High Density Cell Respirator (HDCR) for the production of vectors, viruses and vaccines

项目概要/摘要 此I/II期STTR快速通道提案响应了2018/2019 NCATS SBIR/STTR的号召 研究优先发展技术，以便“新的治疗方法和疾病的治愈方法可以提供给 患者更快”。生产改变生命的基因编辑载体，杀死癌症的病毒，以及生命- 保存疫苗目前依赖于传统的细胞培养技术。一些基于病毒和细胞- 基础疗法已经成为癌症、遗传性失明、 免疫缺陷和先天性代谢缺陷。在这个令人兴奋的领域，许多正在开发的疗法 都在等待测试的名单上然而，目前的基于细胞培养的生产是昂贵的，并且参与过程缓慢。 现有需求。例如，基于AAV的基因编辑的临床试验需要10个病毒颗粒， 目前需要一年的生产和成本1-2百万美元的数量。因此，40万美元的费用， 最近批准的基因药物每名患者140万美元并不奇怪。这些价格标签根本不是 可持续的社会。如果发生大流行，需要数年时间才能生产出足够剂量的疫苗 以保护世界上70亿人口的现有生产方式。因此，提高了效率， 生产细胞系的培养速度是制造基因编辑载体、溶瘤载体和细胞培养载体的共同目标。 病毒和疫苗。我们在XDemics公司、加州理工学院的联合研究工作， 和希望之城国家医疗中心，已经导致了一种改进的细胞培养方法。基于 已知的事实是，氧气输送是增加细胞密度、活力和病毒的最限速的过程 我们从高密度细胞呼吸器（HDCR）（美国专利号10，053，660）中创造了一种新的高密度细胞呼吸器（HDCR）。 一种可渗透氧气的材料，它可以廉价地模制成大薄片，并具有集成的细胞保持功能 结构，用于贴壁或悬浮细胞的有效膜氧合。我们的假设是 消除剪切应力和通过HDCR的低流量介质输送，这是通过以下解耦实现的： 通过细胞的膜氧合进行气体交换，将允许提高产率，降低成本，并增加 治疗性病毒载体和病毒的生产速度。我们有初步数据证实这一点 假设，并产生了原型优化。在此，我们提出了I期研究，以优化 设计用于细胞生长的HDCR并证明病毒生产。拟定的II期研究将包括 研究和开发多种病毒载体的生产工艺，包括AAV和免疫- 溶瘤痘病毒/疫苗。我们预计，HDCR将破坏载体和病毒生产领域， 允许>10倍的效率和>2-10倍的生产速度。我们的长期目标是 加快生产临床相关数量的病毒药物和疫苗， 个月降低基因治疗载体、基于细胞的免疫疗法和疫苗的成本将 加速开发治疗癌症、基因缺陷和传染病的新疗法。

PROJECT SUMMARY/ABSTRACT This Phase I/II STTR Fast Track proposal responds to the call from the 2018/2019 NCATS SBIR/STTR Research Priorities to develop technologies so that “new treatments and cures for disease can be delivered to patients more quickly”. The production of life-altering gene editing vectors, cancer killing viruses, and life- saving vaccines currently depends on traditional cell culture techniques. A number of virus-based and cell- based therapies have become clinical treatments for cancer, for genetically-related blindness, for immunodeficiency, and for inborn errors of metabolism. In this exciting field, many therapies being developed are on waitlists to be tested. However, current cell culture-based production is costly and slow to attend the existing demand. For instance, a clinical trial for AAV-based gene editing requires 10 viral particles, a quantity currently requiring a year for production and costing 1-2 million dollars. Thus, the cost of $400,000 to $1,400,000 per patient for recently approved gene medicines is not surprising. These price tags simply are not sustainable for society. In the event of a pandemic, it would take years to generate sufficient doses of vaccines to protect the 7 billion world population by current production methods. Thus, increasing the efficiency and speed of culture of production cell lines are common goals for manufacturing of gene editing vectors, oncolytic viruses, and vaccines. Our joint research efforts at XDemics Corporation, the California Institute of Technology, and the City of Hope National Medical Center, have resulted in an improved method of cell culture. Based on a known fact that oxygen delivery is the most rate-limiting process for increasing cell density, viability, and virus production we created a novel high density cell respirator (HDCR) (US Patent no. 10,053,660) from highly oxygen permeable material that can be inexpensively molded into large sheets, with integrated cell retention architecture, for efficient membrane oxygenation of adherent or suspension cells. Our hypothesis is that elimination of shear stress and the low flow media delivery through the HDCR, enabled by the decoupling of gas exchange via membrane oxygenation of cells, will allow for improved yield, decreased cost, and increased speed of production of therapeutic viral vectors and viruses. We have preliminary data confirming this hypothesis and have produced prototypes for optimization. Herein we propose Phase I studies to optimize the design of the HDCR for cell growth and demonstrate virus production. Proposed Phase II studies will consist of research and development of production processes for multiple viral vectors, including AAV and immuno- oncolytic poxvirus/vaccine. We expect that the HDCR will disrupt the field of vector and virus production, by allowing >10 times greater efficiency and >2-10 times greater speed of production. Our long-term goal is to speed up production of clinically-relevant quantities of viral medicines and vaccines from years down to months. Decreasing the cost of gene therapy vectors, cell-based immunotherapies, and vaccines will accelerate development of novel therapies for treating cancers, gene defects, and infectious diseases.