Defects of mitochondria-lysosomes communication induce secretion of mitochondria-derived vesicles and drive chemoresistance in ovarian cancer cells
Abstract Background Among the mechanisms of mitochondrial quality control (MQC), generation of mitochondria-derived vesicles (MDVs) is a process to avoid complete failure of mitochondria determining lysosomal degradation of mitochondrial damaged proteins. In this context, RAB7, a late endocytic smal...
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| Format: | Article |
| Language: | English |
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BMC
2024-03-01
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| Series: | Cell Communication and Signaling |
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| Online Access: | https://doi.org/10.1186/s12964-024-01507-y |
| _version_ | 1797266683843313664 |
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| author | Sinforosa Gagliardi Marco Mitruccio Riccardo Di Corato Roberta Romano Alessandra Aloisi Rosaria Rinaldi Pietro Alifano Flora Guerra Cecilia Bucci |
| author_facet | Sinforosa Gagliardi Marco Mitruccio Riccardo Di Corato Roberta Romano Alessandra Aloisi Rosaria Rinaldi Pietro Alifano Flora Guerra Cecilia Bucci |
| author_sort | Sinforosa Gagliardi |
| collection | DOAJ |
| description | Abstract Background Among the mechanisms of mitochondrial quality control (MQC), generation of mitochondria-derived vesicles (MDVs) is a process to avoid complete failure of mitochondria determining lysosomal degradation of mitochondrial damaged proteins. In this context, RAB7, a late endocytic small GTPase, controls delivery of MDVs to late endosomes for subsequent lysosomal degradation. We previously demonstrated that RAB7 has a pivotal role in response to cisplatin (CDDP) regulating resistance to the drug by extracellular vesicle (EVs) secretion. Methods Western blot and immunofluorescence analysis were used to analyze structure and function of endosomes and lysosomes in CDDP chemosensitive and chemoresistant ovarian cancer cell lines. EVs were purified from chemosensitive and chemoresistant cells by ultracentrifugation or immunoisolation to analyze their mitochondrial DNA and protein content. Treatment with cyanide m-chlorophenylhydrazone (CCCP) and RAB7 modulation were used, respectively, to understand the role of mitochondrial and late endosomal/lysosomal alterations on MDV secretion. Using conditioned media from chemoresistant cells the effect of MDVs on the viability after CDDP treatment was determined. Seahorse assays and immunofluorescence analysis were used to study the biochemical role of MDVs and the uptake and intracellular localization of MDVs, respectively. Results We observed that CDDP-chemoresistant cells are characterized by increased MDV secretion, impairment of late endocytic traffic, RAB7 downregulation, an increase of RAB7 in EVs, compared to chemosensitive cells, and downregulation of the TFEB-mTOR pathway overseeing lysosomal and mitochondrial biogenesis and turnover. We established that MDVs can be secreted rather than delivered to lysosomes and are able to deliver CDDP outside the cells. We showed increased secretion of MDVs by chemoresistant cells ultimately caused by the extrusion of RAB7 in EVs, resulting in a dramatic drop in its intracellular content, as a novel mechanism to regulate RAB7 levels. We demonstrated that MDVs purified from chemoresistant cells induce chemoresistance in RAB7-modulated process, and, after uptake from recipient cells, MDVs localize to mitochondria and slow down mitochondrial activity. Conclusions Dysfunctional MQC in chemoresistant cells determines a block in lysosomal degradation of MDVs and their consequent secretion, suggesting that MQC is not able to eliminate damaged mitochondria whose components are secreted becoming effectors and potential markers of chemoresistance. |
| first_indexed | 2024-04-25T01:04:36Z |
| format | Article |
| id | doaj.art-256abdbb623f4165ac3237c370d5a0a5 |
| institution | Directory Open Access Journal |
| issn | 1478-811X |
| language | English |
| last_indexed | 2024-04-25T01:04:36Z |
| publishDate | 2024-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Cell Communication and Signaling |
| spelling | doaj.art-256abdbb623f4165ac3237c370d5a0a52024-03-10T12:18:16ZengBMCCell Communication and Signaling1478-811X2024-03-0122112610.1186/s12964-024-01507-yDefects of mitochondria-lysosomes communication induce secretion of mitochondria-derived vesicles and drive chemoresistance in ovarian cancer cellsSinforosa Gagliardi0Marco Mitruccio1Riccardo Di Corato2Roberta Romano3Alessandra Aloisi4Rosaria Rinaldi5Pietro Alifano6Flora Guerra7Cecilia Bucci8Department of Biological and Environmental Sciences and Technologies, University of SalentoDepartment of Biological and Environmental Sciences and Technologies, University of SalentoInstitute for Microelectronics and Microsystems (IMM), CNRDepartment of Biological and Environmental Sciences and Technologies, University of SalentoInstitute for Microelectronics and Microsystems (IMM), CNRDepartment of Mathematics and Physics “E. De Giorgi”, University of SalentoDepartment of Biological and Environmental Sciences and Technologies, University of SalentoDepartment of Biological and Environmental Sciences and Technologies, University of SalentoDepartment of Biological and Environmental Sciences and Technologies, University of SalentoAbstract Background Among the mechanisms of mitochondrial quality control (MQC), generation of mitochondria-derived vesicles (MDVs) is a process to avoid complete failure of mitochondria determining lysosomal degradation of mitochondrial damaged proteins. In this context, RAB7, a late endocytic small GTPase, controls delivery of MDVs to late endosomes for subsequent lysosomal degradation. We previously demonstrated that RAB7 has a pivotal role in response to cisplatin (CDDP) regulating resistance to the drug by extracellular vesicle (EVs) secretion. Methods Western blot and immunofluorescence analysis were used to analyze structure and function of endosomes and lysosomes in CDDP chemosensitive and chemoresistant ovarian cancer cell lines. EVs were purified from chemosensitive and chemoresistant cells by ultracentrifugation or immunoisolation to analyze their mitochondrial DNA and protein content. Treatment with cyanide m-chlorophenylhydrazone (CCCP) and RAB7 modulation were used, respectively, to understand the role of mitochondrial and late endosomal/lysosomal alterations on MDV secretion. Using conditioned media from chemoresistant cells the effect of MDVs on the viability after CDDP treatment was determined. Seahorse assays and immunofluorescence analysis were used to study the biochemical role of MDVs and the uptake and intracellular localization of MDVs, respectively. Results We observed that CDDP-chemoresistant cells are characterized by increased MDV secretion, impairment of late endocytic traffic, RAB7 downregulation, an increase of RAB7 in EVs, compared to chemosensitive cells, and downregulation of the TFEB-mTOR pathway overseeing lysosomal and mitochondrial biogenesis and turnover. We established that MDVs can be secreted rather than delivered to lysosomes and are able to deliver CDDP outside the cells. We showed increased secretion of MDVs by chemoresistant cells ultimately caused by the extrusion of RAB7 in EVs, resulting in a dramatic drop in its intracellular content, as a novel mechanism to regulate RAB7 levels. We demonstrated that MDVs purified from chemoresistant cells induce chemoresistance in RAB7-modulated process, and, after uptake from recipient cells, MDVs localize to mitochondria and slow down mitochondrial activity. Conclusions Dysfunctional MQC in chemoresistant cells determines a block in lysosomal degradation of MDVs and their consequent secretion, suggesting that MQC is not able to eliminate damaged mitochondria whose components are secreted becoming effectors and potential markers of chemoresistance.https://doi.org/10.1186/s12964-024-01507-yRAB7Mitochondria-derived vesiclesExtracellular vesiclesCisplatin chemoresistanceOvarian cancer |
| spellingShingle | Sinforosa Gagliardi Marco Mitruccio Riccardo Di Corato Roberta Romano Alessandra Aloisi Rosaria Rinaldi Pietro Alifano Flora Guerra Cecilia Bucci Defects of mitochondria-lysosomes communication induce secretion of mitochondria-derived vesicles and drive chemoresistance in ovarian cancer cells Cell Communication and Signaling RAB7 Mitochondria-derived vesicles Extracellular vesicles Cisplatin chemoresistance Ovarian cancer |
| title | Defects of mitochondria-lysosomes communication induce secretion of mitochondria-derived vesicles and drive chemoresistance in ovarian cancer cells |
| title_full | Defects of mitochondria-lysosomes communication induce secretion of mitochondria-derived vesicles and drive chemoresistance in ovarian cancer cells |
| title_fullStr | Defects of mitochondria-lysosomes communication induce secretion of mitochondria-derived vesicles and drive chemoresistance in ovarian cancer cells |
| title_full_unstemmed | Defects of mitochondria-lysosomes communication induce secretion of mitochondria-derived vesicles and drive chemoresistance in ovarian cancer cells |
| title_short | Defects of mitochondria-lysosomes communication induce secretion of mitochondria-derived vesicles and drive chemoresistance in ovarian cancer cells |
| title_sort | defects of mitochondria lysosomes communication induce secretion of mitochondria derived vesicles and drive chemoresistance in ovarian cancer cells |
| topic | RAB7 Mitochondria-derived vesicles Extracellular vesicles Cisplatin chemoresistance Ovarian cancer |
| url | https://doi.org/10.1186/s12964-024-01507-y |
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