背景:靶向T细胞疗法已成为治疗血液恶性肿瘤的有希望的策略。然而,由于其有限的可及性和异质性,其在实体瘤中的应用提出了重大挑战。使用生物材料局部递送肿瘤特异性T细胞已显示出希望,然而,由于成本和时间限制,遗传修饰和离体产生足够数量的肿瘤特异性T细胞所需的程序仍然是主要障碍。
方法:开发了基于聚乙二醇(PEG)的三维(3D)支架,并使用尿素化学与带正电荷的聚L-赖氨酸(PLL)缀合,以有效加载携带肿瘤抗原特异性T细胞受体(TCR)的慢病毒(LV)。广泛表征了支架的物理和生物学特性。Further,将载有OVA-TCRLV的支架植入表达卵清蛋白的B16F10细胞(B16-OVA)肿瘤模型中,以评估抗肿瘤应答和转导T细胞的存在.
结果:我们的发现表明,支架在皮下植入时不会诱导任何全身性炎症,并有效地将T细胞募集到该部位。在B16-OVA黑色素瘤荷瘤小鼠中,支架有效地转导具有OVA特异性TCR的宿主T细胞。这些基因修饰的T细胞表现出对肿瘤和次级淋巴器官的归巢能力,导致肿瘤大小的显着减少和抗肿瘤细胞因子的系统性增加。免疫细胞谱分析显示,在植入这些支架的小鼠的肿瘤内,转导的T细胞的百分比明显较高,抑制免疫细胞的减少。
结论:我们的基于支架的T细胞治疗为编程T细胞靶向实体瘤提供了一种创新的原位定位方法。这种方法为T细胞的体外操作提供了可行的替代方法,避免了大规模体外产生和培养肿瘤特异性T细胞的需要。它提供了一个现成的替代品,便于使用宿主细胞而不是同种异体细胞,因此,克服一个主要障碍。
BACKGROUND: Targeted T-cell therapy has emerged as a promising strategy for the treatment of hematological malignancies. However, its application to solid tumors presents significant challenges due to the limited accessibility and heterogeneity. Localized delivery of tumor-specific T-cells using biomaterials has shown promise, however, procedures required for genetic modification and generation of a sufficient number of tumor-specific T-cells ex vivo remain major obstacles due to cost and time constraints.
METHODS: Polyethylene glycol (PEG)-based three-dimensional (3D) scaffolds were developed and conjugated with positively charged poly-L-lysine (PLL) using carbamide chemistry for efficient loading of lentiviruses (LVs) carrying tumor antigen-specific T-cell receptors (TCRs). The physical and biological properties of the scaffold were extensively characterized. Further, the scaffold loaded with OVA-TCR LVs was implanted in B16F10 cells expressing ovalbumin (B16-OVA) tumor model to evaluate the anti-tumor response and the presence of transduced T-cells.
RESULTS: Our findings demonstrate that the scaffolds do not induce any systemic inflammation upon subcutaneous implantation and effectively recruit T-cells to the site. In B16-OVA melanoma tumor-bearing mice, the scaffolds efficiently transduce host T-cells with OVA-specific TCRs. These genetically modified T-cells exhibit homing capability towards the tumor and secondary lymphoid organs, resulting in a significant reduction of tumor size and systemic increase in anti-tumor cytokines. Immune cell profiling revealed a significantly high percentage of transduced T-cells and a notable reduction in suppressor immune cells within the tumors of mice implanted with these scaffolds.
CONCLUSIONS: Our scaffold-based T-cell therapy presents an innovative in situ localized approach for programming T-cells to target solid tumors. This approach offers a viable alternative to in vitro manipulation of T-cells, circumventing the need for large-scale in vitro generation and culture of tumor-specific T-cells. It offers an off-the-shelf alternative that facilitates the use of host cells instead of allogeneic cells, thereby, overcoming a major hurdle.