Risk Factors for Intracranial Aneurysm Rupture Using Metabolomics and Cytokine Profiling Analysis

doi:10.3969/j.issn.1673-5765.2023.04.013

Abstract

Abstract: Objectives To investigate the metabolite and cytokine biomarkers related to intracranial aneurysm rupture using the metabolomics and cytokine profiling analysis.
Methods The patients undergoing surgery for single intracranial aneurysm from January 2020 to January 2022 were prospectively enrolled in this study. The aneurysm tissues and serum samples of patients were collected. According to intracranial aneurysm state at admission, all patients were divided into the ruptured aneurysm group and unruptured aneurysm group. The targeted metabolomics was used to detect the abnormal metabolites in aneurysm tissue and serum, and the 46-cytokine profiling analysis was used to detect the abnormal cytokines. The Spearman correlation analysis was used to analyze the correlation of these metabolites and cytokines with aneurysm rupture in aneurysm tissues and serum. Receiver operating characteristic (ROC) curve was used to evaluate the clinical value of these risk factors for aneurysm rupture, and multivariate logistic regression analysis was used to determine the risk factors for aneurysm rupture.
Results This study included 25 cases with ruptured aneurysm and 31 cases of unruptured aneurysm. The targeted metabolomics and cytokine profiling analysis found 43 metabolites and 5 cytokines related to aneurysm rupture (all P<0.05) . Enrichment analysis showed that these metabolites and cytokines are mainly lipids and lipid analogs, which involve the signaling pathways related with inflammation. The levels of serum oleic acid [22.0 (12.9-23.5) μmol/mL vs. 41.5 (40.5-43.8) ] μmol/mL, P<0.001] and IL-1 receptor antagonist [398.6 (356.6-730.1) pg/mL vs. 1589.5 (1580.3-1595.5) pg/mL, P<0.001] decreased in ruptured aneurysm group compared to that in unruptured aneurysm group, while the levels of arachnoid acid [59.9 (26.7-76.6) μmol/mL vs. 12.7 (10.7-16.9) μmol/mL, P<0.001], IL-1β [28.8 (28.8-157.5) pg/mL vs. 46.7 (17.2-61.1) pg/mL, P=0.010], monocyte chemoattractant protein (MCP-1) [7.9 (3.0-15.3) pg/mL vs. 0.7 (0.5-1.1) pg/mL, P<0.001], IL-6 [9.8 (3.9-15.3) pg/mL vs. 2.8 (2.6-3.0) pg/mL, P<0.001] and TNF-α [141.9 (37.1-555.7) pg/mL vs. 345.1 (307.8-384.5) pg/mL, P=0.006] increased in ruptured aneurysm group compared to that in unruptured aneurysm groups. The correlation analysis showed that the expression levels of oleic acid (r2=0.554, P<0.001) , arachnoid acid (r2=0.527, P<0.001) and IL-1β (r2=0.592, P<0.001) in serum and aneurysm tissue had a positive correlation. The ROC curve analysis showed that the levels of oleic acid, arachnoid acid and IL-1β in serum and aneurysm tissue had a good diagnosis value for aneurysm rupture (AUC>0.7) . Multivariate logistic regression analysis showed that serum oleic acid (OR 0.74, 95% CI 0.60-0.87, P=0.001) was an independent risk factor for aneurysm rupture.
Conclusions The levels of oleic acid and IL-1 receptor antagonist decreased, and the levels of arachnoid acid, IL-1β, MCP-1 and TNF-α increased in serum and aneurysm tissue in patients with ruptured intracranial aneurysm, compared to that in patients with unruptured aneurysm. The decreased level of serum oleic acid was an independent risk factor for intracranial aneurysm rupture.

Key words: Intracranial aneurysm; Rupture; Metabolite; Cytokine; Biomarker

摘要： 目的探讨血清与颅内动脉瘤组织中与动脉瘤破裂相关的代谢物和细胞因子标志物。
方法前瞻性纳入2020年1月—2022年1月行单发颅内动脉瘤手术治疗的患者。收集患者的动脉瘤组织和血清样本。根据患者入院时是否发生动脉瘤破裂分为破裂颅内动脉瘤和未破裂颅内动脉瘤组。采用靶向代谢组学的方法检测动脉瘤组织和血清中的代谢物，采用46种细胞因子芯片的方法检测细胞因子。运用Spearman相关性分析血清与颅内动脉瘤组织中与动脉瘤破裂相关的代谢物和细胞因子。运用ROC检测代谢物和细胞因子能否区分破裂和未破裂的颅内动脉瘤。运用logistic回归分析探讨和颅内动脉瘤破裂相关的因素。
结果研究纳入25例破裂和31例未破裂颅内动脉瘤患者。非靶向代谢组学和细胞因子谱学分析显示，在破裂和未破裂颅内动脉瘤组织之间，有43个代谢物和5个细胞因子的差异具有统计学意义（均P<0.05）。富集分析显示这些差异代谢物和细胞因子主要为脂质和脂质类似物，并参与炎症相关的信号通路。靶向代谢组学和细胞因子谱学分析显示颅内动脉瘤破裂患者血清中油酸[22.0（12.9～23.5）μmol/mL vs. 41.5（40.5～43.8）μmol/mL，P<0.001]和I L-1受体拮抗剂（I L-1 receptor antagonist，I L-1ra）[398.6（356.6～730.1）pg/mL vs. 1589.5（1580.3～1595.5）pg/mL，P<0.001]较未破裂颅内动脉瘤患者更低，而花生四烯酸[59.9（26.7～76.6）μmol/mL vs. 12.7（10.7～16.9）μmol/mL，
P<0.001]、IL-1β[28.8（28.8～157.5）pg/mL vs. 46.7（17.2～61.1）pg/mL，P=0.010]、单核细胞趋化蛋白（monocyte chemoattractant protein，MCP-1）[7.9（3.0～15.3）pg/mL vs. 0.7（0.5～1.1）pg/mL，
P<0.001]、IL-6[9.8（3.9～15.3）pg/mL vs. 2.8（2.6～3.0）pg/mL，P<0.001]和TNF-α[141.9（37.1～555.7）pg/mL vs. 345.1（307.8～384.5）pg/mL，P=0.006]的表达水平在颅内动脉瘤破裂患者中更高。相关性分析显示血清中油酸（r2=0.554，P<0.001）、花生四烯酸（r2=0.527，P<0.001）和IL-1β（r2=0.592，P<0.001）与动脉瘤组织中的表达水平存在正相关性。进一步ROC分析显示，动脉瘤组织中和血清中油酸、花生四烯酸和IL-1β的表达水平能很好地区分破裂和未破裂颅内动脉瘤（AUC>0.7）。多因素logistic回归分析显示，血清油酸水平（OR 0.74，95%CI 0.60～0.87，P=0.001）是颅内动脉瘤破裂的独立相关因素。
结论相比于未破裂颅内动脉瘤，血清和颅内动脉瘤组织中油酸和IL-1ra的表达水平在破裂颅内动脉瘤中显著降低，而花生四烯酸、IL-1β、MCP-1和TNF-α的表达水平在破裂颅内动脉瘤中显著升高。血清油酸降低是颅内动脉瘤破裂的独立危险因素。

关键词: 颅内动脉瘤; 破裂; 代谢物; 细胞因子; 生物标志物

LIU Qingyuan, JIANG Pengjun, WANG Shuo, WU Jun.

Risk Factors for Intracranial Aneurysm Rupture Using Metabolomics and Cytokine Profiling Analysis

[J]. Chinese Journal of Stroke, 2023, 18(04): 463-471.

刘清源, 姜朋军, 王硕, 吴俊. 代谢物组学和细胞因子谱学探讨颅内动脉瘤破裂的危险因素研究 [J]. 中国卒中杂志, 2023, 18(04): 463-471.

References

[1] CLAASSEN J，PARK S. Spontaneous subarachnoid haemorrhage [J]. Lancet，2022，400（10355）：846-862.
[2] CHALOUHI N，HOH B L，HASAN D. Review of cerebral aneurysm formation，growth，and rupture[J]. Stroke，2013，44（12）：3613-3622.
[3] SIGNORELLI F，SELA S，GESUALDO L，et al. Hemodynamic stress，inflammation，and intracranial aneurysm development and rupture：a systematic review[J/OL]. World Neurosurg，2018，115：234-244[2022-11-01]. https://doi.org/10.1016/j.wneu.2018.04.143.
[4] FRÖSEN J，TULAMO R，PAETAU A，et al. Saccular intracranial aneurysm：pathology and mechanisms[J]. Acta Neuropathol，2012，123（6）：773-786.
[5] BRINJIKJI W，ZHU Y Q，LANZINO G，et al. Risk factors for growth of intracranial aneurysms：a systematic review and meta-analysis[J]. AJNR Am J Neuroradiol，2016，37（4）：615-620.
[6] FEGHALI C A，WRIGHT T M. Cytokines in acute and chronic inflammation[J/OL]. Front Biosci，1997，2：d12-d26[2022-11-01]. https://doi.org/10.2741/a171.
[7] GARLANDA C，DINARELLO C A，MANTOVANI A. The interleukin-1 family：back to the future[J]. Immunity，2013，39（6）：1003-1018.
[8] MAI W Q，LIAO Y H. Targeting IL-1β in the treatment of atherosclerosis[J/OL]. Front Immunol，2020，11：589654[2022-11-01]. https://doi.org/10.3389/fimmu.2020.589654.
[9] YANG S Z，LIU Q Y，YANG J H，et al. Increased levels of serum IL-15 and TNF-β indicate the progression of human intracranial aneurysm[J/OL]. Front Aging Neurosci，2022，14：903619[2022-11-01]. https://doi.org/10.3389/fnagi.2022.903619.
[10] LIU Q Y，ZHANG Y S，ZHU C C，et al. Serum IL-1，pyroptosis and intracranial aneurysm wall enhancement：analysis integrating radiology，serum cytokines and histology[J/OL]. Front Cardiovasc Med，2022，9：818789[2022-11-01]. https://doi.org/10.3389/fcvm.2022.818789.
[11] 张鸿祺，杨新健，屈延，等. 中国颅内未破裂动脉瘤诊疗指南2021 [J]. 中国脑血管病杂志，2021，18（9）：31.
[12] CAO H H，ZHU Y J，HU G F，et al. Gut microbiome and metabolites，the future direction of diagnosis and treatment of atherosclerosis? [J/OL]. Pharmacol Res，2023，187：106586[2022-11-01]. https://doi.org/10.1016/j.phrs.2022.106586.
[13] FRÖSEN J，PIIPPO A，PAETAU A，et al. Remodeling of saccular cerebral artery aneurysm wall is associated with rupture：histological analysis of 24 unruptured and 42 ruptured cases[J]. Stroke，2004，35（10）：2287-2293.
[14] MEHTA A，SHAPIRO M D. Apolipoproteins in vascular biology and atherosclerotic disease[J]. Nat Rev Cardiol，2022，19（3）：168-179.
[15] BJÖRKEGREN J L M，LUSIS A J. Atherosclerosis：recent developments[J]. Cell，2022，185（10）：1630-1645.
[16] RODEMERK J，JUNKER A，CHEN B，et al. Pathophysiology of intracranial aneurysms：COX-2 expression，iron deposition in aneurysm wall，and correlation with magnetic resonance imaging[J]. Stroke，2020，51（8）：2505-2513.
[17] SAMSON F P，PATRICK A T，FABUNMI T E，et al. Oleic acid，cholesterol，and linoleic acid as angiogenesis initiators[J]. ACS Omega，2020，5（32）：20575-20585.
[18] DINARELLO C A. A clinical perspective of IL-1β as the gatekeeper of inflammation[J]. Eur J Immunol，2011，41（5）：1203-1217.
[19] MORIWAKI T，TAKAGI Y，SADAMASA N，et al. Impaired progression of cerebral aneurysms in interleukin-1beta-deficient mice[J]. Stroke，2006，37（3）：900-905.
[20] MENG H，TUTINO V M，XIANG J，et al. High WSS or low WSS? Complex interactions of hemodynamics with intracranial aneurysm initiation，growth，and rupture：toward a unifying hypothesis[J]. AJNR Am J Neuroradiol，2014，35（7）：1254-1262.
[21] PARKER H，ELLISON S M，HOLLEY R J，et al. Haematopoietic stem cell gene therapy with IL-1Ra rescues cognitive loss in mucopolysaccharidosis IIIA[J/OL]. EMBO Mol Med，2020，12（3）：e11185[2022-11-01]. https://doi.org/10.15252/emmm.201911185.
[22] KOCH M，ACHARJEE A，AMENT Z，et al. Machine learning-driven metabolomic evaluation of cerebrospinal fluid：insights into poor outcomes after aneurysmal subarachnoid hemorrhage[J]. Neurosurgery，2021，88（5）：1003-1011.