Abstract
Subarachnoid hemorrhage (SAH) is a devastating disease. Cerebral vasospasm is still an important cause of post-SAH poor outcomes, but its mechanisms remain unveiled. Activation of epidermal growth factor receptor (EGFR) is suggested to cause vasoconstriction in vitro, but no report has demonstrated the involvement of EGFR in vasospasm development after SAH in vivo. Cross-talk of EGFR and vascular endothelial growth factor (VEGF) receptor, which may affect post-SAH vasospasm, was also reported in cancer cells, but has not been demonstrated in post-SAH vasospasm. The aim of this study was to investigate whether EGFR as well as EGFR-VEGF receptor cross-talk engage in the development of cerebral vasospasm in a mouse SAH model. C57BL6 mice underwent endovascular perforation SAH or sham modeling. At 30 min post-modeling, mice were randomly administrated vehicle or 2 doses of selective EGFR inhibitors intracerebroventricularly. A higher dose of the inhibitor significantly prevented post-SAH neurological impairments at 72 h and vasospasm at 24 h associated with suppression of post-SAH activation of EGFR and extracellular signal-regulated kinase (ERK) 1/2 in the cerebral artery wall, especially in the smooth muscle cell layers. Anti-EGFR neutralizing antibody also showed similar effects. However, neither expression levels of VEGF nor activation levels of a major receptor of VEGF, VEGF receptor-2, were affected by SAH and two kinds of EGFR inactivation. Thus, this study first showed that EGFR-ERK1/2 pathways may be involved in post-SAH vasospasm development, and that EGFR-VEGF receptor cross-talk may not play a significant role in the development of vasospasm in mice.
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References
Suzuki H, Shiba M, Nakatsuka Y, Nakano F, Nishikawa H (2017) Higher cerebrospinal fluid pH may contribute to the development of delayed cerebral ischemia after aneurysmal subarachnoid hemorrhage. Transl Stroke Res 8:165–173
Fujimoto M, Shiba M, Kawakita F, Liu L, Shimojo N, Imanaka-Yoshida K, Yoshida T, Suzuki H (2018) Effects of tenascin-C knockout on cerebral vasospasm after experimental subarachnoid hemorrhage in mice. Mol Neurobiol 55:1951–1958
Borel CO, McKee A, Parra A, Haglund MM, Solan A, Prabhakar V, Sheng H, Warner DS et al (2003) Possible role for vascular cell proliferation in cerebral vasospasm after subarachnoid hemorrhage. Stroke 34:427–433
McGirt MJ, Lynch JR, Blessing R, Warner DS, Friedman AH, Laskowitz DT (2002) Serum von Willebrand factor, matrix metalloproteinase-9, and vascular endothelial growth factor levels predict the onset of cerebral vasospasm after aneurysmal subarachnoid hemorrhage. Neurosurgery 51:1128–1135
Yan J, Chen C, Lei J, Yang L, Wang K, Liu J, Zhou C (2006) 2-methoxyestradiol reduces cerebral vasospasm after 48 hours of experimental subarachnoid hemorrhage in rats. Exp Neurol 202:348–356
Midwood KS, Hussenet T, Langlois B, Orend G (2011) Advances in tenascin-C biology. Cell Mol Life Sci 68:3175–3199
Suzuki H, Kinoshita N, Imanaka-Yoshida K, Yoshida T, Taki W (2008) Cerebrospinal fluid tenascin-C increased preceding the development of chronic shunt-dependent hydrocephalus after subarachnoid hemorrhage. Stroke 39:1610–1612
Fujimoto M, Shiba M, Kawakita F, Liu L, Nakasaki A, Shimojo N, Imanaka-Yoshida K, Yoshida T et al (2016) Epidermal growth factor-like repeats of tenascin-C-induced constriction of cerebral arteries via activation of epidermal growth factor receptors in rats. Brain Res 1642:436–444
Kawanabe Y, Masaki T, Hashimoto N (2004) Involvement of epidermal growth factor receptor-protein tyrosine kinase transactivation in endothelin-1-induced vascular contraction. J Neurosurg 100:1066–1071
Liu L, Fujimoto M, Kawakita F, Nakano F, Imanaka-Yoshida K, Yoshida T, Suzuki H (2016) Anti-vascular endothelial growth factor treatment suppressed early brain injury after subarachnoid hemorrhage in mice. Mol Neurobiol 53:4529–4538
Ray L, Mathieu M, Jespers P, Hadad I, Mahmoudabady M, Pensis A, Motte S, Peters IR et al (2008) Early increase in pulmonary vascular reactivity with overexpression of endothelin-1 and vascular endothelial growth factor in canine experimental heart failure. Exp Physiol 93:434–442
Adeoye OO, Butler SM, Hubbell MC, Semotiuk A, Williams JM, Pearce WJ (2013) Contribution of increased VEGF receptors to hypoxic changes in fetal ovine carotid artery contractile proteins. Am J Physiol Cell Physiol 304:C656–C665
Larsen AK, Ouaret D, El Ouadrani K, Petitprez A (2011) Targeting EGFR and VEGF(R) pathway cross-talk in tumor survival and angiogenesis. Pharmacol Ther 131:80–90
Shim JW, Madsen JR (2018) VEGF signaling in neurological disorders. Int J Mol Sci 19:275
Shim JW, Sandlund J, Hameed MQ, Blazer-Yost B, Zhou FC, Klagsbrun M, Madsen JR (2016) Excess HB-EGF, which promotes VEGF signaling, leads to hydrocephalus. Sci Rep 6:26794
Li XM, Su F, Ji MH, Zhang GF, Qiu LL, Jia M, Gao J, Xie Z et al (2014) Disruption of hippocampal neuregulin 1-ErbB4 signaling contributes to the hippocampus-dependent cognitive impairment induced by isoflurane in aged mice. Anesthesiology 121:79–88
Formisano L, D’Amato V, Servetto A, Brillante S, Raimondo L, Di Mauro C, Marciano R, Orsini RC et al (2015) Src inhibitors act through different mechanisms in non-small cell lung cancer models depending on EGFR and RAS mutational status. Oncotarget 6:26090–26103
Kawakita F, Fujimoto M, Liu L, Nakano F, Nakatsuka Y, Suzuki H (2017) Effects of Toll-like receptor 4 antagonists against cerebral vasospasm after experimental subarachnoid hemorrhage in mice. Mol Neurobiol 54:6624–6633
Sabri M, Ai J, Knight B, Tariq A, Jeon H, Shang X, Marsden PA, Loch Macdonald R (2011) Uncoupling of endothelial nitric oxide synthase after experimental subarachnoid hemorrhage. J Cereb Blood Flow Metab 31:190–199
Schneider CA, Rasband WS, Eliceiri KW (2012) NIH image to image J: 25 years of image analysis. Nat Methods 9:671–675
Karkkainen MJ, Petrova TV (2000) Vascular endothelial growth factor receptors in the regulation of angiogenesis and lymphangiogenesis. Oncogene 19:5598–5605
Wang WM, Zhao ZL, Ma SR, Yu GT, Liu B, Zhang L, Zhang WF, Kulkarni AB et al (2015) Epidermal growth factor receptor inhibition reduces angiogenesis via hypoxia-inducible factor-1α and Notch1 in head neck squamous cell carcinoma. PLoS One 10:e0119723
Ledón N, Casacó A, Casanova E, Beausoleil I (2011) Comparative analysis of binding affinities to epidermal growth factor receptor of monoclonal antibodies nimotuzumab and cetuximab using different experimental animal models. Placenta 32:531–534
Zervas NT, Liszczak TM, Mayberg MR, Black PM (1982) Cerebrospinal fluid may nourish cerebral vessels through pathways in the adventitia that may be analogous to systemic vasa vasorum. J Neurosurg 56:475–481
Langheinrich AC, Michniewicz A, Bohle RM, Ritman EL (2007) Vasa vasorum neovascularization and lesion distribution among different vascular beds in ApoE-/-/LDL-/- double knockout mice. Atherosclerosis 191:73–81
Bedussi B, van Lier MG, Bartstra JW, de Vos J, Siebes M, VanBavel E, Bakker EN (2015) Clearance from the mouse brain by convection of interstitial fluid towards the ventricular system. Fluids Barriers CNS 12:23
Bedussi B, van der Wel NN, de Vos J, van Veen H, Siebes M, VanBavel E, Bakker EN (2017) Paravascular channels, cisterns, and the subarachnoid space in the rat brain: a single compartment with preferential pathways. J Cereb Blood Flow Metab 37:1374–1385
Abbott NJ (2004) Evidence for bulk flow of brain interstitial fluid: significance for physiology and pathology. Neurochem Int 45:545–552
Makki N, Thiel KW, Miller FJ Jr (2013) The epidermal growth factor receptor and its ligands in cardiovascular disease. Int J Mol Sci 14:20597–20613
Kalmes A, Daum G, Clowes AW (2001) EGFR transactivation in the regulation of SMC function. Ann N Y Acad Sci 947:42–55
Kim J, Lee CK, Park HJ, Kim HJ, So HH, Lee KS, Lee HM, Roh HY et al (2006) Epidermal growth factor induces vasoconstriction through the phosphatidylinositol 3-kinase-mediated mitogen-activated protein kinase pathway in hypertensive rats. J Pharmacol Sci 101:135–143
Chansel D, Ciroldi M, Vandermeersch S, Jackson LF, Gomez AM, Henrion D, Lee DC, Coffman TM et al (2006) Heparin binding EGF is necessary for vasospastic response to endothelin. FASEB J 20:1936–1938
Koide M, Penar PL, Tranmer BI, Wellman GC (2007) Heparin-binding EGF-like growth factor mediates oxyhemoglobin-induced suppression of voltage-dependent potassium channels in rabbit cerebral artery myocytes. Am J Physiol Heart Circ Physiol 293:H1750–H1759
Koide M, Wellman GC (2015) SAH-induced MMP activation and K V current suppression is mediated via both ROS-dependent and ROS-independent mechanisms. Acta Neurochir Suppl 120:89–94
Koide M, Wellman GC (2013) SAH-induced suppression of voltage-gated K(+) (K(V)) channel currents in parenchymal arteriolar myocytes involves activation of the HB-EGF/EGFR pathway. Acta Neurochir Suppl 115:179–184
Friedrich B, Müller F, Feiler S, Schöller K, Plesnila N (2012) Experimental subarachnoid hemorrhage causes early and long-lasting microarterial constriction and microthrombosis: an in-vivo microscopy study. J Cereb Blood Flow Metab 32:447–455
Liu H, Dienel A, Schöller K, Schwarzmaier SM, Nehrkorn K, Plesnila N, Terpolilli NA (2018) Microvasospasms after experimental subarachnoid hemorrhage do not depend on endothelin A receptors. Stroke 49:693–699
Suzuki H, Hasegawa Y, Kanamaru K, Zhang JH (2010) Mechanisms of osteopontin-induced stabilization of blood-brain barrier disruption after subarachnoid hemorrhage in rats. Stroke 41:1783–1790
Macdonald RL, Higashida RT, Keller E, Mayer SA, Molyneux A, Raabe A, Vajkoczy P, Wanke I et al (2011) Clazosentan, an endothelin receptor antagonist, in patients with aneurysmal subarachnoid hemorrhage undergoing surgical clipping: a randomised, double-blind, placebo-controlled phase 3 trial (CONSCIOUS-2). Lancet Neurol 10:618–625
Pore N, Jiang Z, Gupta A, Cerniglia G, Kao GD, Maity A (2006) EGFR tyrosine kinase inhibitors decrease VEGF expression by both hypoxia-inducible factor (HIF)-1-independent and HIF-1-dependent mechanisms. Cancer Res 66:3197–3204
Miller CA, Lombard FW, Wu CT, Hubbard CJ, Silbajoris L, Borel CO, Niklason LE (2006) Role of vascular mitogens in subarachnoid hemorrhage-associated cerebral vasculopathy. Neurocrit Care 5:215–221
Acknowledgments
We thank Ms. Chiduru Yamamoto-Nakamura (Department of Neurosurgery, Mie University Graduate School of Medicine) for her assistance with administrative support.
Funding
This study was funded by a grant-in-aid for Scientific Research from Japan Society for the Promotion of Science to Drs. Suzuki [17K10825] and Shiba [17K16640].
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The authors declare that they have no conflict of interest.
Research Involving Animals
All procedures were approved by the Animal Ethics Review Committee of Mie University and were carried out according to the institution’s Guidelines for Animal Experiments.
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Nakano, F., Kawakita, F., Liu, L. et al. Anti-vasospastic Effects of Epidermal Growth Factor Receptor Inhibitors After Subarachnoid Hemorrhage in Mice. Mol Neurobiol 56, 4730–4740 (2019). https://doi.org/10.1007/s12035-018-1400-6
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DOI: https://doi.org/10.1007/s12035-018-1400-6