The heparan sulfate (HS) 6-O-endosulfatases or the Sulfs (Sulf1 and Sulf2) are the only known enzymes that can modify HS sulfation status extracellularly and have been shown to regulate diverse biological processes. The role of the Sulfs in bone marrow (BM) hematopoiesis is not known. In this study, we generated a novel mouse line with myeloid-specific deletion of the Sulfs by crossing Sulf1/2 double floxed mice with the LysM-cre line. The LysM-Sulf knockout (KO) male mice exhibited age-dependent expansion of hematopoietic stem cells and the granulocyte-monocyte lineages in the BM, whereas common lymphoid progenitors and B lymphocyte populations were significantly reduced. Although megakaryocytic and erythroid progenitors were not reduced in the BM, the LysM-Sulf KO males suffered age-dependent reduction of red blood cells (RBCs) and platelets in the peripheral blood, suggesting that the production of RBCs and platelets was arrested at later stages. In addition, LysM-Sulf KO males displayed progressive splenomegaly with extramedullary hematopoiesis. Compared to males, LysM-Sulf KO females exhibited a much-reduced phenotype, and ovariectomy had little effect. Mechanistically, reduced TGF-β/Smad2 but enhanced p53/p21 signaling were observed in male but not female LysM-Sulf KO mice. Finally, HS disaccharide analysis via LC-MS/MS revealed increased HS 6-O-sulfation in the BM from both male and female LysM-Sulf KO mice, however, the distribution of 6-O-sulfated motifs were different between the sexes with compensatory increase in Sulf1 expression observed only in LysM-Sulf KO females. In conclusion, our study reveals that myeloid deficiency of the Sulfs leads to multilineage abnormalities in BM hematopoiesis in an age- and sex-dependent manner.

中文翻译：

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硫酸乙酰肝素 6-O-硫代硫酸酶的髓系缺乏症损害骨髓造血功能


硫酸乙酰肝素 （HS） 6-O-硫乙酰氨基糖酶或硫磺酸酯（Sulf1 和 Sulf2）是唯一已知的可以在细胞外改变 HS 硫酸化状态的酶，并已被证明可以调节多种生物过程。Sulfs 在骨髓 （BM） 造血中的作用尚不清楚。在这项研究中，我们通过将 Sulf1/2 双絮凝小鼠与 LysM-cre 系杂交，生成了一种具有髓系特异性 Sulfs 缺失的新型小鼠系。LysM-Sulf 敲除 （KO） 雄性小鼠表现出造血干细胞和 BM 中粒细胞-单核细胞谱系的年龄依赖性扩增，而共同淋巴祖细胞和 B 淋巴细胞群显着减少。尽管 BM 中的巨核细胞和红细胞祖细胞没有减少，但 LysM-Sulf KO 男性外周血中红细胞 （RBC） 和血小板的年龄依赖性减少，表明红细胞和血小板的产生在后期停止。此外，LysM-Sulf KO 男性表现出进行性脾肿大伴髓外造血。与雄性相比，LysM-Sulf KO 雌性表现出大大减少的表型，卵巢切除术几乎没有影响。从机制上讲，在雄性而非雌性 LysM-Sulf KO 小鼠中观察到 TGF-β/Smad2 降低但 p53/p21 信号增强。最后，通过 LC-MS/MS 进行的 HS 二糖分析显示，雄性和雌性 LysM-Sulf KO 小鼠在 BM 中的 HS 6-O-硫酸化增加，然而，6-O-硫酸化基序的分布在两性之间是不同的，仅在 LysM-Sulf KO 雌性中观察到 Sulf1 表达的代偿性增加。总之，我们的研究表明，Sulfs 的髓系缺乏症以年龄和性别依赖性方式导致 BM 造血多谱系异常。