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

基本信息

批准号：
10256524
负责人：
Riya Muckom
金额：
$ 42.22万
依托单位：
AXENT BIOSCIENCES INC.
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2023-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10256524
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

项目摘要

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.

项目摘要帕金森氏病（PD）影响美国超过100万成年人，细胞替代疗法（CRT）使用人类胚胎干细胞（HESC）衍生的中脑多巴胺能（DA）神经元显示在减轻PD症状的动物研究中的巨大希望。但是，当前的制造策略细胞疗法受行业标准格式的约束 - 单层，二维（2D），细胞培养 - 此外，它也遭受了动物来源材料（例如Matrigel以及苛刻的酶细胞检索方法。 Axent Biosciences Inc.正在开发专有的3维（3D）基于水凝胶的培养方法为细胞在生产过程中扩展和从而显着增加每个培养体积产生的细胞数量。此外，我们的专有水凝胶技术是完全合成的，避免了动物衍生的产品，其温度响应相位变化可以通过冷却来轻轻，高的生存能力收获。我们已经证明了我们的3D文化方法实现了hESC的高度可扩展的扩展，然后将3D分化与DA神经元分化。尤其，我们实现了使用3D产生的潜在DA神经元的25倍与标准2D格式相比，培养格式。向前迈进，我们的总体目标是扩展我们的3D 细胞制造过程，以产生高纯度，功能性hESC衍生的DA神经元并验证它们体外和体内功能，以开发我们的化学，制造和控制（CMC）研究新药（IND）应用。在AIM 1中，我们将扩大hESC衍生的DA神经元的生产对于能够在I期临床试验中吸收足够数量的细胞的试点生物反应器。在AIM 2中，我们将制定纯化策略，以去除残留的污染细胞类型。在AIM 3中，我们将在体内验证在3D生物反应器过程中产生的DA神经元的功能，并将对DA神经元进行基准测试以常规2D生产格式生成。这些目标的成功完成将为 CGMP符合CGMP的制造平台，能够产生更高数量的功能性DA神经元与当前的2D培养格式相比，培养量，从而克服了重要的瓶颈 PD细胞疗法的开发。此外，我们的制造过程是可以推广的，可以调整用hESC制造许多细胞类型。最后，净化过程将使最终配方中的污染细胞类型，从而提高了我们候选治疗的安全性为了准备FDA互动会议，并随后提交IND申请。为此建立在II期SBIR提案中进一步的目标，我们将包括成立的临床前研究，尤其是9-- 为了验证安全性的月份动物研究 - 进步我们的DA神经元治疗PD到诊所的翻译。