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Abstract
Breast cancer, a leading cause of female mortality due to delayed detection owing to asymptomatic nature and limited early diagnostic tools, was investigated using a multi-modal approach. Plasma-derived small EVs from breast cancer patients (BrCa, n = 74) and healthy controls (HC, n = 30) were analyzed. Small EVs (n = 104), isolated through chemical precipitation, underwent characterization via transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA). Validation involved antibody-based tests (TSG101, CD9, CD81, CD63). Infrared spectra of small EVs were obtained, revealing significant differences in lipid acyl chains, particularly in the C–H stretching of CH3. The study focused on the lipid region (3050–2900 cm−1), identifying peaks (3015 cm−1, 2960 cm−1, 2929 cm−1) as distinctive lipid characteristics. Spectroscopic lipid-to-lipid ratios [(I3015/I2929), (I2960/I2929)] emerged as prominent breast cancer markers. Exploration of protein, nucleic acid, and carbohydrate ratios indicated variations in alpha helices, asymmetric C–H stretching vibrations, and C–O stretching at 1033 cm−1. Principal component analysis (PCA) successfully differentiated BrCa and HC small EVs, and heatmap analysis and receiver operating characteristic (ROC) curve evaluations underscored the discriminatory power of lipid ratios. Notably, (I2960/I2929) exhibited 100% sensitivity and specificity, highlighting its potential as a robust BrCa sEV marker for breast cancer detection.
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1 All India Institute of Medical Sciences, Department of Biophysics, New Delhi, India (GRID:grid.413618.9) (ISNI:0000 0004 1767 6103)
2 All India Institute of Medical Sciences, Department of Pathology, New Delhi, India (GRID:grid.413618.9) (ISNI:0000 0004 1767 6103)
3 All India Institute of Medical Sciences, Department of Surgical Disciplines, New Delhi, India (GRID:grid.413618.9) (ISNI:0000 0004 1767 6103)
4 All India Institute of Medical Sciences, Department of Medical Oncology, New Delhi, India (GRID:grid.413618.9) (ISNI:0000 0004 1767 6103)
5 Indian Institute of Technology (BHU), Department of Pharmaceutical Engineering & Technology, Varanasi, India (GRID:grid.467228.d) (ISNI:0000 0004 1806 4045)
6 Lulea University of Technology, Department of Health, Education, and Technology, Luleå, Sweden (GRID:grid.6926.b) (ISNI:0000 0001 1014 8699)
7 All India Institute of Medical Sciences, Department of Biophysics, New Delhi, India (GRID:grid.413618.9) (ISNI:0000 0004 1767 6103); Lulea University of Technology, Department of Health, Education, and Technology, Luleå, Sweden (GRID:grid.6926.b) (ISNI:0000 0001 1014 8699)