TY - JOUR
T1 - Isolation and mutational assessment of pancreatic cancer extracellular vesicles using a microfluidic platform
AU - Kamyabi, Nabiollah
AU - Abbasgholizadeh, Reza
AU - Maitra, Anirban
AU - Ardekani, Arezoo
AU - Biswal, Sibani L.
AU - Grande-Allen, K. Jane
N1 - Funding Information:
Scanning electron microscopy was performed at the High-Resolution Electron Microscopy Facility, while nanoString analysis was performed at the Sequencing and Microarray facility (SMF) in MD Anderson Cancer Center (Houston, TX), both of which are supported by the NCI Cancer Center Support Grant (P30CA016672). N.K. was supported by the CPRIT Research Training Program (RP170067).
Publisher Copyright:
© 2020, Springer Science+Business Media, LLC, part of Springer Nature.
PY - 2020/6/1
Y1 - 2020/6/1
N2 - Cancer cells release extracellular vesicles known as extracellular vesicles (EVs), containing tumor-derived DNA, RNA and proteins within their cargo, into the circulation. Circulating tumor-derived extracellular vesicles (TEV) can be used in the context of serial “liquid biopsies” for early detection of cancer, for monitoring disease burden in patients, and for assessing recurrence in the post-resection setting. Nonetheless, isolating sufficient TEV by ultracentrifugation-based approaches, in order to enable molecular assessment of EVs cargo, can be an arduous, time-consuming process and is inconsistent in the context of yield and purity among institutions. Herein, we describe a microfluidic platform, which we have named MITEV (Microfluidic Isolation of Tumor-derived Extracellular Vesicles) for the rapid isolation of TEV from the plasma of pancreatic cancer patients. The device, which has ~100,000 pillars placed in a zigzag pattern and is coated with antibodies against generic EV surface proteins (anti-CD63, -CD9, and -CD81 antibodies) or the TEV specific anti-Epithelial Cell Adhesion Molecule (EpCAM) antibody, is capable of high-throughput EVs isolation and yields sufficient DNA (total of ~2–14 ng from 2-ml plasma) for downstream genomic analysis. Using two independent quantitative platforms, droplet digital polymerase chain reaction (ddPCR) and molecular barcoding using nanoString nCounter® technology, we can reliably identify KRAS mutations within isolated TEV of treatment-naïve metastatic pancreatic cancer patients. Our study suggests that the MITEV device can be used for point-of-care applications, such as in the context of monitoring residual or recurrent tumor presence in pancreatic cancer patients undergoing therapy.
AB - Cancer cells release extracellular vesicles known as extracellular vesicles (EVs), containing tumor-derived DNA, RNA and proteins within their cargo, into the circulation. Circulating tumor-derived extracellular vesicles (TEV) can be used in the context of serial “liquid biopsies” for early detection of cancer, for monitoring disease burden in patients, and for assessing recurrence in the post-resection setting. Nonetheless, isolating sufficient TEV by ultracentrifugation-based approaches, in order to enable molecular assessment of EVs cargo, can be an arduous, time-consuming process and is inconsistent in the context of yield and purity among institutions. Herein, we describe a microfluidic platform, which we have named MITEV (Microfluidic Isolation of Tumor-derived Extracellular Vesicles) for the rapid isolation of TEV from the plasma of pancreatic cancer patients. The device, which has ~100,000 pillars placed in a zigzag pattern and is coated with antibodies against generic EV surface proteins (anti-CD63, -CD9, and -CD81 antibodies) or the TEV specific anti-Epithelial Cell Adhesion Molecule (EpCAM) antibody, is capable of high-throughput EVs isolation and yields sufficient DNA (total of ~2–14 ng from 2-ml plasma) for downstream genomic analysis. Using two independent quantitative platforms, droplet digital polymerase chain reaction (ddPCR) and molecular barcoding using nanoString nCounter® technology, we can reliably identify KRAS mutations within isolated TEV of treatment-naïve metastatic pancreatic cancer patients. Our study suggests that the MITEV device can be used for point-of-care applications, such as in the context of monitoring residual or recurrent tumor presence in pancreatic cancer patients undergoing therapy.
KW - Extracellular vesicles
KW - Microfluidics
KW - Pancreatic cancer
KW - ddPCR
UR - http://www.scopus.com/inward/record.url?scp=85081735858&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85081735858&partnerID=8YFLogxK
U2 - 10.1007/s10544-020-00483-7
DO - 10.1007/s10544-020-00483-7
M3 - Article
C2 - 32162067
AN - SCOPUS:85081735858
SN - 1387-2176
VL - 22
JO - Biomedical Microdevices
JF - Biomedical Microdevices
IS - 2
M1 - 23
ER -