Photodynamic therapy (PDT) is a relatively safe and clinically promising treatment to combat primary tumors, especially epidermal carcinoma, while has negligible effects on distant metastasis. Therefore, this work reports a multifunctional nanosystem (HPR@CCP) exerting a combined photodynamic and immunotherapy to amplify the therapeutic effect on primary tumors and distant metastasis. Specifically, this nanosystem was obtained by electrostatic adsorption of a negatively charged hyaluronic acid “shell” with a positively charged “core” consisting of the CRISPR-Cas9 system targeting the Ptpn2 gene (Cas9-Ptpn2) and a modified mitochondria-targeting chlorin e6 (TPP-PEI-Ce6). Cell experiments demonstrated that the HPR@CCP nanoparticles possessed very high transfection efficiency on B16F10 cells, and TPP-PEI-Ce6 in the nanoparticles resulted in a significant PDT efficacy due to the efficient singlet oxygen generation in mitochondria under laser-irradiation. The accumulation of the nanoparticles in the tumor by active and passive tumor-targeting in vivo led to the disruption of the Ptpn2 gene by the Cas9-Ptpn2 plasmids in the nanocarriers, thus sensitizing tumors to immunotherapy by the increase of the IFN-γ and TNF-α signaling and the promotion of the proliferation of CD8⁺ T cells. In addition, Hyaluronidase was administered in advance to destroy the hyaluronic acid in the condensed extracellular matrix and to remove the hyaluronic acid “shell” from the nanosystem, subsequently leading to an enhanced penetration of oxygen and therapeutic agents. Fortunately, the primary and distant tumors in the experimental animals were remarkably inhibited after the combination of PDT-immunotherapy, thus, this easy-to-built nanomedicine could be used as a potential combination therapy against tumors.

光动力疗法（PDT）是一种相对安全且在临床上有希望的疗法，可抗击原发性肿瘤，尤其是表皮癌，而对远处转移的影响可忽略不计。因此，这项工作报告了一种多功能纳米系统（HPR @ CCP），该系统发挥了光动力和免疫疗法的结合，以增强对原发肿瘤和远处转移的治疗作用。具体而言，该纳米系统是通过带负电的透明质酸“壳”与带正电的“核心”的静电吸附而获得的，该核由靶向Ptpn2基因的CRISPR-Cas9系统（Cas9-Ptpn2）和靶向线粒体的二氢二氢卟酚e6（ TPP-PEI-Ce6）。细胞实验表明，HPR @ CCP纳米颗粒对B16F10细胞具有很高的转染效率，纳米粒子中的TPP-PEI-Ce6和TPP-PEI-Ce6产生了显着的PDT功效，这是由于在激光照射下线粒体中有效产生单线态氧。通过主动和被动靶向肿瘤在肿瘤中纳米颗粒的积累体内导致纳米载体中的Cas9-Ptpn2质粒破坏Ptpn2基因，从而通过增加IFN-γ和TNF-α信号以及促进CD8 ⁺ T细胞的增殖来使肿瘤对免疫疗法敏感。另外，预先施用透明质酸酶以破坏浓缩的细胞外基质中的透明质酸并从纳米系统中去除透明质酸“壳”，随后导致增强的氧气和治疗剂的渗透。幸运的是，PDT免疫疗法联合后，实验动物的原发性和远处肿瘤得到了显着抑制，因此，这种易于构建的纳米药物可以用作潜在的抗肿瘤联合疗法。