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The Role of Chaperone-Mediated Autophagy in Bortezomib Resistant Multiple Myeloma

Background: Multiple myeloma (MM) remains incurable despite high-dose chemotherapy, autologous stem cell transplants and novel agents. Even with the improved survival of MM patients treated with novel agents, including bortezomib (Bz), the therapeutic options in relapsed/refractory MM remain limited...

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Autores principales: Nikesitch, Nicholas, Rebeiro, Patricia, Ho, Lye Lin, Pothula, Srinivasa, Wang, Xin Maggie, Khong, Tiffany, Quek, Hazel, Spencer, Andrew, Lee, Cheok Soon, Roberts, Tara L., Ling, Silvia C. W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8700264/
https://www.ncbi.nlm.nih.gov/pubmed/34943972
http://dx.doi.org/10.3390/cells10123464
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author Nikesitch, Nicholas
Rebeiro, Patricia
Ho, Lye Lin
Pothula, Srinivasa
Wang, Xin Maggie
Khong, Tiffany
Quek, Hazel
Spencer, Andrew
Lee, Cheok Soon
Roberts, Tara L.
Ling, Silvia C. W.
author_facet Nikesitch, Nicholas
Rebeiro, Patricia
Ho, Lye Lin
Pothula, Srinivasa
Wang, Xin Maggie
Khong, Tiffany
Quek, Hazel
Spencer, Andrew
Lee, Cheok Soon
Roberts, Tara L.
Ling, Silvia C. W.
author_sort Nikesitch, Nicholas
collection PubMed
description Background: Multiple myeloma (MM) remains incurable despite high-dose chemotherapy, autologous stem cell transplants and novel agents. Even with the improved survival of MM patients treated with novel agents, including bortezomib (Bz), the therapeutic options in relapsed/refractory MM remain limited. The majority of MM patients eventually develop resistance to Bz, although the mechanisms of the resistance are poorly understood. Methods: Lysosomal associated membrane protein 2A (LAMP2A) mRNA and protein expression levels were assessed in ex vivo patient samples and a Bz-resistant MM cell line model by in real-rime PCR, western blotting and immunohistochemistry. In vitro modelling of chaperone-mediated autophagy (CMA) activity in response to ER stress were assessed by western blotting and confocal microscopy. The effects of CMA inhibition on MM cell viability and Bz sensitivity in MM cells were assessed by Annexin V/7AAD apoptosis assays using flow cytometry. Results: In this study, there is evidence that CMA, a chaperone-mediated protein degradation pathway, is upregulated in Bz-resistant MM and the inhibition of CMA sensitises resistant cells to Bz. The protein levels of LAMP2A, the rate-limiting factor of the CMA pathway, are significantly increased in MM patients resistant to Bz and within our Bz-resistant cell line model. Bz-resistant cell lines also possessed higher basal CMA activity than the Bz-sensitive parent cell line. In MM cell lines, CMA activity was upregulated in response to ER stress induced by Bz. The inhibition of CMA sensitises Bz-resistant cells to Bz and the combination of CMA inhibition and Bz in vitro had a more cytotoxic effect on myeloma cells than Bz alone. Conclusion: In summary, the upregulation of CMA is a potential mechanism of resistance to Bz and a novel target to overcome Bz-resistant MM.
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spelling pubmed-87002642021-12-24 The Role of Chaperone-Mediated Autophagy in Bortezomib Resistant Multiple Myeloma Nikesitch, Nicholas Rebeiro, Patricia Ho, Lye Lin Pothula, Srinivasa Wang, Xin Maggie Khong, Tiffany Quek, Hazel Spencer, Andrew Lee, Cheok Soon Roberts, Tara L. Ling, Silvia C. W. Cells Article Background: Multiple myeloma (MM) remains incurable despite high-dose chemotherapy, autologous stem cell transplants and novel agents. Even with the improved survival of MM patients treated with novel agents, including bortezomib (Bz), the therapeutic options in relapsed/refractory MM remain limited. The majority of MM patients eventually develop resistance to Bz, although the mechanisms of the resistance are poorly understood. Methods: Lysosomal associated membrane protein 2A (LAMP2A) mRNA and protein expression levels were assessed in ex vivo patient samples and a Bz-resistant MM cell line model by in real-rime PCR, western blotting and immunohistochemistry. In vitro modelling of chaperone-mediated autophagy (CMA) activity in response to ER stress were assessed by western blotting and confocal microscopy. The effects of CMA inhibition on MM cell viability and Bz sensitivity in MM cells were assessed by Annexin V/7AAD apoptosis assays using flow cytometry. Results: In this study, there is evidence that CMA, a chaperone-mediated protein degradation pathway, is upregulated in Bz-resistant MM and the inhibition of CMA sensitises resistant cells to Bz. The protein levels of LAMP2A, the rate-limiting factor of the CMA pathway, are significantly increased in MM patients resistant to Bz and within our Bz-resistant cell line model. Bz-resistant cell lines also possessed higher basal CMA activity than the Bz-sensitive parent cell line. In MM cell lines, CMA activity was upregulated in response to ER stress induced by Bz. The inhibition of CMA sensitises Bz-resistant cells to Bz and the combination of CMA inhibition and Bz in vitro had a more cytotoxic effect on myeloma cells than Bz alone. Conclusion: In summary, the upregulation of CMA is a potential mechanism of resistance to Bz and a novel target to overcome Bz-resistant MM. MDPI 2021-12-08 /pmc/articles/PMC8700264/ /pubmed/34943972 http://dx.doi.org/10.3390/cells10123464 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Nikesitch, Nicholas
Rebeiro, Patricia
Ho, Lye Lin
Pothula, Srinivasa
Wang, Xin Maggie
Khong, Tiffany
Quek, Hazel
Spencer, Andrew
Lee, Cheok Soon
Roberts, Tara L.
Ling, Silvia C. W.
The Role of Chaperone-Mediated Autophagy in Bortezomib Resistant Multiple Myeloma
title The Role of Chaperone-Mediated Autophagy in Bortezomib Resistant Multiple Myeloma
title_full The Role of Chaperone-Mediated Autophagy in Bortezomib Resistant Multiple Myeloma
title_fullStr The Role of Chaperone-Mediated Autophagy in Bortezomib Resistant Multiple Myeloma
title_full_unstemmed The Role of Chaperone-Mediated Autophagy in Bortezomib Resistant Multiple Myeloma
title_short The Role of Chaperone-Mediated Autophagy in Bortezomib Resistant Multiple Myeloma
title_sort role of chaperone-mediated autophagy in bortezomib resistant multiple myeloma
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8700264/
https://www.ncbi.nlm.nih.gov/pubmed/34943972
http://dx.doi.org/10.3390/cells10123464
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