T-ALL来源的骨髓基质细胞通过FGF2-FGFR2通路促进T-ALL增殖

doi:10.11958/20240355

天津医药 ›› 2025, Vol. 53 ›› Issue (1): 29-34.doi: 10.11958/20240355

T-ALL来源的骨髓基质细胞通过FGF2-FGFR2通路促进T-ALL增殖

杨健¹(), 李敏², 李越洋², 田晨²^,^△()

1 青岛市中心（肿瘤）医院肿瘤外科（邮编266042）
2 天津医科大学肿瘤医院血液科，国家恶性肿瘤临床医学研究中心，天津市恶性肿瘤临床医学研究中心，天津市肿瘤防治重点实验室

收稿日期:2024-03-23 修回日期:2024-09-11 出版日期:2025-01-15 发布日期:2025-02-06
通讯作者: ^△E-mail：tianchen@tjmuch.com
作者简介:杨健（1983），男，主治医师，主要从事胃肠道肿瘤、消化道常见疾病及普通外科方面研究。E-mail:446926319@qq.com
基金资助:
新疆维吾尔自治区自然科学基金资助项目(2022D01A09)

T-ALL derived bone marrow stromal stem cells promote T-ALL proliferation through the FGF2-FGFR2 pathway

YANG Jian¹(), LI Min², LI Yueyang², TIAN Chen²^,^△()

1 Department of Surgical Oncology, Qingdao Central Cancer Hospital, Qingdao 266042, China
2 Department of Hematology, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin Clinical Research Center for Cancer

Received:2024-03-23 Revised:2024-09-11 Published:2025-01-15 Online:2025-02-06
Contact: ^△E-mail：tianchen@tjmuch.com

杨健, 李敏, 李越洋, 田晨. T-ALL来源的骨髓基质细胞通过FGF2-FGFR2通路促进T-ALL增殖[J]. 天津医药, 2025, 53(1): 29-34.

YANG Jian, LI Min, LI Yueyang, TIAN Chen. T-ALL derived bone marrow stromal stem cells promote T-ALL proliferation through the FGF2-FGFR2 pathway[J]. Tianjin Medical Journal, 2025, 53(1): 29-34.

摘要/Abstract

摘要：

目的探究骨髓基质细胞（BM-MSCs）对急性T淋巴细胞白血病（T-ALL）影响的作用机制，寻找靶向BM-MSCs的有效治疗策略。方法构建Notch-1过表达诱导的T-ALL小鼠模型，分选T-ALL中BM-MSCs，与T-ALL细胞系建立体外共培养系统，检测共培养后T-ALL增殖能力变化。运用RNA测序技术寻找不同来源MSCs的差异表达基因，并用PCR验证。在小鼠体内注射BGJ398，检测肿瘤生长情况。结果与T-ALL来源的MSCs共培养后，T-ALL细胞的增殖能力显著增加。RNA测序结果显示，T-ALL来源的MSCs成纤维细胞生长因子2（FGF2）分泌增加，与T-ALL细胞上的成纤维细胞生长因子2受体（FGFR2）结合可激活T-ALL细胞中PI3K/AKT/mTOR信号通路。加入BGJ398阻断FGF2和FGFR2之间的相互作用可以抑制小鼠T-ALL肿瘤的生长。结论 BM-MSCs可通过FGF2/FGFR2通路促进T-ALL肿瘤生长，阻断FGF2/FGFR2通路是克服BM-MSCs介导T-ALL进展的有效策略。

关键词: 前体细胞淋巴母细胞白血病淋巴瘤, 成纤维细胞生长因子2, 细胞增殖, 骨髓基质细胞, BGJ398

Abstract:

Objective To elucidate the mechanistic role of bone marrow mesenchymal stromal cells (BM-MSCs) in T-cell acute lymphoblastic leukemia (T-ALL), and to find effective therapeutic strategies targeting BM-MSCs.Methods A T-ALL mouse model induced by Notch-1 overexpression was constructed. An in vitro co-culture system was established to investigate the proliferative capacity of T-ALL cells upon co-culturing with leukemia-derived MSCs. RNA sequencing was performed to identify key differentially expressed genes, which were further validated by PCR. BGJ398 was injected into mice to detect tumor growth.Results Co-culturing with T-ALL-derived MSCs resulted in a significant increase in T-ALL cell proliferation. RNA sequencing results revealed that the secretion of fibroblast growth factor 2 (FGF2) from T-ALL-derived MSCs was increased, which binds to fibroblast growth factor 2 receptor (FGFR2) on T-ALL cells, activating the PI3K/AKT/mTOR signaling pathway. Blocking the interaction between FGF2 and FGFR2 using BGJ398 inhibited the growth of T-ALL tumors in mice.Conclusion BM-MSCs can promote T-ALL tumor growth through FGF2/FGFR2 pathway, and blocking FGF2/FGFR2 pathway is an effective strategy to overcome BM-MSCS-mediated T-ALL progression.

Key words: precursor cell lymphoblastic leukemia-lymphoma, fibroblast growth factor 2, cell proliferation, bone marrow mesenchymal stromal cells, BGJ398

中图分类号:

R733.71

图/表 7

图1 T-ALL小鼠模型构建流程图

Fig.1 Flow chart of T-ALL mouse model construction

表1 PCR引物序列

Tab.1 Primer sequence for PCR

基因名称	引物序列（5'→3'）	产物大小/bp
ICN1	上游：GATGGCCTCAATGGGTACAAG	74
ICN1	下游：TCGTTGTTGTTGATGTCACAGT	74
Csf2	上游：TTCAAGAAGCTAACATGTGTGC	159
Csf2	下游：GGTAACTTGTGTTTCACAGTCC	159
Fgf2	上游：AGTTGTGTCTATCAAGGGAGTG	244
Fgf2	下游：CATTGGAAGAAACAGTATGGCC	244
Igf2	上游：GTTGGTGCTTCTCATCTCTTTG	103
Igf2	下游：AAACTGAAGCGTGTCAACAAG	103
Ctf1	上游：CTCATTCCTACCCCATTTGGAG	96
Ctf1	下游：ACGTATTCCTCCAGAAGTTGTT	96
Hgf	上游：ACCTACAGGAAAACTACTGTCG	123
Hgf	下游：TGCATTCAACTTCTGAACACTG	123
Lif	上游：GCTACTATAGACGTCATGAGGG	140
Lif	下游：CAACCCAACTTTTTCCTTTGGA	140
Fgfr1	上游：GAGGACAACGTGATGAAGATCG	82
Fgfr1	下游：GGTTGGTCGTCTTCTTGTAGTA	82
Fgfr2	上游：CTAAAGGCAACCTCCGAGAATA	164
Fgfr2	下游：ACATTTTTGGGAAGCCAAGTAC	164
Fgfr3	上游：GCCTGCGTGCTAGTGTTCT	217
Fgfr3	下游：TACCATCCTTAGCCCAGACCG	217
Fgfr4	上游：TCCATGACCGTCGTACACAAT	193
Fgfr4	下游：ATTTGACAGTATTCCCGGCAG	193

图2 T-ALL小鼠模型的验证 A：T-ALL组及Control组小鼠脾脏大小；B：荧光显微镜观察GFP+ T-ALL细胞；C、D：流式细胞术检测GFP+细胞中CD3的表达。

Fig.2 Validation of the T-ALL mouse model

图3 T-ALL来源BM-MSCs对T-ALL细胞的作用 A：与T-ALL组及Control组BM-MSCs共培养后T-ALL细胞的增殖情况；B：与T-ALL组及Control组BM-MSCs共培养后T-ALL细胞的凋亡情况；C：与T-ALL组及Control组BM-MSCs共培养后T-ALL细胞的细胞周期，t G0/G1=2.353，t S=2.776，t G2/M=3.747，均P<0.05。

Fig.3 T-ALL-derived BM-MSCs on T-ALL cells

图4 T-ALL来源的BM-MSCs异常激活FGF2/FGFR2通路 A：T-ALL组与Control组BM-MSCs的RNA-seq；B：T-ALL组和Control组差异表达基因的验证，tCsf2=0.238，P=0.647；t Ctf1=1.156，P=0.259；t Fgfr=21.949，P<0.001；t Hgf=2.775，P=0.025；t Igf2 =7.104，P=0.002；t Lif=1.943，P=0.068；C：T-ALL组和Control组FGFR各亚型表达水平比较；t Fgfr1=6.170，P=0.003；t Fgfr2=59.157，P<0.001；t Fgfr3=0.976，P=0.384；t Fgfr4=1.099，P=0.199。＊P<0.05，＊＊P<0.01。

Fig.4 T-ALL-derived BM-MSCs aberrantly activate the FGF2/FGFR2 pathway

图5 FGF2/FGFR2对PI3K/AKT/mTOR通路的作用 A：抑制FGF2/FGFR2通路后T-ALL细胞中PI3K/AKT/mTOR通路蛋白表达情况，1—4分别为Control组、T-ALL组、BGJ398组及sh2组；B：蛋白表达水平的比较结果；F p-PI3K=361.752，F p-AKT=377.657，F p-mTOR=254.832，均P<0.01；a与Control组比较；b与T-ALL组比较，P<0.05。

Fig.5 Role of FGF2/FGFR2 on the PI3K/AKT/mTOR pathway

图6 PBS组与BGJ398组T-ALL小鼠肿瘤大小

Fig.6 Tumor size of T-ALL mice in the PBS group and the BGJ398 group

参考文献 21

[1]	ZHU H, DONG B, ZHANG Y, et al. Integrated genomic analyses identify high-risk factors and actionable targets in T-cell acute lymphoblastic leukemia[J]. Blood Sci, 2022, 4(1):16-28. doi:10.1097/BS9.0000000000000102.
[2]	COLMONE A, AMORIM M, PONTIER A L, et al. Leukemic cells create bone marrow niches that disrupt the behavior of normal hematopoietic progenitor cells[J]. Science, 2008, 322(5909):1861-1865. doi:10.1126/science.1164390.
[3]	YIN X, HU L, ZHANG Y, et al. PDGFB-expressing mesenchymal stem cells improve human hematopoietic stem cell engraftment in immunodeficient mice[J]. Bone Marrow Transplant, 2020, 55(6):1029-1040. doi:10.1038/s41409-019-0766-z.
[4]	SINGH A K, PRASAD P, CANCELAS J A. Mesenchymal stromal cells, metabolism, and mitochondrial transfer in bone marrow normal and malignant hematopoiesis[J]. Front Cell Dev Biol, 2023,11:1325291. doi:10.3389/fcell.2023.1325291.
[5]	WU C H, WENG T F, LI J P, et al. Biology and therapeutic properties of mesenchymal stem cells in leukemia[J]. Int J Mol Sci, 2024, 25(5):2527. doi:10.3390/ijms25052527.
[6]	NWABO KAMDJE A H, SEKE ETET P F, TAGNE SIMO R, et al. Emerging data supporting stromal cell therapeutic potential in cancer: reprogramming stromal cells of the tumor microenvironment for anti-cancer effects[J]. Cancer Biol Med, 2020, 17(4):828-841. doi:10.20892/j.issn.2095-3941.2020.0133.
[7]	PARK C S, YOSHIHARA H, GAO Q, et al. Stromal-induced epithelial-mesenchymal transition induces targetable drug resistance in acute lymphoblastic leukemia[J]. Cell Rep, 2023, 42(7):112804. doi:10.1016/j.celrep.2023.112804.
[8]	SøNDERGAARD R H, HøJGAARD L D, REESE-PETERSEN A L, et al. Adipose-derived stromal cells increase the formation of collagens through paracrine and juxtacrine mechanisms in a fibroblast co-culture model utilizing macromolecular crowding[J]. Stem Cell Res Ther, 2022, 13(1):250. doi:10.1186/s13287-022-02923-y.
[9]	TIAN C, ZHENG G, CAO Z, et al. Hes1 mediates the different responses of hematopoietic stem and progenitor cells to T cell leukemic environment[J]. Cell Cycle, 2013, 12(2):322-331. doi:10.4161/cc.23160.
[10]	MAJUMDAR M K, THIEDE M A, MOSCA J D, et al. Phenotypic and functional comparison of cultures of marrow-derived mesenchymal stem cells (MSCs) and stromal cells[J]. J Cell Physiol, 1998, 176(1):57-66. doi:10.1002/(SICI)1097-4652(199807)176:1<57::AID-JCP7>3.0.CO;2-7.
[11]	邢逸, 窦一鸣, 王敏, 等. 骨髓间充质干细胞来源外泌体对炎症微环境中巨噬细胞表型及软骨细胞的调控作用[J]. 天津医药, 2022, 50(4):343-349.
	XING Y, DOU Y M, WANG M, et al. The regulating effect of bone marrow mesenchymal stem cell-derived exosomes on macrophage phenotype and chondrocytes in the inflammatory microenvironmen[J]. Tianjin Med J, 2022, 50(4):343-349. doi:10.11958/20212366.
[12]	WANG J, LIU X, QIU Y, et al. Cell adhesion-mediated mitochondria transfer contributes to mesenchymal stem cell-induced chemoresistance on T cell acute lymphoblastic leukemia cells[J]. J Hematol Oncol, 2018, 11(1):11. doi:10.1186/s13045-018-0554-z.
[13]	CAI J, WANG J, HUANG Y, et al. ERK/Drp1-dependent mitochondrial fission is involved in the MSC-induced drug resistance of T-cell acute lymphoblastic leukemia cells[J]. Cell Death Dis, 2016, 7(11):e2459. doi:10.1038/cddis.2016.370.
[14]	JIA R, SUN T, ZHAO X, et al. DEX-induced SREBF1 promotes BMSCs differentiation into adipocytes to attract and protect residual T-cell acute lymphoblastic leukemia cells after chemotherapy[J]. Adv Sci(Weinh), 2023, 10(19):e2205854. doi:10.1002/advs.202205854.
[15]	IM J H, BUZZELLI J N, JONES K, et al. FGF2 alters macrophage polarization, tumour immunity and growth and can be targeted during radiotherapy[J]. Nat Commun, 2020, 11(1):4064. doi:10.1038/s41467-020-17914-x.
[16]	HOSAKA K, YANG Y, SEKI T, et al. Therapeutic paradigm of dual targeting VEGF and PDGF for effectively treating FGF-2 off-target tumors[J]. Nat Commun, 2020, 11(1):3704. doi:10.1038/s41467-020-17525-6.
[17]	ZHOU Z, LIU Z, OU Q, et al. Targeting FGFR in non-small cell lung cancer: implications from the landscape of clinically actionable aberrations of FGFR kinases[J]. Cancer Biol Med, 2021, 18(2):490-501. doi:10.20892/j.issn.2095-3941.2020.0120.
[18]	NEW J, ARNOLD L, ANANTH M, et al. Secretory autophagy in cancer-associated fibroblasts promotes head and neck cancer progression and offers a novel therapeutic target[J]. Cancer Res, 2017, 77(23):6679-6691. doi:10.1158/0008-5472.CAN-17-1077.
[19]	TRAER E, JAVIDI-SHARIFI N, AGARWAL A, et al. Ponatinib overcomes FGF2-mediated resistance in CML patients without kinase domain mutations[J]. Blood, 2014, 123(10):1516-1524. doi:10.1182/blood-2013-07-518381.
[20]	SHAH C A, BEI L, WANG H, et al. HoxA10 protein regulates transcription of gene encoding fibroblast growth factor 2 (FGF2) in myeloid cells[J]. J Biol Chem, 2012, 287(22):18230-18248. doi:10.1074/jbc.M111.328401.
[21]	MELNICK A F, MULLIN C, LIN K, et al. Cdc73 protects Notch-induced T-cell leukemia cells from DNA damage and mitochondrial stress[J]. Blood, 2023, 142(25):2159-2174. doi:10.1182/blood.2023020144.

T-ALL来源的骨髓基质细胞通过FGF2-FGFR2通路促进T-ALL增殖

T-ALL derived bone marrow stromal stem cells promote T-ALL proliferation through the FGF2-FGFR2 pathway

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