Hematopoietic Cell Maturation Is Regulated by Maintaining a Balance against Cytokine Induced-Cell Proliferation - Clinical Application for Differentiation-inducing Therapy of Leukemia-

通过维持细胞因子诱导的细胞增殖的平衡来调节造血细胞成熟 - 白血病分化诱导治疗的临床应用 -

• 批准号: 09671135
• 负责人: MIYAZAWA Keisuke
• 金额: $ 1.15万
• 依托单位: Tokyo Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671135/
• 关键词: Hematopoiesis; Cell Growth; Differentiation; Cytokine; Leukemia; Differentiation-inducing therapy; Hematopoietic growth factor

Using a factor-dependent cell line MO7ER, which contains a stably transduced human erythropoietin (EPO) receptor gene in human megakaryoblastic cell line MO7e and which resulted in concomitant expression of EPO receptor, c-Mpl and c-Kit, we investigated the biological effects of these cytokines in terms of cell growth and differentiation. Thrombopoietin (TPO), EPO and Steel factor (SLF) all stimulated MO7ER cell proliferation in a dose-dependent manner. Combined stimulation of cells with SLF plus either TPO or EPO resulted in striking synergistic enhancement of MO7ER cell growth as compared with each cytokine alone, whereas combination of TPO plus EPO showed only an additive effect on cell proliferation. With regards * cell differentiation, either TPO or EPO treatment induced enhancement of platelet glycoprotein (GP) IIb/IIIa and GPIb expression. SLF induced GPIIb/IIIa and GPIb expression, but the effect was much weaker than that of EPO or TPO. However, addition of SLF to either TPO- o … More r EPO-containing cultures (which induced potent mitogenesis in MO7ER cells) resulted in suppression of these megakaryocyte specific antigens. Addition of low-dose cytosine arabinoside (Ara-C)(1 to 10 ng/ml) enhanced TPO- or EPO- induced megakaryocytic differentiation in MO7ER cells while mildly suppressing cell growth. Treatment the cells with low-dose Ara-C plus TPO plus SLF overrode the proliferative enhancing effects of SLF and induced GPIIb/IIIa and GPIb expression as efficient as TPO alone. Retardation of TPO-induced megakaryocytic maturation was also observed in normal murine bone marrow cells by combined stimulation with TPO and SLF as assessed by the numbers of acetylcholinesterase staining-positive cells and megakaryocyte nuclear polyploidy. In addition, combination of low-dose Ara-C plus G-CSF enhanced granulocytic differentiation in WEHI-3B cells, as compared with the cells treated with G-CSF alone. This phenomenon was also reproduced using the primary cultured leukemia cells which were isolated from the patients with various types of acute myelogenous leukemias,These results suggest that hematopoietic cell maturation is, at least in part, regulated by countering cytokine-induced cell proliferation. Less

在人巨核母细胞MO7e中含有一个稳定转导的人促红细胞生成素（EPO）受体基因，并导致EPO受体、c-Mpl和c-Kit的同时表达，我们利用因子依赖性细胞系MO7ER，研究了这些细胞因子在细胞生长和分化方面的生物学效应。血小板生成素（TPO）、促红细胞生成素（EPO）和钢铁因子（SLF）均以剂量依赖性方式刺激MO7ER细胞增殖。与单独使用每种细胞因子相比，SLF加TPO或EPO联合刺激细胞可显著增强MO7ER细胞的生长，而TPO加EPO联合刺激细胞仅显示出累加效应。在*细胞分化方面，TPO和EPO均可诱导血小板糖蛋白（GP） IIb/IIIa和GPIb表达增强。SLF诱导GPIIb/IIIa和GPIb表达，但作用明显弱于EPO和TPO。然而，将SLF添加到TPO- o或含有更多epo的培养物（在MO7ER细胞中诱导有效的有丝分裂发生）导致这些巨核细胞特异性抗原的抑制。添加低剂量的阿拉伯糖胞嘧啶（Ara-C）（1 ~ 10 ng/ml）可增强TPO或EPO诱导的MO7ER细胞的巨核细胞分化，同时轻度抑制细胞生长。用低剂量Ara-C + TPO + SLF处理细胞，可以抑制SLF的增殖增强作用，诱导GPIIb/IIIa和GPIb的表达与单独使用TPO一样有效。通过观察乙酰胆碱酯酶染色阳性细胞和巨核细胞核多倍体的数量，TPO和SLF联合刺激正常小鼠骨髓细胞也观察到TPO诱导的巨核细胞成熟迟缓。此外，与单独使用G-CSF的细胞相比，低剂量Ara-C与G-CSF联合使用可增强WEHI-3B细胞的粒细胞分化。从不同类型的急性骨髓性白血病患者中分离出的原代培养白血病细胞也能重现这一现象。这些结果表明，造血细胞的成熟，至少在一定程度上是通过对抗细胞因子诱导的细胞增殖来调节的。少

项目成果

Yaguchi M, Miyazawa K, Katagiri T, Nishimaki J, Kizaki M, Tohyama K, Toyama K: "Vitamin K2 and its derivatives induce apoptosis in leukemia cells and enhance of all-trans retinoic acid. 17."Leukemia. 11. 779-787 (1997)

Yaguchi M、Miyazawa K、Katagiri T、Nishimaki J、Kizaki M、Tohyama K、Toyama K：“维生素 K2 及其衍生物诱导白血病细胞凋亡并增强全反式视黄酸。17.”白血病。

Yaguchi M, Miyazawa K, Otawa M, Ito Y, Kawanishi N, Toyama K: "Vitamin K2 therapy for a patient with myelodysplastic syndrome"Leukmeia. 13. 144-145 (1999)

Yaguchi M、Miyazawa K、Otawa M、Ito Y、Kawanishi N、Toyama K：“维生素 K2 治疗骨髓增生异常综合征患者”Leukmeia。

Miyazawa K.: "Hematopoietic stem cells" Int.J.Hematol.68・1. 337-338 (1998)

宫泽K.：“造血干细胞” Int.J.Hematol.68・1.337-338（1998）

Katagiri T, Miyazawa K, Uchida Y, Shigehumi H, Iwama H, Shyoji N, Kawakubo K, Shimamoto T, Inatomi Y, Kuriyama Y, Yaguchi M, Nehashi Y, Ohyashiki K, Toyama K: "Induction of Ph-negative clone and long-term survival by combined treatment with G-CSF plus mid

Katagiri T、Miyazawa K、Uchida Y、Shigehumi H、Iwama H、Shyoji N、Kawakubo K、Shimamoto T、Inatomi Y、Kuriyama Y、Yaguchi M、Nehashi Y、Ohyashiki K、Toyama K：“Ph 阴性克隆的诱导和

Yaguchi M,Miyazawa K et al: "Vitemin K2 therapy for a patient with myelodysplastic syndrome" Leukemia. 13・1. 144-145 (1999)

Yaguchi M、Miyazawa K 等：“骨髓增生异常综合征患者的维生素 K2 治疗”白血病 13・1（1999）。