Adoptive T-cell therapy against solid tumours is limited by the apoptosis resistance mechanisms of tumour cells and by the extracellular, immunosuppressive tumour microenvironment. Here we report a temperature-sensitive genome-editing nanodevice that can deliver a Cas9 editor with an external trigger which can be used to edit the genome of tumour cells to reduce resistance to apoptosis and modulate the tumour microenvironment via a mild heating trigger. After local or systemic delivery of Cas9, mild heating is induced by non-invasive near-infrared (NIR) light or focused ultrasound (FUS) to activate Cas9, which initiates simultaneous genome editing of HSP70 (HSPA1A) and BAG3 in tumour cells. This disrupts the apoptotic resistance machinery of the tumour cells against adoptive T cells. At the same time, an NIR- or FUS-induced mild thermal effect reshapes the extracellular tumour microenvironment by disrupting the physical barriers and immune suppression. This facilitates the infiltration of adoptive T cells and enhances their therapeutic activity. Mild thermal Cas9 delivery is demonstrated in different murine tumour models which mimic a range of clinical indications, including a tumour model based on humanized patient-derived xenografts. As a result, the non-invasive thermal delivery of Cas9 significantly enhances the therapeutic efficacies of tumour-infiltrating lymphocytes and chimeric antigen receptor T and shows potential for clinical application.

针对实体瘤的过继性 T 细胞疗法受到肿瘤细胞的凋亡抵抗机制和细胞外免疫抑制肿瘤微环境的限制。在这里，我们报道了一种温度敏感的基因组编辑纳米器件，它可以提供带有外部触发器的Cas9编辑器，可用于编辑肿瘤细胞的基因组，以减少对细胞凋亡的抵抗，并通过温和的加热触发器调节肿瘤微环境。Cas9 局部或全身递送后，通过非侵入性近红外 (NIR) 光或聚焦超声 (FUS) 诱导轻微加热来激活 Cas9，从而启动肿瘤细胞中HSP70 ( HSPA1A ) 和BAG3的同时基因组编辑。这破坏了肿瘤细胞对过继性 T 细胞的凋亡抵抗机制。同时，NIR 或 FUS 诱导的温和热效应通过破坏物理屏障和免疫抑制来重塑细胞外肿瘤微环境。这有利于过继性 T 细胞的浸润并增强其治疗活性。Cas9 的温和热传递在不同的小鼠肿瘤模型中得到了证实，这些模型模拟了一系列临床适应症，包括基于人源化患者异种移植物的肿瘤模型。因此，Cas9的非侵入性热传递显着增强了肿瘤浸润淋巴细胞和嵌合抗原受体T的治疗效果，并显示出临床应用的潜力。