靶向CD74的嵌合抗原受体T细胞的抗白血病细胞作用研究

doi:10.19586/j.2095-2341.2024.0016

生物技术进展 ›› 2024, Vol. 14 ›› Issue (3): 501-505.DOI: 10.19586/j.2095-2341.2024.0016

• 研究论文 • 上一篇

靶向CD74的嵌合抗原受体T细胞的抗白血病细胞作用研究

刘一鸣¹^,²(), 谢乐灵¹^,², 邢海燕¹^,², 唐克晶¹^,², 王敏¹^,², 饶青¹^,²()

^1.中国医学科学院血液病医院（中国医学科学院血液学研究所）北京协和医学院，血液与健康全国重点实验室，国家血液系统疾病临床医学研究中心，天津市血液病细胞治疗研究重点实验室，细胞生态海河实验室，天津 300020
^2.天津医学健康研究院，天津 301600

收稿日期:2024-01-31 接受日期:2024-03-21 出版日期:2024-05-25 发布日期:2024-06-18
通讯作者: 饶青
作者简介:刘一鸣 E-mail: liuyiming@ihcams.ac.cn；
基金资助:
国家自然科学基金项目(81770106)

Anti-leukemia Effect of Chimeric Antigen Receptor T Cells Targeting CD74 Positive Leukemia Cells

Yiming LIU¹^,²(), Leling XIE¹^,², Haiyan XING¹^,², Kejing TANG¹^,², Min WANG¹^,², Qing RAO¹^,²()

^1.State Key Laboratory of Experimental Hematology，National Clinical Research Center for Blood Diseases，Tianjin Key Laboratory of Cell Therapy for Blood Diseases，Haihe Laboratory of Cell Ecosystem，Institute of Hematology & Blood Diseases Hospital，Chinese Academy of Medical Sciences & Peking Union Medical College，Tianjin 300020，China
^2.Tianjin Institutes of Health Science，Tianjin 301600，China

Received:2024-01-31 Accepted:2024-03-21 Online:2024-05-25 Published:2024-06-18
Contact: Qing RAO

摘要/Abstract

摘要：

嵌合抗原受体T细胞（chimeric antigen receptor T cell，CAR-T）疗法目前已应用于血液肿瘤的治疗，前期研究发现CD74在白血病患者白血病干细胞中高表达，为了探索其对白血病细胞的杀伤作用，通过CD74 CAR表达载体的构建、慢病毒载体的包装及T细胞的感染，制备CD74 CAR-T细胞。利用体外杀伤实验及细胞因子的释放检测确定CAR-T细胞对CD74⁺的白血病细胞的特异性杀伤作用。结果显示，CD74 CAR-T分别与CD74⁺的THP-1及CD74^-的K562白血病细胞共培养时，THP-1细胞可被清除，而对CD74^-的K562细胞无明显杀伤作用。CD74 CAR-T细胞可被CD74⁺的THP-1细胞有效激活，通过细胞因子释放途径有效杀伤白血病细胞。研究成功构建了靶向CD74⁺的白血病细胞的 CAR-T细胞，为白血病的免疫治疗提供了新的靶点和免疫治疗策略。

关键词: 嵌合抗原受体修饰的T细胞, CD74, 急性髓系白血病, 白血病干细胞

Abstract:

Chimeric antigen receptor T cell （CAR-T） therapy has been used in treatment of hematological malignancy. The previous study has found that CD74 is highly expressed in leukemia stem cells. In order to explore its killing effect on leukemia cells， CD74 CAR-T cells were prepared by constructing CD74 CAR expression vector， packing lentivirus and infecting T cells. The specific killing effect of CAR-T cells on CD74 positive leukemia cells was determined by killing experiment in vitro and cytokine release detection. The results showed that when CD74 CAR-T was co-cultured with CD74⁺ THP-1 or CD74^- K562 leukemia cells， THP-1 cells could be eliminated， but there was no obvious killing effect on CD74^- K562 cells. CD74⁺THP-1 cells could effectively activate CAR-T cells， and activated anti-CD74 CAR-T cells could effectively leukemia cells through cytotoxicity pathways. The study successfully established CAR-T cells targeting CD74⁺leukemia cells，thus providing new target and immunotherapy strategy for leukemia immunotherapy.

Key words: chimeric antigen receptor T cell, CD74, acute myeloid leukemia, leukemia stem cell

中图分类号:

Q291

刘一鸣, 谢乐灵, 邢海燕, 唐克晶, 王敏, 饶青. 靶向CD74的嵌合抗原受体T细胞的抗白血病细胞作用研究[J]. 生物技术进展, 2024, 14(3): 501-505.

Yiming LIU, Leling XIE, Haiyan XING, Kejing TANG, Min WANG, Qing RAO. Anti-leukemia Effect of Chimeric Antigen Receptor T Cells Targeting CD74 Positive Leukemia Cells[J]. Current Biotechnology, 2024, 14(3): 501-505.

图/表 4

图1 CD74 CAR-T细胞的建立A：CD74 CAR-T结构图；B：CD74 CAR-T和VEC-T细胞感染效率流式图及统计图；scFv—单链抗体可变区；GFP—绿色荧光蛋白

Fig. 1 Preparation of CD74 CAR-T cells

图2 IFN-γ诱导的THP-1细胞和K562细胞表面CD74的表达

Fig. 2 The expression of CD74 in THP-1 and K562 cells

图3 CD74 CAR-T细胞与靶细胞共培养24、48 h后相对残留靶细胞比例注：**和****分别表示在P<0.01和P<0.000 1水平上具有统计学意义。

Fig. 3 Relative proportion of residual target cells after co culturing of CD74 CAR-T cells with target cell for 24 and 48 h

图4 CD74 CAR-T或VEC-T 和靶细胞共培养上清中各类细胞因子分泌水平注：*、**、***分别表示在P<0.05、P<0.01、P<0.001水平上具有统计学意义；ns表示无统计学意义。

Fig. 4 Production of cytokines by VEC-T or CD74 CAR-T cells after co-culturing with target cells for 48 h

参考文献 19

1	STELMACH P, TRUMPP A. Leukemic stem cells and therapy resistance in acute myeloid leukemia[J]. Haematologica, 2023, 108(2): 353-366.
2	SU H, NA N, ZHANG X, et al.. The biological function and significance of CD74 in immune diseases[J]. Inflamm. Res., 2017, 66(3): 209-216.
3	SANCHEZ-NIÑO M D, SANZ A B, RUIZ-ANDRES O, et al.. MIF, CD74 and other partners in kidney disease: tales of a promiscuous couple[J]. Cytokine Growth Factor Rev., 2013, 24(1): 23-40.
4	KASHIMA Y, SHIBAHARA D, SUZUKI A, et al.. Single-cell analyses reveal diverse mechanisms of resistance to EGFR tyrosine kinase inhibitors in lung cancer[J]. Cancer Res., 2021, 81(18): 4835-4848.
5	LASKIN J, LIU S V, TOLBA K, et al.. NRG1 fusion-driven tumors: biology, detection, and the therapeutic role of afatinib and other ErbB-targeting agents[J]. Ann. Oncol., 2020, 31(12): 1693-1703.
6	D·M·戈登伯格,H·汉森,S·-O·梁,等. 内在化抗-CD74抗体和使用方法: CN 03809863.6[P].2011-04-13.
7	范蓓,王园园,赵云霞,等.免疫系统人源化小鼠模型构建技术探讨[J].生物技术进展,2022,12(6):922-928.
	FAN B, WANG Y Y, ZHAO Y X, et al.. Discussion on the construction technology of mouse model with mumanized immune[J]. Curr. Biotechnol., 2022, 12(6): 922-928.
8	LIU H. Emerging agents and regimens for AML[J/OL]. J. Hematol. Oncol., 2021, 14(1): 49[2024-04-10]. .
9	张鹏晓,胡念.黑色素瘤免疫治疗作用机制研究进展[J].生物技术进展,2023,13(6):900-906.
	ZHANG P X, HU N. The research progress on action mechanism of melanoma immunotherapy[J]. Curr. Biotechnol., 2023, 13(6): 900-906.
10	高朗,于思雪,袁春森,等.黏蛋白在肿瘤免疫治疗中的研究进展[J].生物技术进展,2023,13(3):390-398.
	GAO L, YU S X, YUAN C S, et al.. Research progress of mucin in tumor immunotherapy[J]. Curr. Biotechnol., 2023, 13(3): 390-398.
11	MILES R R, CAIRO M S, SATWANI P, et al.. Immunophenotypic identification of possible therapeutic targets in paediatric non-hodgkin lymphomas: a children's oncology group report[J]. Br. J. Haematol., 2007, 138(4): 506-512.
12	BURTON J D, ELY S, REDDY P K, et al.. CD74 is expressed by multiple myeloma and is a promising target for therapy[J]. Clin. Cancer Res., 2004, 10(19): 6606-6611.
13	MCCLELLAND M, ZHAO L, CARSKADON S, et al.. Expression of CD74, the receptor for macrophage migration inhibitory factor, in non-small cell lung cancer[J]. Am. J. Pathol., 2009, 174(2): 638-646.
14	CONROY H, MAWHINNEY L, DONNELLY S C. Inflammation and cancer: macrophage migration inhibitory factor (MIF): the potential missing link[J]. QJM-Int.J.Med., 2010, 103(11): 831-836.
15	STEIN R, MATTES M J, CARDILLO T M, et al.. CD74: a new candidate target for the immunotherapy of B-cell neoplasms[J]. Clin. Cancer Res., 2007, 13(18): 5556-5563.
16	OCHAKOVSKAYA R, OSORIO L, GOLDENBERG D M, et al.. Therapy of disseminated B-cell lymphoma xenografts in severe combined immunodeficient mice with an anti-CD74 antibody conjugated with (111)indium, (67)gallium, or (90).ttrium[J]. Clin. Cancer Res., 2001, 7(6): 1505-1510.
17	SAPRA P, STEIN R, PICKETT J, et al.. Anti-CD74 antibody-doxorubicin conjugate, IMMU-110, in a human multiple myeloma xenograft and in monkeys[J]. Clin. Cancer Res., 2005, 11(14): 5257-5264.
18	CHANG C H, SAPRA P, VANAMA S S, et al.. Effective therapy of human lymphoma xenografts with a novel recombinant ribonuclease/anti-CD74 humanized IgG4 antibody immunotoxin[J]. Blood, 2005, 106(13): 4308-4314.
19	BORGHESE F, CLANCHY F I. CD74: an emerging opportunity as a therapeutic target in cancer and autoimmune disease[J]. Expert Opin. Ther. Targets, 2011, 15(3): 237-251.

[1]	黄琬玲, 朱文琦, 郭妮妮, 王楠, 任倩, 马小彤. NRG4基因对急性髓系白血病细胞增殖、凋亡及周期的影响[J]. 生物技术进展, 2023, 13(2): 305-310.
[2]	张月明, 罗雅琴, 郑伟, 徐杰, 董雪燕, 李善琛, 王敬毅. 急性髓系白血病mRNA与非编码RNA差异表达谱及CeRNA调控网络分析[J]. 生物技术进展, 2023, 13(1): 146-153.
[3]	许杰,王可飞,魏晓晶,龚莉欣,焦阳,邱录贵,郝牧. 基于生物信息学分析SCHIP1在急性髓系白血病中的表达及其临床意义[J]. 生物技术进展, 2020, 10(4): 417-425.

靶向CD74的嵌合抗原受体T细胞的抗白血病细胞作用研究

Anti-leukemia Effect of Chimeric Antigen Receptor T Cells Targeting CD74 Positive Leukemia Cells

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

图/表 4

参考文献 19

相关文章 3

编辑推荐

Metrics

本文评价