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    • 21. 发明申请
    • HIMF AND BTK IN PULMONARY, CARDIAC, AND INFLAMMATION DISORDERS
    • HIMF和BTK在肺,心脏和炎症疾病中的作用
    • WO2008077022A2
    • 2008-06-26
    • PCT/US2007/087899
    • 2007-12-18
    • THE JOHNS HOPKINS UNIVERSITYJOHNS, Roger, A.SU, QingningCHAMPION, Hunter, Clay
    • JOHNS, Roger, A.SU, QingningCHAMPION, Hunter, Clay
    • A61K39/395
    • G01N33/6884A61K2039/505C07K14/4702C07K16/18C07K16/22C07K16/40C12N9/1205C12Y207/10002G01N33/54306G01N33/74G01N2333/47G01N2333/4703G01N2333/912G01N2500/02G01N2800/12G01N2800/122G01N2800/7038
    • Hypoxia induced mitogenic factor (HIMF) is a member of the "found in inflammatory zone" (FIZZ)/resistin family of proteins and has potent mitogenic, angiogenic, and vasoconstrictive effects in the lung vasculature. The receptor/binding partners for this family of proteins have been largely unknown. We identified Bruton's tyrosine kinase (BTK) as a functional HIMF binding partner through GST-HIMF pull-downs and mass spectrometry. Using primary cultured HIMF-stimulated murine bone marrow cells, we demonstrated that BTK was recruited to the leading edge of the cells. We also demonstrated that BTK and the closely related tyrosine kinase Fyn, colocalized at the growth cone process in these cells. HIMF stimulation induced BTK autophosphorylation, which peaked at 2.5 minutes. A transwell migration assay showed that treatment with recombinant murine HIMF induced migration of primary cultured bone marrow cells, which was completely blocked by the BTK inhibitor, LFM-A13. In vivo studies, using the rat hindlimb ischemia model, revealed that HIMF can stimulate angiogenesis in the hypoxic tissue probably through inducing the migration of endothelial progenitor cells (EPCs) to areas of active angiogenesis. Our results indicate that HIMF may acts as a chemotactic molecule in stimulating the migration of leukocytes/EPCs from bone marrow to targeted tissues through activation of the BTK pathway.
    • 缺氧诱导的促有丝分裂因子(HIMF)是“在炎症区中发现的”的成员。 (FIZZ)/抵抗素家族的蛋白质,并在肺血管系统中具有有效的促有丝分裂,血管生成和血管收缩作用。 这个蛋白质家族的受体/结合伴侣在很大程度上是未知的。 我们通过GST-HIMF pull-downs和质谱法将Bruton酪氨酸激酶(BTK)鉴定为功能性HIMF结合配偶体。 使用原代培养的HIMF刺激的小鼠骨髓细胞,我们证明BTK被募集到细胞的前沿。 我们还证明了BTK和密切相关的酪氨酸激酶Fyn在这些细胞的生长锥过程中共定位。 HIMF刺激诱导BTK自磷酸化,其在2.5分钟达到峰值。 transwell迁移试验表明,用重组鼠HIMF处理诱导了原代培养的骨髓细胞的迁移,其被BTK抑制剂LFM-A13完全阻断。 在体内研究中,使用大鼠后肢缺血模型显示HIMF可能通过诱导内皮祖细胞(EPCs)迁移到活性血管生成区域来刺激缺氧组织中的血管生成。 我们的研究结果表明HIMF可能作为一种趋化分子,通过激活BTK途径刺激白血球/ EPCs从骨髓迁移至靶向组织。