CAR-T（CAR-T）细胞疗法在非肿瘤性疾病中的应用
Chimeric antigen receptor T (CAR-T) cell therapy in non-oncological diseases.
CAR-T（CAR-T）细胞在血液系统恶性肿瘤中的应用推动了这种免疫治疗形式的显著进展。
CELL INTELLIGENCE · 肿瘤细胞治疗研究
肿瘤细胞治疗研究
一体化 CRISPR/Cas 工程化糖皮质激素受体敲除 EBV-gp350-CAR 敲入 T 细胞强效且对地塞米松耐药
英文原题：All-in-one CRISPR/Cas-engineered glucocorticoid-receptor knock-out EBV-gp350-CAR knock-in T cells are potent and resistant to dexamethasone.
查看英文原题
All-in-one CRISPR/Cas-engineered glucocorticoid-receptor knock-out EBV-gp350-CAR knock-in T cells are potent and resistant to dexamethasone.
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研究概要
该研究为糖皮质激素耐药 CAR-T 细胞产品的无病毒一体化 GMP 生产奠定了概念验证。
中文摘要
背景
免疫功能低下患者及移植后出现 Epstein-Barr 病毒（EBV）再激活，与发病、死亡及多种恶性疾病发生有关。过继 T 细胞疗法已成为有前景的治疗选择，但移植后的免疫抑制会损害过继转移 T 细胞提供的抗 EBV 保护性免疫监视。
方法
研究者采用一体化 CRISPR/Cas 方法，在符合药品生产质量管理规范（GMP）的平台上，将抗 EBV（gp350）CAR 敲入 T 细胞受体（TRAC）位点，并同时敲除糖皮质激素受体（GR）。
结果
在原代人 T 细胞中，从基因和蛋白水平均确认 CAR 敲入（CAR KI），平均效率为 41%。在 CAR KI 细胞中，额外敲除 GR 的效率很高，达到 83%。在功能层面，CAR KI、GR 敲除 T 细胞对靶点具有特异性效力，表现为细胞因子分泌模式、增殖能力，以及针对表达 gp350 靶细胞的细胞毒活性。CAR KI、GR 敲除 T 细胞还对地塞米松处理不敏感，并保持 T 细胞功能。相反，这些细胞仍对不依赖 GR 的免疫抑制剂环孢素 A（CsA）敏感，因此出现安全问题时仍可对患者采取补救治疗。
结论
本研究提供了概念验证，表明可采用无病毒的一体化方法按 GMP 要求生产耐糖皮质激素 CAR-T 产品。此外，耐糖皮质激素 gp350-CAR-T 细胞未来可能成为移植后 EBV 再激活高危患者，或需接受类固醇治疗的 EBV 相关病变患者的一种治疗选择。
展开英文摘要原文
BACKGROUND
Epstein-Barr virus (EBV) reactivation in immunocompromised patients and post-transplantation is associated with morbidity, mortality and with the onset of a variety of malignant diseases. Adoptive T-cell therapies have emerged as promising therapeutic options, but post-transplant immunosuppression jeopardizes the protective anti-EBV immune surveillance by adoptively transferred T cells.
METHODS
Using an all-in-one CRISPR/Cas-mediated approach, we inserted an anti-EBV (gp350) CAR into the T-cell receptor (TRAC) locus and simultaneously knocked-out the glucocorticoid receptor (GR) on a good manufacturing practice (GMP)-compatible platform.
RESULTS
CAR knock-in (CAR KI ) was confirmed in primary human T cells on genetic and on protein level with a mean efficiency of 41%. With 83%, additional GR knock-out was highly efficient in CAR KI cells. On a functional level CAR KI GR KO T cells showed target-specific potency in terms of cytokine secretion patterns, proliferative capacity and cytotoxic activity against gp350-expressing target cells. Further, CAR KI GR KO T cells were insensitive to dexamethasone treatment and maintained T-cell functionality. In contrast, CAR KI GR KO T cells were sensitive to the GR-independent immunosuppressant cyclosporine A (CsA), thereby providing a rescue treatment for patients in case of safety issues.
CONCLUSIONS
The study lays the proof-of-concept for virus-free all-in-one GMP-manufacturing of glucocorticoid-resistant CAR T-cell products. Further, the glucocorticoid-resistant gp350-CAR T cells can provide a future therapeutic option for high-risk post-transplant patients with EBV-reactivations or patients with EBV-associated pathologies requiring steroid treatment.
论文信息
- 作者
- Kaeuferle T、Zwermann M、Stoll N、Ferrada-Ernst P、Jablonowski L、Zeidler R、Willier S、Stenger D
- 单位
- Department of Pediatric Hematology, Oncology, Hemostaseology and Stem Cell Transplantation, Dr. von Hauner Children's Hospital, University Hospital LMU Munich, Munich, Germany. Theresa.Kaeuferle@uniklinik-freiburg.de.Germany
- 期刊
- Experimental hematology & oncology2025 Mar 19
- 原文标识
- PubMed 40108708 · DOI 10.1186/s40164-025-00631-w
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