Sensitizing drug-resistant cancer cells from blood using microfluidic electroporator.
Direct assessment of patient samples holds unprecedented potential in the treatment of cancer. Circulating tumor cells (CTCs) in liquid biopsies are a rapidly evolving source of primary cells in the clinic and are ideal candidates for functional assays to uncover real-time tumor information in real-...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2022-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0264907 |
| _version_ | 1818138831259435008 |
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| author | Hyun Woo Sung Sung-Eun Choi Chris H Chu Mengxing Ouyang Srivathsan Kalyan Nathan Scott Soojung Claire Hur |
| author_facet | Hyun Woo Sung Sung-Eun Choi Chris H Chu Mengxing Ouyang Srivathsan Kalyan Nathan Scott Soojung Claire Hur |
| author_sort | Hyun Woo Sung |
| collection | DOAJ |
| description | Direct assessment of patient samples holds unprecedented potential in the treatment of cancer. Circulating tumor cells (CTCs) in liquid biopsies are a rapidly evolving source of primary cells in the clinic and are ideal candidates for functional assays to uncover real-time tumor information in real-time. However, a lack of routines allowing direct and active interrogation of CTCs directly from liquid biopsy samples represents a bottleneck for the translational use of liquid biopsies in clinical settings. To address this, we present a workflow for using a microfluidic vortex-assisted electroporation system designed for the functional assessment of CTCs purified from blood. Validation of this approach was assessed through drug response assays on wild-type (HCC827 wt) and gefitinib-resistant (HCC827 GR6) non-small cell lung cancer (NSCLC) cells. HCC827 cells trapped within microscale vortices were electroporated to sequentially deliver drug agents into the cytosol. Electroporation conditions facilitating multi-agent delivery were characterized for both cell lines using an automatic single-cell image fluorescence intensity algorithm. HCC827 GR6 cells spiked into the blood to emulate drug-resistant CTCs were able to be collected with high purity, demonstrating the ability of the device to minimize background cell impact for downstream sensitive cell assays. Using our proposed workflow, drug agent combinations to restore gefitinib sensitivity reflected the anticipated cytotoxic response. Taken together, these results represent a microfluidics multi-drug screening panel workflow that can enable functional interrogation of patient CTCs in situ, thereby accelerating the clinical standardization of liquid biopsies. |
| first_indexed | 2024-12-11T10:18:27Z |
| format | Article |
| id | doaj.art-f78604c7e21f46b4bf48c7b7d36581a9 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-11T10:18:27Z |
| publishDate | 2022-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-f78604c7e21f46b4bf48c7b7d36581a92022-12-22T01:11:33ZengPublic Library of Science (PLoS)PLoS ONE1932-62032022-01-01173e026490710.1371/journal.pone.0264907Sensitizing drug-resistant cancer cells from blood using microfluidic electroporator.Hyun Woo SungSung-Eun ChoiChris H ChuMengxing OuyangSrivathsan KalyanNathan ScottSoojung Claire HurDirect assessment of patient samples holds unprecedented potential in the treatment of cancer. Circulating tumor cells (CTCs) in liquid biopsies are a rapidly evolving source of primary cells in the clinic and are ideal candidates for functional assays to uncover real-time tumor information in real-time. However, a lack of routines allowing direct and active interrogation of CTCs directly from liquid biopsy samples represents a bottleneck for the translational use of liquid biopsies in clinical settings. To address this, we present a workflow for using a microfluidic vortex-assisted electroporation system designed for the functional assessment of CTCs purified from blood. Validation of this approach was assessed through drug response assays on wild-type (HCC827 wt) and gefitinib-resistant (HCC827 GR6) non-small cell lung cancer (NSCLC) cells. HCC827 cells trapped within microscale vortices were electroporated to sequentially deliver drug agents into the cytosol. Electroporation conditions facilitating multi-agent delivery were characterized for both cell lines using an automatic single-cell image fluorescence intensity algorithm. HCC827 GR6 cells spiked into the blood to emulate drug-resistant CTCs were able to be collected with high purity, demonstrating the ability of the device to minimize background cell impact for downstream sensitive cell assays. Using our proposed workflow, drug agent combinations to restore gefitinib sensitivity reflected the anticipated cytotoxic response. Taken together, these results represent a microfluidics multi-drug screening panel workflow that can enable functional interrogation of patient CTCs in situ, thereby accelerating the clinical standardization of liquid biopsies.https://doi.org/10.1371/journal.pone.0264907 |
| spellingShingle | Hyun Woo Sung Sung-Eun Choi Chris H Chu Mengxing Ouyang Srivathsan Kalyan Nathan Scott Soojung Claire Hur Sensitizing drug-resistant cancer cells from blood using microfluidic electroporator. PLoS ONE |
| title | Sensitizing drug-resistant cancer cells from blood using microfluidic electroporator. |
| title_full | Sensitizing drug-resistant cancer cells from blood using microfluidic electroporator. |
| title_fullStr | Sensitizing drug-resistant cancer cells from blood using microfluidic electroporator. |
| title_full_unstemmed | Sensitizing drug-resistant cancer cells from blood using microfluidic electroporator. |
| title_short | Sensitizing drug-resistant cancer cells from blood using microfluidic electroporator. |
| title_sort | sensitizing drug resistant cancer cells from blood using microfluidic electroporator |
| url | https://doi.org/10.1371/journal.pone.0264907 |
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