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Conference Paper: Developmental delays and subcellular stress as downstream effects of sonoporation

TitleDevelopmental delays and subcellular stress as downstream effects of sonoporation
Authors
Issue Date2013
PublisherISTU2013
Citation
The 13th International Symposium on Therapeutic Ultrasound (ISTU 2013), Shanghai, China, 12-16 May 2013. In 13th ISTU Final Programme, 2013, p. 150 How to Cite?
AbstractOBJECTIVES: The biological impact of sonoporation has often been overlooked. Here we seek to obtain insight into the cytotoxic impact of sonoporation by gaining new perspectives on anti-proliferative characteristics that may emerge within sonoporated cells. We particularly focused on investigating the cell-cycle progression kinetics of sonoporated cells and identifying organelles that may be stressed in the recovery process. METHODS: In line with recommendations on exposure hardware design, an immersion-based ultrasound platform has been developed. It delivers 1 MHz ultrasound pulses (100 cycles; 1 kHz PRF; 60 s total duration) with 0.45 MPa peak negative pressure to a cell chamber that housed HL-60 leukemia cells and lipid-shelled microbubbles at a 10:1 cell-tobubble ratio (for 1e6/ml cell density). Calcein was used to facilitate tracking of sonoporated cells with enhanced uptake of exogenous molecules. The developmental trend of sonoporated cells was quantitatively analyzed using BrdU/DNA flow cytometry that monitors the cell population’s DNA synthesis kinetics. This allowed us to measure the temporal progression of DNA synthesis of sonoporated cells. To investigate whether sonoporation would upset subcellular homeostasis, post-exposure cell samples were also assayed for various proteins using Western blot analysis. Analysis focus was placed on the endoplasmic reticulum (ER): an important organelle with multi-faceted role in cellular functioning. The post-exposure observation time spanned between 0-24 h. RESULTS: Despite maintaining viability, sonoporated cells were found to exhibit delays in cell-cycle progression. Specifically, their DNA synthesis time was lengthened substantially (for HL-60 cells: 8.7 h for control vs 13.4 h for the sonoporated group). This indicates that sonoporated cells were under stress: a phenomenon that is supported by our Western blot assays showing upregulation of ER-resident enzymes (PDI, Ero1), ER stress sensors (PERK, IRE1), and ER-triggered pro-apoptotic signals (CHOP, JNK). CONCLUSIONS: Sonoporation, whilst being able to facilitate internalization of exogenous molecules, may inadvertently elicit a cellular stress response. These findings seem to echo recent calls for reconsideration of efficiency issues in sonoporation-mediated drug delivery. Further efforts would be necessary to improve the efficiency of sonoporation-based biomedical applications where cell death is not desirable.
DescriptionPosters: no. 2
Control ID: 1672434
Persistent Identifierhttp://hdl.handle.net/10722/189833

 

DC FieldValueLanguage
dc.contributor.authorChen, Xen_US
dc.contributor.authorZhong, Wen_US
dc.contributor.authorYu, ACHen_US
dc.contributor.authorWan, JMFen_US
dc.date.accessioned2013-09-17T15:00:46Z-
dc.date.available2013-09-17T15:00:46Z-
dc.date.issued2013en_US
dc.identifier.citationThe 13th International Symposium on Therapeutic Ultrasound (ISTU 2013), Shanghai, China, 12-16 May 2013. In 13th ISTU Final Programme, 2013, p. 150en_US
dc.identifier.urihttp://hdl.handle.net/10722/189833-
dc.descriptionPosters: no. 2-
dc.descriptionControl ID: 1672434-
dc.description.abstractOBJECTIVES: The biological impact of sonoporation has often been overlooked. Here we seek to obtain insight into the cytotoxic impact of sonoporation by gaining new perspectives on anti-proliferative characteristics that may emerge within sonoporated cells. We particularly focused on investigating the cell-cycle progression kinetics of sonoporated cells and identifying organelles that may be stressed in the recovery process. METHODS: In line with recommendations on exposure hardware design, an immersion-based ultrasound platform has been developed. It delivers 1 MHz ultrasound pulses (100 cycles; 1 kHz PRF; 60 s total duration) with 0.45 MPa peak negative pressure to a cell chamber that housed HL-60 leukemia cells and lipid-shelled microbubbles at a 10:1 cell-tobubble ratio (for 1e6/ml cell density). Calcein was used to facilitate tracking of sonoporated cells with enhanced uptake of exogenous molecules. The developmental trend of sonoporated cells was quantitatively analyzed using BrdU/DNA flow cytometry that monitors the cell population’s DNA synthesis kinetics. This allowed us to measure the temporal progression of DNA synthesis of sonoporated cells. To investigate whether sonoporation would upset subcellular homeostasis, post-exposure cell samples were also assayed for various proteins using Western blot analysis. Analysis focus was placed on the endoplasmic reticulum (ER): an important organelle with multi-faceted role in cellular functioning. The post-exposure observation time spanned between 0-24 h. RESULTS: Despite maintaining viability, sonoporated cells were found to exhibit delays in cell-cycle progression. Specifically, their DNA synthesis time was lengthened substantially (for HL-60 cells: 8.7 h for control vs 13.4 h for the sonoporated group). This indicates that sonoporated cells were under stress: a phenomenon that is supported by our Western blot assays showing upregulation of ER-resident enzymes (PDI, Ero1), ER stress sensors (PERK, IRE1), and ER-triggered pro-apoptotic signals (CHOP, JNK). CONCLUSIONS: Sonoporation, whilst being able to facilitate internalization of exogenous molecules, may inadvertently elicit a cellular stress response. These findings seem to echo recent calls for reconsideration of efficiency issues in sonoporation-mediated drug delivery. Further efforts would be necessary to improve the efficiency of sonoporation-based biomedical applications where cell death is not desirable.-
dc.languageengen_US
dc.publisherISTU2013-
dc.relation.ispartof13th International Symposium on Therapeutic Ultrasound (ISTU 2013) Final Programmeen_US
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.titleDevelopmental delays and subcellular stress as downstream effects of sonoporationen_US
dc.typeConference_Paperen_US
dc.identifier.emailYu, ACH: alfred.yu@hku.hken_US
dc.identifier.emailWan, JMF: jmfwan@hku.hken_US
dc.identifier.authorityYu, ACH=rp00657en_US
dc.identifier.authorityWan, JMF=rp00798en_US
dc.description.naturepostprint-
dc.identifier.hkuros222138en_US
dc.identifier.spage150-
dc.identifier.epage150-
dc.publisher.placeChina-

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