Growth factors and cytokines upregulate gelatinase expression in bone marrow CD34+ cells and their transmigration through reconstituted basement membrane
Growth factors and cytokines upregulate gelatinase expression in bone marrow CD34+ cells and their transmigration through reconstituted basement membrane
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DOI：
10.1182/blood.v93.10.3379.410k09_3379_3390
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发表时间：
1999-05-15
期刊：
影响因子：
20.3
通讯作者：
Turner, AR
中科院分区：
文献类型：
--
作者：
Janowska-Wieczorek, A;Marquez, LA;Turner, AR
干细胞/祖细胞从骨髓释放到血液循环的机制尚不清楚。我们假设基质金属蛋白酶（MMPs），特别是明胶酶，被认为参与基底膜的蛋白水解和白细胞的迁移，可能促进这一过程。首先，我们研究了CD34（+）干细胞/祖细胞是否表达明胶酶A （MMP-2）和/或B (MMP-9)，以及生长因子和细胞因子（粒细胞集落刺激因子[G-CSF]、粒细胞-巨噬细胞集落刺激因子[GM-CSF]、干细胞因子[SCF]、巨噬细胞集落刺激因子[M-CSF]、白细胞介素-3 [IL-3]、IL-6、IL-8和肿瘤坏死因子- α [tnf - α]）是否能够调节它们的表达。接下来，我们研究了这些干细胞/祖细胞通过重组基底膜（Matrigel）的迁移及其受生长因子和细胞因子的调节。CD34（+）细胞来自稳态骨髓和外周血（来自稳态造血或G-CSF联合化疗或单独G-CSF动员后收集的白细胞分离产物）。我们发现外周血CD34（+）细胞，无论它们是否被动员，都强烈表达明胶酶（MMP-5和MMP-9），而稳定状态的骨髓CD34（+）细胞则没有。然而，所有的生长因子和细胞因子均能诱导后者细胞分泌MMP-2和MMP-9。此外，G-CSF和SCF对MMP-5和MMP-9分泌的刺激作用在骨髓分离的CD34（+）细胞中明显高于外周血分离的CD34（+）细胞。此外，tnf - α、GM-CSF和IL-6增加了部分活性形式的MMP-2的分泌。骨髓CD34（+）细胞通过Matrigel的基础转运低于外周血CD34（+）细胞（P < 0.0001），而生长因子和细胞因子使其增加了50% ~ 150%。明胶酶的表达与CD34（+）细胞迁移呈正相关（r < 0.05）。G-CSF对骨髓CD34（+）细胞迁移的刺激作用明显大于外周血CD34（+）细胞迁移的刺激作用（P = 0.004）。此外，通过MMP-2和MMP-9、金属蛋白酶组织抑制剂（rhTIMP-1和-2）和邻菲罗啉的抗体，CD34（+）细胞迁移减少到约50%。tnf α诱导的CD34（+）细胞和克隆祖细胞（集落形成单位-粒细胞-巨噬细胞[CFU-GM]、突发性形成单位-红细胞[BFU-E]、集落形成单位粒细胞、红细胞、单核细胞、巨核细胞[CFU-GEMM]和集落形成单位-巨核细胞[CFU-MK]）明胶酶的分泌和迁移呈剂量依赖性。因此，本研究表明，在外周血中循环的CD34（+）细胞同时表达MMP-5和MMP-9，并通过Matrigel转运。相比之下，来自稳态骨髓的CD34（+）细胞在暴露于生长因子和细胞因子后获得类似的特性，当这些细胞进入血液时，它们上调明胶酶的表达和转运。因此，我们认为生长因子和细胞因子通过明胶酶信号通路诱导干细胞/祖细胞在动员和稳态造血过程中从骨髓释放到外周血中。(C) 1999年由美国血液病学会出版。
The mechanism(s) underlying the release of stem/progenitor cells from bone marrow into the circulation is poorly understood. We hypothesized that matrix metalloproteinases (MMPs), especially gelatinases, which are believed to participate in the proteolysis of basement membranes and in the migration of leukocytes, may facilitate this process. First, we investigated whether CD34(+) stem/progenitor cells express gelatinases A (MMP-2) and/or B (MMP-9) and whether growth factors and cytokines (granulocyte colony-stimulating factor [G-CSF], granulocyte-macrophage colony-stimulating factor [GM-CSF], stem cell factor [SCF], macrophage colony-stimulating factor [M-CSF], interleukin-3 [IL-3], IL-6, IL-8, and tumor necrosis factor-alpha [TNF-alpha]) are able to modulate their expression. Next, we examined the transmigration of these stem/progenitor cells through reconstituted basement membrane (Matrigel) and its modulation by growth factors and cytokines. CD34(+) cells were obtained from steady-state bone marrow and peripheral blood (from leukapheresis products collected either in steady-state hematopoiesis or after mobilization with G-CSF plus chemotherapy or G-CSF alone). We found that peripheral blood CD34(+) cells, regardless of whether they were mobilized or not, strongly expressed both gelatinases (MMP-5 and MMP-9) in contrast to steady-state bone marrow CD34(+) cells, which did not. However, all the growth factors and cytokines tested could induce MMP-2 and MMP-9 secretion by the latter cells. Moreover, the stimulatory effects of G-CSF and SCF on both MMP-5 and MMP-9 secretion were found to be significantly higher in CD34(+) cells isolated from bone marrow than in those from peripheral blood. In addition TNF-alpha, GM-CSF, and IL-6 increased the secretion of a partially active form of MMP-2. Basal transmigration of bone marrow CD34(+) cells through Matrigel was lower than that of peripheral blood CD34(+) cells (P < .0001), but growth factors and cytokines increased it by 50% to 150%. Positive correlations were established between expression of gelatinases and CD34(+) cell migration (r > .9). The stimulatory effect of G-CSF was significantly greater on the migration of CD34(+) cells from bone marrow than on those from peripheral blood (P = .004). Moreover, CD34(+) cell migration was reduced to approximately 50% by antibodies to MMP-2 and MMP-9, tissue inhibitors of metalloproteinases (rhTIMP-1 and -2), and o-phenanthroline. TNF-alpha-induced gelatinase secretion and migration of CD34(+) cells and of clonogenic progenitors (colony-forming unit-granulocyte-macrophage [CFU-GM], burst-forming unit-erythroid [BFU-E], colony-forming unit granulocyte, erythroid, monocyte, megakaryocyte [CFU-GEMM], and colony-forming unit-megakaryocyte [CFU-MK]) were dose-dependent. Therefore, this study demonstrated that CD34(+) cells that are circulating in peripheral blood express both MMP-5 and MMP-9 and transmigrate through Matrigel. In contrast, CD34(+) cells from steady-state bone marrow acquire similar properties after exposure to growth factors and cytokines, which upregulate expression of gelatinases and transmigration of these cells when they enter the bloodstream. Hence, we suggest that growth factors and cytokines induce release of stem/progenitor cells from bone marrow into peripheral blood during mobilization, as well as during steady-state hematopoiesis, by signaling through gelatinase pathways. (C) 1999 by The American Society of Hematology.