纳米工程化T细胞体系的构建及其对口腔鳞状细胞癌的体外治疗研究

doi:10.3969/j.issn.1674-8115.2025.07.008

上海交通大学学报（医学版） ›› 2025, Vol. 45 ›› Issue (7): 866-873.doi: 10.3969/j.issn.1674-8115.2025.07.008

纳米工程化T细胞体系的构建及其对口腔鳞状细胞癌的体外治疗研究

孟靖¹, 谢玉婷², 左佳鑫¹, 熊屏¹()

^1.上海交通大学医学院附属第九人民医院超声医学科，上海 200011
^2.福建医科大学附属福建省妇幼保健院妇产科，福州 335001

收稿日期:2025-01-08 接受日期:2025-04-11 出版日期:2025-07-28 发布日期:2025-07-28
通讯作者: 熊　屏，主任医师，博士；电子信箱：xiongpingxp@163.com。
作者简介:第一联系人：孟靖、谢玉婷为共同第一作者(co-first authors)。
基金资助:
上海市科学技术委员会基金(23ZR1438000);上海交通大学医学院附属第九人民医院学科群建设项目(JYJC202132)

Nanoengineered T cell system for the in vitro treatment of oral squamous cell carcinoma

MENG Jing¹, XIE Yuting², ZUO Jiaxin¹, XIONG Ping¹()

^1.Department of Ultrasound, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China
^2.Department of Obstetrics and Gynecology, Fujian Maternal and Child Health Hospital affiliated to Fujian Medical University, Fuzhou 335001, China

Received:2025-01-08 Accepted:2025-04-11 Online:2025-07-28 Published:2025-07-28
Contact: XIONG Ping, E-mail: xiongpingxp@163.com.
Supported by:
Project of Shanghai Science and Technology Commission(23ZR1438000);Project for Disciplinary Group Construction of Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine(JYJC202132)

摘要/Abstract

摘要：

目的·构建基于纳米工程化的T细胞体系并探究其体外治疗口腔鳞状细胞癌的协同效果，为口腔鳞状细胞癌的治疗提供新策略。方法·制备高亲水性透明质酸（hyaluronic acid，HA）涂层的锆基金属有机框架（Zr-metal-organic framework，Zr-MOF）纳米颗粒（nanoparticle，NP）PCN224-HA NPs，活化其结构上的羧基，使其可以通过酰胺反应与抗CD45抗体偶联。从小鼠脾脏中提取高纯度CD8⁺ T细胞，并利用T细胞表面抗原CD45负载上PCN224-HA，利用生物电镜和共聚焦显微镜观察偶联情况，并检测在超声辐照下自由基的生成。酶标仪记录体外治疗口腔鳞癌Cal27细胞活性变化，探究体外实验的治疗效果。结果·成功合成PCN224-HA NPs，并在生物电镜和共聚焦显微镜上验证其成功偶联上CD8⁺ T细胞的表面，构建纳米工程化T细胞体系（T细胞@PCN224-HA NPs，简称PH T细胞）；在超声辐照后监测到自由基产生，进行体外协同治疗后Cal27细胞活性低至31.70%。结论·通过初步实验证明，构建的体系中纳米颗粒能成功负载于T细胞的表面，同时具备纳米颗粒本身的声动力效能和T细胞的免疫杀伤作用的多重功能，可为口腔鳞癌治疗提供新策略，有望用于进一步的体内肿瘤杀伤实验和疾病治疗。

关键词: 口腔鳞状细胞癌, 声动力治疗, 免疫治疗, 纳米工程化T细胞体系, PCN224-HA纳米颗粒, CD8? T细胞, 协同治疗

Abstract:

Objective ·To construct a nanoengineered T cell system and explore its synergistic efficacy in the in vitro treatment of oral squamous cell carcinoma, and provide a new strategy for this disease. Methods ·Zr-MOF nanoparticles with a highly hydrophilic hyaluronic acid (HA) coating (PCN224-HA NPs) were prepared, and their carboxyl groups were activated to enable coupling with anti-CD45 antibodies through an amide reaction. High-purity CD8⁺ T cells were then extracted from the spleens of mice, and the PCN224-HA NPs were loaded onto the T cells via the CD45 antigens on their surface. The coupling was observed using bioelectron microscopy and confocal microscopy, and free radical generation was detected under ultrasound irradiation. The viability of Cal27 cells was recorded by a microplate reader to explore therapeutic effects in vitro. Results ·PCN224-HA NPs were successfully synthesized and confirmed to be coupled onto the surface of CD8⁺ T cells, constructing a nanoengineered T cell system (T cells@PCN224-HA NPs). Free radical production was monitored after ultrasound irradiation and the viability of Cal27 cells decreased to 31.70% after in vitro treatment. Conclusions ·Preliminary experiments demonstrated that the nanoparticles can be successfully loaded onto the surface of T cells, providing combined sonodynamic therapeutic effects from the nanoparticles and immunocidal effects from the T cells. This offers a novel strategy for the treatment of oral squamous cell carcinoma, with potential for further in vivo anti-tumor experiments and therapeutic applications.

Key words: oral squamous cell carcinoma, sonodynamic therapy, immunotherapy, nanoengineered T cells, PCN224-HA nanoparticle, CD8⁺ T cell, synergistic therapy

中图分类号:

R730.5

孟靖, 谢玉婷, 左佳鑫, 熊屏. 纳米工程化T细胞体系的构建及其对口腔鳞状细胞癌的体外治疗研究[J]. 上海交通大学学报（医学版）, 2025, 45(7): 866-873.

MENG Jing, XIE Yuting, ZUO Jiaxin, XIONG Ping. Nanoengineered T cell system for the in vitro treatment of oral squamous cell carcinoma[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2025, 45(7): 866-873.

图/表 5

图1 具有声动力活性的T细胞@PCN224-HA系统的合成示意

Fig 1 Schematic representation of the synthesis of the T cell @PCN224-HA system with sonodynamic activity

图2 PCN224和PCN224- HA纳米粒的结构表征Note： A. TEM structure diagrams of PCN224 and PCN224-HA, each with a scale bar of 100 nm. B. Electron microscope image of the elemental distribution of PCN224, with a scale bar of 50 nm. C. Electron microscope image of the elemental distribution of PCN224-HA, with a scale bar of 50 nm. D. Hydrodynamic diameter distribution graphs of PCN224 and PCN224-HA. E. Zeta potential graphs of PCN224 and PCN224-HA.

Fig 2 Structural characterization of PCN224 and PCN224-HA nanoparticles

图3 PCN224-HA纳米颗粒与T细胞表面偶联情况Note： A. Elemental distribution maps of T cells and PH T cells obtained by biological electron microscopy, both with a scale bar of 2 nm. B. Laser confocal image of T-cell conjugated PCN224-HA NPs. a—control group, T cells without PCN224-HA NPs (labeled with green fluorescence), with a scale bar of 5 μm; b—PCN224-HA NPs (labeled with red fluorescence) bound to T cells (labeled with green fluorescence), with a scale bar of 5 μm; c—PCN224-HA NPs (labeled with red fluorescence) bound to T cells (labeled with green fluorescence), with a scale bar of 2 μm.

Fig 3 Observation of PCN224-HA nanoparticles coupled to the surface of T cells

图4 PCN224-HA和PH T细胞的声动力学性能Note： A. Ultraviolet absorption spectra of the DPBF probe after ultrasonic irradiation of PCN224-HA for different durations. B. Electron spin resonance (ESR) spectra of the singlet oxygen probe TEMP after various treatments, showing characteristic peaks of generated free radicals.

Fig 4 Sonodynamic performance of PCN224-HA and PH T cells

图5 PH T细胞对Cal27细胞的体外治疗实验Note： A. Cytotoxicity of PCN224-HA nanoparticles at different concentrations. B/C. Laser confocal images showing Cal27 cell viability and live/dead fluorescence staining after different treatments. Green fluorescence represents live cells and red fluorescence represents dead cells. The treatment conditions were as follows: a—control group, no treatment; b—PCN224-HA only; c—ultrasonic irradiation only; d—CD8⁺ T cells only; e—PH T cells only; f—PH T cells + ultrasonic irradiation. ①P=0.014, ②P<0.001.

Fig 5 In vitro therapeutic experiments of PH T cells against Cal27 cells

参考文献 31

[1]	JOHNSON D E, BURTNESS B, LEEMANS C R, et al. Head and neck squamous cell carcinoma[J]. Nat Rev Dis Primers, 2020, 6(1): 92.
[2]	CHOW L Q M. Head and neck cancer[J]. N Engl J Med, 2020, 382(1): 60-72.
[3]	TABERNA M, MENA M, PAVÓN M A, et al. Human papillomavirus-related oropharyngeal cancer[J]. Ann Oncol, 2017, 28(10): 2386-2398.
[4]	GRABOYES E M, KOMPELLI A R, NESKEY D M, et al. Association of treatment delays with survival for patients with head and neck cancer: a systematic review[J]. JAMA Otolaryngol Head Neck Surg, 2019, 145(2): 166-177.
[5]	SOLOMON B, YOUNG R J, RISCHIN D. Head and neck squamous cell carcinoma: genomics and emerging biomarkers for immunomodulatory cancer treatments[J]. Semin Cancer Biol, 2018, 52(Pt 2): 228-240.
[6]	HANNA G J, O'NEILL A, SHIN K Y, et al. Neoadjuvant and adjuvant nivolumab and lirilumab in patients with recurrent, resectable squamous cell carcinoma of the head and neck[J]. Clin Cancer Res, 2022, 28(3): 468-478.
[7]	MACHIELS J P, RENÉ LEEMANS C, GOLUSINSKI W, et al. Squamous cell carcinoma of the oral cavity, larynx, oropharynx and hypopharynx: EHNS-ESMO-ESTRO Clinical Practice Guidelines for diagnosis, treatment and follow-up[J]. Ann Oncol, 2020, 31(11): 1462-1475.
[8]	CRAMER J D, BURTNESS B, LE Q T, et al. The changing therapeutic landscape of head and neck cancer[J]. Nat Rev Clin Oncol, 2019, 16(11): 669-683.
[9]	TANG Z M, ZHAO P R, WANG H, et al. Biomedicine meets Fenton chemistry[J]. Chem Rev, 2021, 121(4): 1981-2019.
[10]	LIN X H, SONG J B, CHEN X Y, et al. Ultrasound-activated sensitizers and applications[J]. Angew Chem Int Ed, 2020, 59(34): 14212-14233.
[11]	XU M M, ZHOU L Q, ZHENG L, et al. Sonodynamic therapy-derived multimodal synergistic cancer therapy[J]. Cancer Lett, 2021, 497: 229-242.
[12]	ZHANG Y, ZHANG C F, QIAN W L, et al. Recent advances in MOF-based nanozymes: synthesis, activities, and bioapplications[J]. Biosens Bioelectron, 2024, 263: 116593.
[13]	HU Y H, ZHENG Y J, LIU C, et al. Mitochondrial MOF regulates energy metabolism in heart failure via ATP5B hyperacetylation[J]. Cell Rep, 2024, 43(10): 114839.
[14]	MALEKI A, SEYEDHAMZEH M, YUAN M, et al. Titanium-based nanoarchitectures for sonodynamic therapy-involved multimodal treatments[J]. Small, 2023, 19(12): e2206253.
[15]	XIE X X, ZHANG J X, WANG Y, et al. Nanomaterials augmented bioeffects of ultrasound in cancer immunotherapy[J]. Mater Today Bio, 2023, 24: 100926.
[16]	ZHANG N S, ZENG W L, XU Y X, et al. Pyroptosis induction with nanosonosensitizer-augmented sonodynamic therapy combined with PD-L1 blockade boosts efficacy against liver cancer[J]. Adv Healthc Mater, 2024, 13(7): e2302606.
[17]	JIANG H, YANG K W, ZHAO X X, et al. Highly stable Zr(IV)-based metal-organic frameworks for chiral separation in reversed-phase liquid chromatography[J]. J Am Chem Soc, 2021, 143(1): 390-398.
[18]	PARK J, JIANG Q, FENG D W, et al. Size-controlled synthesis of porphyrinic metal-organic framework and functionalization for targeted photodynamic therapy[J]. J Am Chem Soc, 2016, 138(10): 3518-3525.
[19]	LÁZARO I A, WELLS C J R, FORGAN R S. Multivariate modulation of the Zr MOF UiO-66 for defect-controlled combination anticancer drug delivery[J]. Angew Chem Int Ed, 2020, 59(13): 5211-5217.
[20]	HAO F, YAN Z Y, YAN X P. Intracellular fate and immune response of porphyrin-based nano-sized metal-organic frameworks[J]. Chemosphere, 2022, 307: 135680.
[21]	SNETKOV P, ZAKHAROVA K, MOROZKINA S, et al. Hyaluronic acid: the influence of molecular weight on structural, physical, physico-chemical, and degradable properties of biopolymer[J]. Polymers (Basel), 2020, 12(8): 1800.
[22]	GHOSH B, BISWAS S. Polymeric micelles in cancer therapy: state of the art[J]. J Control Release, 2021, 332: 127-147.
[23]	KYU SHIM M, YANG S, SUN I C, et al. Tumor-activated carrier-free prodrug nanoparticles for targeted cancer Immunotherapy: preclinical evidence for safe and effective drug delivery[J]. Adv Drug Deliv Rev, 2022, 183: 114177.
[24]	LIU Y T, SUN Z J. Turning cold tumors into hot tumors by improving T-cell infiltration[J]. Theranostics, 2021, 11(11): 5365-5386.
[25]	LIU C P, WANG Y C, LI L M, et al. Engineered extracellular vesicles and their mimetics for cancer immunotherapy[J]. J Control Release, 2022, 349: 679-698.
[26]	HIAM-GALVEZ K J, ALLEN B M, SPITZER M H. Systemic immunity in cancer[J]. Nat Rev Cancer, 2021, 21(6): 345-359.
[27]	HUANG Z C, LIU L P, ZHU Z Y, et al. Tuning surface valences of nanoengagers to enhance their structural advantages for efficiently redirecting T cells against solid tumors[J]. ACS Nano, 2025, 19(1): 381-395.
[28]	MIN H, WANG J, QI Y Q, et al. Biomimetic metal-organic framework nanoparticles for cooperative combination of antiangiogenesis and photodynamic therapy for enhanced efficacy[J]. Adv Mater, 2019, 31(15): e1808200.
[29]	WANG Y F, SHI J Q, XIN M H, et al. Cell-drug conjugates[J]. Nat Biomed Eng, 2024, 8(11): 1347-1365.
[30]	GAO H B, SUN L, WANG H, et al. In situ non-canonical activation and sensitization of cGAS-STING pathway with manganese telluride nanosheets[J]. Biomaterials, 2025, 318: 123170.
[31]	SUN Z H, LIU J, LI Y Y, et al. Aggregation-induced-emission photosensitizer-loaded nano-superartificial dendritic cells with directly presenting tumor antigens and reversed immunosuppression for photodynamically boosted immunotherapy[J]. Adv Mater, 2023, 35(3): e2208555.

纳米工程化T细胞体系的构建及其对口腔鳞状细胞癌的体外治疗研究

Nanoengineered T cell system for the in vitro treatment of oral squamous cell carcinoma

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

图/表 5

参考文献 31

相关文章 15

编辑推荐

Metrics

[1]	陈子旋, 刘敏. 肾细胞癌免疫细胞治疗的研究进展[J]. 上海交通大学学报（医学版）, 2025, 45(7): 916-925.
[2]	张涤凡, 王明慧, 赵洁, 万江波, 黄方, 郝思国. B7基因修饰的白血病细胞外泌体的抗白血病效应[J]. 上海交通大学学报（医学版）, 2025, 45(2): 129-137.
[3]	唐珺倩, 李本尚. 儿童高危细胞遗传学B系急性淋巴细胞白血病治疗新进展[J]. 上海交通大学学报（医学版）, 2025, 45(10): 1390-1399.
[4]	何蕊, 颜克鹏, 王静. 靶向髓源性抑制细胞的叶酸循环增强肿瘤免疫治疗效果研究[J]. 上海交通大学学报（医学版）, 2024, 44(8): 1011-1022.
[5]	胡飞, 蔡晓涵, 程睿, 季诗雨, 苗嘉欣, 朱晏, 范广建. 骨肉瘤免疫微环境及其免疫治疗临床转化研究进展[J]. 上海交通大学学报（医学版）, 2024, 44(7): 814-821.
[6]	李虎虓, 李笑甜, 赵旭日, 张桓瑜, 周薇, 宋忠臣. 牙龈素提取物对口腔鳞癌细胞HN6生物学特性的影响[J]. 上海交通大学学报（医学版）, 2024, 44(2): 161-168.
[7]	周海霞, 张靖. m⁶A甲基化修饰调控肿瘤免疫的研究进展[J]. 上海交通大学学报（医学版）, 2024, 44(1): 137-144.
[8]	于莉, 苏显都, 张敏, 李雅慧, 王乐. 基于生物学分析构建及验证棕榈酰化相关酶长链非编码RNA的肝癌预后风险模型[J]. 上海交通大学学报（医学版）, 2023, 43(6): 747-754.
[9]	杨琳, 王晶波, 黄小娟, 任继亮, 袁瑛, 陶晓峰. 荧光探针cMBP⁃ICG检测口腔鳞状细胞癌颈部淋巴结转移及包膜外侵犯的探索性研究[J]. 上海交通大学学报（医学版）, 2022, 42(9): 1296-1302.
[10]	卢雨, 王昊, 巴乾. 肠道菌群在肝癌发生发展及治疗中的作用研究进展[J]. 上海交通大学学报（医学版）, 2022, 42(7): 939-944.
[11]	凌徐心仪, 张瑶, 钟华. 非小细胞肺癌免疫治疗获益人群筛选的研究进展[J]. 上海交通大学学报(医学版), 2021, 41(8): 1114-1119.
[12]	李静威, 王俐文, 蒋玲曦, 詹茜, 陈皓, 沈柏用. 胰腺癌免疫抑制性肿瘤微环境研究综述[J]. 上海交通大学学报(医学版), 2021, 41(8): 1103-1108.
[13]	马韵芳, 潘丽娜, 李圳, 高蓓莉, 胡家安, 徐志红. 司美替尼下调KRAS G12V突变型非小细胞肺癌细胞PD-L1水平的探索性研究[J]. 上海交通大学学报(医学版), 2021, 41(6): 741-748.
[14]	何春明，尹航，郑佳杰，唐健，傅于捷，赵晓菁. 肺癌免疫治疗：免疫抑制细胞和肺内免疫[J]. 上海交通大学学报(医学版), 2020, 40(8): 1137-1142.
[15]	刘洁，仇晓春. 乳腺肿瘤干细胞研究热点及趋势分析[J]. 上海交通大学学报(医学版), 2020, 40(7): 881-888.