Ptbp1在小鼠急性T淋巴细胞白血病中的作用研究

doi:10.19586/j.2095-2341.2024.0182

生物技术进展 ›› 2025, Vol. 15 ›› Issue (2): 341-348.DOI: 10.19586/j.2095-2341.2024.0182

• 研究论文 • 上一篇

Ptbp1在小鼠急性T淋巴细胞白血病中的作用研究

朱旭¹(), 张英楠¹, 马金法¹, 石莉红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），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，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:2024-11-20 接受日期:2025-01-21 出版日期:2025-03-25 发布日期:2025-04-29
通讯作者: 石莉红
作者简介:朱旭 E-mail: zhuxu@ihcams.ac.cn；
基金资助:
国家重点研发计划项目(2022YFA1103503);国家自然科学基金项目(82225003);中国医学科学院医学与健康科技创新工程项目(2023-I2M-007);细胞生态海河实验室“揭榜挂帅”项目(24HHXBSS00013);津门医学英才项目(TJSJMYXYC-D2-022);天津市科技计划项目(24ZXRKSY00010)

The Role of Ptbp1 in T Cell Acute Lymphoblastic Leukemia Mice

Xu ZHU¹(), Yingnan ZHANG¹, Jinfa MA¹, Lihong SHI¹^,²()

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

Received:2024-11-20 Accepted:2025-01-21 Online:2025-03-25 Published:2025-04-29
Contact: Lihong SHI

摘要/Abstract

摘要：

研究旨在探究经典的RNA结合蛋白多聚嘧啶束结合蛋白1（PTBP1）在急性T淋巴细胞白血病（T cell acute lymphoblastic leukemia，T-ALL）发生及发展中的作用。通过Cre;Flox系统，构建了在造血系统中特异性敲除Ptbp1的Mx1-Cre; Ptbp1^fl/fl小鼠模型，并利用Notch1的逆转录病毒感染小鼠原代骨髓lin^-细胞，通过移植实验，建立敲除Ptbp1的T-ALL疾病模型。研究发现Ptbp1的缺失加速了T-ALL的发展，表现为Ptbp1敲除组小鼠骨髓中Notch1-GFP⁺细胞比例更高，提示疾病发展速度更快。此外，Ptbp1敲除后的白血病细胞表现为增殖能力增强以及凋亡水平降低，这与小鼠体内实验观察到疾病快速进展相吻合。这些结果提示Ptbp1在T-ALL发生发展中的重要作用，为Ptbp1作为T-ALL潜在生物标志物提供了更多认识。

关键词: Ptbp1, T-ALL, 小鼠模型, 细胞增殖

Abstract:

The aim of this study is to explore the role of the classical RNA binding protein polypyrimidine bundle binding protein 1 （PTBP1） in the occurrence and development of T cell acute lymphoblastic leukemia （T-ALL）. Generating Mx1-Cre；Ptbp1^fl/fl mouse model that hematopoietic system specific knockout of Ptbp1 through Cre；Flox system. Mouse bone marrow lin^- cells were infected with Notch1 retrovirus， and the Ptbp1 knockout T-ALL model was established by transplantation experiments. The deletion of Ptbp1 was found to accelerate the disease progression of T-ALL， as indicated by a higher proportion of Notch1-GFP⁺ cells in the bone marrow of Ptbp1 knockout mice， suggesting faster disease progression. In addition， Ptbp1 knockout leukemia cells showed enhanced proliferation and decreased apoptosis， which was consistent with the rapid disease progression observed in mice. These results suggest the important role of Ptbp1 in the occurrence and development of T-ALL， and provide more understanding of Ptbp1 as a potential biomarker for T-ALL.

Key words: Ptbp1, T-ALL, mouse model, cell proliferation

中图分类号:

Q812

朱旭, 张英楠, 马金法, 石莉红. Ptbp1在小鼠急性T淋巴细胞白血病中的作用研究[J]. 生物技术进展, 2025, 15(2): 341-348.

Xu ZHU, Yingnan ZHANG, Jinfa MA, Lihong SHI. The Role of Ptbp1 in T Cell Acute Lymphoblastic Leukemia Mice[J]. Current Biotechnology, 2025, 15(2): 341-348.

图/表 5

图1 Ptbp1-Flox构建示意图

Fig. 1 Construction of Ptbp1-Flox

图2 移植实验流程

Fig. 2 Flow chart of transplantation experiment

图3 建立了造血系统中特异性敲除Ptbp1的小鼠模型A:Western Blot实验显示Mx1-Cre-;Ptbp1fl/fl对照组和Mx1-Cre+;Ptbp1fl/fl敲除组小鼠骨髓细胞中的Ptbp1表达量；B:Image J对照组和敲除组PTBP1蛋白定量；C:RT-qPCR检测对照组和敲除组Ptbp1 mRNA的相对表达量。*,***分别表示不同处理在P<0.05、P<0.001水平具有统计学差异。

Fig. 3 The mouse model with specific knockout of Ptbp1 in the hematopoietic system

图4 Ptbp1敲除组小鼠T-ALL的疾病进展A：第一次移植后对照组和Ptbp1敲除组小鼠生存曲线，n=6； B：第二次移植后对照组和Ptbp1敲除组的小鼠生存曲线，n=6； C：流式细胞术分析第一次移植后对照组和敲除组小鼠骨髓中GFP+的细胞比例； D：对照组和敲除组GFP+细胞的差异，n=3； *,***分别表示不同处理在P<0.05、P<0.001水平具有统计学差异。

Fig. 4 Progression of T-ALL disease in Ptbp1 knockout mice

图5 敲除Ptbp1对小鼠骨髓中T-ALL肿瘤细胞周期和凋亡的影响A：对照组和Ptbp1敲除组小鼠骨髓中T-ALL的细胞周期检测流式图，G0期细胞—Ki67-Hoechst-；G1期细胞—Ki67+Hoechst-；S/G2M期细胞—Ki67+Hoechst+；死细胞及细胞碎片—Ki67- Hoechst+。B：对照组和Ptbp1敲除组细胞周期检测统计图，n=3；C：对照组和Ptbp1敲除组小鼠骨髓中T-ALL的细胞凋亡流式图，活细胞—Annexin V-DAPI-；早期凋亡细胞—Annexin V+DAPI-；晚期凋亡细胞—Annexin V+DAPI+；死细胞及细胞碎片—Annexin-DAPI+。D：对照组和Ptbp1敲除组细胞凋亡检测统计图，n=3。*, **, ***分别表示不同处理在P<0.05、P<0.01、P<0.001水平具有统计学差异。

Fig. 5 Effects of knockout of Ptbp1 on the cell cycle and apotosis of T-ALL tumor cells in mouse

参考文献 27

1	LEE Y, RIO D C. Mechanisms and regulation of alternative pre-mRNA splicing[J]. Annu. Rev. Biochem., 2015, 84: 291-323.
2	MERKIN J, RUSSELL C, CHEN P, et al.. Evolutionary dynamics of gene and isoform regulation in mammalian tissues[J]. Science, 2012, 338(6114): 1593-1599.
3	MILLEVOI S, DECORSIÈRE A, LOULERGUE C, et al.. A physical and functional link between splicing factors promotes pre-mRNA 3' end processing[J]. Nucleic Acids Res., 2009, 37(14): 4672-4683.
4	CASTELO-BRANCO P, FURGER A, WOLLERTON M, et al.. Polypyrimidine tract binding protein modulates efficiency of polyadenylation[J]. Mol. Cell. Biol., 2004, 24(10): 4174-4183.
5	JACKSON R J, KAMINSKI A. Internal initiation of translation in eukaryotes: the picornavirus paradigm and beyond[J]. RNA, 1995, 1(10): 985-1000.
6	KAMINSKI A, HUNT S L, PATTON J G, et al.. Direct evidence that polypyrimidine tract binding protein (PTB) is essential for internal initiation of translation of encephalomyocarditis virus RNA[J]. RNA, 1995, 1(9): 924-938.
7	GAMA-CARVALHO M, BARBOSA-MORAIS N L, BRODSKY A S, et al.. Genome-wide identification of functionally distinct subsets of cellular mRNAs associated with two nucleocytoplasmic-shuttling mammalian splicing factors[J/OL]. Genome Biol., 2006, 7(11): R113[2024-12-10]. .
8	COELHO M B, ASCHER D B, GOODING C, et al.. Functional interactions between polypyrimidine tract binding protein and PRI peptide ligand containing proteins[J]. Biochem. Soc. Trans., 2016, 44(4): 1058-1065.
9	LIU J, LI Y, TONG J, et al.. Long non-coding RNA-dependent mechanism to regulate heme biosynthesis and erythrocyte development[J/OL]. Nat. Commun., 2018, 9: 4386[2024-12-10]. .
10	SUCKALE J, WENDLING O, MASJKUR J, et al.. PTBP1 is required for embryonic development before gastrulation[J/OL]. PLoS One, 2011, 6(2): e16992[2024-12-10]. .
11	SHIBAYAMA M, OHNO S, OSAKA T, et al.. Polypyrimidine tract-binding protein is essential for early mouse development and embryonic stem cell proliferation[J]. FEBS J., 2009, 276(22): 6658-6668.
12	REHN M, WENZEL A, A-KFRANK, et al.. PTBP1 promotes hematopoietic stem cell maintenance and red blood cell development by ensuring sufficient availability of ribosomal constituents[J/OL]. Cell Rep., 2022, 39(6): 110793[2024-12-10]. .
13	NEWTON K, WICKLIFFE K E, DUGGER D L, et al.. Cleavage of RIPK1 by caspase-8 is crucial for limiting apoptosis and necroptosis[J]. Nature, 2019, 574(7778): 428-431.
14	IWAMORI N, TOMINAGA K, SATO T, et al.. MRG15 is required for pre-mRNA splicing and spermatogenesis[J/OL]. Proc. Natl. Acad. Sci. USA, 2016, 113(37): 1611995113 [2024-12-10]. .
15	TAHMASEBI S, JAFARNEJAD S M, TAM I S, et al.. Control of embryonic stem cell self-renewal and differentiation via coordinated alternative splicing and translation of YY2[J]. Proc. Natl. Acad. Sci. USA, 2016, 113(44): 12360-12367.
16	ALDAVE G, GONZALEZ-HUARRIZ M, RUBIO A, et al.. The aberrant splicing of BAF45d links splicing regulation and transcription in glioblastoma[J]. Neuro Oncol., 2018, 20(7): 930-941.
17	GRIDLEY T, GROVES A K. Overview of genetic tools and techniques to study Notch signaling in mice[J]. Methods Mol. Biol., 2014, 1187: 47-61.
18	WENDORFF A A, FERRANDO A A. Modeling NOTCH1 driven T-cell acute lymphoblastic leukemia in mice[J/OL]. Bio. Protoc., 2020, 10(10): e3620[2024-12-10]. .
19	XING L, GUO X, ZHANG X, et al.. PTBP1 is a potential indicator of disease progression in acute myeloid leukaemia[J/OL]. J. Investig. Med. Off. Publ. Am. Fed. Clin. Res., 2024, 10815589241264783[2024-12-15]. .
20	ZIEGLER N, CORTÉS-LÓPEZ M, ALT F, et al.. Analysis of RBP expression and binding sites identifies PTBP1 as a regulator of CD19 expression in B-ALL[J/OL]. Oncoimmunology, 2023, 12(1): 2184143[2024-12-10]. .
21	黄琬玲,朱文琦,郭妮妮,等.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.
	HUANG W L, ZHU W Q, GUO N N, et al.. Effects of NRG4 on proliferation, apoptosis and cell cycle of acute myeloid leukemia cells[J]. Curr. Biotechnol., 2023, 13(2): 305-310.
22	LA PORTA J, MATUS-NICODEMOS R, VALENTÍN-ACEVEDO A, et al.. The RNA-binding protein, polypyrimidine tract-binding protein 1 (PTBP1) is a key regulator of CD4 T cell activation[J/OL]. PLoS One, 2016, 11(8): e0158708[2024-12-10]. .
23	MARASCA S S, VADALA R, et al.. LINE1 are spliced in non-canonical transcript variants to regulate T cell quiescence and exhaustion[J]. Nat. Genet., 2022, 54:180-193.
24	LI T, GAO R, XU K, et al.. BCL7A inhibits the progression and drug-resistance in acute myeloid leukemia[J/OL]. Drug Resist. Updat., 2024, 76: 101120[2024-12-10]. .
25	ZHANG Q X, PAN Y M, XIAO H L, et al.. Alternative splicing analysis showed the splicing factor polypyrimidine tract-binding protein 1 as a potential target in acute myeloid leukemia therapy[J]. Neoplasma, 2022, 69(5): 1198-1208.
26	WANG L, YANG L, YANG Z, et al.. Glycolytic enzyme PKM2 mediates autophagic activation to promote cell survival in NPM1-mutated leukemia[J]. Int. J. Biol. Sci., 2019, 15(4): 882-894.
27	BAI H, CHEN B. Abnormal PTBP1 expression sustains the disease progression of multiple myeloma[J/OL]. Dis. Markers, 2020, 2020: 4013658[2024-12-10]. .

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Ptbp1在小鼠急性T淋巴细胞白血病中的作用研究

The Role of Ptbp1 in T Cell Acute Lymphoblastic Leukemia Mice

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