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

摘要 CAR-T疗法是治疗癌症的一种快速新兴疗法，两家公司获得了FDA 2017年首次获得CAR-T疗法的批准。这些疗法使用称为T细胞的免疫细胞，无论是来自 病人自己，或者健康的捐赠者，并通过基因工程来杀死肿瘤细胞。遗传 工程化涉及将核酸直接递送到这些细胞，这是具有挑战性的，因为目前的递送 这些方法要么缺乏效率，要么对细胞非常有害，或者两者兼而有之。这项工程是在一个案例中进行的- 这种方法是基于个案的，成本高，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事件。目前正在进行一项 顺序方式，这增加了最终产品的患者时间。为了使CAR-T疗法 为了使更多的患者能够以具有成本效益、安全和及时的方式获得这些药物，需要开发新的方法。 目前的研究正在通过开发新的细胞治疗技术来解决这一需求， 处理细胞的恢复和扩增时间，允许在一次注射中递送多种分子，并且可以 集成到一个封闭的、可随时生产的系统中。另一个研究重点是健康工程 供体T细胞将用于CAR-T疗法，这将允许现成的细胞治疗。为此 要实现的产品类型，需要开发能够处理> 1010细胞，同时还保持高细胞活力和递送效率。 OpenCell Technologies开发了一种专有技术波罗斯，用于递送大分子，如 DNA，RNA和蛋白质到各种各样的细胞类型。波罗斯使用声学驱动器推动细胞通过 喷嘴阵列一次一个细胞，从而产生穿透细胞的剪切力。在这个SBIR计划中， OpenCell正在通过开发更高容量的波罗斯设备来扩展其POROS平台的功能。 方法：在这个第二阶段项目中，OpenCell将开发一个高容量版本的波罗斯，能够 治疗更多的细胞以使CAR-T能够发育。我们将开发一个原型波罗斯设备， 能够在10分钟内处理1-5 × 108个细胞，并证明T细胞中的基因编辑是 概念实验波罗斯设备的开发将为开发一种大型的、一体化的、 符合GMP的POROS-giga设备，用于生产现成的细胞治疗剂。另夕h 二期项目中开发的波罗斯装置也将是一个具有预期应用的可销售装置 在研发、个性化用药和小批量生产细胞为自体细胞 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.