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mayumi hirano Last modified date:2019.06.17

Assistant Professor / Science for Biological Information
Department of Basic Medicine
Faculty of Medical Sciences


Graduate School
Other Organization


Homepage
http://www.molcar.med.kyushu-u.ac.jp/
Division of Molecular Cardiology, Research Institute of Angiocardiology,
Graduate School of Medical Sciences, Kyushu University .
Phone
092-642-5550
Fax
092-642-5552
Academic Degree
PhD
Country of degree conferring institution (Overseas)
No
Field of Specialization
Cell Biology
Total Priod of education and research career in the foreign country
00years00months
Outline Activities
Elucidation of the intracellular signal networks involved in the growth arrest induced by Cell-cell contact in vascular endothelial cells.
Role of endothelial proteinase-activated receptors in the early phase of the development of vascular lesions.
Research
Research Interests
  • A study elucidating the molecular mechanism of the growth arrest by the formation of cell-cell contact in vascular endothelial cells.
    keyword : Endothelial cells, Cell-cell contact, Growth arrest, Cell cycle, Transcriptional up-regulation
    1997.04~2021.03I am clarifying the molecular mechanism for the cell contact-induced growth inhibition of the vascular endothelial cells. I found that p27Kip1 was up-regulated by transcription upon formation of cell-cell contact in enfothelial cells..
  • Elucidation of the molecular mechanism underlying the vascular dysfunction mediated by proteinase-activated receptor 1
    keyword : Endothelial cells, Proteinase activated receptor-1, vascular dysfenction, endothelial barrier disruption
    2015.04~2021.03I am clarifying the molecular mechanism for the cell contact-induced growth inhibition of the vascular endothelial cells. I found that p27Kip1 was up-regulated by transcription upon formation of cell-cell contact in enfothelial cells..
  • A study elucidating the molecular mechanism for the activation of vascular endothelial cells by thrombin
    keyword : Endothelial cells, MLC phosphorylation, Myosin phophatase, Actin stress fiber,Permeability
    2010.04~2021.03.
  • A study elucidated the role of proteinase-activated receptors(PARs) in the early phase of the development of atherosclerotic vascular lesions
    keyword : Endothelial cells, Thrombin, Proteinase-activated receptor, Ateriosclerosis, NO
    2012.04I.
  • A study elucidating the mechanisms regulating the transcription of the cell cycle regulatory protein p27Kip1 in the cell-cell contact-induced growth arrest of vascular endothelial cells.
    keyword : Endothelial cells, Cell-cell contact, Growth arrest, Cell cycle, Transcription, Rac1
    2001.04~2010.03I.
  • A study elucidating the mechanism underlying the production of nitric oxide by thrombin in vascular endothelial cells
    keyword : Endothelial cells, Proteinase-activated receptor, Thrombin, NO, Ca
    2003.04~2012.03.
  • Stromal interaction molecule-1 (STIM1) plays an important role in the thrombin-induced Ca2+ signal and Ca2+-dependent NO production in endothelial cells.
    keyword : Endothelial cells, Ca influx, Ca Store
    2007.04I.
Current and Past Project
  • Elucidation of the intracellular signal networks involved in the growth arrest induced by cell-cell contact in vascular endothelial cells.
  • Effect of gravitational alteration on the structure and function of vascular system
Academic Activities
Books
1. Hirano K, Hirano M, Abe S and Kanaide H., Cytosolic calcium transients in vascular smooth muscle. in Ion Channels of Vascular Smooth Muscle Cells and Endothelial Cells, eds Sperelakis N and Kuriyama H., Elsivier, New York,, 93-105,, 1991.01.
Papers
1. Mayumi Hirano, Katsuya Hirano, Myosin di-phosphorylation and peripheral actin bundle formation as initial events during endothelial barrier disruption , Scientific Report , 10.1038/srep20989, 6: 20989, 2016.02, The phosphorylation of the 20-kD myosin light chain (MLC) and actin filament formation play a key role in endothelial barrier disruption. MLC is either mono- or di-phosphorylated (pMLC and ppMLC) at T18 or S19. The present study investigated whether there are any distinct roles of pMLC and ppMLC in barrier disruption induced by thrombin. Thrombin induced a modest bi-phasic increase in pMLC and a robust mono-phasic increase in ppMLC. pMLC localized in the perinuclear cytoplasm during the initial phase, while ppMLC localized in the cell periphery, where actin bundles were formed. Later, the actin bundles were rearranged into stress fibers, where pMLC co-localized. Rho-kinase inhibitors inhibited thrombin-induced barrier disruption and peripheral localization of ppMLC and actin bundles. The double, but not single, mutation of phosphorylation sites abolished the formation of peripheral actin bundles and the barrier disruption, indicating that mono-phosphorylation of MLC at either T18 or S19 is functionally sufficient for barrier disruption. Namely, the peripheral localization, but not the degree of phosphorylation, is suggested to be essential for the functional effect of ppMLC. These results suggest that MLC phosphorylation and actin bundle formation in cell periphery are initial events during barrier disruption. .
2. Toshiro Saito, mayumi hirano, Kenji Sunagawa, Katsuya Hirano, Pivotal Role of Rho-Associated Kinase 2 in Generating the Intrinsic Circadian Rhythm of Vascular Contractility. , Circulation 127(1), 10.1161/CIRCULATIONAHA.112.135608 , 127, 104-114, 2013.01, Background—The circadian variation in the incidence of cardiovascular events may be attributable to the circadian changes in vascular contractility. The circadian rhythm of vascular contractility is determined by the interplay between the central and peripheral clocks. However, the molecular mechanism of the vascular intrinsic clock that generates the circadian rhythm of vascular contractility still remains largely unknown.
Methods and Results—The agonist-induced phosphorylation of myosin light chain in cultured smooth muscle cells synchronized by dexamethasone pulse treatment exhibited an apparent circadian oscillation, with a 25.4-hour cycle length. The pharmacological inhibition and knockdown of Rho-associated kinase 2 (ROCK2) abolished the circadian rhythm of myosin light chain phosphorylation. The expression and activity of ROCK2 exhibited a circadian rhythm in phase with that of myosin light chain phosphorylation. A clock gene, RORα, activated the promoter of the ROCK2 gene, whereas its knockdown abolished the rhythmic expression of ROCK2. In the mouse aorta, ROCK2 expression exhibited the circadian oscillation, with a peak at Zeitgeber time 0/24 and a nadir at Zeitgeber time 12. The myofilament Ca2+ sensitization induced by GTPγS and U46619, a thromboxane A2 analog, at Zeitgeber time 0/24 was greater than that seen at Zeitgeber time 12. The circadian rhythm of ROCK2 expression and myofilament Ca2+ sensitivity was abolished in staggerer mutant mice, which lack a functional RORα.
Conclusions—ROCK2 plays a pivotal role in generating the intrinsic circadian rhythm of vascular contractility by receiving a cue from RORα. The ROCK2-mediated intrinsic rhythm of vascular contractility may underlie the diurnal variation of the incidence of cardiovascular diseases..
3. Yuichiro Kikkawa, S Matsuo, Katsuharu Kameda, Katsuya Hirano, A Nakamizo, mayumi hirano, Tomio Sasaki, Mechanisms underlying potentiation of endothelin-1-induced myofilament Ca2+ sensitization after subarachnoid hemorrhage.
, J Cereb Blood Flow Metab , 32, 341-352, 2012.03, Increased vascular smooth muscle contractility has an important role in the development of cerebral vasospasm after subarachnoid hemorrhage (SAH). Myofilament Ca2+ sensitivity is a major determinant of smooth muscle contractility. We investigated changes in the Ca2+-sensitizing effect of endothelin-1 (ET-1) and the mechanisms underlying ET-1-induced Ca2+ sensitization after SAH using a rabbit SAH model. After SAH, the contractile response to ET-1 was enhanced, and the ETA receptor expression was upregulated in the basilar artery. In a-toxin-permeabilized preparations, ET-1 induced enhanced and prolonged contraction after SAH, suggesting that ET-1-induced Ca2+ sensitization is potentiated after SAH. Endothelin-1-induced Ca2+ sensitization became less sensitive to inhibitors of Rho-associated coiled-coil protein kinase (ROCK) and protein kinase C (PKC) after SAH. The expression of PKCa, ROCK2, PKC-potentiated phosphatase inhibitor of 17 kDa (CPI-17) and myosin phosphatase target subunit 1 (MYPT1) was upregulated, and the level of phosphorylation of CPI-17 and MYPT1 was elevated after SAH. This study demonstrated for the first time that the Ca2+-sensitizing effect of ET-1 on myofilaments is potentiated after SAH. The increased expression and activity of PKCa, ROCK2, CPI-17, and MYPT1, as well as the upregulation of ETA receptor expression are suggested to underlie the enhanced and prolonged Ca2+sensitization induced by ET-1..
4. Katsuharu Kameda, Yuichiro Kikkawa, mayumi hirano, S Matsuo, Tomio Sasaki, Hideo Kanaide, Combined argatroban and anti-oxidative agents prevents increased vascular contractility to thrombin and other ligands after subarachnoid hemorrhage2+-sensitivity and induces vasoconstriction in porcine pulmonary arteries., Br J Pharmacol, 165 , 106-119, 2012.01.
5. Hirano K, Hirano M, Hanada A:, Involvement of STIM1 in the proteinase-activated receptor 1-mediated Ca2+ influx in vascular endothelial cells., J Cell Biochem, 108, 499-507, 2009.09.
6. Maeda Y, Hirano K, Hirano M, Kikkawa Y, Kameda K, Sasaki T, Kanaide H, Enhanced contractile response of the basilar artery to platelet-derived growth factor in subarachnoid hemorrhage., Stroke 40, 591-596, 2009, 40, 591-596, 2009.02.
7. Hirano M, Kanaide H, Hirano K, Rac1-dependent transcriptional up-regulation of p27Kip1 by homophilic cell-cell contact in vascular endothelial cells., Biochim Biophys Acta -Mol Cell Res 1773, 1500-1510, 2007, 1773, 1500-1510, 2007
, 2007.08.
8. Yufu T, Hirano K, Bi D, Hirano M, Nishimura J, Iwamoto Y, Kanaide H, Rac1 regulation of the surface expression of PAR1 and responsiveness to thrombin in vascular smooth muscle cells, Arterioscler Thromb Vasc Biol, 10.1161/01.ATV.0000168418.10276.f0, 25, 7, 1506-1511, vol. 25, 1506-1511, 2005.01.
9. Shiga N, Hirano K, Hirano M, Nishimura J, Nawata H, Kanaide H, Long-term inhibition of RhoA attenuates vascular contractility by enhancing endothelial NO production in an intact rabbit mesenteric artery, Cir Res, 10.1161/01.RES.0000165483.34603.91, 96, 9, 1014-1021, vol. 96, 1014-1021, 2005.01.
10. Koga M, Hirano K, Hirano M, Nishimura J, Kanaide H:, Akt plays a central role in the anti-apoptotic effect of estrogen in endothelial cells, Biochem Biophys Res Commun, 10.1016/j.bbrc.2004.09.060, 324, 1, 321-325, vol. 324, 321-325, 2004.01.
11. Hirano K, Ihara E, Hirano M, Nishimura J, Nawata H, Kanaide H:, Facilitation of proteasomal degradation of p27Kip1 by N-terminal cleavage, and their sequence requirements, FEBS Lett, 10.1016/j.febslet.2004.08.014, 574, 1-3, 111-115, vol. 574, 111-115, 2004.01.
12. Hirano K, Hirano M, Nishimura J, Kanaide H:, A critical period requiring Rho proteins for cell cycle progression uncovered by reversible protein transduction in endothelial cells, FEBS Lett, 10.1016/j.febslet.2004.05.084, 570, 1-3, 149-154, vol. 570, 149-154, 2004.01.
13. Nakayama T, Hirano K, Hirano M. Nishimura J, Kuga H, Nakamura K, Takahashi S, Kanaide H:, Inactivation of protease-activated receptor-1 by proteolytic removal of the ligand region in vascular endothelial cells, Biochem Pharmacol, 10.1016/j.bcp.2004.03.005, 68, 1, 23-32, vol. 68, 23-32, 2004.01.
14. Hirano K, Derkach DN, Hirano M, Nishimura J, Takahashi S, Kanaide H, Transduction of the N-terminal fragments of MYPT1 enhances myofilament Ca2+ sensitivity in an intact coronary artery, Arterioscler Thromb Vasc Biol, 10.1161/01.ATV.0000116028.42230.4c, 24, 3, 464-469, vol. 24, 464-469, 2004.01.
15. Eto W, Hirano K, Hirano M, Nishimura J, Kanaide H:, Intracellular alkalinization induces Ca2+ influx via non-voltage-operated Ca2+ channels in the rat aortic smooth muscle cells, Cell Calcium, 10.1016/S0143-4160(03)00151-9, 34, 6, 477-484, vol. 34, 477-484, 2003.01.
16. Hirano K, Zeng Y, Hirano M, Nishismura J, Kanaide H, Sequence requirement for nuclear localization and growth inhibition of p27Kip1R, a degradation-resistant isoform of p27Kip1, J Cell Biochem, 10.1002/jcb.10499, 89, 1, 191-202, vol. 89, 191-202, 2003.01.
17. Ihara E, Hirano K, Hirano M, Nishimura J, Nawata H, Kanaide H, The mechanism of down-regulation of L-type Ca2+ channel in the proliferating smooth muscle cells of rat aorta, J Cell Biochem 87, 242-251, vol. 87, 242-251, 2002.01.
18. Hirano M, Hirano K, Nishimura J, Kanaide H, Transcriptional up-regulation of p27Kip1 during contact-induced growth arrest in vascular endothelial cells, Exp Cell Res, 10.1006/excr.2001.5384, 271, 2, 356-367, vol. 271, 356-367, 2001.01.
19. Hirano K, Hirano M, Zeng Y, Nishimura J, Hara K, Muta K, Nawata H, Kanaide H, Cloning and functional expression of a degradation-resistant novel isoform of p27Kip1, Biochem J, 10.1042/0264-6021:3530051, 353, 51-57, vol. 353, 51-57, 2001.01.
20. Hirano K, Hirano M, Eto W, Nishimura J, Kanaide H, Mitogen-induced upregulation of non-smooth muscle isoform of a-tropomyosin in rat aortic smooth muscle cells, Eur J Pharmacol, 10.1016/S0014-2999(00)00681-6, 406, 2, 209-218, vol. 406, 209-218, 2000.01.
21. Hirano M, Niiro N, Hirano K, Nishimura J, Hartshorne DJ, Kanaide H: , Expression, subcellular localization and cloning of the 130 kDa regulatory subunit of myosin phosphatase in porcine aortic endothelial cells.
, Biochem. Biophys. Res. Commun. , 10.1006/bbrc.1998.9973, 254, 2, 490-496, 1999.01.
Presentations
1. Mayumi Hirano, Katsuya Hirano, Submembranous di-phosphorylation of myosin light chain and actin fiber formation play a critical role as an initial event during endothelial barrier disruption, 11th International Symposium on Resistance Arteries ISRA, 2014.09, The phosphorylation of myosin light chain (MLC) plays an essential role in endothelial barrier disruption. MLC is phosphorylated at Thr18 and Ser19. We elucidated distinct role of mono- and di-phosphorylation of MLC during barrier disruption in porcine aortic endothelial cells. Thrombin decreased trans-endothelial electrical resistance (TEER) with a bottom at 3-5 min. Phos-tag SDS-PAGE analysis revealed that 25% and 2% of MLC were mono- and di-phosphorylated (pMLC and ppMLC) before stimulation. pMLC slightly increased in the cytoplasmic region 3 min after stimulation, while ppMLC increased to 35% in the submembranous region, where actin fiber formation was also induced. Thereafter, ppMLC returned to the resting level, while pMLC reached a second heave after 15 min, when stress fiber formation was observed. Rho-kinase inhibitor abolished thrombin-induced decrease in TEER, with inhibition of the submembranous ppMLC and actin fiber formation at 3 min as well as pMLC at 15 min. Myosin ATPase inhibitor, blebbistatin, inhibited the decrease in TEER with no effect on MLC phosphorylation. Adenoviral expression of a mutant of MLC with substitution of Thr18 and Ser19 to Ala (MLCT18A+S19A) abolished the thrombin-induced submembranous ppMLC and actin fiber formation as well as the decrease in TEER. However, those events were observed in the cells expressing MLCT18A or MLCS19A as in the cells expressing wild-type MLC. In conclusion, the submembranous ppMLC and actin fiber formation and the resultant circumferential contraction serve as an initial event during endothelial barrier disruption. Mono-phosphorylation of MLC is functionally sufficient for submembranous actin fiber formation and barrier disruption..
2. Mayumi Hirano, Katsuya Hirano, Di-phosphorylation of myosin light chain and actin fiber formation in the submembranous region at inter-endothelial junction play a crucial role in thrombin-induced endothelial barrier disruption, American Heart Association Scientific Session, 2013, 2013.11.
3. Mayumi Hirano, Katsuya Hirano, Myosin light chain di-phosphorylation at sub-membranous region plays a critical role during thrombin-induced barrier disruption in vascular endothelial cells., International Symposium on Regulatory circuits in cell motility, honoring Dave Hartshorne, 2013.10.
4. Katsuya Hirano, Katsuharu Kameda, Akiko Hanada, mayumi hirano, Oxidative stress and Extracellular Signal-regulated Kinase (ERK) Play a Key Role in Impairment of Receptor Desensitization and Prolongation of Vascular Reactivity in the Cerebral Artery after Subarachnoid Hemorrhage., International Stroke Conference 2013, 2013.02.
5. mayumi hirano, Akiko Hanada, Katsuya Hirano, Rho Kinase-mediated Di-phosphorylation of Myosin Light Chain in the Sub-membranous Regions and Circumferential Actomyosin Contraction Mediate Thrombin-induced Barrier Disruption in Vascular Endothelial Cells., International Stroke Conference 2013, 2013.02.
6. mayumi hirano, Akiko Hanada, Katsuya Hirano, Oxidative Stress and Rho-associated Coiled-coil Protein Kinase (ROCK)-mediated Double Phosphorylation of Myosin Light Chain in the Submembranous Region Plays a Key Role in Thrombin-induced Barrier Dysfunction in Vascular Endothelial Cells, Scientific Sessions 2012 of the American Heart Association, 2012.11.
7. Toshiro Saito, mayumi hirano, Tomomi Ide, Toshihiro Ichiki, Noriyuki Koibuchi, Kenji Sunagawa, Katsuya Hirano, A Clock Gene ROR-mediated Regulation of the Activity of Rho-associated Coiled-coil Protein Kinase 2 (ROCK2) Plays a Key Role in Generating Vascular Intrinsic Circadian Rhythm of Myofilament Ca2+ Sensitivity and Vascular Contractility, Scientific Sessions 2012 of the American Heart Association, 2012.11.
8. Katsuharu kameda, Yuichiro Kikkawa, mayumi hirano, O Matsuo, Tomio Sasaki, Katsuya Hirano, Oxidative Stress Impairs Desensitization of the G-protein Coupled Receptors (GPCRs) and Increases Vascular Reactivity by Prolonging the Contractile Responses in the Cerebral Arteries after Subarachnoid Hemorrhage., Scientific Sessions 2011 of the American Heart Association, 2011.11.
9. mayumi hirano, Akiko Hanada, Katsuya Hirano, Depletion of Intracellular Ca2+ Stores Induces Phosphorylation of Stromal Interaction Molecular 1 (STIM1), which Contributes to the Sustained Phase of Store-operated Ca2+ Influx in Vascular Endothelial Cells
, Scientific Sessions 2011 of the American Heart Association, 2011.11.
10. Thrombin increases myosin light chain (MLC) phosphorylation, thereby increasing vascular permeability. This study investigated the regulatory mechanism of the thrombin-induced MLC phosphorylation in porcine aortic endothelial cells, by using a Phos-tag SDS-PAGE analysis. Unstimulated cells contained 25.4% mono-phosphorylated (MLC-P) and 2.3% di-phosphorylated (MLC-PP) MLC. MLC-PP transiently increased upon thrombin stimulation with a peak (33.3%) at 3min, while MLC-P showed a modest change; 28.9% at 3 min and 33.8% at 20 min. Removal of external Ca2+ decreased MLC-P at both 3 and 20min, while having no effect on MLC-PP. BAPTA loading had no effect on MLC-P at 3 min, while decreasing MLC-P at 20min and MLC-PP. Rho kinase inhibitors, Y27632 and H1152, substantially abolished the thrombin-induced MLC-PP, while they suppressed the MLC-P at 20min. However, MLC-P at 3min was suppressed by H1152 but not Y27632. The results suggest that the thrombin-induced MLC-P and MLC-PP was differentially regulated. MLC-PP was external Ca2+-independent and Rho kinase-dependent. MLC-P at an early phase was external Ca2+-dependent, while MLC-P at a late phase was Ca2+-dependent and Rho kinase-dependent..
11. Myosin light chain (MLC) phosphorylation plays an essential role not only in smooth muscle contraction, but also in endothelial barrier function. Thrombin has been shown to increase MLC phosphorylation, thereby increasing the endothelial permeability. However, the mechanism of the thrombin-induced MLC phosphorylation still remains to be determined. This study investigated the roles of intra- and extra-cellular Ca2+ sources in the thrombin-induced MLC phosphorylation in the porcine aortic endothelial cells. The levels of mono- and di-phosphorylated forms of MLC (MLC-P and MLC-PP, respectively) were separately evaluated by utilizing Phos-tagTM SDS-PAGE and an immunoblot analysis. The unstimulated cells contained 25.4% MLC-P and 2.3% MLC-PP. Upon stimulation with 1 U/mL thrombin, MLC-PP increased to the peak of 33% at 3-5 min, and thereafter declined to the level close to the resting level. However, no significant changes in the level of MLC-P were observed. In the absence of the extracellular Ca2+, the thrombin-induced increase in MLC-PP remained unaffected, while the level of MLC-P was decreased. When a Ca2+ chelator BAPTA was loaded, the resting level of MLC-P significantly decreased. Upon thrombin stimulation, MLC-P increased to the level similar to that seen in the control by 10 min after the stimulation, while it thereafter gradually decreased to the pre-stimulation level. On the other hand, the increase in MLC-PP was substantially but not completely suppressed by BAPTA loading. These observations suggest that most parts of MLC-P and MLC-PP are Ca2+-dependent. However, MLC-PP is totally dependent on the intracellular Ca2+ source, while MLC-P is dependent on the extracellular source..
Membership in Academic Society
  • American Heart Association
  • The Japanese Circulation Society
  • physiological Society of Japan
  • The Japaneese Pharmacological Society
  • The Molecular Biology Society of Japan
  • Japan Society of Smooth Muscle Research
  • Japanese Society for Circulation Research
Educational
Other Educational Activities
  • 2014.08.
  • 2013.10.
  • 2013.03.
  • 2013.01.
  • 2012.01.
  • 2011.07.
  • 2010.11.
  • 2010.11.
  • 2009.11.
  • 2007.07.
  • 2005.03.
  • 2004.03.