure; Modeling; Motor; Neurites; Neuroglia; Oxidopamine; Parkinson Disease; Patients; Phase; Phase I Clinical Trials; Platelet-Derived Growth Factor alpha Receptor; Pluripotent Stem Cells; Population; Preparation; Process; Production; Protocols documentation; Rattus; Residual state; Retrieval; Risk; Rotation; Running; Safety; Small Business Innovation Research Grant; Sorting - Cell Movement; System; Technology; Temperature; Time; Tissues; Touch sensation; Translations; Undifferentiated; base; cell dimension; cell replacement therapy; cell type; clinical development; combinatorial; commercialization; density; design; dopaminergic differentiation; dopaminergic neuron; experimental study; fetal; flasks; human embryonic stem cell; implantation; in vivo; innovation; manufacturing process; matrigel; meetings; monolayer; motor deficit; patient population; phase change; preclinical study; reduce symptoms; risk minimization; scale up; stem cell therapy; two-dimensional

PROJECT SUMMARY Parkinson's Disease (PD) affects more than 1 million adults in the U.S, and cell replacement therapy (CRT) using human embryonic stem cell (hESC) derived midbrain dopaminergic (DA) neurons has shown great promise in animal studies in alleviating symptoms of PD. However, current manufacturing strategies for cell therapies are constrained by the industry standard format – monolayer, 2-dimensional (2D), cell culture – that additionally suffer from the lot-to-lot variability of animal-derived materials such as Matrigel as well as harsh enzymatic cell retrieval methods. Axent Biosciences Inc. is developing a proprietary 3-dimensional (3D) hydrogel based culture method to offer a critical extra dimension for cells to expand during production and thereby significantly increase the cell quantity produced per culture volume. Furthermore, our proprietary hydrogel technology is fully synthetic and avoids animal derived products, and its temperature responsive phase change enables gentle, high viability cell harvesting by cooling. We have shown that our 3D culture approach enables highly scalable expansion of hESCs, followed by 3D differentiation to DA neurons. Notably, we have achieved up to 25 times the quantity of potent DA neurons produced per culture volume with our 3D culture format compared to standard 2D formats. Moving forward, our overall objective is to scale up our 3D cell manufacturing process to produce high-purity, functional hESC-derived DA neurons and validate their in vitro and in vivo functionality to develop our Chemistry, Manufacturing, and Controls (CMC) towards filing an Investigational New Drug (IND) application. In Aim 1, we will scale up production of hESC-derived DA neurons to a pilot bioreactor capable of suppling sufficient numbers of cells for a Phase I clinical trial. In Aim 2, we will develop a purification strategy to remove residual contaminating cell types. In Aim 3, we will validate in vivo functionality of DA neurons produced in 3D bioreactor process and will benchmark against DA neurons generated in a conventional 2D production format. Successful completion of these aims will yield the basis for a cGMP-compliant manufacturing platform capable of producing higher quantities of functional DA neurons per culture volume compared to current 2D culture formats and thereby overcome a significant bottleneck for the development of PD cell therapies. Furthermore, our manufacturing process is generalizable and can be tuned to manufacture numerous cell types from hESCs. Finally, the purification process will minimize the risk of contaminating cell types in the final formulation, thereby increasing the safety profile of our candidate therapy in preparation for an FDA INTERACT meeting and subsequent filing of an IND application. To build toward this goal further in a Phase II SBIR proposal, we will include IND-enabling preclinical studies – particularly a 9- month animal study to validate safety – to progress translation of our DA neuron therapy for PD to the clinic.

项目概要 帕金森病 (PD) 影响美国超过 100 万成年人，细胞替代疗法 (CRT) 使用人胚胎干细胞 (hESC) 衍生的中脑多巴胺能 (DA) 神经元显示 在动物研究中，该药物在减轻帕金森病症状方面有着巨大的前景。然而，当前的制造策略 细胞疗法受到行业标准格式的限制——单层、二维（2D）、细胞培养—— 此外，还受到动物源性材料（例如 Matrigel 和 Matrigel）的批次间差异的影响 苛刻的酶促细胞修复方法。 Axent Biosciences Inc. 正在开发一种专有的 3 维 (3D) 基于水凝胶的培养方法为细胞在生产过程中扩展提供了关键的额外维度 从而显着增加单位培养体积产生的细胞数量。此外，我们专有的 水凝胶技术是完全合成的，避免动物源性产品，并且其温度响应 相变可通过冷却实现温和、高活力的细胞收获。我们已经证明我们的 3D 文化 该方法能够高度扩展 hESC 的扩增，然后进行 3D 分化为 DA 神经元。尤其， 我们的 3D 技术使每个培养体积产生的有效 DA 神经元数量增加了 25 倍 文化格式与标准 2D 格式相比。展望未来，我们的总体目标是扩大我们的 3D 细胞制造工艺，生产高纯度、功能性 hESC 衍生的 DA 神经元，并验证其在 开发我们的化学、制造和控制 (CMC) 的体外和体内功能，以提交申请 研究性新药 (IND) 申请。在目标 1 中，我们将扩大 hESC 衍生的 DA 神经元的生产规模 能够为第一阶段临床试验提供足够数量的细胞的中试生物反应器。在目标 2 中，我们将 制定纯化策略以去除残留的污染细胞类型。在目标 3 中，我们将在体内验证 3D 生物反应器过程中产生的 DA 神经元的功能，并将以 DA 神经元为基准 以传统的 2D 制作格式生成。成功完成这些目标将为 符合 cGMP 的制造平台，能够生产更多数量的功能性 DA 神经元 与当前的 2D 培养格式相比，培养体积更大，从而克服了 PD细胞疗法的发展。此外，我们的制造工艺是通用的并且可以调整 从 hESC 制造多种细胞类型。最后，净化过程将最大限度地降低风险 最终配方中污染细胞类型，从而提高我们候选疗法的安全性 准备 FDA INTERACT 会议和随后提交 IND 申请。为实现这一目标而建设 在第二阶段 SBIR 提案中进一步实现目标，我们将包括支持 IND 的临床前研究——特别是 9- 为期一个月的动物研究来验证安全性——以推进我们用于 PD 的 DA 神经元疗法向临床的转化。