Fluorescence Super-Resolution Imaging Chip for Gene Silencing Exosomes
Tumor cell-derived extracellular vesicles and their cargo of bioactive substances have gradually been recognized as novel biomarkers for cancer diagnosis. Meanwhile, the PD-L1 (Programmed Death-Ligand 1) protein, as an immune checkpoint molecule, is highly expressed on certain tumor cells and holds...
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Published in | Sensors (Basel, Switzerland) Vol. 24; no. 1; p. 173 |
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Main Authors | , , , , , , |
Format | Journal Article |
Language | English |
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28.12.2023
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Abstract | Tumor cell-derived extracellular vesicles and their cargo of bioactive substances have gradually been recognized as novel biomarkers for cancer diagnosis. Meanwhile, the PD-L1 (Programmed Death-Ligand 1) protein, as an immune checkpoint molecule, is highly expressed on certain tumor cells and holds significant potential in immune therapy. In comparison to PD-L1 monoclonal antibodies, the inhibitory effect of PD-L1 siRNA (small interfering RNA) is more advantageous. In this article, we introduced a microfluidic chip integrating cell cultivation and exosome detection modules, which were intended for the investigation of the gene silencing effect of PD-L1 siRNA. Basically, cells were first cultured with PD-L1 siRNA in the chip. Then, the secreted exosomes were detected via super-resolution imaging, to validate the inhibitory effect of siRNA on PD-L1 expression. To be specific, a "sandwich" immunological structure was employed to detect exosomes secreted from HeLa cells. Immunofluorescence staining and DNA-PAINT (DNA Point Accumulation for Imaging in Nanoscale Topography) techniques were utilized to quantitatively analyze the PD-L1 proteins on HeLa exosomes, which enabled precise structural and content analysis of the exosomes. Compared with other existing PD-L1 detection methods, the advantages of our work include, first, the integration of microfluidic chips greatly simplifying the cell culture, gene silencing, and PD-L1 detection procedures. Second, the utilization of DNA-PAINT can provide an ultra-high spatial resolution, which is beneficial for exosomes due to their small sizes. Third, qPAINT could allow quantitative detection of PD-L1 with better precision. Hence, the combination of the microfluidic chip with DNA-PAINT could provide a more powerful integrated platform for the study of PD-L1-related tumor immunotherapy. |
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AbstractList | Tumor cell-derived extracellular vesicles and their cargo of bioactive substances have gradually been recognized as novel biomarkers for cancer diagnosis. Meanwhile, the PD-L1 (Programmed Death-Ligand 1) protein, as an immune checkpoint molecule, is highly expressed on certain tumor cells and holds significant potential in immune therapy. In comparison to PD-L1 monoclonal antibodies, the inhibitory effect of PD-L1 siRNA (small interfering RNA) is more advantageous. In this article, we introduced a microfluidic chip integrating cell cultivation and exosome detection modules, which were intended for the investigation of the gene silencing effect of PD-L1 siRNA. Basically, cells were first cultured with PD-L1 siRNA in the chip. Then, the secreted exosomes were detected via super-resolution imaging, to validate the inhibitory effect of siRNA on PD-L1 expression. To be specific, a "sandwich" immunological structure was employed to detect exosomes secreted from HeLa cells. Immunofluorescence staining and DNA-PAINT (DNA Point Accumulation for Imaging in Nanoscale Topography) techniques were utilized to quantitatively analyze the PD-L1 proteins on HeLa exosomes, which enabled precise structural and content analysis of the exosomes. Compared with other existing PD-L1 detection methods, the advantages of our work include, first, the integration of microfluidic chips greatly simplifying the cell culture, gene silencing, and PD-L1 detection procedures. Second, the utilization of DNA-PAINT can provide an ultra-high spatial resolution, which is beneficial for exosomes due to their small sizes. Third, qPAINT could allow quantitative detection of PD-L1 with better precision. Hence, the combination of the microfluidic chip with DNA-PAINT could provide a more powerful integrated platform for the study of PD-L1-related tumor immunotherapy. |
Audience | Academic |
Author | Yin, Gaoqiang Qi, Tongsheng Wei, Jinxiu Wang, Tingyu Wang, Zhuyuan Zong, Shenfei Cui, Yiping |
AuthorAffiliation | Advanced Photonics Center, School of Electronic Science & Engineering, Southeast University, Nanjing 210096, China; 220211711@seu.edu.cn (G.Y.); 213162551@seu.edu.cn (T.Q.); 230208633@seu.edu.cn (J.W.); 230208632@seu.edu.cn (T.W.); wangzy@seu.edu.cn (Z.W.); cyp@seu.edu.cn (Y.C.) |
AuthorAffiliation_xml | – name: Advanced Photonics Center, School of Electronic Science & Engineering, Southeast University, Nanjing 210096, China; 220211711@seu.edu.cn (G.Y.); 213162551@seu.edu.cn (T.Q.); 230208633@seu.edu.cn (J.W.); 230208632@seu.edu.cn (T.W.); wangzy@seu.edu.cn (Z.W.); cyp@seu.edu.cn (Y.C.) |
Author_xml | – sequence: 1 givenname: Gaoqiang surname: Yin fullname: Yin, Gaoqiang organization: Advanced Photonics Center, School of Electronic Science & Engineering, Southeast University, Nanjing 210096, China – sequence: 2 givenname: Tongsheng surname: Qi fullname: Qi, Tongsheng organization: Advanced Photonics Center, School of Electronic Science & Engineering, Southeast University, Nanjing 210096, China – sequence: 3 givenname: Jinxiu surname: Wei fullname: Wei, Jinxiu organization: Advanced Photonics Center, School of Electronic Science & Engineering, Southeast University, Nanjing 210096, China – sequence: 4 givenname: Tingyu surname: Wang fullname: Wang, Tingyu organization: Advanced Photonics Center, School of Electronic Science & Engineering, Southeast University, Nanjing 210096, China – sequence: 5 givenname: Zhuyuan surname: Wang fullname: Wang, Zhuyuan organization: Advanced Photonics Center, School of Electronic Science & Engineering, Southeast University, Nanjing 210096, China – sequence: 6 givenname: Yiping surname: Cui fullname: Cui, Yiping organization: Advanced Photonics Center, School of Electronic Science & Engineering, Southeast University, Nanjing 210096, China – sequence: 7 givenname: Shenfei surname: Zong fullname: Zong, Shenfei organization: Advanced Photonics Center, School of Electronic Science & Engineering, Southeast University, Nanjing 210096, China |
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Keywords | PD-L1 microfluidic chip integrated platform DNA-PAINT exosomes immunotherapy |
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SubjectTerms | B7-H1 Antigen - genetics Biopsy Cell culture DNA DNA-PAINT Exosomes Fluorescence Genes Genetic engineering HeLa Cells Humans Immune system Immunotherapy integrated platform Localization Lymphocytes microfluidic chip Microfluidics Monoclonal antibodies PD-L1 Proteins RNA RNA, Small Interfering - genetics |
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Title | Fluorescence Super-Resolution Imaging Chip for Gene Silencing Exosomes |
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