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

用于精密 CAR-T 细胞制造的单锥形管装置

• 批准号: 10115651
• 负责人: Yuguo Lei
• 金额: $ 35.61万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-19 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10115651
• 关键词: 3-Dimensional; Address; Adopted; Alginates; Allogenic; Autologous; B lymphoid malignancy; CAR T cell therapy; Caliber; Cancer Patient; Cell Aggregation; Cell Culture Techniques; Cell Death; Cells; Clinical Research; Cultured Cells; Data; Data Analyses; Development; Device Designs; Devices; Diagnosis; Diffusion; Disease; Dose; Ensure; Environmental Risk Factor; Gammaretrovirus; Genetic; Genetic Engineering; Geometry; Goals; Human; Hydrogels; Lentivirus; Malignant Neoplasms; Methods; Outcome; Patients; Population; Process; Production; Protocols documentation; Quality of life; Radial; Reproducibility; Solid Neoplasm; Statistical Data Interpretation; Stress; Suspension Culture; T-Lymphocyte; Technology; Time; Translations; Treatment Cost; Tube; Tubular formation; Variant; Virus; Work; base; cancer therapy; cancer type; cell growth; chimeric antigen receptor; chimeric antigen receptor T cells; clinically relevant; cost; cost effective; design; exome sequencing; improved; in vivo; in vivo evaluation; instrument; manufacturing process; mathematical model; new technology; novel; operation; preclinical study; prototype; scale up; shear stress; sound; success; therapy development; transcriptome sequencing; transduction efficiency; tumor

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.

项目总结/摘要 嵌合抗原受体T细胞（CAR T细胞）在治疗B细胞白血病方面取得了非凡的成功， 恶性肿瘤，并已显示出治疗实体瘤的高潜力。然而，它们的广泛使用 受到制造这种电池的许多困难的限制。目前的制造工艺复杂， 成本高，只能为小群体生产细胞。此外，它们的生产差异很大。作为 因此，不同的患者接受非常不同的产品或治疗。长期目标是开发一部小说 技术，以解决汽车制造的挑战。为了实现这一目标，该团队开发了一个证明- 概念技术称为无应力管内细胞培养技术（SFIT）。SFIT提供了均匀的细胞， 高度可再生、可控和细胞友好的微环境，导致极高的培养 效率和一致性。SFIT通过一个小锥形管为每个患者生产细胞， 降低生产成本，提高生产能力。该项目的目的是进一步发展 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。 利用良好的初步研究和多元化的专家团队，具体目标是：（1）验证SFIT 用于培养来自广泛范围的供体的T细胞和用于培养各种T细胞亚型，（2）进一步开发和 验证在SFIT中用慢病毒和γ-逆转录病毒转导T细胞的方法，以及（3）进一步开发和 验证用于产生自体CAR T细胞的基于SFIT的单锥形管装置。该项目将建设 该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、exome-Seq和RNA-Seq数据分析、体内测试、统计分析和比较，以及 与ADS BIOTEC合作，ADS BIOTEC是一家设计、制造和销售用于处理人体细胞的仪器的公司。 随着这个项目的完成，简单，一次性，负担得起的设备的可扩展性，成本效益和 预期自体CAR T细胞的一致产生。这项技术也可以扩大规模， 如果需要的话，大规模生产同种异体CAR T细胞。这项技术不仅可以制造CAR T细胞， 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%的人类在其一生中将被诊断患有某种类型的癌症。 目前，只有大约67%的癌症患者在诊断后能存活五年。CAR T细胞疗法是 有望大大增加他们的生存机会。该提案中开发的技术将使 CAR T细胞可用于许多患者。

项目成果

Isolating and cryopreserving pig skin cells for single-cell RNA sequencing study.

• DOI: 10.1371/journal.pone.0263869
• 发表时间: 2022
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Han L;Jara CP;Wang O;Shi Y;Wu X;Thibivilliers S;Wóycicki RK;Carlson MA;Velander WH;Araújo EP;Libault M;Zhang C;Lei Y
• 通讯作者: Lei Y