Establishing the POROS Intracellular Delivery System as a Platform for Cellular Therapeutics

摘要 CAR-T疗法是一种迅速崛起的癌症治疗方法，有两家公司接受FDA 他们的第一个CAR-T疗法于2017年获得批准。这些疗法使用称为T细胞的免疫细胞，要么来自 患者自己，或健康的捐赠者，并通过基因工程来杀死肿瘤细胞。基因 工程学涉及到将核酸直接输送到这些细胞，这与目前的输送方式相比具有挑战性 这些方法要么缺乏效率，要么对细胞造成极大的破坏，或者两者兼而有之。这项工程是在一个案例上进行的- 这是一种基于个案的方法，费用昂贵，耗时长。下一代CAR-T疗法将包括 更复杂的基因工程，需要两个或更多的编辑活动。这项工作目前正在一个 顺序方式，这增加了最终产品的耐心时间。为了让CAR-T疗法 为了以具有成本效益、安全和及时的方式为更多的患者提供服务，需要开发新的方法。 目前的研究正在通过开发新的细胞处理技术来满足这一需求，这种技术可以更快地 经过处理的细胞的恢复和扩张时间，允许一次传递多个分子，这可以 集成到一个封闭的、可用于制造的系统中。另一个研究的焦点是工程健康 捐献的T细胞将用于CAR-T治疗，这将允许现成的细胞治疗。为了这个 要实现的产品类型，需要开发能够处理的高容量处理方法&燃气轮机； 1010细胞，同时保持较高的细胞存活率和输送效率。 OpenCell Technologies开发了一项名为Poros的专有技术，以交付以下大分子 DNA、RNA和蛋白质到各种各样的细胞类型。Poros使用声学驱动器推动细胞通过 一次一个单元的喷嘴阵列，从而产生使单元穿孔的剪切力。在这个SBIR计划中， OpenCell正在通过开发更高容量的POROS设备来扩展其POROS平台的能力。 方法：在这个第二阶段的项目中，OpenCell将开发一个高容量版本的POROS，能够 治疗更多的细胞以促进CAR-T的发展。我们将开发一个POROS设备的原型， 能够在10分钟内处理1-5×108个细胞，并演示T细胞的基因编辑 概念实验。Poros设备的开发将为开发一款大型、一体化、 符合GMP的Poros-Giga设备，用于制造现成的细胞治疗药物。此外， 在此二期项目中开发的POROS单元也将是一个可销售的单元，具有预期的应用 在研发、个性化药物和小批量生产自体细胞方面 治疗和临床试验。

Abstract CAR-T therapy is a rapidly emerging therapy for the treatment of cancer, with two companies receiving FDA approval for their first CAR-T therapies in 2017. These therapies use immune cells called T cells, either from the patient themselves, or a healthy donor, and genetically engineer them to kill the tumor cells. The genetic engineering involves direct delivery of nucleic acids to these cells, which is challenging as current delivery methods either lack efficiency, are very damaging to cells, or both. This engineering is performed on a case- by-case basis and the methods are costly and time-consuming. Next generation CAR-T therapies will include more complex genetic engineering, with two or more editing events required. This is currently being done in a sequential fashion, which increases the time-to-patient of the final product. In order to make CAR-T therapies available to more patients in a cost-effective, safe and timely manner, new approaches need to be developed. Current research is addressing this need by developing new technologies for cell treatment that allow faster recovery and expansion time of the treated cells, allow delivery of multiple molecules in one shot and that can be integrated into a closed, manufacturing-ready system. Another focus of research is engineering healthy donor T cells to be used in CAR-T therapy, which would allow for an off-the-shelf cellular therapeutic. For this type of product to be realized, high capacity treatment methods need to be developed capable of treating > 1010 cells while also maintaining high cell viability and delivery efficiency. OpenCell Technologies has developed a proprietary technology, POROS, to deliver macromolecules such as DNA, RNA and protein to a wide variety of cell types. POROS uses an acoustic drive to push cells through an array of nozzles one cell at a time, thus creating shear force that porates the cells. In this SBIR program, OpenCell is expanding the capabilities of its POROS platform by developing higher capacity POROS devices. Approach: In this Phase II project, OpenCell will develop a high capacity version of POROS, capable of treating a larger number of cells to enable CAR-T development. We will develop a prototype POROS device, capable of treating 1-5 × 108 cells in under 10 minutes, and demonstrate gene editing in T cells as a proof-of- concept experiment. The POROS device development will pave the way for development of a large, all-in-one, GMP-compliant POROS-giga device, for manufacturing of off-the-shelf cellular therapeutics. Additionally, the POROS unit developed in this Phase II project will also be a salable unit of its own with anticipated applications in research and development, personalized medicine and small batch production of cells for autologous cell therapies and for clinical trials.