Assessment of a Size-Based Method for Enriching Circulating Tumour Cells in Colorectal Cancer
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Ethical Approval
2.2. Cell Culture
2.3. Immunostaining to Assess MetaCell Recovery Rates and WBC Depletion Rates
2.4. Gene Expression Analysis for Assessing Recovery and WBC Depletion Rates for MetaCell
2.5. Patient Sampling and Blood Processing
2.6. Statistical Analysis
3. Results
3.1. Qualitative Analysis Demonstrates MetaCell’s Ability for Capturing CRC Cells from Blood Spiked with High CRC Cell Numbers
3.2. Gene Expression Analysis Confirmed MetaCells’s Ability to Highly Enrich for CRC Cells
3.3. Quantitative Assessment Demonstrates That MetaCell Recovery Rates Were Not Affected by Low Cell Numbers
3.4. Gene Expression Analysis Revealed Different CRC Cell Numbers Did Not Have a Significant Effect on MetaCell Recovery Rates
3.5. Estimating WBC Depletion Rate Revealed That MetaCell Significantly Depleted the WBC Population
3.6. Patient Characteristics and CTC Detection
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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Characteristics | Number of Patients (%) |
---|---|
Age (years) | |
40–70 | 11 (58.8) |
71–80 | 6 (35.2) |
Gender | |
Male | 7 (41.2) |
Female | 10 (58.8) |
Ethnicity | |
New Zealand European | 15 (88.2) |
Other European | 2 (11.8) |
Clinical Stage (AJCC 8th edition) | |
Early Stage (stage I and II) | 7 (41.2) |
Late Stage (stage III and IV) | 10 (58.8) |
Tumour Site | |
Left sided CRC | 10 (58.8) |
Right sided CRC | 7 (41.2) |
Metastasis | |
No distant metastases | 11 (64.7) |
Distant metastases present | 6 (35.3) |
MMR (IHC) | |
MMR proficient | 11 (64.7) |
MMR deficient | 6 (35.3) |
BRAF (IHC) | |
BRAF V600E −ve | 5 (29.4) |
BRAF V600E +ve | 7 (41.2) |
ND | 5 (29.4) |
Clinical or Pathological Variables | n | % CTC +ve Patients (n) | % CTC −ve Patients (n) | p-Value | % cCTC +ve Patients (n) | % cCTC −ve Patients (n) | p-Value |
---|---|---|---|---|---|---|---|
Age (years) | |||||||
40–70 | 11 | 54.5 (6) | 45.5 (5) | >0.99 | 18.2 (2) | 81.8 (9) | 0.58 |
71–85 | 6 | 50 (3) | 50 (3) | 33.3 (2) | 66.7 (4) | ||
Gender | |||||||
Male | 7 | 71.4 (5) | 28.6 (2) | 0.33 | 42.8 (3) | 57.2 (4) | 0.25 |
Female | 10 | 40 (4) | 60 (6) | 10 (1) | 90 (9) | ||
Clinical stage | |||||||
Early stage (I & II) | 7 | 71.4 (5) | 28.6 (2) | 0.33 | 14.3 (1) | 85.7 (6) | 0.60 |
Late stage (III & IV) | 10 | 40 (4) | 60 (6) | 30 (3) | 70 (7) | ||
Tumour site | |||||||
Left side | 10 | 60 (6) | 40 (4) | 0.63 | 30 (3) | 70 (7) | 0.60 |
Right side | 7 | 42.8 (3) | 57.2 (4) | 14.3 (1) | 85.7 (6) | ||
Metastasis | |||||||
No distant metastases | 11 | 54.5 (6) | 45.5 (5) | >0.99 | 18.2 (2) | 81.8 (9) | 0.58 |
Distant metastases present | 6 | 50 (3) | 50 (3) | 33.3 (2) | 66.7 (4) | ||
MMR (IHC) | |||||||
MMR proficient | 11 | 63.6 (7) | 36.4 (4) | 0.33 | 27.2 (3) | 72.8 (8) | >0.99 |
MMR deficient | 6 | 33.3 (2) | 66.7 (4) | 16.6 (1) | 83.4 (5) | ||
BRAF V600E (IHC) | |||||||
BRAF −ve | 5 | 40 (2) | 60 (3) | >0.99 | 40 (2) | 60 (3) | 0.52 |
BRAF +ve | 7 | 42.8 (3) | 57.2 (4) | 14.3 (1) | 85.7 (6) | ||
ND | 5 | 80 (4) | 20 (1) | 20 (1) | 80 (4) |
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Vasantharajan, S.S.; Barnett, E.; Gray, E.S.; McCall, J.L.; Rodger, E.J.; Eccles, M.R.; Munro, F.; Pattison, S.; Chatterjee, A. Assessment of a Size-Based Method for Enriching Circulating Tumour Cells in Colorectal Cancer. Cancers 2022, 14, 3446. https://0-doi-org.brum.beds.ac.uk/10.3390/cancers14143446
Vasantharajan SS, Barnett E, Gray ES, McCall JL, Rodger EJ, Eccles MR, Munro F, Pattison S, Chatterjee A. Assessment of a Size-Based Method for Enriching Circulating Tumour Cells in Colorectal Cancer. Cancers. 2022; 14(14):3446. https://0-doi-org.brum.beds.ac.uk/10.3390/cancers14143446
Chicago/Turabian StyleVasantharajan, Sai Shyam, Edward Barnett, Elin S. Gray, John L. McCall, Euan J. Rodger, Michael R. Eccles, Fran Munro, Sharon Pattison, and Aniruddha Chatterjee. 2022. "Assessment of a Size-Based Method for Enriching Circulating Tumour Cells in Colorectal Cancer" Cancers 14, no. 14: 3446. https://0-doi-org.brum.beds.ac.uk/10.3390/cancers14143446