Acute ischemic stroke (AIS) requires detailed etiology information to guide optimal management. Given the pivotal role of lipids in AIS, we conducted a comprehensive lipidomics analysis of paired thrombi and plasma from AIS patients, correlating the findings with stroke etiology. Patients were recruited across four etiologies: cardioembolism (CE), large artery atherosclerosis (LAA), active cancer (Cancer), and undetermined. Plasma and thrombi were collected before and during endovascular thrombectomy and analyzed using in-house targeted lipidomics. Among 51 patients (37 CE, 7 LAA, 4 Cancer, and 3 undetermined), we identified 37 and 70 lipid species significantly different between thrombi in CE and LAA, and CE and Cancer, respectively (FDR-corrected P < 0.05). No significant differences were observed in plasma. Notably, 21 diacylglycerols and 11 polyunsaturated triacylglycerols were depleted (2.5 to 12 folds) in LAA compared to CE, while 10 ceramides and 57 glycerophospholipids were elevated in Cancer. With 80% validation accuracy, 29 and 59 lipids distinguished LAA and Cancer from CE, respectively. A neural network model using these lipids effectively classified undetermined patients. This study emphasizes the significance of thrombus lipids in distinguishing between LAA, CE, and Cancer etiologies in AIS, enhancing our understanding of stroke pathophysiology and informing future clinical managements.
JohnsonCONguyenMRothGA, et al.
Global, regional, and national burden of stroke, 1990–2016: a systematic analysis for the global burden of disease study 2016. Lancet Neurol2019;
18: 439–458.
2.
BenjaminEJMuntnerPAlonsoA, et al.
Heart disease and stroke statistics-2019 update a report from the American heart association. Circulation2019;
139: E56–E528.
3.
AdamsHPJrBendixenBHKappelleLJ, et al.
Classification of subtype of acute ischemic stroke. Definitions for use in a multicenter clinical trial. TOAST. Trial of org 10172 in acute stroke treatment. Stroke1993;
24: 35–41.
4.
SaccoRLKasnerSEBroderickJP, et al.
An updated definition of stroke for the 21st century a statement for healthcare professionals from the American Heart Association/American Stroke Association. Stroke2013;
44: 2064–2089.
5.
HaeuslerKGTütüncüSSchnabelRB.Detection of atrial fibrillation in cryptogenic stroke. Curr Neurol Neurosci Rep2018;
18: 66.
6.
WangHChenSHanZ, et al.
Screening of phospholipids in plasma of large-artery atherosclerotic and cardioembolic stroke patients with hydrophilic interaction chromatography-mass spectrometry. Front Mol Biosci2022;
9: 794057.
7.
WangXZhangLSunW, et al.
Changes of metabolites in acute ischemic stroke and its subtypes. Front Neurosci2020;
14: 580929.
8.
SunDTiedtSYuB, et al.
A prospective study of serum metabolites and risk of ischemic stroke. Neurology2019;
92: e1890–e1898.
9.
BhaskarSSaabJCappelen-SmithC, et al.
Clot histopathology in ischemic stroke with infective endocarditis. Can J Neurol Sci2019;
46: 331–336.
10.
BhaskarSCordatoDCappelen-SmithC, et al.
Clarion call for histopathological clot analysis in “cryptogenic” ischemic stroke: implications for diagnosis and treatment. Ann Clin Transl Neurol2017;
4: 926–930.
11.
FuCHChenCHLinYH, et al.
Fibrin and Platelet-Rich composition in retrieved thrombi hallmarks stroke with active cancer. Stroke2020;
51: 3723–3727.
12.
SuissaLGuigonisJMGraslinF, et al.
Combined omic analyzes of cerebral thrombi: a new molecular approach to identify cardioembolic stroke origin. Stroke2021;
52: 2892–2901.
13.
LiWBaiXHaoJ, et al.
Thrombosis origin identification of cardioembolism and large artery atherosclerosis by distinct metabolites. J Neurointerv Surg2023;
15: 701–707.
14.
TonksKTCosterACFChristopherMJ, et al.
Skeletal muscle and plasma lipidomic signatures of insulin resistance and overweight/obesity in humans. Obesity (Silver Spring)2016;
24: 908–916.
15.
HuynhKBarlowCKJayawardanaKS, et al.
high-throughput plasma lipidomics: detailed mapping of the associations with cardiometabolic risk factors. Cell Chem Biol2019;
26: 71–84.e4.
16.
LappasMMundraPAWongG, et al.
The prediction of type 2 diabetes in women with previous gestational diabetes mellitus using lipidomics. Diabetologia2015;
58: 1436–1442.
17.
Del GobboLCImamuraFAslibekyanS, et al.
ω-3 polyunsaturated fatty acid biomarkers and coronary heart disease pooling project of 19 cohort studies. Jama Intern Med2016;
176: 1155–1166.
18.
NakamuraAOtaniKShichitaT.Lipid mediators and sterile inflammation in ischemic stroke. Int Immunol2020;
32: 719–725.
19.
RinkTJSanchezAHallamTJ.Diacylglycerol and phorbol ester stimulate secretion without raising cytoplasmic free calcium in human platelets. Nature1983;
305: 317–319.
20.
PowersWJRabinsteinAAAckersonT, et al.
2018 Guidelines for the early management of patients with acute ischemic stroke: a guideline for healthcare professionals from the American Heart Association/American Stroke Association. Stroke2018;
49: e46–e110.
21.
BenjaminiYHochbergY.Controlling the false discovery rate: a practical and powerful approach to multiple testing. Journal of the Royal Statistical Society Series B (Methodological)1995;
57: 289–300.
22.
BlaiseBJCorreiaGDSHaggartGA, et al.
Statistical analysis in metabolic phenotyping. Nat Protoc2021;
16: 4299–4326.
23.
TirandiASguraCCarboneF, et al.
Inflammatory biomarkers of ischemic stroke. Intern Emerg Med2023;
18: 723–732.
24.
DeLongJHOhashiSNO'ConnorKC, et al.
Inflammatory responses after ischemic stroke. Semin Immunopathol2022;
44: 625–648.
25.
Boeckh-BehrensTKleineJFZimmerC, et al.
Thrombus histology suggests cardioembolic cause in cryptogenic stroke. Stroke2016;
47: 1864–1871.
26.
MaegerleinCFriedrichBBerndtM, et al.
Impact of histological thrombus composition on preinterventional thrombus migration in patients with acute occlusions of the Middle cerebral artery. Interv Neuroradiol2018;
24: 70–75.
27.
ShimizuHHatakeyamaKSaitoK, et al.
Age and composition of the thrombus retrieved by mechanical thrombectomy from patients with acute ischemic stroke are associated with revascularization and clinical outcomes. Thromb Res2022;
219: 60–69.
28.
HuangJKillingsworthMCBhaskarSMM.Is composition of brain clot retrieved by mechanical thrombectomy associated with stroke aetiology and clinical outcomes in acute ischemic stroke? – a systematic review and meta-analysis. Neurol Int2022;
14: 748–770.
29.
BacigaluppiMSemeranoAGullottaGS, et al.
Insights from thrombi retrieved in stroke due to large vessel occlusion. J Cereb Blood Flow Metab2019;
39: 1433–1451.
30.
KimSKYoonWKimTS, et al.
Histologic analysis of retrieved clots in acute ischemic stroke: Correlation with stroke etiology and Gradient-Echo MRI. AJNR Am J Neuroradiol2015;
36: 1756–1762.
31.
GoldbergIJTrentCMSchulzePC.Lipid metabolism and toxicity in the heart. Cell Metab2012;
15: 805–812.
32.
ChiuHCKovacsABlantonRM, et al.
Transgenic expression of fatty acid transport protein 1 in the heart causes lipotoxic cardiomyopathy. Circ Res2005;
96: 225–233.
33.
XieXTieYFLaiS, et al.
Cardiac-specific CGI-58 deficiency activates the ER stress pathway to promote heart failure in mice. Cell Death Dis2021;
12: 1003.
34.
LeeHCChengWCMaWL, et al.
Association of lipid composition and unsaturated fatty acids of VLDL with atrial remodeling in metabolic syndrome. Sci Rep2023;
13: 6575.
35.
AdkinsYKelleyDS.Mechanisms underlying the cardioprotective effects of omega-3 polyunsaturated fatty acids. J Nutr Biochem2010;
21: 781–792.
36.
PavlovicDNiciforovicDMarkovicM, et al.
Cancer-associated thrombosis: epidemiology, pathophysiological mechanisms, treatment, and risk assessment. Clin Med Insights Oncol2023;
17: 11795549231220297.
37.
ZhangCYangZZhouP, et al.
Phosphatidylserine-exposing tumor-derived microparticles exacerbate coagulation and cancer cell transendothelial migration in triple-negative breast cancer. Theranostics2021;
11: 6445–6460.
38.
SzlasaWZendranIZalesińskaA, et al.
Lipid composition of the cancer cell membrane. J Bioenerg Biomembr2020;
52: 321–342.
39.
BirgeRBBoeltzSKumarS, et al.
Phosphatidylserine is a global immunosuppressive signal in efferocytosis, infectious disease, and cancer. Cell Death Differ2016;
23: 962–978.
40.
SharmaBKanwarSS.Phosphatidylserine: a cancer cell targeting biomarker. Semin Cancer Biol2018;
52: 17–25.
41.
LiuQPLuoQHalimA, et al.
Targeting lipid metabolism of cancer cells: a promising therapeutic strategy for cancer. Cancer Lett2017;
401: 39–45.
42.
BrachtendorfSEl-HindiKGröschS.Ceramide synthases in cancer therapy and chemoresistance. Prog Lipid Res2019;
74: 160–185.
43.
ChenLChenHLiY, et al.
Endocannabinoid and ceramide levels are altered in patients with colorectal cancer. Oncol Rep2015;
34: 447–454.
44.
KarahataySThomasKKoybasiS, et al.
Clinical relevance of ceramide metabolism in the pathogenesis of human head and neck squamous cell carcinoma (HNSCC): attenuation of C(18)-ceramide in HNSCC tumors correlates with lymphovascular invasion and nodal metastasis. Cancer Lett2007;
256: 101–111.
45.
SchiffmannSSandnerJBirodK, et al.
Ceramide synthases and ceramide levels are increased in breast cancer tissue. Carcinogenesis2009;
30: 745–752.
46.
WajapeyeeNBeamonTCGuptaR.Roles and therapeutic targeting of ceramide metabolism in cancer. Mol Metab2024;
83: 101936.
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