GRP75-driven, cell-cycle-dependent macropinocytosis of Tat/pDNA-Ca2+ nanoparticles underlies distinct gene therapy effect in ovarian cancer
Abstract Practice of tumor-targeted suicide gene therapy is hampered by unsafe and low efficient delivery of plasmid DNA (pDNA). Using HIV-Tat-derived peptide (Tat) to non-covalently form Tat/pDNA complexes advances the delivery performance. However, this innovative approach is still limited by intr...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2022-07-01
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| Series: | Journal of Nanobiotechnology |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12951-022-01530-6 |
| _version_ | 1811277735230701568 |
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| author | Linjia Su Zhe Sun Fangzheng Qi Huishan Su Luomeng Qian Jing Li Liang Zuo Jinhai Huang Zhilin Yu Jinping Li Zhinan Chen Sihe Zhang |
| author_facet | Linjia Su Zhe Sun Fangzheng Qi Huishan Su Luomeng Qian Jing Li Liang Zuo Jinhai Huang Zhilin Yu Jinping Li Zhinan Chen Sihe Zhang |
| author_sort | Linjia Su |
| collection | DOAJ |
| description | Abstract Practice of tumor-targeted suicide gene therapy is hampered by unsafe and low efficient delivery of plasmid DNA (pDNA). Using HIV-Tat-derived peptide (Tat) to non-covalently form Tat/pDNA complexes advances the delivery performance. However, this innovative approach is still limited by intracellular delivery efficiency and cell-cycle status. In this study, Tat/pDNA complexes were further condensed into smaller, nontoxic nanoparticles by Ca2+ addition. Formulated Tat/pDNA-Ca2+ nanoparticles mainly use macropinocytosis for intercellular delivery, and their macropinocytic uptake was persisted in mitosis (M-) phase and highly activated in DNA synthesis (S-) phase of cell-cycle. Over-expression or phosphorylation of a mitochondrial chaperone, 75-kDa glucose-regulated protein (GRP75), promoted monopolar spindle kinase 1 (MPS1)-controlled centrosome duplication and cell-cycle progress, but also driven cell-cycle-dependent macropinocytosis of Tat/pDNA-Ca2+ nanoparticles. Further in vivo molecular imaging based on DF (Fluc-eGFP)-TF (RFP-Rluc-HSV-ttk) system showed that Tat/pDNA-Ca2+ nanoparticles exhibited highly suicide gene therapy efficiency in mouse model xenografted with human ovarian cancer. Furthermore, arresting cell-cycle at S-phase markedly enhanced delivery performance of Tat/pDNA-Ca2+ nanoparticles, whereas targeting GRP75 reduced their macropinocytic delivery. More importantly, in vivo targeting GRP75 combined with cell-cycle or macropinocytosis inhibitors exhibited distinct suicide gene therapy efficiency. In summary, our data highlight that mitochondrial chaperone GRP75 moonlights as a biphasic driver underlying cell-cycle-dependent macropinocytosis of Tat/pDNA-Ca2+ nanoparticles in ovarian cancer. |
| first_indexed | 2024-04-13T00:22:04Z |
| format | Article |
| id | doaj.art-47c34d7741b94ee0b201803d80bfcabd |
| institution | Directory Open Access Journal |
| issn | 1477-3155 |
| language | English |
| last_indexed | 2024-04-13T00:22:04Z |
| publishDate | 2022-07-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Nanobiotechnology |
| spelling | doaj.art-47c34d7741b94ee0b201803d80bfcabd2022-12-22T03:10:44ZengBMCJournal of Nanobiotechnology1477-31552022-07-0120112010.1186/s12951-022-01530-6GRP75-driven, cell-cycle-dependent macropinocytosis of Tat/pDNA-Ca2+ nanoparticles underlies distinct gene therapy effect in ovarian cancerLinjia Su0Zhe Sun1Fangzheng Qi2Huishan Su3Luomeng Qian4Jing Li5Liang Zuo6Jinhai Huang7Zhilin Yu8Jinping Li9Zhinan Chen10Sihe Zhang11Department of Cell Biology, School of Medicine, Nankai University, Nankai DistrictSchool of Life Sciences, Tianjin UniversityDepartment of Cell Biology, School of Medicine, Nankai University, Nankai DistrictDepartment of Cell Biology, School of Medicine, Nankai University, Nankai DistrictDepartment of Cell Biology, School of Medicine, Nankai University, Nankai DistrictDepartment of Cell Biology, School of Medicine, Nankai University, Nankai DistrictDepartment of Cell Biology, School of Medicine, Nankai University, Nankai DistrictSchool of Life Sciences, Tianjin UniversityState Key Laboratory of Medicinal Chemical Biology, College of Chemistry, Nankai UniversityDepartment of Medical Biochemistry and Microbiology, Uppsala UniversityNational Translational Science Center for Molecular Medicine, Department of Cell Biology, State Key Laboratory of Cancer Biology, Fourth Military Medical UniversityDepartment of Cell Biology, School of Medicine, Nankai University, Nankai DistrictAbstract Practice of tumor-targeted suicide gene therapy is hampered by unsafe and low efficient delivery of plasmid DNA (pDNA). Using HIV-Tat-derived peptide (Tat) to non-covalently form Tat/pDNA complexes advances the delivery performance. However, this innovative approach is still limited by intracellular delivery efficiency and cell-cycle status. In this study, Tat/pDNA complexes were further condensed into smaller, nontoxic nanoparticles by Ca2+ addition. Formulated Tat/pDNA-Ca2+ nanoparticles mainly use macropinocytosis for intercellular delivery, and their macropinocytic uptake was persisted in mitosis (M-) phase and highly activated in DNA synthesis (S-) phase of cell-cycle. Over-expression or phosphorylation of a mitochondrial chaperone, 75-kDa glucose-regulated protein (GRP75), promoted monopolar spindle kinase 1 (MPS1)-controlled centrosome duplication and cell-cycle progress, but also driven cell-cycle-dependent macropinocytosis of Tat/pDNA-Ca2+ nanoparticles. Further in vivo molecular imaging based on DF (Fluc-eGFP)-TF (RFP-Rluc-HSV-ttk) system showed that Tat/pDNA-Ca2+ nanoparticles exhibited highly suicide gene therapy efficiency in mouse model xenografted with human ovarian cancer. Furthermore, arresting cell-cycle at S-phase markedly enhanced delivery performance of Tat/pDNA-Ca2+ nanoparticles, whereas targeting GRP75 reduced their macropinocytic delivery. More importantly, in vivo targeting GRP75 combined with cell-cycle or macropinocytosis inhibitors exhibited distinct suicide gene therapy efficiency. In summary, our data highlight that mitochondrial chaperone GRP75 moonlights as a biphasic driver underlying cell-cycle-dependent macropinocytosis of Tat/pDNA-Ca2+ nanoparticles in ovarian cancer.https://doi.org/10.1186/s12951-022-01530-6MacropinocytosisCell-cycleGlucose-regulated proteinTat/pDNA-Ca2+ nanoparticleSuicide gene therapyOvarian cancer |
| spellingShingle | Linjia Su Zhe Sun Fangzheng Qi Huishan Su Luomeng Qian Jing Li Liang Zuo Jinhai Huang Zhilin Yu Jinping Li Zhinan Chen Sihe Zhang GRP75-driven, cell-cycle-dependent macropinocytosis of Tat/pDNA-Ca2+ nanoparticles underlies distinct gene therapy effect in ovarian cancer Journal of Nanobiotechnology Macropinocytosis Cell-cycle Glucose-regulated protein Tat/pDNA-Ca2+ nanoparticle Suicide gene therapy Ovarian cancer |
| title | GRP75-driven, cell-cycle-dependent macropinocytosis of Tat/pDNA-Ca2+ nanoparticles underlies distinct gene therapy effect in ovarian cancer |
| title_full | GRP75-driven, cell-cycle-dependent macropinocytosis of Tat/pDNA-Ca2+ nanoparticles underlies distinct gene therapy effect in ovarian cancer |
| title_fullStr | GRP75-driven, cell-cycle-dependent macropinocytosis of Tat/pDNA-Ca2+ nanoparticles underlies distinct gene therapy effect in ovarian cancer |
| title_full_unstemmed | GRP75-driven, cell-cycle-dependent macropinocytosis of Tat/pDNA-Ca2+ nanoparticles underlies distinct gene therapy effect in ovarian cancer |
| title_short | GRP75-driven, cell-cycle-dependent macropinocytosis of Tat/pDNA-Ca2+ nanoparticles underlies distinct gene therapy effect in ovarian cancer |
| title_sort | grp75 driven cell cycle dependent macropinocytosis of tat pdna ca2 nanoparticles underlies distinct gene therapy effect in ovarian cancer |
| topic | Macropinocytosis Cell-cycle Glucose-regulated protein Tat/pDNA-Ca2+ nanoparticle Suicide gene therapy Ovarian cancer |
| url | https://doi.org/10.1186/s12951-022-01530-6 |
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