Research Advances and Clinical Application Prospects of in Vivo CAR-T Therapy

doi:10.19586/j.2095-2341.2025.0159

Current Biotechnology ›› 2026, Vol. 16 ›› Issue (4): 909-918.DOI: 10.19586/j.2095-2341.2025.0159

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Research Advances and Clinical Application Prospects of in Vivo CAR-T Therapy

Mingdi ZHANG¹(), Qigai LI²(), Lu WANG², Hui OUYANG², Jinghua LIAN², Fei XIE¹()

^1.College of Chemistry and Life Science，Beijing University of Technology，Beijing 100124，China
^2.Huizhou High School，Guangdong Province，Guangdong Huizhou 516001，China

Received:2025-11-17 Accepted:2026-01-21 Online:2026-07-25 Published:2026-09-11
Contact: Fei XIE

Abstract

Abstract:

Chimeric antigen receptor T-cell （CAR-T） therapy， as a revolutionary immunotherapy method， has demonstrated remarkable clinical efficacy in the treatment of hematological malignancies. However， the traditional in vitro CAR-T therapy is limited in wide application due to numerous problems. In recent years， the research on in vivo CAR-T therapy， which directly modifies T cells in the body， holds the promise of overcoming the limitations of traditional therapies. This article reviewed the technical principles，latest research progress， clinical application potential， safety issues， and future development directions of CAR-T therapy in vivo， aiming to provide comprehensive references for clinical research on malignant tumors and promote the further optimization and transformation of innovative therapy.

Key words: CAR-T therapy, in vivo gene editing, immunotherapy, malignant tumors, safety

CLC Number:

Q291

Mingdi ZHANG, Qigai LI, Lu WANG, Hui OUYANG, Jinghua LIAN, Fei XIE. Research Advances and Clinical Application Prospects of in Vivo CAR-T Therapy[J]. Current Biotechnology, 2026, 16(4): 909-918.

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Figures/Tables 3

References 67

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载体类型	核心特性	在体内CAR-T中的优势	局限性
腺相关病毒（AAV）	非致病性、单链DNA、多数无整合能力、血清型特异性靶向（如AAV6靶向T细胞）	安全性高、T细胞靶向性强、表达稳定性好	装载容量有限（约4.7 kb）、免疫原性问题（如预存抗体）
慢病毒（LV）	逆转录病毒科、整合型、感染分裂与非分裂细胞、整合位点随机	长期表达能力强、装载容量较大（约 8 kb）、感染效率高	插入突变风险、生产难度大、免疫原性较强
逆转录病毒（RV）	仅感染分裂期细胞、整合效率高、整合位点偏好转录起始区域	整合效率高、技术成熟度高	细胞周期依赖性强、插入致癌风险高、装载容量有限（约6 kb）

载体类型	核心特性	在体内CAR-T中的优势	局限性
腺相关病毒（AAV）	非致病性、单链DNA、多数无整合能力、血清型特异性靶向（如AAV6靶向T细胞）	安全性高、T细胞靶向性强、表达稳定性好	装载容量有限（约4.7 kb）、免疫原性问题（如预存抗体）
慢病毒（LV）	逆转录病毒科、整合型、感染分裂与非分裂细胞、整合位点随机	长期表达能力强、装载容量较大（约 8 kb）、感染效率高	插入突变风险、生产难度大、免疫原性较强
逆转录病毒（RV）	仅感染分裂期细胞、整合效率高、整合位点偏好转录起始区域	整合效率高、技术成熟度高	细胞周期依赖性强、插入致癌风险高、装载容量有限（约6 kb）

载体类型	核心特性	在体内CAR-T中的优势	局限性
脂质纳米颗粒（LNP）	核-壳结构，包裹mRNA/DNA；通过内吞进入细胞，酸性环境中结构变化实现内体逃逸^[31]	生物相容性好、递送效率较高、技术成熟度高	靶向性有限、表达周期短、规模化生产成本较高
聚合物纳米载体	阳离子聚合物与核酸静电结合；通过内吞进入，部分聚合物可促进内体逃逸^[32]	负载能力强、结构可设计性高、生产成本低	细胞毒性较高、递送效率不稳定、内体逃逸效率低
DNA纳米载体	DNA自组装形成纳米结构，可包裹CAR基因；尺寸可控，生物相容性好	靶向精度高、生物安全性优、功能模块化	制备难度大、体内稳定性差、转染效率低

载体类型	核心特性	在体内CAR-T中的优势	局限性
脂质纳米颗粒（LNP）	核-壳结构，包裹mRNA/DNA；通过内吞进入细胞，酸性环境中结构变化实现内体逃逸^[31]	生物相容性好、递送效率较高、技术成熟度高	靶向性有限、表达周期短、规模化生产成本较高
聚合物纳米载体	阳离子聚合物与核酸静电结合；通过内吞进入，部分聚合物可促进内体逃逸^[32]	负载能力强、结构可设计性高、生产成本低	细胞毒性较高、递送效率不稳定、内体逃逸效率低
DNA纳米载体	DNA自组装形成纳米结构，可包裹CAR基因；尺寸可控，生物相容性好	靶向精度高、生物安全性优、功能模块化	制备难度大、体内稳定性差、转染效率低

	体外CAR-T疗法	体内CAR-T疗法
技术流程	①白细胞分离术抽取患者外周血；②免疫磁珠分选分离T细胞；③GMP实验室病毒/非病毒载体介导CAR基因转染；④细胞因子（IL-2/IL-7）体外扩增（10⁵→10⁹~10¹⁰个）；⑤质量检测（CAR表达率、活性、内毒素）；⑥静脉回输患者体内	①规模化生产携带CAR基因的递送载体（AAV/LNP等）；②载体质量检测（靶向性、基因包封率）；③静脉/局部注射载体至患者体内；④载体体内靶向T细胞并实现CAR基因原位表达与T细胞改造
治疗周期	从抽血到回输需2~4周，T细胞数量不足/功能弱时可延长至6周，其间需桥接治疗控制肿瘤	载体可提前储存，患者确诊后1~2 d完成给药前评估，给药后1~3 d检测到CAR-T活性，总周期≤1周
治疗成本	单例成本30万~50万美元，核心成本：白细胞分离术（≈2万美元）、GMP细胞培养（≈10万美元）、病毒载体制备（≈15万美元）、质量检测（≈5万美元）	单例成本有望降至5 万~10万美元，核心成本：规模化载体生产（LNP：1 000~5 000美元·剂^-1；AAV：1万~3万美元·剂^-1）；无细胞分离/培养成本
T细胞功能特性	体外培养易致T细胞耗竭：PD-1/LAG-3阳性率 60%~80%（新鲜T细胞仅10%~20%）；体内存活时间2~4周；记忆T细胞比例 15%~20%，长期抗肿瘤记忆弱	体内生理环境保留T细胞功能，无明显耗竭标志物高表达；体内存活时间8~12周；记忆T细胞比例35%~45%，可动态改造新生T细胞，维持长期活性^[39]
适用人群	仅30%~40%患者符合标准，排除以下人群：T细胞数量<1×10⁵个·mL^-1（老年/多次化疗患者）；合并严重感染（活动性乙肝）、器官衰竭（肝功能Child-Pugh C级）；无法耐受白细胞分离术	适用人群大幅拓展，可覆盖T细胞数量不足/功能弱的患者；合并轻度器官功能不全/感染的患者；实体瘤患者（肿瘤微环境内直接改造T细胞，减少迁移损耗）
安全性风险	①主要毒性：CRS（3级及以上发生率20%~30%）、ICANS（3级及以上10%~15%）、B细胞耗竭；②毒性管理成熟：CRS用托珠单抗，ICANS用糖皮质激素，B细胞耗竭用免疫球蛋白^[40]；③死亡率<1%	①新型风险：载体脱靶感染（肝细胞/红细胞，致肝损伤等）；基因编辑脱靶（潜在二次肿瘤风险）；CAR-T活性难调控（可能引发过度炎症）；②临床前数据：诱导CRS和CAR-T相关脑病综合征（神经毒性），长期安全性待验证^[41]
技术成熟度	已实现临床转化，全球多款产品获批（如阿基仑赛、瑞基奥仑赛），临床数据丰富，质控标准明确	处于临床前/早期临床试验阶段，无获批产品，核心技术（载体靶向性、表达效率）仍需优化，长期疗效与安全性数据不足

Research Advances and Clinical Application Prospects of in Vivo CAR-T Therapy

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