Small Extracellular Vesicles in Pre-Therapy Plasma Predict Clinical Outcome in Non-Small-Cell Lung Cancer Patients
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Patients and Plasma Samples
2.2. Protein Measurement Using the BCA Protein Assay
2.3. Exosome Size and Concentration Assessment by Tunable Resistive Pulse Sensing (TRPS)
2.4. Transmission Electron Microscopy (TEM)
2.5. Detection of the Immune Cargo in Isolated sEV by Western Blots
2.6. Flow Cytometry for Immunophenotypic Analysis and Enumeration of Immune Cells
2.7. Statistical Analysis
3. Results
3.1. Clinicopathological Characteristics of NSCLC Patients
3.2. Characterization of Exosomes Isolated from the Plasma of NSCLC Patients
3.3. Association of TEP Levels with the Clinicopathological Characteristics of Chemotherapy-Naïve NSCLC Patients
3.4. Prognostic Role of TEP Levels in Patients with NSCLC
3.5. Quantitative Distribution of Peripheral Immune Effector Cells in NSCLC Patients with High and Low TEP Levels
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Patients (n = 76) Levels of Exosomes | ||||
---|---|---|---|---|
Parameters | n (%) | Low | High | p Value |
Age (years) | Median 65 (range, 44–84) | 0.037 | ||
<65 | 38 (50) | 12 | 22 | |
≥65 | 38 (50) | 26 | 16 | |
Gender | 0.544 | |||
Male | 63 (82.9) | 30 | 33 | |
Female | 13 (17.1) | 8 | 5 | |
Stage | 0.318 | |||
Stage IIIA/B | 23 (30.3) | 14 | 9 | |
Stage IV | 53 (69.7) | 24 | 29 | |
Histology | 0.100 | |||
Squamous | 37 (48.7) | 19 | 18 | |
Non-squamous | 39 (51.3) | 19 | 20 | |
EGFR status | 0.484 | |||
Wild-type | 30 (39.5) | 16 | 14 | |
Mutant | 2 (2.6) | 0 | 2 | |
NE | 44 (57.9) | 22 | 22 | |
Platinum-based chemotherapy regimen | 0.725 | |||
Taxanes | 35 (47.9) | 18 | 17 | |
Pemetrexed | 18 (24.7) | 8 | 10 | |
Gemcitabine | 18 (24.7) | 10 | 8 | |
Vinorelbine | 2 (2.7) | 1 | 1 | |
Response to therapy (after 3 cycles) | 0.001 | |||
PR | 19 (25) | 14 | 5 | |
SD | 20 (26.3) | 15 | 5 | |
PD | 35 (46) | 8 | 27 | |
NE | 2 (2.7) | 1 | 1 |
Univariate Analysis | PROGRESSION-FREE SURVIVAL | OVERALL SURVIVAL | ||
Hazard Ratio (95% CI) | p Value | Hazard Ratio (95% CI) | p Value | |
Age (≥65 vs. <65) | 1.351 (0.835–2.187) | 0.220 | 1.028 (0.624–1.696) | 0.912 |
Gender (Male vs. Female) | 1.190 (0.633–2.237) | 0.386 | 1.320 (0.669–2.607) | 0.424 |
Histology (SQ vs. Non-SQ) | 1.345 (0.828–2.183) | 0.231 | 1.286 (0.782–2.114) | 0.322 |
EGFR status (WT vs. Mutant) | 1.657 (0.383–7.164) | 0.499 | 2.707 (0.591–12.404) | 0.200 |
Stage (IV vs. III) | 2.223 (1.265–3.908) | 0.005 | 1.176 (0.689–2.007) | 0.552 |
Exosomes (high vs. low) | 3.074 (1.856–5.089) | <0.0001 | 3.755 (2.215–6.365) | <0.0001 |
Multivariate Analysis | PROGRESSION-FREE SURVIVAL | OVERALL SURVIVAL | ||
Hazard Ratio (95% CI) | p Value | Hazard Ratio (95% CI) | p Value | |
Stage (IV vs. III) | 1.955 (1.113–3.435) | 0.020 | N/A | N/A |
Exosomes (high vs. low) | 2.852 (1.714–4.744) | <0.0001 | 3.755 (2.215–6.365) | <0.0001 |
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Vetsika, E.-K.; Sharma, P.; Samaras, I.; Markou, A.; Georgoulias, V.; Whiteside, T.L.; Kotsakis, A. Small Extracellular Vesicles in Pre-Therapy Plasma Predict Clinical Outcome in Non-Small-Cell Lung Cancer Patients. Cancers 2021, 13, 2041. https://doi.org/10.3390/cancers13092041
Vetsika E-K, Sharma P, Samaras I, Markou A, Georgoulias V, Whiteside TL, Kotsakis A. Small Extracellular Vesicles in Pre-Therapy Plasma Predict Clinical Outcome in Non-Small-Cell Lung Cancer Patients. Cancers. 2021; 13(9):2041. https://doi.org/10.3390/cancers13092041
Chicago/Turabian StyleVetsika, Eleni-Kyriaki, Priyanka Sharma, Ioannis Samaras, Alexandra Markou, Vassilis Georgoulias, Theresa L. Whiteside, and Athanasios Kotsakis. 2021. "Small Extracellular Vesicles in Pre-Therapy Plasma Predict Clinical Outcome in Non-Small-Cell Lung Cancer Patients" Cancers 13, no. 9: 2041. https://doi.org/10.3390/cancers13092041