MECHANISMS OF HEMORRHAGE AND EDEMA IN TPA STROKE THERAPY

Summary

Principal Investigator: ENG LO
Abstract: A major problem with tissue plasminogen activator (tPA) therapy for ischemic stroke is the complication of hemorrhage and edema. In our previous grant, we established a quantitative model of tPA-induced hemorrhage in rodent embolic (clot-based) focal ischemia, and showed an overall role for oxidative damage. In this renewal application, we focus on novel signaling mechanisms that are involved. Our overall hypothesis is that after stroke, tPA binds the Low-density-lipoprotein Related Protein (LRP). LRP is a receptor present on neurons; astrocytes and endothelial cells which can trigger MAP kinase signaling cascades, resulting in upregulation of matrix metalloproteinase-9 (MMP-9) in the neurovascular unit (vascular/ astrocytic/neuronal compartments). MMP-9 degrades critical neurovascular matrix substrates that weaken blood vessel integrity and lead to leakage and rupture. To test this overall hypothesis, we will pursue 3 Specific Aims. In Aim 1, we will examine LRP signaling pathways that upregulate MMP-9 activity after tPA therapy. To do this, we will use in vitro and in vivo models to document the following events: increased levels of LRP after hypoxia/ischemia, tPA-LRP triggering of MAP kinase, activation of AP1 promoter transcription of MMP-9. In vitro experiments will use neurons, astrocytes, and cerebral endothelial cells. In vivo experiments will use the tPA-induced hemorrhage model in rats. In Aim 2, we wiI1 document a specific role for MMP-9. After focal ischemia, we will compare the severity of tPA-induced edema and hemorrhage in wildtype mice versus MMP-2 knockouts, MMP-7 knockouts, MMP-9 knockouts, and transgenic overexpressors of TIMP1 (an inhibitor of MMPs). In Aim 3, we develop novel imaging methods to correlate the effects of tPA, MMP-9 activation, and vascular leakage. We will construct an MMP-9 specific probe that enables near-infrared fluorescence imaging of MMP-9 in vivo after tPA. Cranial window imaging experiments in rat and mouse embolic focal ischemia will correlate in real-time MMP-9 upregulation in the neurovascular unit with the onset of vascular leakage. At the NINDS Stroke Program Review Group, proteolytic pathology in the neurovascular unit was identified as a high priority research area. This proposal should fit the goals of the Stroke PRG because it examines novel mechanisms that link tPA, cell signaling, MMPs, and hemorrhagic transformation in the neurovascular unit.
Funding Period: 1998-01-01 - 2008-09-25
more information: NIH RePORT

Top Publications

  1. pmc Proteasome inhibition protects HT22 neuronal cells from oxidative glutamate toxicity
    Klaus van Leyen
    Neuroprotection Research Laboratory, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA 02129, USA
    J Neurochem 92:824-30. 2005
  2. pmc Experimental model of warfarin-associated intracerebral hemorrhage
    Christian Foerch
    Neuroprotection Research Laboratory, Massachusetts General Hospital, Harvard Medical School, Charlestown, Masschusetts 02129, USA
    Stroke 39:3397-404. 2008
  3. pmc Vascular endothelial growth factor regulates the migration of oligodendrocyte precursor cells
    Kazuhide Hayakawa
    Neuroprotection Research Laboratory, Department of Radiology, Massachusetts General Hospital and Harvard Medical School, Charlestown, Massachusetts 02129, USA
    J Neurosci 31:10666-70. 2011
  4. pmc Matrix metalloprotease regulation of neuropathic pain
    Ru Rong Ji
    Pain Research Center, Department of Anesthesiology, Brigham and Women s Hospital and Harvard Medical School, Boston, MA 02115, USA
    Trends Pharmacol Sci 30:336-40. 2009
  5. pmc Edaravone, a free radical scavenger, protects components of the neurovascular unit against oxidative stress in vitro
    Brian J Lee
    Neuroprotection Research Laboratory, Department of Radiology, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA
    Brain Res 1307:22-7. 2010
  6. pmc Annexin A2 combined with low-dose tPA improves thrombolytic therapy in a rat model of focal embolic stroke
    Haihao Zhu
    Department of Neurology, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts 02129, USA
    J Cereb Blood Flow Metab 30:1137-46. 2010
  7. pmc Brain angiogenesis in developmental and pathological processes: neurovascular injury and angiogenic recovery after stroke
    Ken Arai
    Neuroprotection Research Laboratory, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA 02129, USA
    FEBS J 276:4644-52. 2009
  8. pmc Distinct roles of matrix metalloproteases in the early- and late-phase development of neuropathic pain
    Yasuhiko Kawasaki
    Pain Research Center, Department of Anesthesiology, Brigham and Women s Hospital and Harvard Medical School, 75 Francis Street, Medical Research Building, Room 604, Boston, Massachusetts 02115, USA
    Nat Med 14:331-6. 2008
  9. ncbi Beyond NMDA and AMPA glutamate receptors: emerging mechanisms for ionic imbalance and cell death in stroke
    Elaine Besancon
    Neuroprotection Research Laboratory, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02129, USA
    Trends Pharmacol Sci 29:268-75. 2008
  10. pmc Effects of neuroglobin overexpression on acute brain injury and long-term outcomes after focal cerebral ischemia
    Xiaoying Wang
    Neuroprotection Research Laboratory, Departments of Neurology and Radiology, Massachusetts General Hospital, 149 13th Street, Room 2411A, Charlestown, MA 02129, USA
    Stroke 39:1869-74. 2008

Scientific Experts

  • Josephine M Lok
  • Xiaoying Wang
  • Ru Rong Ji
  • C Foerch
  • Michael Whalen
  • Peter J Kelly
  • ENG LO
  • Aneesh Singhal
  • Klaus van Leyen
  • Ken Arai
  • Bing Qiao Zhao
  • Emiri Tejima
  • Sun Ryung Lee
  • Shuzhen Guo
  • Kiyoshi Tsuji
  • Brian J Lee
  • Guang Jin
  • Changhong Xing
  • Yoshihiro Murata
  • Elaine Besancon
  • Woo Jean Kim
  • A Rosell
  • Joan Montaner
  • Sophia Wang
  • Kazuhide Hayakawa
  • Haihao Zhu
  • Yong Guang Yang
  • Young R Kim
  • Yasuhiko Kawasaki
  • Kenneth J Rhodes
  • Robert H Scannevin
  • Anna Rosell
  • Y Murata
  • Bruce R Rosen
  • S R Lee
  • Hwa Kyoung Shin
  • Shu Zhen Guo
  • Seong Ryong Lee
  • Hahn Young Kim
  • M Ning
  • Wellington Pham
  • Angel T Som
  • Loc Duyen D Pham
  • Katherine A Hajjar
  • Jianxiang Liu
  • Xiang Fan
  • Yasuhiro Egi
  • Jie Lu
  • Zhanyang Yu
  • Song Zhao
  • Sunryung Lee
  • Xunming Ji
  • Zhen Zhong Xu
  • Shoukat Dedhar
  • Kristine Roy
  • Ping Heng Tan
  • Yoshi Murata
  • Gabriel Corfas
  • Zhi Ye Zhuang
  • Maurits P A van Meer
  • Yong Jing Gao
  • Michael Tymianski
  • Jong Yeon Park
  • De Maw Chuang
  • Joseph B Mandeville
  • Bing Hao Luo
  • Monique F Stins
  • George Dai
  • R Gilberto Gonzalez
  • Cenk Ayata
  • P L Huang
  • Phillip B Jones
  • Brian C Magliaro
  • H Y Kim
  • M A Moskowitz
  • David A Boas
  • D Atochin
  • C Ayata
  • Michael A Moskowitz
  • Andrew K Dunn
  • W J Koroshetz
  • David J Mooney
  • Hannah Storrie
  • Pradeep Bhide
  • K L Furie
  • M Zhu
  • M Lederer
  • A G Sorensen
  • H Lee
  • Jack M Parent

Detail Information

Publications36

  1. pmc Proteasome inhibition protects HT22 neuronal cells from oxidative glutamate toxicity
    Klaus van Leyen
    Neuroprotection Research Laboratory, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA 02129, USA
    J Neurochem 92:824-30. 2005
    ..These findings suggest that caspases can be decoupled from oxidative stress under some conditions, and implicate the ubiquitin/proteasome pathway in neuronal cell death caused by oxidative glutamate toxicity...
  2. pmc Experimental model of warfarin-associated intracerebral hemorrhage
    Christian Foerch
    Neuroprotection Research Laboratory, Massachusetts General Hospital, Harvard Medical School, Charlestown, Masschusetts 02129, USA
    Stroke 39:3397-404. 2008
    ..This study describes the development of a mouse model of W-ICH in which hematoma growth and outcomes can be correlated with anticoagulation parameters...
  3. pmc Vascular endothelial growth factor regulates the migration of oligodendrocyte precursor cells
    Kazuhide Hayakawa
    Neuroprotection Research Laboratory, Department of Radiology, Massachusetts General Hospital and Harvard Medical School, Charlestown, Massachusetts 02129, USA
    J Neurosci 31:10666-70. 2011
    ..Our findings demonstrate that VEGF-A can induce OPC migration via an ROS- and FAK-dependent mechanism, and suggest a novel role for VEGF-A in white-matter maintenance and homeostasis...
  4. pmc Matrix metalloprotease regulation of neuropathic pain
    Ru Rong Ji
    Pain Research Center, Department of Anesthesiology, Brigham and Women s Hospital and Harvard Medical School, Boston, MA 02115, USA
    Trends Pharmacol Sci 30:336-40. 2009
    ..Inhibition of MMP-9 or MMP-2 might provide a new strategy for the prevention and treatment of neuropathic pain...
  5. pmc Edaravone, a free radical scavenger, protects components of the neurovascular unit against oxidative stress in vitro
    Brian J Lee
    Neuroprotection Research Laboratory, Department of Radiology, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA
    Brain Res 1307:22-7. 2010
    ..Edaravone significantly ameliorated this response. These data suggest that free radical scavengers are effective in all cell types of the neurovascular unit, and should still be considered as a potential therapeutic approach for stroke...
  6. pmc Annexin A2 combined with low-dose tPA improves thrombolytic therapy in a rat model of focal embolic stroke
    Haihao Zhu
    Department of Neurology, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts 02129, USA
    J Cereb Blood Flow Metab 30:1137-46. 2010
    ..As a result, brain hemorrhage and infarction are reduced, and the time window for stroke reperfusion prolonged. Our present findings provide a promising new approach for enhancing tPA-based thrombolytic stroke therapy...
  7. pmc Brain angiogenesis in developmental and pathological processes: neurovascular injury and angiogenic recovery after stroke
    Ken Arai
    Neuroprotection Research Laboratory, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA 02129, USA
    FEBS J 276:4644-52. 2009
    ..Understanding how neurovascular signals and substrates make the transition from initial injury to angiogenic recovery will be important if we are to find new therapeutic approaches for stroke...
  8. pmc Distinct roles of matrix metalloproteases in the early- and late-phase development of neuropathic pain
    Yasuhiko Kawasaki
    Pain Research Center, Department of Anesthesiology, Brigham and Women s Hospital and Harvard Medical School, 75 Francis Street, Medical Research Building, Room 604, Boston, Massachusetts 02115, USA
    Nat Med 14:331-6. 2008
    ..Inhibition of MMP may provide a novel therapeutic approach for the treatment of neuropathic pain at different phases...
  9. ncbi Beyond NMDA and AMPA glutamate receptors: emerging mechanisms for ionic imbalance and cell death in stroke
    Elaine Besancon
    Neuroprotection Research Laboratory, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02129, USA
    Trends Pharmacol Sci 29:268-75. 2008
    ..Further in vivo validation of these pathways could ultimately lead us to new therapeutic targets for stroke, trauma and neurodegeneration...
  10. pmc Effects of neuroglobin overexpression on acute brain injury and long-term outcomes after focal cerebral ischemia
    Xiaoying Wang
    Neuroprotection Research Laboratory, Departments of Neurology and Radiology, Massachusetts General Hospital, 149 13th Street, Room 2411A, Charlestown, MA 02129, USA
    Stroke 39:1869-74. 2008
    ..Emerging data suggest that neuroglobin (Ngb) may protect against hypoxic/ischemic neuronal insults. However, the underlying mechanisms in vivo and implications for long-term outcomes are still not well understood...
  11. pmc Neuroprotection via matrix-trophic coupling between cerebral endothelial cells and neurons
    Shuzhen Guo
    Neuroprotection Research Laboratory, Departments of Neurology and Radiology, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA 02129
    Proc Natl Acad Sci U S A 105:7582-7. 2008
    ..Targeting these signals of matrix and trophic coupling between endothelium and neurons may provide new therapeutic opportunities for stroke and other CNS disorders...
  12. ncbi Targeting extracellular matrix proteolysis for hemorrhagic complications of tPA stroke therapy
    Xiaoying Wang
    Department of Neurology, Massachusetts General Hospital, Charlestown, MA 02129, USA
    CNS Neurol Disord Drug Targets 7:235-42. 2008
    ..The data strongly suggest that targeting the extracellular matrix proteolytic imbalance within the neurovascular unit may provide new approaches for improving the safety and efficacy of thrombolytic reperfusion therapy of stroke...
  13. pmc Neuregulin-1 signaling in brain endothelial cells
    Josephine Lok
    Neuroprotection Research Laboratory, Department of Radiology, Massachusetts General Hospital, Charlestown, MA 02129, USA
    J Cereb Blood Flow Metab 29:39-43. 2009
    ..These data suggest that NRG1 signaling is functional and cytoprotective in BMECs...
  14. pmc Extension of the thrombolytic time window with minocycline in experimental stroke
    Yoshihiro Murata
    Neuroprotection Res Lab, Charlestown, MA 02129, USA
    Stroke 39:3372-7. 2008
    ..In this study, we ask whether minocycline can prevent tPA-associated cerebral hemorrhage and extend the reperfusion window in an experimental stroke model in rats...
  15. pmc Role of oxidative stress and caspase 3 in CD47-mediated neuronal cell death
    Changhong Xing
    Neuroprotection Research Laboratory, Department of Radiology and Neurology, Massachusetts General Hospital, Harvard Medical School, MA 02129, USA
    J Neurochem 108:430-6. 2009
    ..We conclude that CD47 mediates neuronal cell death through caspase-dependent and caspase-independent pathways...
  16. ncbi Mechanisms and markers for hemorrhagic transformation after stroke
    A Rosell
    Neuroprotection Research Laboratory, Department of Radiology, Massachusetts General Hospital, and Program in Neuroscience, Harvard Medical School, Boston, MA, USA
    Acta Neurochir Suppl 105:173-8. 2008
    ....
  17. pmc CD47 gene knockout protects against transient focal cerebral ischemia in mice
    Guang Jin
    Department of Radiology and Neurology, Neuroprotection Research Laboratory, Massachusetts General Hospital, Harvard Medical School, Massachusetts 02129, USA
    Exp Neurol 217:165-70. 2009
    ..We conclude that CD47 is broadly involved in neuroinflammation, and this integrin-associated-protein plays a role in promoting MMP-9 upregulaton, neutrophil extravasation, brain swelling and progression of acute ischemic brain injury...
  18. pmc Functional MRI of delayed chronic lithium treatment in rat focal cerebral ischemia
    Young R Kim
    Athinoula Martinos Center for Biomedical Imaging Massachusetts General Hospital, 149 13th Street, Room 2301, Charlestown, MA 02129, USA
    Stroke 39:439-47. 2008
    ..We further investigated neurohemodynamic aspects of the treatment-associated recovery by assessing the therapeutic efficacy of delayed chronic lithium treatment using functional MRI...
  19. pmc Advances in emerging nondrug therapies for acute stroke 2007
    Aneesh B Singhal
    Neuroprotection Research Laboratory, Harvard Medical School, Department of Neurology, Massachusetts General Hospital, Boston, MA 02114, USA
    Stroke 39:289-91. 2008
  20. ncbi Crossing the blood-brain barrier: a potential application of myristoylated polyarginine for in vivo neuroimaging
    Wellington Pham
    MGH MIT HMS Athinoula A Martinos Center for Biomedical Imaging, Department of Radiology, Massachusetts General Hospital and Harvard Medical School, Charlestown, 02129, USA
    Neuroimage 28:287-92. 2005
    ..We conclude that this approach can be used for the delivery of imaging probes and potentially targeted therapeutics across the BBB...
  21. ncbi Tissue plasminogen activator promotes matrix metalloproteinase-9 upregulation after focal cerebral ischemia
    Kiyoshi Tsuji
    Department of Neurology, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA, USA
    Stroke 36:1954-9. 2005
    ..Here, we used a combination of pharmacological and genetic approaches to show that tPA promotes MMP-9 levels in stroke in vivo...
  22. ncbi Role of matrix metalloproteinases in delayed cortical responses after stroke
    Bing Qiao Zhao
    Neuroprotection Research Laboratory, Department of Radiology, Massachusetts General Hospital, MGH East 149 2401, Charlestown, Massachusetts 02129, USA
    Nat Med 12:441-5. 2006
    ..These data suggest that, contrary to MMP inhibitor therapies for acute stroke, strategies that modulate MMPs may be needed for promoting stroke recovery...
  23. ncbi Involvement of matrix metalloproteinase in neuroblast cell migration from the subventricular zone after stroke
    Seong Ryong Lee
    Department of Neurology and Radiology, Massachusetts General Hospital, Boston, Massachusetts 02129, USA
    J Neurosci 26:3491-5. 2006
    ..These data suggest that MMPs are involved in endogenous mechanisms of neurogenic migration as the brain seeks to heal itself after injury...
  24. ncbi Association between tPA therapy and raised early matrix metalloproteinase-9 in acute stroke
    M Ning
    Stroke Service, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA
    Neurology 66:1550-5. 2006
    ..The authors investigated plasma MMP9 and its endogenous inhibitor, tissue inhibitor of metalloproteinase (TIMP1), in tPA-treated and -untreated stroke patients...
  25. ncbi Reduction of tissue plasminogen activator-induced matrix metalloproteinase-9 by simvastatin in astrocytes
    Sophia Wang
    Neuroprotection Research Laboratory, Department of Radiology, Massachusetts General Hospital, and Program in Neuroscience, Harvard Medical School, Boston, Mass 02129, USA
    Stroke 37:1910-2. 2006
    ..Here, we test the idea that statins may directly ameliorate tPA-induced MMP-9 dysregulation...
  26. ncbi Astrocytic induction of matrix metalloproteinase-9 and edema in brain hemorrhage
    Emiri Tejima
    Neuroprotection Research Laboratory, Department of Radiology, Massachusetts General Hospital, and Program in Neuroscience, Harvard Medical School, Boston, MA, USA
    J Cereb Blood Flow Metab 27:460-8. 2007
    ....
  27. ncbi Neurovascular proteases in brain injury, hemorrhage and remodeling after stroke
    Bing Qiao Zhao
    Neuroprotection Research Laboratory, Department of Radiology, Massachusetts General Hospital, Charlestown, MA 02129, USA
    Stroke 38:748-52. 2007
    ..Acute versus chronic protease profiles within the neurovascular unit are likely to underlie critical responses to stroke, therapy, and recovery...
  28. ncbi Induction of matrix metalloproteinase, cytokines and chemokines in rat cortical astrocytes exposed to plasminogen activators
    Sun Ryung Lee
    Neuroprotection Research Laboratory, Departments of Radiology and Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA
    Neurosci Lett 417:1-5. 2007
    ..These data suggest that plasminogen activators may trigger selected pro-inflammatory responses at the neurovascular interface. Whether these effects influence thrombolytic stroke therapy warrants further investigation...
  29. ncbi Cell-cell signaling in the neurovascular unit
    Josephine Lok
    Neuroprotection Research Laboratory, Department of Radiology, Massachusetts General Hospital, MGH East 149 2401, Charlestown, MA 02129, USA
    Neurochem Res 32:2032-45. 2007
    ....
  30. pmc Normobaric hyperoxia improves cerebral blood flow and oxygenation, and inhibits peri-infarct depolarizations in experimental focal ischaemia
    Hwa Kyoung Shin
    Stroke and Neurovascular Regulation Laboratory, Department of Radiology, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA 02129, USA
    Brain 130:1631-42. 2007
    ..Our data show that normobaric hyperoxia increases tissue O2 delivery, and that novel mechanisms such as CBF augmentation, and suppression of PIDs may afford neuroprotection during hyperoxia...
  31. pmc Acute plasmalemma permeability and protracted clearance of injured cells after controlled cortical impact in mice
    Michael J Whalen
    Neuroscience Center, Massachusetts General Hospital, Charlestown, MA 02129, USA
    J Cereb Blood Flow Metab 28:490-505. 2008
    ..The data suggest that plasmalemma damage is a fundamental marker of cellular injury after CCI; some injured cells might have an extended window for potential rescue by neuroprotective strategies...
  32. pmc Reduction of hippocampal cell death and proteolytic responses in tissue plasminogen activator knockout mice after transient global cerebral ischemia
    S R Lee
    Neuroprotection Research Laboratory, Department of Radiology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02129, USA
    Neuroscience 150:50-7. 2007
    ..Taken together, these data suggest that endogenous tPA contributes to hippocampal injury after cerebral ischemia, and these pathophysiologic pathways may involve links to aberrant activation of caspases and MMPs...
  33. ncbi Oxidative stress and matrix metalloproteinase-9 in acute ischemic stroke: the Biomarker Evaluation for Antioxidant Therapies in Stroke (BEAT-Stroke) study
    Peter J Kelly
    Neurovascular Clinical Science Unit, Mater University Hospital and University College Dublin, Ireland
    Stroke 39:100-4. 2008
    ..We aimed to determine the change in plasma F2IP levels over time and relationship with plasma MMP-9 in tPA-treated and tPA-untreated stroke patients...
  34. pmc Experimental models, neurovascular mechanisms and translational issues in stroke research
    E H Lo
    Neuroprotection Research Laboratory, Department of Radiology and Neurology, Massachusetts General Hospital, Charlestown, MA, USA
    Br J Pharmacol 153:S396-405. 2008
    ..Ultimately, both bench-to-bedside and bedside-back-to-bench interactions may be required to overcome the translational hurdles for this challenging disease...
  35. ncbi Effect of neuregulin-1 on histopathological and functional outcome after controlled cortical impact in mice
    Josephine Lok
    Neuroprotection Research Laboratory, Department of Radiology, Massachusetts General Hospital, and Program in Neuroscience, Harvard Medical School, Boston, Massachusetts, USA
    J Neurotrauma 24:1817-22. 2007
    ..Further studies are therefore warranted to more carefully explore molecular mechanisms, dose-responses, and relationships between morphological outcome and long-term recovery...