Biomaterial Scaffolds for In Vivo CAR T Cell Manufacture

项目总结 CAR-T细胞疗法彻底改变了包括白血病和淋巴瘤在内的液体肿瘤的治疗方法， 也为实体癌症的治疗带来了巨大的希望。然而，尽管他们史无前例的临床 这种疗法的成功和广泛应用受到了漫长和劳动密集型制造的阻碍 程序。CAR-T细胞的制造既费力又耗时，导致非常高的治疗成本 (约50万美元)。较长的制造时间造成给患者输注CAR-T细胞的延迟数周或数月 随着疾病的迅速发展。广泛的体外细胞操作创造了具有异质性的细胞产品 限制CAR-T细胞植入和持久性的成分和终末分化。努力克服 这些限制主要集中在封闭和自动化的制造设备上，以遏制所需的劳动力 体外制造CAR-T细胞，并提出同种异体现成的CAR-T细胞来克服 需要为每个患者制造CAR-T细胞。这些技术前景看好，但减少了 制造或完全消除体外程序的时间、成本和监管负担仍然是关键 未得到满足的需求。体内产生自体CAR-T细胞将消除体外程序，防止 体外扩增的CAR-T细胞的终末分化及确保自体T细胞的效力和寿命 与同种异体CAR-T细胞产品相比，CAR-T细胞产品被广泛操纵以防止排斥反应和 移植物抗宿主病。这份提案概述了高度创新的高风险/高回报努力的第一步 开发生物指导性生物材料支架，在患者体内完全产生CAR-T细胞并产生 提高了疗效和持久性的CAR-T细胞。我们的努力建立在重要的出版和初步研究的基础上- 几乎没有数据表明，我们的生物材料支架已经有效地激活和介导了CAR-T细胞的反式反应。 体外诱导并在体内高效募集和释放CAR-T细胞，减少CAR-T制造次数 从几周到一天。我们认为，生物相容性藻酸盐生物材料支架可以被修饰成 包裹吸引T细胞的趋化因子，将T细胞招募到支架上。招募后，生物材料SCAF- FLODS将提供αCD3/CD2 8信号来激活T细胞。激活后，T细胞特异性病毒颗粒 已经存在于生物材料中或作为单独步骤给药的生物材料将转导T细胞， 在原位产生肿瘤特异性CAR-T细胞，其方式与放射性淋巴枯竭相容。最后， 支架中的白介素信号将扩大和促进形成的CAR-T细胞的释放，以实现全身疗效。 这种方法可以显著降低治疗成本并显著降低治疗成本，从而产生巨大的临床影响 扩大受益于CAR-T细胞疗法的患者群体。我们预计这些研究将提供 CAR-T细胞制造的基础技术，并促进患者的广泛接触。除了……之外 然而，这种合理的、基于材料的细胞制造方法在癌症中的明确应用可能会 可用于实体肿瘤和其他疾病的治疗性淋巴细胞编程。

PROJECT SUMMARY CAR-T cell therapy has revolutionized the treatment of liquid tumors, including leukemia and lymphoma, and hold enormous promise for treatment of solid cancers as well. However, despite their unprecedented clinical success, widespread utilization of this therapy is hampered by the lengthy and labor-intensive manufacturing procedures. CAR-T cell manufacturing is both laborious and time-consuming, results in very high costs of therapy (~$500,000). The long manufacturing time creates delays of weeks or months to infuse CAR-T cells to patients with rapidly progressing disease. Extensive ex vivo cell manipulation creates cell products with heterogeneous composition and terminal differentiation that limit CAR-T cell engraftment and persistence. Effort to overcome these limitations have focused on closed and automatic manufacturing devices to contain the labor needed to manufacture CAR-T cells ex vivo, and allogeneic off-the-shelf CAR-T cells have been proposed to overcome the need of CAR-T cell manufacturing for each single patient. These technologies are promising, but reducing the time, costs and regulatory burden of manufacturing or eliminating ex vivo procedures entirely remains a critical unmet need. In vivo generation of autologous CAR-T cells would eliminate the ex vivo procedures, prevent the terminal differentiation of ex vivo expanded CAR-T cells and ensure the potency and longevity of autologous T cells as compared to allogeneic CAR-T cell products that are extensively manipulated to prevent rejection and graft-versus-host disease. This proposal outlines the first steps in a highly innovative high-risk/high-reward effort to develop bioinstructive biomaterials scaffolds that generate CAR-T cells entirely within the patient and produce CAR-T cells with improved efficacy and persistence. Our endeavor is built on significant published and prelimi- nary data demonstrating that our biomaterial scaffolds already efficiently activate and mediate CAR-T cell trans- duction in vitro and efficiently recruit and release CAR-T cells in vivo and reduce CAR-T manufacturing times from weeks to a single day. We propose that biocompatible alginate biomaterial scaffolds can be modified to encapsulate T cell-attracting chemokines to recruit T cells to the scaffold. After recruitment, the biomaterial scaf- folds will provide αCD3/CD28 signaling to activate the T cells. After activation, T cell-specific viral particles either already present in the biomaterial or administered to the biomaterial as a separate step will transduce the T cells, generating tumor-specific CAR-T cells in situ in manner compatible with irradiative lymphodepletion. Finally, interleukin signaling in the scaffold will expand and promote release of formed CAR-T cells for systemic efficacy. This approach could have enormous clinical impact by significantly reducing therapy costs and dramatically expanding the patient population benefiting from CAR-T-cell therapy. We expect that these studies will provide a foundational technology for CAR-T cells manufacturing and promote widespread patient access. In addition to the clear application in cancer, however, this rational, materials-based approach for cellular manufacturing could be adopted to program therapeutic lymphocytes in solid tumors and for other diseases.