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Proteomic analysis of protein changes in plasma by balloon test occlusion
Journal of Clinical Neuroscience xxx (xxxx) xxx
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Experimental study
Proteomic analysis of protein changes in plasma by balloon test occlusion Tomohiko Ozaki a,b,c, Rieko Muramatsu a,d,e,⇑, Hajime Nakamura b, Manabu Kinoshita c, Haruhiko Kishima b, Toshihide Yamashita a,d,f a
Department of Molecular Neuroscience, Graduate School of Medicine, Osaka University, Osaka, Japan Department of Neurosurgery, Graduate School of Medicine, Osaka University, Japan Department of Neurosurgery, Osaka International Cancer Institute, Osaka, Japan d WPI Immunology Frontier Research Center, Osaka University, Japan e Department of Molecular Pharmacology, National Institute of Neuroscience, National Center of Neurology and Neurology and Psychiatry, Kodaira, Japan f Department of Molecular Neuroscience, Graduate School of Frontier Biosciences, Osaka University, Japan b c
a r t i c l e
i n f o
Article history: Received 31 August 2019 Accepted 1 December 2019 Available online xxxx Keywords: Ischemic preconditioning Stroke Human Vascular endothelial cell Neuron
a b s t r a c t Transient ischemia provides the tolerance against prolonged ischemia in the brain. In mouse experimental model, transient ischemia changes the composition ratio of circulating proteins, which associate with neuroprotection; however, the human evidence is lacking. Here we mimicked balloon test occlusion (BTO) of carotid artery as a transient ischemia and investigated the change of composition ratio of the circulating protein in the human plasma. We collected blood samples from nine patients (5 men and 4 women; mean age 64.2 years; range 45 to 77 years) before and 48 h after BTO and investigated the changes of circulating molecules level in the proteome using LC-MS/MS analysis. Leucine-rich alpha-2glycoprotein and serum amyloid A-1 increased and protein AMBP decreased in the blood samples after BTO. Transient change of blood flow in the brain alters molecular expression in the plasma. Because the alteration of plasma protein composition is involved in ischemic tolerance in animal models, the proteins whose level was changed by BTO may be also involved in neuroprotection against ischemia in human. Ó 2019 Published by Elsevier Ltd.
1. Introduction The absolute number of new stroke events was approximately 11.6 million in 2010, [1] and the number of ischemic stroke events is expected to increase to 23 million by 2030. [2] Although the therapeutic approaches, such as administration of tissue-type plasminogen activator (tPA) and endovascular techniques, are approved for acute stroke treatment [3,4], these therapies has serious time limitation (within hours after onset of stroke) and more 50% patients with stroke are not able to achieve the benefits [5]. Therefore, developing the pharmacological interventions which aim to reduce neuronal damage have been attributed from pharmacological companies [6,7]. Some of the drugs were aimed trapping free radical after stroke and had been reached to clinical trial, but with little proof of effectiveness [8]. Therefore, the other mech-
⇑ Corresponding author at: Department of Molecular Pharmacology, National Institute of Neuroscience, National Center of Neurology and Neurology and Psychiatry, Kodaira, Japan. E-mail address: [email protected] (R. Muramatsu).
anisms which exert neuroprotection may be useful for developing the therapy for stroke. Ischemic preconditioning (IPC) is an endogenous protective mechanism and is proved at mammalian experimental models [9,10] and is considered conserved in human [11]. Short sublethal ischemia exposure is known as one of the IPC, and its renders the brain more resistance against prolonged ischemia. Because of the ethical issue, the underlying the molecular mechanism of IPC has been anticipated to apply the drug development. In the molecular mechanism of IPC, almost of all research have been focusing on the role of the molecular mechanims which is detectable in the central nervous system, such as astrocytic P2 receptor activation-mediated neuroprotective functions [12]. In contrast, it is reported that the IPC-mediated neuroprotection depends on the change of expression of vascular-derived osteopontin release in mouse experimental model, and this change was also detectable in the plasma of the mice [13]. In addition, change of vascular-derived molecules release was also detected in circulation in the plasma obtained from the mice after transient ischemia. Therefore, in this study,
https://doi.org/10.1016/j.jocn.2019.12.005 0967-5868/Ó 2019 Published by Elsevier Ltd.
Please cite this article as: T. Ozaki, R. Muramatsu, H. Nakamura et al., Proteomic analysis of protein changes in plasma by balloon test occlusion, Journal of Clinical Neuroscience, https://doi.org/10.1016/j.jocn.2019.12.005
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T. Ozaki et al. / Journal of Clinical Neuroscience xxx (xxxx) xxx
we challenged wheatear our observation obtained from mouse were also conserved in human. Here we used the blood sample from the patients with BTO, a procedure to stop blood flow by a balloon catheter in an artery. We mimicked the BTO as a transient ischemia and investigated the changes of composition ratio of circulating protein before and after BTO. We found that transient hypoperfusion by BTO causes changes of protein levels. We also analyzed the protein level in the plasma by the difference of sex and age. 2. Methods 2.1. Patients We collected data from 9 serial cancer patients (5 men and 4 women; mean age 64.2 years; range 45 to 77 years) who underwent BTO for presurgical evaluation between Feb 2015 and Jan 2017. The patients’ clinical diagnoses comprise 8 neck cancer (smooth muscle sarcoma; n = 1, thyroid cancer; n = 5, paraganglioma; n = 1, nasopharyngeal cancer; n = 1) and 1 oral cancer. Six patients underwent BTO at common carotid artery (CCA), 1 patient at internal carotid artery (ICA), and 2 patients at CCA and ICA concurrently. Mean occlusion time was 24.4 ± 6.8 (Mean ± SD) minutes. Oral and written informed consent was obtained from all study participants. Patient preradiosurgical demographic information is shown in Table 1. Patient #1 showed the transient flittering scotoma during BTO, but the symptom was completely resolved after balloon deflation, and the other patients had no symptom. This study was approved by the clinical research committee of Osaka University Hospital (approval number: 14182) and Osaka International Cancer Institute (approval number: 1604159014). Oral and written informed consent was obtained from all study participants. 2.2. Balloon test occlusion procedure The BTO procedure was performed under local anesthesia. In case of BTO at CCA, a 7 Fr balloon catheter (OPTIMO, Tokai Medical Products) was positioned in the CCA. The balloon was inflated under fluoroscopic visualization until the contrast injection from the catheter demonstrated complete flow arrest. In case of BTO at ICA, a 5.2 Fr balloon catheter (Selecon MP catheter, Terumo Clinical Supply) was positioned in the cervical portion of the ICA. In case of BTO at ICA and CCA concurrently, a 7 Fr balloon catheter (OPTIMO) was positioned in the CCA, and a 3.5 Fr double-lumen silicone balloon catheter (MASAMUNE, Fuji Systems) was advanced into cervical portion of the ICA. Temporary occlusion was continued for 20 to 35 min, and angiogram of the contralateral ICA and the dominant vertebral artery were performed to evaluate
collateral circulation with a second 4 Fr catheter (JB2 100 cm, Terumo Clinical Supply). Neurologic examination and measurement of arterial blood pressure were performed every 5 min. ICA stump pressure was measured by clamping the CCA, and mean of ICA stump pressure was 64.7 mm Hg. 2.3. Blood sampling To prepare plasma, peripheral blood samples were obtained from the patients using EDTA containing tube before BTO and 48 h after BTO. Samples were centrifuged for 15 min at 1000 g within 30 min at 4°C. The supernatants were immediately aliquoted and stored at 80 °C. 2.4. Proteome analysis To remove high abundance proteins (albumin, transferrin, haptoglobin, a-1-antitrypsin, IgG, IgA and fibrinogen) in the plasma, we used Multiple Affinity Removal Spin Cartridge Human 7 (Agilent Technologies). The protein concentrations of samples were determined using a bicinchoninic acid (BCA) Protein Assay (Pierce Chemical). LC-MS/MS analysis was performed by UltiMate 3000 Nano LC systems (Thermo Fisher Scientific) coupled to Q Exactive mass spectrometer (Thermo Fisher Scientific), including electrospray ionization (ESI). The solvent gradient was composed of Solvent A (Water containing 0.1% formic acid) and Solvent B (Acetonitrile containing 0.1% formic acid). Flow rate was 300 nL/ min over 90 min. Total time for an LC MS/MS run was approximately 40 min. The proteins were identified using Mascot Distiller 2.5 and Mascot Server 2.5 (Matrix Science). Among the 18 samples (2 samples of each 9 cases), we exclude the 3 samples to analysis due to the failure of protein detection. Protein were then validated using Scaffold Q + software 4.4.3 (Proteome Software Inc.), and were investigated the characterization by UniProt (release 2017_02: February 5, 2017, http://www.uniprot.org/) and UniProt-GOA (Ver. 159, http://www.ebi.ac.uk/GOA). 2.5. Statistical analysis For quantitative analysis, we excluded the date of the sample which did not detect proteins. The significance of the differences between samples was examined using paired t-test, Student’s ttest, or multiple linear regression analysis. In the multiple linear regression analysis, objective variable is defined that counter number of Post BTO divided by that of Pre BTO in each samples. Explanatory variable is defined in sex (female: 1, male: 0), in ages (over 65 years old: 1, under 65 years old: 0). P values of < 0.05 were considered to be significant.
Table 1 Summary of clinical characteristics and results in 9 patients. Pt No.
Age
Sex
Desease
Occlusion location
Occlusion time
Neurological Results
Stump pressure ratio
1 2 3 4 5 6 7 8 9 Maen
56 64 63 77 62 70 67 45 74 64.2
F F M M F F M M M
leiomyosarcoma of neck thyroid cancer thyroid cancer thyroid cancer paraganglioma thyroid cancer thyroid cancer P54haryngeal cancer oral cancer
CCA CCA CCA CCA ICA + CCA CCA CCA ICA ICA + CCA
20 min 20 min 20 min 30 min 20 min 35 min 35 min 20 min 20 min 24.4
Transient visual field defect (–) (–) (–) (–) (–) (–) (–) (–)
0.67 0.63 0.63 0.67 0.52 0.54 0.52 0.39 0.73 0.59
Stump pressure ratio was defined as the mean stump pressure divided by the mean blood pressure. F: female; M: male; CCA: common carotid artery; ICA: internal cerebral artery.
Please cite this article as: T. Ozaki, R. Muramatsu, H. Nakamura et al., Proteomic analysis of protein changes in plasma by balloon test occlusion, Journal of Clinical Neuroscience, https://doi.org/10.1016/j.jocn.2019.12.005
T. Ozaki et al. / Journal of Clinical Neuroscience xxx (xxxx) xxx
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3. Results
4. Discussion
3.1. Comparison between pre and post BTO
Ischemic preconditioning is considered an emerging concept for stroke treatment, therefore, underling the molecular mechanism of IPC has been awaited to contribute develop the treatment for ischemia. In the mouse after IPC, the level of circulating molecule was changed and this change relates to neuroprotection after prolonged ischemia. In this study, we investigated the changes of circulating molecules level in the human by BTO and found that some of the molecules which level were changed after BTO have reportedly neuroprotective function by direct and indirect mechanisms. LGR1, which level in circulation was increased by the BTO, is known to promote angiogenesis through the transforming growth factor (TGF)-b1 signaling pathway [19]. In general, vascularderived factors provide trophic supports to the neuronal survival [20]. Furthermore, microvessel density in the ischemic border correlates with the survival periods after stroke in the patients [21]. These reports support the hypothesis that the changes of circulating molecule level in response to transient ischemia contribute to control ischemic tolerance via neovascularization. In the same direction, serum amyloid A-1 protein, which level was also changed by BTO, also reportedly induce angiogenesis through Toll-like receptor 2 [22]. AMBP, which is not regulates angiogenesis by itself, binds to adrenomedullin resulting in exert neuroprotection following brain injury [23]. Transthyretin increases activity of Na/H exchanger and reduces intracellular acid load by ischemia, resulting in exerts neuroprotective function [24]. Therefore, circulating molecules which level is changed by transient ischemia may be just marker which shows obtaining ischemic tolerance but also work on neuroprotective effect by itself. We also found that the change of the level of circulating molecules were different between age and sex. Fibrinogen gamma chain in plasma significantly increased in the younger age group more than in older age group. About relation between age and fibrinogen gamma, fibrinogen gamma increased significantly with increased
In this experiment, we collected blood samples from each patients pre BTO and 48 h after BTO, and identified 224 kinds of proteins in the samples. To investigate the changes of protein concentration in the blood by BTO, we used the number of spectral counts which infer the relative protein concentrations in the samples. We compared the difference of the count number in each sample between pre and post BTO. Among the detectable proteins, the count number of leucine-rich alpha-2-glycoprotein 1 (LRG1) and serum amyloid A-1 protein (SAA1) were significantly increased after BTO (Fig. 1a, b). In contrast, the count number of protein AMBP was decreased after BTO (Fig. 1c). These data indicate that BTO affects the circulating protein concentration in human with transient blood flow change.
3.2. Focused on sexes and ages We next investigated whether the change of protein concentration in plasma by BTO depends on the sex and age differences, because sex differences affect clinical outcomes after stroke [14,15,16], clinical outcome by stroke got worse with age [17,18]. We divided the results of count numbers by sex and two age groups over 65 years or not and compared the difference of the count number in each sample between pre and post BTO by multiple linear regression analysis. Fibrinogen gamma chain was significantly changed comparing with ages (estimate value 0.32, p < 0.05). Apolipoprotein E was significantly changed comparing with sexes (estimate value 0.21, p < 0.01) and ages (estimate value 0.15, p < 0.05). Apolipoprotein M (ApoM) was also significantly changed comparing with sexes (estimate value 0.38, p < 0.01) and ages (estimate value 0.52, p < 0.01) (Fig. 2).
Fig. 1. The effect of BTO in circulating protein levels. (a-c) The change of total spectrum counts of indicated proteins between pre (just before BTO) and 48 h after BTO. High abundance proteins (albumin, transferrin, haptoglobin, a-1-antitrypsin, IgG, IgA and fibrinogen) were excluded from the sample before proteomics analysis. Comparison analysis were performed for the all detectable proteins in the samples. n = 9, *p < 0.05, paired t-test.
Please cite this article as: T. Ozaki, R. Muramatsu, H. Nakamura et al., Proteomic analysis of protein changes in plasma by balloon test occlusion, Journal of Clinical Neuroscience, https://doi.org/10.1016/j.jocn.2019.12.005
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T. Ozaki et al. / Journal of Clinical Neuroscience xxx (xxxx) xxx
Fig. 2. The effect of age in the circulating protein level in the patients with BTO. The rate of total spectrum counts of Fibrinogen gamma chain of pre BTO divided that of post BTO in the younger age and older age groups (**p < 0.01, Student’s t-test). The data represents the mean ± s.e.m. (n = 5 each). (b) The rate of total spectrum counts of Apolipoprotein M of pre BTO divided that of post BTO (*p < 0.05, Student’s t-test). The data represents the mean ± s.e.m. (n = 5 each).
age [25,26]. Although, the reason why fibrinogen gamma increased in the younger age group than that of older age after the ischemic stimulation to the brain has never been reported, it may be that fibrinogen gamma of the elderly has already high in the normal condition and remaining capacity to increase in the ischemic condition is smaller than younger age. Fibrinogen gamma has both antithrombotic feature binding sites for thrombin and disruption of platelet binding to fibrinogen and prothrombotic feature binding site for the factor XIII B subunit [27]. Ratio of fibrinogen gamma to total fibrinogen is increased in the acute phase of ischemic stroke, which may reflect an antithrombotic defense mechanism of the human body [28]. Considering this result in addition with our result, fibrinogen gamma may be one of the neurotrophic molecules against ischemic stroke. In this research, ApoM in plasma significantly increased in the younger age group more than in older age group. ApoM, one of the latest additions to the apolipoprotein family, was identified in 1999 [29]. Zhao et al. reported that ApoM T-778C and C-1065 A single-nucleotide polymorphisms were associated with increased risk of ischemic stroke [30]. On the other hand, the ApoM is critical in the formation of pre-b-high-density lipoprotein (HDL) and cholesterol efflux to HDL from foam cells in atherosclerotic lesions which suggests that ApoM may protect against the formation of atherosclerosis [31,32]. From these aspects, ApoM have the potential to be neuroprotective molecule against ischemic stroke. About correlation between age and ApoM levels, Yoshida et al. reported that ApoM levels were positively correlated with age [33]. So, same as fibrinogen gamma, APoM of the elderly has already high in the normal condition and remaining capacity to increase in the ischemic condition is smaller than younger age. As above mentioned, we identified several molecules which expression was changed by transient ischemia in human. However, we should note that the sample we used were obtained from the patients who has tumor. Because tumor cells release diffusible factor and exosome which capable of activation of circulating cells including T cells [34], our identified molecules may strongly link to the tumor environments. Therefore, we carefully evaluate the contribution of our identified molecules to the neuroprotection against ischemia, and the present study just provide the evidence that transient ischemia in the brain changes the circulating molecules composition in human. 5. Conclusions Transient change of blood flow in the brain alters molecular expression in the plasma. Because the alteration of plasma protein
composition is involved in ischemic tolerance in animal models, the proteins whose level was changed by BTO may be also involved in neuroprotection against ischemia in human. Declaration of competing interest The authors report no conflict of interest concerning the materials or methods used in this study or the findings specified in the present paper. Acknowledgments This work was supported by a Grant-in-Aid for Early-Career Scientists (B) from the Japan Society for the Promotion of Science to TO (18 K16582), and a Grant-in-Aid for Scientific Research (B) from the Japan Society for the Promotion of Science to R.M (16H04672, 19H03554). References [1] Krishnamurthi RV, Feigin VL, Forouzanfar MH, Mensah GA, Connor M, Bennett DA, et al. Global and regional burden of first-ever ischaemic and haemorrhagic stroke during 1990–2010: findings from the Global Burden of Disease Study 2010. Lancet Global Health 2013;1:e259–81. https://doi.org/10.1016/S2214109X(13)70089-5. [2] Kuklina EV, Tong X, George MG, Bansil P. Epidemiology and prevention of stroke: a worldwide perspective. Expert Rev Neurother 2014;12:199–208. https://doi.org/10.1586/ern.11.99. [3] Goyal M, Menon BK, van Zwam WH, Dippel DWJ, Mitchell PJ, Demchuk AM, et al. Endovascular thrombectomy after large-vessel ischaemic stroke: a metaanalysis of individual patient data from five randomised trials. Lancet 2016;387:1723–31. https://doi.org/10.1016/S0140-6736(16)00163-X. [4] Hacke W, Kaste M, Bluhmki E, Brozman M, Dávalos A, Guidetti D, et al. Thrombolysis with alteplase 3 to 4.5 hours after acute ischemic stroke. N Engl J Med 2008;359:1317–29. https://doi.org/10.1056/NEJMoa0804656. [5] Fonarow GC, Smith EE, Saver JL, Reeves MJ, Hernandez AF, Peterson ED, et al. Improving door-to-needle times in acute ischemic stroke: the design and rationale for the American Heart Association/American Stroke Association’s Target: Stroke initiative. Stroke 2011;42:2983–9. https://doi.org/10.1161/ STROKEAHA.111.621342. [6] Shuaib A, Lees KR, Lyden P, Grotta J, Davalos A, Davis SM, et al. NXY-059 for the treatment of acute ischemic stroke. N Engl J Med 2007;357:562–71. https:// doi.org/10.1056/NEJMoa070240. [7] Tirilazad Mesylate in Acute Ischemic Stroke. Stroke 2000;31:2257–65. https:// doi.org/10.1161/01.str.31.9.2257. [8] Dirnagl U, Becker K, Meisel A. Preconditioning and tolerance against cerebral ischaemia: from experimental strategies to clinical use. Lancet Neurol 2009;8:398–412. https://doi.org/10.1016/S1474-4422(09)70054-7. [9] Zhang J, Yang ZJ, Klaus JA, Koehler RC, Huang J. Delayed tolerance with repetitive transient focal ischemic preconditioning in the mouse. Stroke 2008;39:967–74. https://doi.org/10.1161/STROKEAHA.107.497412. [10] Simon RP, Niiro M, Gwinn R. Prior ischemic stress protects against experimental stroke. Neurosci Lett 1993;163:135–7. https://doi.org/10.1016/ 0304-3940(93)90364-Q.
Please cite this article as: T. Ozaki, R. Muramatsu, H. Nakamura et al., Proteomic analysis of protein changes in plasma by balloon test occlusion, Journal of Clinical Neuroscience, https://doi.org/10.1016/j.jocn.2019.12.005
T. Ozaki et al. / Journal of Clinical Neuroscience xxx (xxxx) xxx [11] Wegener S. Transient ischemic attacks before ischemic stroke: Preconditioning the human brain?: A multicenter magnetic resonance imaging study. Stroke 2004;35:616–21. https://doi.org/10.1161/01.STR.0000115767.17923.6A. [12] Hirayama Y, Ikeda-Matsuo Y, Notomi S, Enaida H, Kinouchi H, Koizumi S. Astrocyte-mediated ischemic tolerance. J Neurosci 2015;35:3794–805. https://doi.org/10.1523/JNEUROSCI.4218-14.2015. [13] Ozaki T, Muramatsu R, Sasai M, Yamamoto M, Kubota Y, Fujinaka T, et al. The P2X4 receptor is required for neuroprotection via ischemic preconditioning. Nature Publishing Group 2016;6:25893. https://doi.org/10.1038/srep25893. [14] Appelros P, Stegmayr B, Terent A. Sex differences in stroke epidemiology: a systematic review. Stroke 2009;40:1082–90. https://doi.org/10.1161/ STROKEAHA.108.540781. [15] Reeves MJ, Bushnell CD, Howard G, Gargano JW, Duncan PW, Lynch G, et al. Sex differences in stroke: epidemiology, clinical presentation, medical care, and outcomes. Lancet Neurol 2008;7:915–26. https://doi.org/10.1016/S14744422(08)70193-5. [16] Kapral MK, Fang J, Hill MD, Silver F, Richards J, Jaigobin C, et al. Sex differences in stroke care and outcomes: results from the Registry of the Canadian Stroke Network. Stroke 2005;36:809–14. https://doi.org/10.1161/01. STR.0000157662.09551.e5. [17] Knoflach M, Matosevic B, Rücker M, Furtner M, Mair A, Wille G, et al. Functional recovery after ischemic stroke–a matter of age: data from the Austrian Stroke Unit Registry. Neurology 2012;78:279–85. https://doi.org/ 10.1212/WNL.0b013e31824367ab. [18] Nakayama H, Jørgensen HS, Raaschou HO, Olsen TS. The influence of age on stroke outcome. The Copenhagen Stroke Study. Stroke 1994;25:808–13. [19] Wang X, Abraham S, McKenzie JAG, Jeffs N, Swire M, Tripathi VB, et al. LRG1 promotes angiogenesis by modulating endothelial TGF-b signalling. Nature 2013;499:306–11. https://doi.org/10.1038/nature12345. [20] Tovar-Y-Romo LB, Ramírez-Jarquín UN, Lazo-Gómez R, Tapia R. Trophic factors as modulators of motor neuron physiology and survival: implications for ALS therapy. Front Cell Neurosci 2014;8:61. https://doi.org/10.3389/ fncel.2014.00061. [21] Krupinski J, Kaluza J, Kumar P, Kumar S, Wang JM. Role of angiogenesis in patients with cerebral ischemic stroke. Stroke 1994;25:1794–8. https://doi. org/10.1161/01.STR.25.9.1794. [22] Connolly M, Marrelli A, Blades M, McCormick J, Maderna P, Godson C, et al. Acute serum amyloid A induces migration, angiogenesis, and inflammation in synovial cells in vitro and in a human rheumatoid arthritis/SCID mouse chimera model. J Immunol 2010;184:6427–37. https://doi.org/10.4049/ jimmunol.0902941.
5
[23] Cheyuo C, Yang W-L, Wang P. The critical role of adrenomedullin and its binding protein, AMBP-1, in neuroprotection. Biol Chem 2012;393:429–39. https://doi.org/10.1515/hsz-2012-0103. [24] Lin H-Y, Davis FB, Luidens MK, Mousa SA, Cao JH, Zhou M, et al. Molecular basis for certain neuroprotective effects of thyroid hormone. Front Mol Neurosci 2011;4. https://doi.org/10.3389/fnmol.2011.00029. [25] Lovely RS, Kazmierczak SC, Massaro JM, D’Agostino RB, O’Donnell CJ, Farrell DH. Fibrinogen: evaluation of a new assay for study of associations with cardiovascular disease. Clin Chem 2010;56:781–8. https://doi.org/ 10.1373/clinchem.2009.138347. [26] Appiah D, Schreiner PJ, MacLehose RF, Folsom AR. Association of plasma c’ fibrinogen with incident cardiovascular disease: the atherosclerosis risk in communities (ARIC) study. Arterioscler Thromb Vasc Biol 2015;35:2700–6. https://doi.org/10.1161/ATVBAHA.115.306284. [27] Mosesson MW. Fibrinogen and fibrin structure and functions. J Thromb Haemost 2005;3:1894–904. https://doi.org/10.1111/j.1538-7836.2005.01365. x. [28] Cheung EYL, de Willige SU, Vos HL, Leebeek FWG, Dippel DWJ, Bertina RM, et al. Fibrinogen c0 in Ischemic Stroke. Stroke 2008;39:1033–5. https://doi.org/ 10.1161/STROKEAHA.107.495499. [29] Xu N, Dahlbäck B. A novel human apolipoprotein (apoM). J Biol Chem 1999;274:31286–90. https://doi.org/10.1074/jbc.274.44.31286. [30] Zhao D, He Z, Qin X, Li L, Liu F, Deng S. Association of apolipoprotein M gene polymorphisms with ischemic stroke in a han chinese population. J Mol Neurosci 2010;43:370–5. https://doi.org/10.1007/s12031-010-9453-7. [31] Wolfrum C, Poy MN, Stoffel M. Apolipoprotein M is required for prebeta-HDL formation and cholesterol efflux to HDL and protects against atherosclerosis. Nat Med 2005;11:418–22. https://doi.org/10.1038/nm1211. [32] Christoffersen C, Nielsen LB, Axler O, Andersson A, Johnsen AH, Dahlbäck B. Isolation and characterization of human apolipoprotein M-containing lipoproteins. J Lipid Res 2006;47:1833–43. https://doi.org/10.1194/jlr. M600055-JLR200. [33] Yoshida K, Kimura T, Shimizu T, Sato K, Aoki T, Sumino H, et al. Serum apolipoprotein M levels are correlated with biomarkers of coagulation. Kitakanto Med J 2016;66:1–6. https://doi.org/10.2974/kmj.66.1. [34] Kucharzewska P, Christianson HC, Welch JE, Svensson KJ, Fredlund E, Ringnér M, et al. Exosomes reflect the hypoxic status of glioma cells and mediate hypoxia-dependent activation of vascular cells during tumor development. Proc Natl Acad Sci USa 2013;110:7312–7. https://doi.org/10.1073/ pnas.1220998110.
Please cite this article as: T. Ozaki, R. Muramatsu, H. Nakamura et al., Proteomic analysis of protein changes in plasma by balloon test occlusion, Journal of Clinical Neuroscience, https://doi.org/10.1016/j.jocn.2019.12.005
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Journal of Clinical Neuroscience journal homepage: www.elsevier.com/locate/jocn
Experimental study
Proteomic analysis of protein changes in plasma by balloon test occlusion Tomohiko Ozaki a,b,c, Rieko Muramatsu a,d,e,⇑, Hajime Nakamura b, Manabu Kinoshita c, Haruhiko Kishima b, Toshihide Yamashita a,d,f a
Department of Molecular Neuroscience, Graduate School of Medicine, Osaka University, Osaka, Japan Department of Neurosurgery, Graduate School of Medicine, Osaka University, Japan Department of Neurosurgery, Osaka International Cancer Institute, Osaka, Japan d WPI Immunology Frontier Research Center, Osaka University, Japan e Department of Molecular Pharmacology, National Institute of Neuroscience, National Center of Neurology and Neurology and Psychiatry, Kodaira, Japan f Department of Molecular Neuroscience, Graduate School of Frontier Biosciences, Osaka University, Japan b c
a r t i c l e
i n f o
Article history: Received 31 August 2019 Accepted 1 December 2019 Available online xxxx Keywords: Ischemic preconditioning Stroke Human Vascular endothelial cell Neuron
a b s t r a c t Transient ischemia provides the tolerance against prolonged ischemia in the brain. In mouse experimental model, transient ischemia changes the composition ratio of circulating proteins, which associate with neuroprotection; however, the human evidence is lacking. Here we mimicked balloon test occlusion (BTO) of carotid artery as a transient ischemia and investigated the change of composition ratio of the circulating protein in the human plasma. We collected blood samples from nine patients (5 men and 4 women; mean age 64.2 years; range 45 to 77 years) before and 48 h after BTO and investigated the changes of circulating molecules level in the proteome using LC-MS/MS analysis. Leucine-rich alpha-2glycoprotein and serum amyloid A-1 increased and protein AMBP decreased in the blood samples after BTO. Transient change of blood flow in the brain alters molecular expression in the plasma. Because the alteration of plasma protein composition is involved in ischemic tolerance in animal models, the proteins whose level was changed by BTO may be also involved in neuroprotection against ischemia in human. Ó 2019 Published by Elsevier Ltd.
1. Introduction The absolute number of new stroke events was approximately 11.6 million in 2010, [1] and the number of ischemic stroke events is expected to increase to 23 million by 2030. [2] Although the therapeutic approaches, such as administration of tissue-type plasminogen activator (tPA) and endovascular techniques, are approved for acute stroke treatment [3,4], these therapies has serious time limitation (within hours after onset of stroke) and more 50% patients with stroke are not able to achieve the benefits [5]. Therefore, developing the pharmacological interventions which aim to reduce neuronal damage have been attributed from pharmacological companies [6,7]. Some of the drugs were aimed trapping free radical after stroke and had been reached to clinical trial, but with little proof of effectiveness [8]. Therefore, the other mech-
⇑ Corresponding author at: Department of Molecular Pharmacology, National Institute of Neuroscience, National Center of Neurology and Neurology and Psychiatry, Kodaira, Japan. E-mail address: [email protected] (R. Muramatsu).
anisms which exert neuroprotection may be useful for developing the therapy for stroke. Ischemic preconditioning (IPC) is an endogenous protective mechanism and is proved at mammalian experimental models [9,10] and is considered conserved in human [11]. Short sublethal ischemia exposure is known as one of the IPC, and its renders the brain more resistance against prolonged ischemia. Because of the ethical issue, the underlying the molecular mechanism of IPC has been anticipated to apply the drug development. In the molecular mechanism of IPC, almost of all research have been focusing on the role of the molecular mechanims which is detectable in the central nervous system, such as astrocytic P2 receptor activation-mediated neuroprotective functions [12]. In contrast, it is reported that the IPC-mediated neuroprotection depends on the change of expression of vascular-derived osteopontin release in mouse experimental model, and this change was also detectable in the plasma of the mice [13]. In addition, change of vascular-derived molecules release was also detected in circulation in the plasma obtained from the mice after transient ischemia. Therefore, in this study,
https://doi.org/10.1016/j.jocn.2019.12.005 0967-5868/Ó 2019 Published by Elsevier Ltd.
Please cite this article as: T. Ozaki, R. Muramatsu, H. Nakamura et al., Proteomic analysis of protein changes in plasma by balloon test occlusion, Journal of Clinical Neuroscience, https://doi.org/10.1016/j.jocn.2019.12.005
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we challenged wheatear our observation obtained from mouse were also conserved in human. Here we used the blood sample from the patients with BTO, a procedure to stop blood flow by a balloon catheter in an artery. We mimicked the BTO as a transient ischemia and investigated the changes of composition ratio of circulating protein before and after BTO. We found that transient hypoperfusion by BTO causes changes of protein levels. We also analyzed the protein level in the plasma by the difference of sex and age. 2. Methods 2.1. Patients We collected data from 9 serial cancer patients (5 men and 4 women; mean age 64.2 years; range 45 to 77 years) who underwent BTO for presurgical evaluation between Feb 2015 and Jan 2017. The patients’ clinical diagnoses comprise 8 neck cancer (smooth muscle sarcoma; n = 1, thyroid cancer; n = 5, paraganglioma; n = 1, nasopharyngeal cancer; n = 1) and 1 oral cancer. Six patients underwent BTO at common carotid artery (CCA), 1 patient at internal carotid artery (ICA), and 2 patients at CCA and ICA concurrently. Mean occlusion time was 24.4 ± 6.8 (Mean ± SD) minutes. Oral and written informed consent was obtained from all study participants. Patient preradiosurgical demographic information is shown in Table 1. Patient #1 showed the transient flittering scotoma during BTO, but the symptom was completely resolved after balloon deflation, and the other patients had no symptom. This study was approved by the clinical research committee of Osaka University Hospital (approval number: 14182) and Osaka International Cancer Institute (approval number: 1604159014). Oral and written informed consent was obtained from all study participants. 2.2. Balloon test occlusion procedure The BTO procedure was performed under local anesthesia. In case of BTO at CCA, a 7 Fr balloon catheter (OPTIMO, Tokai Medical Products) was positioned in the CCA. The balloon was inflated under fluoroscopic visualization until the contrast injection from the catheter demonstrated complete flow arrest. In case of BTO at ICA, a 5.2 Fr balloon catheter (Selecon MP catheter, Terumo Clinical Supply) was positioned in the cervical portion of the ICA. In case of BTO at ICA and CCA concurrently, a 7 Fr balloon catheter (OPTIMO) was positioned in the CCA, and a 3.5 Fr double-lumen silicone balloon catheter (MASAMUNE, Fuji Systems) was advanced into cervical portion of the ICA. Temporary occlusion was continued for 20 to 35 min, and angiogram of the contralateral ICA and the dominant vertebral artery were performed to evaluate
collateral circulation with a second 4 Fr catheter (JB2 100 cm, Terumo Clinical Supply). Neurologic examination and measurement of arterial blood pressure were performed every 5 min. ICA stump pressure was measured by clamping the CCA, and mean of ICA stump pressure was 64.7 mm Hg. 2.3. Blood sampling To prepare plasma, peripheral blood samples were obtained from the patients using EDTA containing tube before BTO and 48 h after BTO. Samples were centrifuged for 15 min at 1000 g within 30 min at 4°C. The supernatants were immediately aliquoted and stored at 80 °C. 2.4. Proteome analysis To remove high abundance proteins (albumin, transferrin, haptoglobin, a-1-antitrypsin, IgG, IgA and fibrinogen) in the plasma, we used Multiple Affinity Removal Spin Cartridge Human 7 (Agilent Technologies). The protein concentrations of samples were determined using a bicinchoninic acid (BCA) Protein Assay (Pierce Chemical). LC-MS/MS analysis was performed by UltiMate 3000 Nano LC systems (Thermo Fisher Scientific) coupled to Q Exactive mass spectrometer (Thermo Fisher Scientific), including electrospray ionization (ESI). The solvent gradient was composed of Solvent A (Water containing 0.1% formic acid) and Solvent B (Acetonitrile containing 0.1% formic acid). Flow rate was 300 nL/ min over 90 min. Total time for an LC MS/MS run was approximately 40 min. The proteins were identified using Mascot Distiller 2.5 and Mascot Server 2.5 (Matrix Science). Among the 18 samples (2 samples of each 9 cases), we exclude the 3 samples to analysis due to the failure of protein detection. Protein were then validated using Scaffold Q + software 4.4.3 (Proteome Software Inc.), and were investigated the characterization by UniProt (release 2017_02: February 5, 2017, http://www.uniprot.org/) and UniProt-GOA (Ver. 159, http://www.ebi.ac.uk/GOA). 2.5. Statistical analysis For quantitative analysis, we excluded the date of the sample which did not detect proteins. The significance of the differences between samples was examined using paired t-test, Student’s ttest, or multiple linear regression analysis. In the multiple linear regression analysis, objective variable is defined that counter number of Post BTO divided by that of Pre BTO in each samples. Explanatory variable is defined in sex (female: 1, male: 0), in ages (over 65 years old: 1, under 65 years old: 0). P values of < 0.05 were considered to be significant.
Table 1 Summary of clinical characteristics and results in 9 patients. Pt No.
Age
Sex
Desease
Occlusion location
Occlusion time
Neurological Results
Stump pressure ratio
1 2 3 4 5 6 7 8 9 Maen
56 64 63 77 62 70 67 45 74 64.2
F F M M F F M M M
leiomyosarcoma of neck thyroid cancer thyroid cancer thyroid cancer paraganglioma thyroid cancer thyroid cancer P54haryngeal cancer oral cancer
CCA CCA CCA CCA ICA + CCA CCA CCA ICA ICA + CCA
20 min 20 min 20 min 30 min 20 min 35 min 35 min 20 min 20 min 24.4
Transient visual field defect (–) (–) (–) (–) (–) (–) (–) (–)
0.67 0.63 0.63 0.67 0.52 0.54 0.52 0.39 0.73 0.59
Stump pressure ratio was defined as the mean stump pressure divided by the mean blood pressure. F: female; M: male; CCA: common carotid artery; ICA: internal cerebral artery.
Please cite this article as: T. Ozaki, R. Muramatsu, H. Nakamura et al., Proteomic analysis of protein changes in plasma by balloon test occlusion, Journal of Clinical Neuroscience, https://doi.org/10.1016/j.jocn.2019.12.005
T. Ozaki et al. / Journal of Clinical Neuroscience xxx (xxxx) xxx
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3. Results
4. Discussion
3.1. Comparison between pre and post BTO
Ischemic preconditioning is considered an emerging concept for stroke treatment, therefore, underling the molecular mechanism of IPC has been awaited to contribute develop the treatment for ischemia. In the mouse after IPC, the level of circulating molecule was changed and this change relates to neuroprotection after prolonged ischemia. In this study, we investigated the changes of circulating molecules level in the human by BTO and found that some of the molecules which level were changed after BTO have reportedly neuroprotective function by direct and indirect mechanisms. LGR1, which level in circulation was increased by the BTO, is known to promote angiogenesis through the transforming growth factor (TGF)-b1 signaling pathway [19]. In general, vascularderived factors provide trophic supports to the neuronal survival [20]. Furthermore, microvessel density in the ischemic border correlates with the survival periods after stroke in the patients [21]. These reports support the hypothesis that the changes of circulating molecule level in response to transient ischemia contribute to control ischemic tolerance via neovascularization. In the same direction, serum amyloid A-1 protein, which level was also changed by BTO, also reportedly induce angiogenesis through Toll-like receptor 2 [22]. AMBP, which is not regulates angiogenesis by itself, binds to adrenomedullin resulting in exert neuroprotection following brain injury [23]. Transthyretin increases activity of Na/H exchanger and reduces intracellular acid load by ischemia, resulting in exerts neuroprotective function [24]. Therefore, circulating molecules which level is changed by transient ischemia may be just marker which shows obtaining ischemic tolerance but also work on neuroprotective effect by itself. We also found that the change of the level of circulating molecules were different between age and sex. Fibrinogen gamma chain in plasma significantly increased in the younger age group more than in older age group. About relation between age and fibrinogen gamma, fibrinogen gamma increased significantly with increased
In this experiment, we collected blood samples from each patients pre BTO and 48 h after BTO, and identified 224 kinds of proteins in the samples. To investigate the changes of protein concentration in the blood by BTO, we used the number of spectral counts which infer the relative protein concentrations in the samples. We compared the difference of the count number in each sample between pre and post BTO. Among the detectable proteins, the count number of leucine-rich alpha-2-glycoprotein 1 (LRG1) and serum amyloid A-1 protein (SAA1) were significantly increased after BTO (Fig. 1a, b). In contrast, the count number of protein AMBP was decreased after BTO (Fig. 1c). These data indicate that BTO affects the circulating protein concentration in human with transient blood flow change.
3.2. Focused on sexes and ages We next investigated whether the change of protein concentration in plasma by BTO depends on the sex and age differences, because sex differences affect clinical outcomes after stroke [14,15,16], clinical outcome by stroke got worse with age [17,18]. We divided the results of count numbers by sex and two age groups over 65 years or not and compared the difference of the count number in each sample between pre and post BTO by multiple linear regression analysis. Fibrinogen gamma chain was significantly changed comparing with ages (estimate value 0.32, p < 0.05). Apolipoprotein E was significantly changed comparing with sexes (estimate value 0.21, p < 0.01) and ages (estimate value 0.15, p < 0.05). Apolipoprotein M (ApoM) was also significantly changed comparing with sexes (estimate value 0.38, p < 0.01) and ages (estimate value 0.52, p < 0.01) (Fig. 2).
Fig. 1. The effect of BTO in circulating protein levels. (a-c) The change of total spectrum counts of indicated proteins between pre (just before BTO) and 48 h after BTO. High abundance proteins (albumin, transferrin, haptoglobin, a-1-antitrypsin, IgG, IgA and fibrinogen) were excluded from the sample before proteomics analysis. Comparison analysis were performed for the all detectable proteins in the samples. n = 9, *p < 0.05, paired t-test.
Please cite this article as: T. Ozaki, R. Muramatsu, H. Nakamura et al., Proteomic analysis of protein changes in plasma by balloon test occlusion, Journal of Clinical Neuroscience, https://doi.org/10.1016/j.jocn.2019.12.005
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T. Ozaki et al. / Journal of Clinical Neuroscience xxx (xxxx) xxx
Fig. 2. The effect of age in the circulating protein level in the patients with BTO. The rate of total spectrum counts of Fibrinogen gamma chain of pre BTO divided that of post BTO in the younger age and older age groups (**p < 0.01, Student’s t-test). The data represents the mean ± s.e.m. (n = 5 each). (b) The rate of total spectrum counts of Apolipoprotein M of pre BTO divided that of post BTO (*p < 0.05, Student’s t-test). The data represents the mean ± s.e.m. (n = 5 each).
age [25,26]. Although, the reason why fibrinogen gamma increased in the younger age group than that of older age after the ischemic stimulation to the brain has never been reported, it may be that fibrinogen gamma of the elderly has already high in the normal condition and remaining capacity to increase in the ischemic condition is smaller than younger age. Fibrinogen gamma has both antithrombotic feature binding sites for thrombin and disruption of platelet binding to fibrinogen and prothrombotic feature binding site for the factor XIII B subunit [27]. Ratio of fibrinogen gamma to total fibrinogen is increased in the acute phase of ischemic stroke, which may reflect an antithrombotic defense mechanism of the human body [28]. Considering this result in addition with our result, fibrinogen gamma may be one of the neurotrophic molecules against ischemic stroke. In this research, ApoM in plasma significantly increased in the younger age group more than in older age group. ApoM, one of the latest additions to the apolipoprotein family, was identified in 1999 [29]. Zhao et al. reported that ApoM T-778C and C-1065 A single-nucleotide polymorphisms were associated with increased risk of ischemic stroke [30]. On the other hand, the ApoM is critical in the formation of pre-b-high-density lipoprotein (HDL) and cholesterol efflux to HDL from foam cells in atherosclerotic lesions which suggests that ApoM may protect against the formation of atherosclerosis [31,32]. From these aspects, ApoM have the potential to be neuroprotective molecule against ischemic stroke. About correlation between age and ApoM levels, Yoshida et al. reported that ApoM levels were positively correlated with age [33]. So, same as fibrinogen gamma, APoM of the elderly has already high in the normal condition and remaining capacity to increase in the ischemic condition is smaller than younger age. As above mentioned, we identified several molecules which expression was changed by transient ischemia in human. However, we should note that the sample we used were obtained from the patients who has tumor. Because tumor cells release diffusible factor and exosome which capable of activation of circulating cells including T cells [34], our identified molecules may strongly link to the tumor environments. Therefore, we carefully evaluate the contribution of our identified molecules to the neuroprotection against ischemia, and the present study just provide the evidence that transient ischemia in the brain changes the circulating molecules composition in human. 5. Conclusions Transient change of blood flow in the brain alters molecular expression in the plasma. Because the alteration of plasma protein
composition is involved in ischemic tolerance in animal models, the proteins whose level was changed by BTO may be also involved in neuroprotection against ischemia in human. Declaration of competing interest The authors report no conflict of interest concerning the materials or methods used in this study or the findings specified in the present paper. Acknowledgments This work was supported by a Grant-in-Aid for Early-Career Scientists (B) from the Japan Society for the Promotion of Science to TO (18 K16582), and a Grant-in-Aid for Scientific Research (B) from the Japan Society for the Promotion of Science to R.M (16H04672, 19H03554). References [1] Krishnamurthi RV, Feigin VL, Forouzanfar MH, Mensah GA, Connor M, Bennett DA, et al. Global and regional burden of first-ever ischaemic and haemorrhagic stroke during 1990–2010: findings from the Global Burden of Disease Study 2010. Lancet Global Health 2013;1:e259–81. https://doi.org/10.1016/S2214109X(13)70089-5. [2] Kuklina EV, Tong X, George MG, Bansil P. Epidemiology and prevention of stroke: a worldwide perspective. Expert Rev Neurother 2014;12:199–208. https://doi.org/10.1586/ern.11.99. [3] Goyal M, Menon BK, van Zwam WH, Dippel DWJ, Mitchell PJ, Demchuk AM, et al. Endovascular thrombectomy after large-vessel ischaemic stroke: a metaanalysis of individual patient data from five randomised trials. Lancet 2016;387:1723–31. https://doi.org/10.1016/S0140-6736(16)00163-X. [4] Hacke W, Kaste M, Bluhmki E, Brozman M, Dávalos A, Guidetti D, et al. Thrombolysis with alteplase 3 to 4.5 hours after acute ischemic stroke. N Engl J Med 2008;359:1317–29. https://doi.org/10.1056/NEJMoa0804656. [5] Fonarow GC, Smith EE, Saver JL, Reeves MJ, Hernandez AF, Peterson ED, et al. Improving door-to-needle times in acute ischemic stroke: the design and rationale for the American Heart Association/American Stroke Association’s Target: Stroke initiative. Stroke 2011;42:2983–9. https://doi.org/10.1161/ STROKEAHA.111.621342. [6] Shuaib A, Lees KR, Lyden P, Grotta J, Davalos A, Davis SM, et al. NXY-059 for the treatment of acute ischemic stroke. N Engl J Med 2007;357:562–71. https:// doi.org/10.1056/NEJMoa070240. [7] Tirilazad Mesylate in Acute Ischemic Stroke. Stroke 2000;31:2257–65. https:// doi.org/10.1161/01.str.31.9.2257. [8] Dirnagl U, Becker K, Meisel A. Preconditioning and tolerance against cerebral ischaemia: from experimental strategies to clinical use. Lancet Neurol 2009;8:398–412. https://doi.org/10.1016/S1474-4422(09)70054-7. [9] Zhang J, Yang ZJ, Klaus JA, Koehler RC, Huang J. Delayed tolerance with repetitive transient focal ischemic preconditioning in the mouse. Stroke 2008;39:967–74. https://doi.org/10.1161/STROKEAHA.107.497412. [10] Simon RP, Niiro M, Gwinn R. Prior ischemic stress protects against experimental stroke. Neurosci Lett 1993;163:135–7. https://doi.org/10.1016/ 0304-3940(93)90364-Q.
Please cite this article as: T. Ozaki, R. Muramatsu, H. Nakamura et al., Proteomic analysis of protein changes in plasma by balloon test occlusion, Journal of Clinical Neuroscience, https://doi.org/10.1016/j.jocn.2019.12.005
T. Ozaki et al. / Journal of Clinical Neuroscience xxx (xxxx) xxx [11] Wegener S. Transient ischemic attacks before ischemic stroke: Preconditioning the human brain?: A multicenter magnetic resonance imaging study. Stroke 2004;35:616–21. https://doi.org/10.1161/01.STR.0000115767.17923.6A. [12] Hirayama Y, Ikeda-Matsuo Y, Notomi S, Enaida H, Kinouchi H, Koizumi S. Astrocyte-mediated ischemic tolerance. J Neurosci 2015;35:3794–805. https://doi.org/10.1523/JNEUROSCI.4218-14.2015. [13] Ozaki T, Muramatsu R, Sasai M, Yamamoto M, Kubota Y, Fujinaka T, et al. The P2X4 receptor is required for neuroprotection via ischemic preconditioning. Nature Publishing Group 2016;6:25893. https://doi.org/10.1038/srep25893. [14] Appelros P, Stegmayr B, Terent A. Sex differences in stroke epidemiology: a systematic review. Stroke 2009;40:1082–90. https://doi.org/10.1161/ STROKEAHA.108.540781. [15] Reeves MJ, Bushnell CD, Howard G, Gargano JW, Duncan PW, Lynch G, et al. Sex differences in stroke: epidemiology, clinical presentation, medical care, and outcomes. Lancet Neurol 2008;7:915–26. https://doi.org/10.1016/S14744422(08)70193-5. [16] Kapral MK, Fang J, Hill MD, Silver F, Richards J, Jaigobin C, et al. Sex differences in stroke care and outcomes: results from the Registry of the Canadian Stroke Network. Stroke 2005;36:809–14. https://doi.org/10.1161/01. STR.0000157662.09551.e5. [17] Knoflach M, Matosevic B, Rücker M, Furtner M, Mair A, Wille G, et al. Functional recovery after ischemic stroke–a matter of age: data from the Austrian Stroke Unit Registry. Neurology 2012;78:279–85. https://doi.org/ 10.1212/WNL.0b013e31824367ab. [18] Nakayama H, Jørgensen HS, Raaschou HO, Olsen TS. The influence of age on stroke outcome. The Copenhagen Stroke Study. Stroke 1994;25:808–13. [19] Wang X, Abraham S, McKenzie JAG, Jeffs N, Swire M, Tripathi VB, et al. LRG1 promotes angiogenesis by modulating endothelial TGF-b signalling. Nature 2013;499:306–11. https://doi.org/10.1038/nature12345. [20] Tovar-Y-Romo LB, Ramírez-Jarquín UN, Lazo-Gómez R, Tapia R. Trophic factors as modulators of motor neuron physiology and survival: implications for ALS therapy. Front Cell Neurosci 2014;8:61. https://doi.org/10.3389/ fncel.2014.00061. [21] Krupinski J, Kaluza J, Kumar P, Kumar S, Wang JM. Role of angiogenesis in patients with cerebral ischemic stroke. Stroke 1994;25:1794–8. https://doi. org/10.1161/01.STR.25.9.1794. [22] Connolly M, Marrelli A, Blades M, McCormick J, Maderna P, Godson C, et al. Acute serum amyloid A induces migration, angiogenesis, and inflammation in synovial cells in vitro and in a human rheumatoid arthritis/SCID mouse chimera model. J Immunol 2010;184:6427–37. https://doi.org/10.4049/ jimmunol.0902941.
5
[23] Cheyuo C, Yang W-L, Wang P. The critical role of adrenomedullin and its binding protein, AMBP-1, in neuroprotection. Biol Chem 2012;393:429–39. https://doi.org/10.1515/hsz-2012-0103. [24] Lin H-Y, Davis FB, Luidens MK, Mousa SA, Cao JH, Zhou M, et al. Molecular basis for certain neuroprotective effects of thyroid hormone. Front Mol Neurosci 2011;4. https://doi.org/10.3389/fnmol.2011.00029. [25] Lovely RS, Kazmierczak SC, Massaro JM, D’Agostino RB, O’Donnell CJ, Farrell DH. Fibrinogen: evaluation of a new assay for study of associations with cardiovascular disease. Clin Chem 2010;56:781–8. https://doi.org/ 10.1373/clinchem.2009.138347. [26] Appiah D, Schreiner PJ, MacLehose RF, Folsom AR. Association of plasma c’ fibrinogen with incident cardiovascular disease: the atherosclerosis risk in communities (ARIC) study. Arterioscler Thromb Vasc Biol 2015;35:2700–6. https://doi.org/10.1161/ATVBAHA.115.306284. [27] Mosesson MW. Fibrinogen and fibrin structure and functions. J Thromb Haemost 2005;3:1894–904. https://doi.org/10.1111/j.1538-7836.2005.01365. x. [28] Cheung EYL, de Willige SU, Vos HL, Leebeek FWG, Dippel DWJ, Bertina RM, et al. Fibrinogen c0 in Ischemic Stroke. Stroke 2008;39:1033–5. https://doi.org/ 10.1161/STROKEAHA.107.495499. [29] Xu N, Dahlbäck B. A novel human apolipoprotein (apoM). J Biol Chem 1999;274:31286–90. https://doi.org/10.1074/jbc.274.44.31286. [30] Zhao D, He Z, Qin X, Li L, Liu F, Deng S. Association of apolipoprotein M gene polymorphisms with ischemic stroke in a han chinese population. J Mol Neurosci 2010;43:370–5. https://doi.org/10.1007/s12031-010-9453-7. [31] Wolfrum C, Poy MN, Stoffel M. Apolipoprotein M is required for prebeta-HDL formation and cholesterol efflux to HDL and protects against atherosclerosis. Nat Med 2005;11:418–22. https://doi.org/10.1038/nm1211. [32] Christoffersen C, Nielsen LB, Axler O, Andersson A, Johnsen AH, Dahlbäck B. Isolation and characterization of human apolipoprotein M-containing lipoproteins. J Lipid Res 2006;47:1833–43. https://doi.org/10.1194/jlr. M600055-JLR200. [33] Yoshida K, Kimura T, Shimizu T, Sato K, Aoki T, Sumino H, et al. Serum apolipoprotein M levels are correlated with biomarkers of coagulation. Kitakanto Med J 2016;66:1–6. https://doi.org/10.2974/kmj.66.1. [34] Kucharzewska P, Christianson HC, Welch JE, Svensson KJ, Fredlund E, Ringnér M, et al. Exosomes reflect the hypoxic status of glioma cells and mediate hypoxia-dependent activation of vascular cells during tumor development. Proc Natl Acad Sci USa 2013;110:7312–7. https://doi.org/10.1073/ pnas.1220998110.
Please cite this article as: T. Ozaki, R. Muramatsu, H. Nakamura et al., Proteomic analysis of protein changes in plasma by balloon test occlusion, Journal of Clinical Neuroscience, https://doi.org/10.1016/j.jocn.2019.12.005