{"id":{"repo_id":"wlv","oai_identifier":"oai:wlv.openrepository.com:2436/624151"},"canonical_url":"https://search.dev.ndltd.org/etd/wlv/oai:wlv.openrepository.com:2436/624151","repository":{"repo_id":"wlv","name":"University of Wolverhampton","base_url":"https://wlv.openrepository.com/server/oai/request"},"display":{"title":"PLGA-DS reverses chemoresistance in malignant mesothelioma by targeting hypoxia induced cancer stem cells","abstract":"Background: Malignant Mesothelioma (MM) is a malignancy related to asbestos exposure which causes a wide variety of molecular aberrations. MM has a very dismal treatment outcome with an overall survival of fewer than 12 months, with less than five drugs available for its treatment. MM recurrence is unavoidable due to chemoresistance. Long-term inflammation triggered by asbestos activates a key transcription factor, nuclear factor-κB (NF-κB), which is further upregulated in cancer stem cells (CSCs) by hypoxia. Both hypoxia and NF-κB pathway plays a pivotal role in the maintenance of stemness in hypoxia-induced CSCs leading to upregulation of anti-apoptotic signalling, chemo-radiation resistance and metastasis. Therefore, the development of drugs targeting hypoxia-NF-κB-CSCs axis is of clinical significance for MM treatment. Our previous studies have shown that Disulfiram (DS), a clinically used anti-alcoholism drug, in combination with Copper (II) (Cu) has substantial toxicity in CSCs in a wide range of cancer types. The clinical application of DS in Cancer is limited by its very short half-life (< 2 minutes) in the bloodstream. MM is Cancer which mainly infiltrates local organs and tissues with rare distant metastasis. Considering this growing feature of MM, we developed a biodegradable and controlled released poly (lactic-co-glycolic acid) microparticle-encapsulate disulfiram (PLGA-DS) for local treatment of MM. This study aims to examine the anti-MM effect of PLGA-DS and elucidate its molecular mechanisms. Methodologies: In order to determine drug sensitivity, stemness, apoptosis, invasiveness and NFκB status, the following methodologies were performed in this study: MTT cytotoxicity assay, flow cytometry, analysis of CSC markers, hypoxic cell cultures, western blot, stable transfection of MM cell line with NFκB, CRISPR-Cas9 knock out of NF-kB-p65, CSC sphere reformation, invasion and migration assay. Results and conclusions: Two MM cell lines were examined and cultured in a hypoxic environment. MM cell lines were highly resistant to Pemetrexed (PMT) and Cisplatin (CIS), the first-line chemotherapeutic agents for MM. Hypoxia cultured MM cells showed high NF-κB activity and CSC markers and manifested strong migration/invasion ability. The NF-κB-p65 over expressed transfected cell lines did not demonstrate CSC traits along with no increase in resistance to first line drugs. PLGA-DS/Cu completely abolished CSC population in a culture which is demonstrated by sphere reformation assays and flow cytometry analysis of CSC markers such as CD24, CD133 and ABCG2. PLGA-DS/Cu also inhibited the hypoxiainduced NF-kB expression and blocked the migration and invasion ability of MM cells. It showed substantial toxicity to MM cell lines and reversed hypoxia-induced chemoresistance. Also, PLGA-DS/Cu potentiated the cytotoxic effect of Cisplatin/Pemetrexed in vitro. Isobologram analysis indicates moderate synergistic effect between PLGA-DS and cisplatin and pemetrexed in MSTO 211 and JU77 cell lines, respectively. As an FDA approved a drug with all preclinical safety data available, further studies may quickly translate it into MM clinical treatment. This is very promising in vitro data and indicate that PLGA-DS could be a promising formulation for localised MM treatment.","abstract_html":"Background: Malignant Mesothelioma (MM) is a malignancy related to asbestos exposure which causes a wide variety of molecular aberrations. MM has a very dismal treatment outcome with an overall survival of fewer than 12 months, with less than five drugs available for its treatment. MM recurrence is unavoidable due to chemoresistance. Long-term inflammation triggered by asbestos activates a key transcription factor, nuclear factor-κB (NF-κB), which is further upregulated in cancer stem cells (CSCs) by hypoxia. Both hypoxia and NF-κB pathway plays a pivotal role in the maintenance of stemness in hypoxia-induced CSCs leading to upregulation of anti-apoptotic signalling, chemo-radiation resistance and metastasis. Therefore, the development of drugs targeting hypoxia-NF-κB-CSCs axis is of clinical significance for MM treatment. Our previous studies have shown that Disulfiram (DS), a clinically used anti-alcoholism drug, in combination with Copper (II) (Cu) has substantial toxicity in CSCs in a wide range of cancer types. The clinical application of DS in Cancer is limited by its very short half-life (&lt; 2 minutes) in the bloodstream. MM is Cancer which mainly infiltrates local organs and tissues with rare distant metastasis. Considering this growing feature of MM, we developed a biodegradable and controlled released poly (lactic-co-glycolic acid) microparticle-encapsulate disulfiram (PLGA-DS) for local treatment of MM. This study aims to examine the anti-MM effect of PLGA-DS and elucidate its molecular mechanisms. Methodologies: In order to determine drug sensitivity, stemness, apoptosis, invasiveness and NFκB status, the following methodologies were performed in this study: MTT cytotoxicity assay, flow cytometry, analysis of CSC markers, hypoxic cell cultures, western blot, stable transfection of MM cell line with NFκB, CRISPR-Cas9 knock out of NF-kB-p65, CSC sphere reformation, invasion and migration assay. Results and conclusions: Two MM cell lines were examined and cultured in a hypoxic environment. MM cell lines were highly resistant to Pemetrexed (PMT) and Cisplatin (CIS), the first-line chemotherapeutic agents for MM. Hypoxia cultured MM cells showed high NF-κB activity and CSC markers and manifested strong migration/invasion ability. The NF-κB-p65 over expressed transfected cell lines did not demonstrate CSC traits along with no increase in resistance to first line drugs. PLGA-DS/Cu completely abolished CSC population in a culture which is demonstrated by sphere reformation assays and flow cytometry analysis of CSC markers such as CD24, CD133 and ABCG2. PLGA-DS/Cu also inhibited the hypoxiainduced NF-kB expression and blocked the migration and invasion ability of MM cells. It showed substantial toxicity to MM cell lines and reversed hypoxia-induced chemoresistance. Also, PLGA-DS/Cu potentiated the cytotoxic effect of Cisplatin/Pemetrexed in vitro. Isobologram analysis indicates moderate synergistic effect between PLGA-DS and cisplatin and pemetrexed in MSTO 211 and JU77 cell lines, respectively. As an FDA approved a drug with all preclinical safety data available, further studies may quickly translate it into MM clinical treatment. This is very promising in vitro data and indicate that PLGA-DS could be a promising formulation for localised MM treatment.","abstract_has_math":false,"creators":["Tyagi, Garima"],"institution":"University of Wolverhampton","degree_name":"MPhil","degree_level":"Masters","degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":["Wang, Weiguang"],"committee_chairs":[],"committee_members":[],"year":2020,"date_issued":"2020-09","date_published":"2020-09","updated_at":"2026-07-24T06:09:41Z","subjects":["PLGA","disulfiram","chemoresistance","mesothelioma","cancer","cancer stem cells","hypoxia","drug repurposing","EMT pathway"],"languages":[],"rights":["Attribution-NonCommercial-NoDerivatives 4.0 International"],"rights_urls":["https://wlv.dspace7.openrepository.com/bitstreams/2f7e0793-2817-4ae4-b711-0bafd41a303e/download"],"identifier_entries":[]},"links":{"outbound_url":null,"outbound_label":null,"outbound_source":null},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Wang, Weiguang"]},{"key":"dc:creator","label":"Author","values":["Tyagi, Garima"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.issued","label":"Date","values":["2020-09"]},{"key":"dc:publisher.institution","label":"Dc Publisher Institution","values":["University of Wolverhampton"]},{"key":"dc:relation.isreferencedby","label":"Dc Relation Isreferencedby","values":["http://hdl.handle.net/2436/624151"]},{"key":"dc:type","label":"Dc Type","values":["Thesis or dissertation"]},{"key":"dc:type.qualificationlevel","label":"Dc Type Qualificationlevel","values":["Masters"]},{"key":"dc:type.qualificationname","label":"Dc Type Qualificationname","values":["MPhil"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["PLGA","disulfiram","chemoresistance","mesothelioma","cancer","cancer stem cells","hypoxia","drug repurposing","EMT pathway"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:rights","label":"Dc Rights","values":["https://wlv.dspace7.openrepository.com/bitstreams/2f7e0793-2817-4ae4-b711-0bafd41a303e/download","Attribution-NonCommercial-NoDerivatives 4.0 International"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.uri","label":"Identifier URI","values":["https://wlv.dspace7.openrepository.com/bitstreams/eb37cfa4-4599-48a9-94ba-a8277e85b26c/download"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Background: Malignant Mesothelioma (MM) is a malignancy related to asbestos exposure which causes a wide variety of molecular aberrations. MM has a very dismal treatment outcome with an overall survival of fewer than 12 months, with less than five drugs available for its treatment. MM recurrence is unavoidable due to chemoresistance. Long-term inflammation triggered by asbestos activates a key transcription factor, nuclear factor-κB (NF-κB), which is further upregulated in cancer stem cells (CSCs) by hypoxia. Both hypoxia and NF-κB pathway plays a pivotal role in the maintenance of stemness in hypoxia-induced CSCs leading to upregulation of anti-apoptotic signalling, chemo-radiation resistance and metastasis. Therefore, the development of drugs targeting hypoxia-NF-κB-CSCs axis is of clinical significance for MM treatment. Our previous studies have shown that Disulfiram (DS), a clinically used anti-alcoholism drug, in combination with Copper (II) (Cu) has substantial toxicity in CSCs in a wide range of cancer types. The clinical application of DS in Cancer is limited by its very short half-life (< 2 minutes) in the bloodstream. MM is Cancer which mainly infiltrates local organs and tissues with rare distant metastasis. Considering this growing feature of MM, we developed a biodegradable and controlled released poly (lactic-co-glycolic acid) microparticle-encapsulate disulfiram (PLGA-DS) for local treatment of MM. This study aims to examine the anti-MM effect of PLGA-DS and elucidate its molecular mechanisms. Methodologies: In order to determine drug sensitivity, stemness, apoptosis, invasiveness and NFκB status, the following methodologies were performed in this study: MTT cytotoxicity assay, flow cytometry, analysis of CSC markers, hypoxic cell cultures, western blot, stable transfection of MM cell line with NFκB, CRISPR-Cas9 knock out of NF-kB-p65, CSC sphere reformation, invasion and migration assay. Results and conclusions: Two MM cell lines were examined and cultured in a hypoxic environment. MM cell lines were highly resistant to Pemetrexed (PMT) and Cisplatin (CIS), the first-line chemotherapeutic agents for MM. Hypoxia cultured MM cells showed high NF-κB activity and CSC markers and manifested strong migration/invasion ability. The NF-κB-p65 over expressed transfected cell lines did not demonstrate CSC traits along with no increase in resistance to first line drugs. PLGA-DS/Cu completely abolished CSC population in a culture which is demonstrated by sphere reformation assays and flow cytometry analysis of CSC markers such as CD24, CD133 and ABCG2. PLGA-DS/Cu also inhibited the hypoxiainduced NF-kB expression and blocked the migration and invasion ability of MM cells. It showed substantial toxicity to MM cell lines and reversed hypoxia-induced chemoresistance. Also, PLGA-DS/Cu potentiated the cytotoxic effect of Cisplatin/Pemetrexed in vitro. Isobologram analysis indicates moderate synergistic effect between PLGA-DS and cisplatin and pemetrexed in MSTO 211 and JU77 cell lines, respectively. As an FDA approved a drug with all preclinical safety data available, further studies may quickly translate it into MM clinical treatment. This is very promising in vitro data and indicate that PLGA-DS could be a promising formulation for localised MM treatment."]},{"key":"dc:format.checksum.md5","label":"Dc Format Checksum Md5","values":["859521bc253f266f2fe7db3dc1399dee","8a4605be74aa9ea9d79846c1fba20a33","6e5b67bd328b26cd537c678d05047381"]},{"key":"dc:title","label":"Title","values":["PLGA-DS reverses chemoresistance in malignant mesothelioma by targeting hypoxia induced cancer stem cells"]}]}],"canonical_facts":{"dc:contributor.advisor":["Wang, Weiguang"],"dc:creator":["Tyagi, Garima"],"dc:date.issued":["2020-09"],"dc:description.abstract":["Background: Malignant Mesothelioma (MM) is a malignancy related to asbestos exposure which causes a wide variety of molecular aberrations. MM has a very dismal treatment outcome with an overall survival of fewer than 12 months, with less than five drugs available for its treatment. MM recurrence is unavoidable due to chemoresistance. Long-term inflammation triggered by asbestos activates a key transcription factor, nuclear factor-κB (NF-κB), which is further upregulated in cancer stem cells (CSCs) by hypoxia. Both hypoxia and NF-κB pathway plays a pivotal role in the maintenance of stemness in hypoxia-induced CSCs leading to upregulation of anti-apoptotic signalling, chemo-radiation resistance and metastasis. Therefore, the development of drugs targeting hypoxia-NF-κB-CSCs axis is of clinical significance for MM treatment. Our previous studies have shown that Disulfiram (DS), a clinically used anti-alcoholism drug, in combination with Copper (II) (Cu) has substantial toxicity in CSCs in a wide range of cancer types. The clinical application of DS in Cancer is limited by its very short half-life (< 2 minutes) in the bloodstream. MM is Cancer which mainly infiltrates local organs and tissues with rare distant metastasis. Considering this growing feature of MM, we developed a biodegradable and controlled released poly (lactic-co-glycolic acid) microparticle-encapsulate disulfiram (PLGA-DS) for local treatment of MM. This study aims to examine the anti-MM effect of PLGA-DS and elucidate its molecular mechanisms. Methodologies: In order to determine drug sensitivity, stemness, apoptosis, invasiveness and NFκB status, the following methodologies were performed in this study: MTT cytotoxicity assay, flow cytometry, analysis of CSC markers, hypoxic cell cultures, western blot, stable transfection of MM cell line with NFκB, CRISPR-Cas9 knock out of NF-kB-p65, CSC sphere reformation, invasion and migration assay. Results and conclusions: Two MM cell lines were examined and cultured in a hypoxic environment. MM cell lines were highly resistant to Pemetrexed (PMT) and Cisplatin (CIS), the first-line chemotherapeutic agents for MM. Hypoxia cultured MM cells showed high NF-κB activity and CSC markers and manifested strong migration/invasion ability. The NF-κB-p65 over expressed transfected cell lines did not demonstrate CSC traits along with no increase in resistance to first line drugs. PLGA-DS/Cu completely abolished CSC population in a culture which is demonstrated by sphere reformation assays and flow cytometry analysis of CSC markers such as CD24, CD133 and ABCG2. PLGA-DS/Cu also inhibited the hypoxiainduced NF-kB expression and blocked the migration and invasion ability of MM cells. It showed substantial toxicity to MM cell lines and reversed hypoxia-induced chemoresistance. Also, PLGA-DS/Cu potentiated the cytotoxic effect of Cisplatin/Pemetrexed in vitro. Isobologram analysis indicates moderate synergistic effect between PLGA-DS and cisplatin and pemetrexed in MSTO 211 and JU77 cell lines, respectively. As an FDA approved a drug with all preclinical safety data available, further studies may quickly translate it into MM clinical treatment. This is very promising in vitro data and indicate that PLGA-DS could be a promising formulation for localised MM treatment."],"dc:format.checksum.md5":["859521bc253f266f2fe7db3dc1399dee","8a4605be74aa9ea9d79846c1fba20a33","6e5b67bd328b26cd537c678d05047381"],"dc:identifier.uri":["https://wlv.dspace7.openrepository.com/bitstreams/eb37cfa4-4599-48a9-94ba-a8277e85b26c/download"],"dc:publisher.institution":["University of Wolverhampton"],"dc:relation.isreferencedby":["http://hdl.handle.net/2436/624151"],"dc:rights":["https://wlv.dspace7.openrepository.com/bitstreams/2f7e0793-2817-4ae4-b711-0bafd41a303e/download","Attribution-NonCommercial-NoDerivatives 4.0 International"],"dc:subject":["PLGA","disulfiram","chemoresistance","mesothelioma","cancer","cancer stem cells","hypoxia","drug repurposing","EMT pathway"],"dc:title":["PLGA-DS reverses chemoresistance in malignant mesothelioma by targeting hypoxia induced cancer stem cells"],"dc:type":["Thesis or dissertation"],"dc:type.qualificationlevel":["Masters"],"dc:type.qualificationname":["MPhil"]},"updated_at":"2026-07-24T06:09:41Z"}