A novel role for peptidylarginine deiminases in microvesicle release reveals therapeutic potential of PAD inhibition in sensitizing prostate cancer cells to chemotherapy
INTRODUCTION: Protein deimination, defined as the post-translational conversion of protein-bound arginine to citrulline, is carried out by a family of 5 calcium-dependent enzymes, the peptidylarginine deiminases (PADs) and has been linked to various cancers. Cellular microvesicle (MV) release, whic...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Co-Action Publishing
2015
|
| Subjects: | |
| Online Access: | https://repository.londonmet.ac.uk/1001/1/JEV%20PAD%20paper%202015.pdf |
| _version_ | 1804071595276238848 |
|---|---|
| author | Kholia, Sharadkumar Rajnikant Jorfi, Samireh Thompson, Paul R. Causey, Corey P. Nicholas, Anthony P. Inal, Jameel Lange, Sigrun |
| author_facet | Kholia, Sharadkumar Rajnikant Jorfi, Samireh Thompson, Paul R. Causey, Corey P. Nicholas, Anthony P. Inal, Jameel Lange, Sigrun |
| author_sort | Kholia, Sharadkumar Rajnikant |
| collection | LMU |
| description | INTRODUCTION:
Protein deimination, defined as the post-translational conversion of protein-bound arginine to citrulline, is carried out by a family of 5 calcium-dependent enzymes, the peptidylarginine deiminases (PADs) and has been linked to various cancers. Cellular microvesicle (MV) release, which is involved in cancer progression, and deimination have not been associated before. We hypothesize that elevated PAD expression, observed in cancers, causes increased MV release in cancer cells and contributes to cancer progression.
BACKGROUND:
We have previously reported that inhibition of MV release sensitizes cancer cells to chemotherapeutic drugs. PAD2 and PAD4, the isozymes expressed in patients with malignant tumours, can be inhibited with the pan-PAD-inhibitor chloramidine (Cl-am). We sought to investigate whether Cl-am can inhibit MV release and whether this pathway could be utilized to further increase the sensitivity of cancer cells to drug-directed treatment.
METHODS:
Prostate cancer cells (PC3) were induced to release high levels of MVs upon BzATP stimulation of P2X7 receptors. Western blotting with the pan-protein deimination antibody F95 was used to detect a range of deiminated proteins in cells stimulated to microvesiculate. Changes in deiminated proteins during microvesiculation were revealed by immunoprecipitation and immunoblotting, and mass spectrometry identified deiminated target proteins with putative roles in microvesiculation.
CONCLUSION:
We report for the first time a novel function of PADs in the biogenesis of MVs in cancer cells. Our results reveal that during the stimulation of prostate cancer cells (PC3) to microvesiculate, PAD2 and PAD4 expression levels and the deimination of cytoskeletal actin are increased. Pharmacological inhibition of PAD enzyme activity using Cl-am significantly reduced MV release and abrogated the deimination of cytoskeletal actin. We demonstrated that combined Cl-am and methotrexate (MTX) treatment of prostate cancer cells increased the cytotoxic effect of MTX synergistically. Refined PAD inhibitors may form part of a novel combination therapy in cancer treatment. |
| first_indexed | 2024-07-09T03:45:45Z |
| format | Article |
| id | oai:repository.londonmet.ac.uk:1001 |
| institution | London Metropolitan University |
| language | English |
| last_indexed | 2024-07-09T03:45:45Z |
| publishDate | 2015 |
| publisher | Co-Action Publishing |
| record_format | eprints |
| spelling | oai:repository.londonmet.ac.uk:10012020-03-18T11:13:28Z http://repository.londonmet.ac.uk/1001/ A novel role for peptidylarginine deiminases in microvesicle release reveals therapeutic potential of PAD inhibition in sensitizing prostate cancer cells to chemotherapy Kholia, Sharadkumar Rajnikant Jorfi, Samireh Thompson, Paul R. Causey, Corey P. Nicholas, Anthony P. Inal, Jameel Lange, Sigrun 610 Medicine & health INTRODUCTION: Protein deimination, defined as the post-translational conversion of protein-bound arginine to citrulline, is carried out by a family of 5 calcium-dependent enzymes, the peptidylarginine deiminases (PADs) and has been linked to various cancers. Cellular microvesicle (MV) release, which is involved in cancer progression, and deimination have not been associated before. We hypothesize that elevated PAD expression, observed in cancers, causes increased MV release in cancer cells and contributes to cancer progression. BACKGROUND: We have previously reported that inhibition of MV release sensitizes cancer cells to chemotherapeutic drugs. PAD2 and PAD4, the isozymes expressed in patients with malignant tumours, can be inhibited with the pan-PAD-inhibitor chloramidine (Cl-am). We sought to investigate whether Cl-am can inhibit MV release and whether this pathway could be utilized to further increase the sensitivity of cancer cells to drug-directed treatment. METHODS: Prostate cancer cells (PC3) were induced to release high levels of MVs upon BzATP stimulation of P2X7 receptors. Western blotting with the pan-protein deimination antibody F95 was used to detect a range of deiminated proteins in cells stimulated to microvesiculate. Changes in deiminated proteins during microvesiculation were revealed by immunoprecipitation and immunoblotting, and mass spectrometry identified deiminated target proteins with putative roles in microvesiculation. CONCLUSION: We report for the first time a novel function of PADs in the biogenesis of MVs in cancer cells. Our results reveal that during the stimulation of prostate cancer cells (PC3) to microvesiculate, PAD2 and PAD4 expression levels and the deimination of cytoskeletal actin are increased. Pharmacological inhibition of PAD enzyme activity using Cl-am significantly reduced MV release and abrogated the deimination of cytoskeletal actin. We demonstrated that combined Cl-am and methotrexate (MTX) treatment of prostate cancer cells increased the cytotoxic effect of MTX synergistically. Refined PAD inhibitors may form part of a novel combination therapy in cancer treatment. Co-Action Publishing 2015-06-22 Article PeerReviewed text en https://repository.londonmet.ac.uk/1001/1/JEV%20PAD%20paper%202015.pdf Kholia, Sharadkumar Rajnikant, Jorfi, Samireh, Thompson, Paul R., Causey, Corey P., Nicholas, Anthony P., Inal, Jameel and Lange, Sigrun (2015) A novel role for peptidylarginine deiminases in microvesicle release reveals therapeutic potential of PAD inhibition in sensitizing prostate cancer cells to chemotherapy. Journal of Extracellular Vesicles, 4 (26192). ISSN 2001-3078 http://www.journalofextracellularvesicles.net/index.php/jev/article/view/26192 10.3402/jev.v4.26192 |
| spellingShingle | 610 Medicine & health Kholia, Sharadkumar Rajnikant Jorfi, Samireh Thompson, Paul R. Causey, Corey P. Nicholas, Anthony P. Inal, Jameel Lange, Sigrun A novel role for peptidylarginine deiminases in microvesicle release reveals therapeutic potential of PAD inhibition in sensitizing prostate cancer cells to chemotherapy |
| title | A novel role for peptidylarginine deiminases in microvesicle release reveals therapeutic potential of PAD inhibition in sensitizing prostate cancer cells to chemotherapy |
| title_full | A novel role for peptidylarginine deiminases in microvesicle release reveals therapeutic potential of PAD inhibition in sensitizing prostate cancer cells to chemotherapy |
| title_fullStr | A novel role for peptidylarginine deiminases in microvesicle release reveals therapeutic potential of PAD inhibition in sensitizing prostate cancer cells to chemotherapy |
| title_full_unstemmed | A novel role for peptidylarginine deiminases in microvesicle release reveals therapeutic potential of PAD inhibition in sensitizing prostate cancer cells to chemotherapy |
| title_short | A novel role for peptidylarginine deiminases in microvesicle release reveals therapeutic potential of PAD inhibition in sensitizing prostate cancer cells to chemotherapy |
| title_sort | novel role for peptidylarginine deiminases in microvesicle release reveals therapeutic potential of pad inhibition in sensitizing prostate cancer cells to chemotherapy |
| topic | 610 Medicine & health |
| url | https://repository.londonmet.ac.uk/1001/1/JEV%20PAD%20paper%202015.pdf |
| work_keys_str_mv | AT kholiasharadkumarrajnikant anovelroleforpeptidylargininedeiminasesinmicrovesiclereleaserevealstherapeuticpotentialofpadinhibitioninsensitizingprostatecancercellstochemotherapy AT jorfisamireh anovelroleforpeptidylargininedeiminasesinmicrovesiclereleaserevealstherapeuticpotentialofpadinhibitioninsensitizingprostatecancercellstochemotherapy AT thompsonpaulr anovelroleforpeptidylargininedeiminasesinmicrovesiclereleaserevealstherapeuticpotentialofpadinhibitioninsensitizingprostatecancercellstochemotherapy AT causeycoreyp anovelroleforpeptidylargininedeiminasesinmicrovesiclereleaserevealstherapeuticpotentialofpadinhibitioninsensitizingprostatecancercellstochemotherapy AT nicholasanthonyp anovelroleforpeptidylargininedeiminasesinmicrovesiclereleaserevealstherapeuticpotentialofpadinhibitioninsensitizingprostatecancercellstochemotherapy AT inaljameel anovelroleforpeptidylargininedeiminasesinmicrovesiclereleaserevealstherapeuticpotentialofpadinhibitioninsensitizingprostatecancercellstochemotherapy AT langesigrun anovelroleforpeptidylargininedeiminasesinmicrovesiclereleaserevealstherapeuticpotentialofpadinhibitioninsensitizingprostatecancercellstochemotherapy AT kholiasharadkumarrajnikant novelroleforpeptidylargininedeiminasesinmicrovesiclereleaserevealstherapeuticpotentialofpadinhibitioninsensitizingprostatecancercellstochemotherapy AT jorfisamireh novelroleforpeptidylargininedeiminasesinmicrovesiclereleaserevealstherapeuticpotentialofpadinhibitioninsensitizingprostatecancercellstochemotherapy AT thompsonpaulr novelroleforpeptidylargininedeiminasesinmicrovesiclereleaserevealstherapeuticpotentialofpadinhibitioninsensitizingprostatecancercellstochemotherapy AT causeycoreyp novelroleforpeptidylargininedeiminasesinmicrovesiclereleaserevealstherapeuticpotentialofpadinhibitioninsensitizingprostatecancercellstochemotherapy AT nicholasanthonyp novelroleforpeptidylargininedeiminasesinmicrovesiclereleaserevealstherapeuticpotentialofpadinhibitioninsensitizingprostatecancercellstochemotherapy AT inaljameel novelroleforpeptidylargininedeiminasesinmicrovesiclereleaserevealstherapeuticpotentialofpadinhibitioninsensitizingprostatecancercellstochemotherapy AT langesigrun novelroleforpeptidylargininedeiminasesinmicrovesiclereleaserevealstherapeuticpotentialofpadinhibitioninsensitizingprostatecancercellstochemotherapy |