A Single Conical Tube Device for Precision CAR-T Cells Manufacturing

项目摘要/摘要 嵌合抗原受体T细胞(CAR T细胞)在治疗B细胞方面取得了非凡的成功 已经显示出治疗实体肿瘤的巨大潜力。然而，它们的广泛使用是 受到制造这种电池的一些困难的限制。目前的制造工艺很复杂， 成本很高，而且只能为少数人生产细胞。此外，它们的产量差异很大。AS 结果，不同的患者接受了非常不同的产品或治疗。长期目标是开发一部小说 技术，以应对汽车T的制造挑战。为了实现这一目标，该团队提出了一个证据-- 概念技术称为无应力肾小管内细胞培养技术(SFIT)。SFIT提供均匀的电池， 高度可重复性、可控性和细胞友好性的微环境，导致极高的培养 效率和一致性。SFIT用一个小圆锥管为每个患者生产细胞，这一点很重要 降低生产成本，提高生产能力。本项目的目的是为了进一步发展 并验证这项技术和相关方法，使其为最终的翻译挑战做好准备。 利用完善的初步研究和不同的专家团队，具体目标是(1)验证SFIT 为了培养来自广泛捐赠者的T细胞和培养各种T细胞亚型，(2)进一步开发和 验证慢病毒和γ逆转录病毒转导T细胞的方法，以及(3)进一步开发和 验证基于SFIT的单锥管装置用于生产自体CAR T细胞。该项目将建立 关于团队强大的前期工作，包括数学建模，原型设备设计，广泛的T 细胞培养，外显子序列和RNA序列数据分析，体内测试，统计分析和比较，以及 与ADS Biotec公司合作，后者是一家设计、制造和销售人体细胞处理仪器的公司。 随着该项目的完成，简单、一次性、负担得起的设备将为可扩展、经济高效和 自体CAR T细胞的稳定生产有望实现。这项技术还可以扩展到 如果需要，可以大规模生产同种异体CAR T细胞。这项技术不仅将使CAR T细胞 可广泛使用，但也使产品更加一致和可预测。根据2012-2014年的数据， 大约38.5%的人口在其有生之年将被诊断出患有某种癌症。 目前，只有大约67%的癌症患者在确诊后能存活五年。CAR T细胞疗法是 预计将显著增加它们的存活机会。这项提案中开发的技术将使 许多患者可以使用CAR T细胞。

PROJECT SUMMARY/ABSTRACT Chimeric-antigen-receptor T cells (CAR T cells) have achieved extraordinary success in treating B cell malignancies and have demonstrated high potential for treating solid tumors. However, their widespread use is limited by a number of difficulties in manufacturing such cells. Current manufacturing processes are complicated, costly, and can only produce cells for small populations. Additionally, they have large production variations. As a result, different patients receive very different products or treatments. The long-term goal is to develop a novel technology to address the CAR T manufacturing challenge. Toward this goal, the team has developed a proof- of-concept technology termed stress-free intra-tubular cell culture technology (SFIT). SFIT provides cells uniform, highly reproducible, controllable, and cell-friendly microenvironments, resulting in extremely high culture efficiency and consistency. SFIT produces cells for each patient with one small conical tube, significantly reducing production cost and increasing production capability. The purpose of this project is to further develop and validate this technology and the associated methods to make it ready for the final translational challenge. Leveraging sound preliminary studies and a diverse team of experts, the specific aims are to (1) validate SFIT for culturing T cells from a wide range of donors and for culturing various T cell subtypes, (2) further develop and validate methods for transducing T cells with lentivirus and γ-retrovirus in SFIT, and (3) further develop and validate the SFIT-based single-conical-tube-device for producing autologous CAR T cells. The project will build on the team’s strong preliminary work and include mathematical modeling, prototype device design, extensive T cell culturing, exome-Seq and RNA-Seq data analyses, in vivo testing, statistical analyses and comparisons, and partnership with ADS BIOTEC, a company that designs, builds, and sells instruments for processing human cells. With the completion of this project, simple, disposable, affordable devices for the scalable, cost-effective and consistent production of autologous CAR T cells are expected. This technology can also be scaled up for producing allogeneic CAR T cells in large scales if needed. This technology will not only make CAR T cells broadly accessible, but also make the product much more consistent and predictable. Based on 2012-2014 data, approximately 38.5% of the human population will be diagnosed with some type of cancer during their lifetime. Currently, only about 67% of cancer patients will survive five years after diagnosis. CAR T cell therapy is expected to significantly increase their chances of survival. The technology developed in this proposal will make CAR T cells available to many patients.