Proteomic characterisation of perhexiline treatment on THP-1 M1 macrophage differentiation
BackgroundDysregulated inflammation is important in the pathogenesis of many diseases including cancer, allergy, and autoimmunity. Macrophage activation and polarisation are commonly involved in the initiation, maintenance and resolution of inflammation. Perhexiline (PHX), an antianginal drug, has b...
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Frontiers Media S.A.
2023-03-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1054588/full |
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| author | Bimala Dhakal Bimala Dhakal Celine Man Ying Li Celine Man Ying Li Mahnaz Ramezanpour Mahnaz Ramezanpour Mahnaz Ramezanpour Ghais Houtak Ghais Houtak Ghais Houtak Runhao Li Runhao Li Runhao Li George Bouras George Bouras George Bouras Alex Collela Nusha Chegeni Tim Kennion Chataway Paul Drew Paul Drew Benedetta C. Sallustio Benedetta C. Sallustio Sarah Vreugde Sarah Vreugde Sarah Vreugde Eric Smith Eric Smith Eric Smith Guy Maddern Guy Maddern Giovanni Licari Giovanni Licari Kevin Fenix Kevin Fenix Kevin Fenix |
| author_facet | Bimala Dhakal Bimala Dhakal Celine Man Ying Li Celine Man Ying Li Mahnaz Ramezanpour Mahnaz Ramezanpour Mahnaz Ramezanpour Ghais Houtak Ghais Houtak Ghais Houtak Runhao Li Runhao Li Runhao Li George Bouras George Bouras George Bouras Alex Collela Nusha Chegeni Tim Kennion Chataway Paul Drew Paul Drew Benedetta C. Sallustio Benedetta C. Sallustio Sarah Vreugde Sarah Vreugde Sarah Vreugde Eric Smith Eric Smith Eric Smith Guy Maddern Guy Maddern Giovanni Licari Giovanni Licari Kevin Fenix Kevin Fenix Kevin Fenix |
| author_sort | Bimala Dhakal |
| collection | DOAJ |
| description | BackgroundDysregulated inflammation is important in the pathogenesis of many diseases including cancer, allergy, and autoimmunity. Macrophage activation and polarisation are commonly involved in the initiation, maintenance and resolution of inflammation. Perhexiline (PHX), an antianginal drug, has been suggested to modulate macrophage function, but the molecular effects of PHX on macrophages are unknown. In this study we investigated the effect of PHX treatment on macrophage activation and polarization and reveal the underlying proteomic changes induced.MethodsWe used an established protocol to differentiate human THP-1 monocytes into M1 or M2 macrophages involving three distinct, sequential stages (priming, rest, and differentiation). We examined the effect of PHX treatment at each stage on the polarization into either M1 or M2 macrophages using flow cytometry, quantitative polymerase chain reaction (qPCR) and enzyme linked immunosorbent assay (ELISA). Quantitative changes in the proteome were investigated using data independent acquisition mass spectrometry (DIA MS).ResultsPHX treatment promoted M1 macrophage polarization, including increased STAT1 and CCL2 expression and IL-1β secretion. This effect occurred when PHX was added at the differentiation stage of the M1 cultures. Proteomic profiling of PHX treated M1 cultures identified changes in metabolic (fatty acid metabolism, cholesterol homeostasis and oxidative phosphorylation) and immune signalling (Receptor Tyrosine Kinase, Rho GTPase and interferon) pathways.ConclusionThis is the first study to report on the action of PHX on THP-1 macrophage polarization and the associated changes in the proteome of these cells. |
| first_indexed | 2024-04-10T04:06:46Z |
| format | Article |
| id | doaj.art-01e53bbe66d247dfb4f326e58ef38816 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-04-10T04:06:46Z |
| publishDate | 2023-03-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-01e53bbe66d247dfb4f326e58ef388162023-03-13T04:57:07ZengFrontiers Media S.A.Frontiers in Immunology1664-32242023-03-011410.3389/fimmu.2023.10545881054588Proteomic characterisation of perhexiline treatment on THP-1 M1 macrophage differentiationBimala Dhakal0Bimala Dhakal1Celine Man Ying Li2Celine Man Ying Li3Mahnaz Ramezanpour4Mahnaz Ramezanpour5Mahnaz Ramezanpour6Ghais Houtak7Ghais Houtak8Ghais Houtak9Runhao Li10Runhao Li11Runhao Li12George Bouras13George Bouras14George Bouras15Alex Collela16Nusha Chegeni17Tim Kennion Chataway18Paul Drew19Paul Drew20Benedetta C. Sallustio21Benedetta C. Sallustio22Sarah Vreugde23Sarah Vreugde24Sarah Vreugde25Eric Smith26Eric Smith27Eric Smith28Guy Maddern29Guy Maddern30Giovanni Licari31Giovanni Licari32Kevin Fenix33Kevin Fenix34Kevin Fenix35Discipline of Surgery, Adelaide Medical School, The University of Adelaide, Adelaide, SA, AustraliaThe Basil Hetzel Institute for Translational Health Research, The Queen Elizabeth Hospital, Adelaide, SA, AustraliaDiscipline of Surgery, Adelaide Medical School, The University of Adelaide, Adelaide, SA, AustraliaThe Basil Hetzel Institute for Translational Health Research, The Queen Elizabeth Hospital, Adelaide, SA, AustraliaDiscipline of Surgery, Adelaide Medical School, The University of Adelaide, Adelaide, SA, AustraliaThe Basil Hetzel Institute for Translational Health Research, The Queen Elizabeth Hospital, Adelaide, SA, AustraliaDepartment of Surgery-Otolaryngology Head and Neck Surgery, Central Adelaide Local Health Network, Adelaide, SA, AustraliaDiscipline of Surgery, Adelaide Medical School, The University of Adelaide, Adelaide, SA, AustraliaThe Basil Hetzel Institute for Translational Health Research, The Queen Elizabeth Hospital, Adelaide, SA, AustraliaDepartment of Surgery-Otolaryngology Head and Neck Surgery, Central Adelaide Local Health Network, Adelaide, SA, AustraliaDiscipline of Surgery, Adelaide Medical School, The University of Adelaide, Adelaide, SA, AustraliaThe Basil Hetzel Institute for Translational Health Research, The Queen Elizabeth Hospital, Adelaide, SA, AustraliaMedical Oncology, The Queen Elizabeth Hospital, Adelaide, SA, AustraliaDiscipline of Surgery, Adelaide Medical School, The University of Adelaide, Adelaide, SA, AustraliaThe Basil Hetzel Institute for Translational Health Research, The Queen Elizabeth Hospital, Adelaide, SA, AustraliaDepartment of Surgery-Otolaryngology Head and Neck Surgery, Central Adelaide Local Health Network, Adelaide, SA, AustraliaFlinders Omics Facility, Department of Human Physiology, Flinders University, Adelaide, SA, AustraliaFlinders Omics Facility, Department of Human Physiology, Flinders University, Adelaide, SA, AustraliaFlinders Omics Facility, Department of Human Physiology, Flinders University, Adelaide, SA, AustraliaDiscipline of Surgery, Adelaide Medical School, The University of Adelaide, Adelaide, SA, AustraliaThe Basil Hetzel Institute for Translational Health Research, The Queen Elizabeth Hospital, Adelaide, SA, AustraliaThe Basil Hetzel Institute for Translational Health Research, The Queen Elizabeth Hospital, Adelaide, SA, AustraliaDiscipline of Pharmacology, School of Biomedicine, The University of Adelaide, Adelaide, SA, AustraliaDiscipline of Surgery, Adelaide Medical School, The University of Adelaide, Adelaide, SA, AustraliaThe Basil Hetzel Institute for Translational Health Research, The Queen Elizabeth Hospital, Adelaide, SA, AustraliaDepartment of Surgery-Otolaryngology Head and Neck Surgery, Central Adelaide Local Health Network, Adelaide, SA, AustraliaDiscipline of Surgery, Adelaide Medical School, The University of Adelaide, Adelaide, SA, AustraliaThe Basil Hetzel Institute for Translational Health Research, The Queen Elizabeth Hospital, Adelaide, SA, AustraliaMedical Oncology, The Queen Elizabeth Hospital, Adelaide, SA, AustraliaDiscipline of Surgery, Adelaide Medical School, The University of Adelaide, Adelaide, SA, AustraliaThe Basil Hetzel Institute for Translational Health Research, The Queen Elizabeth Hospital, Adelaide, SA, AustraliaThe Basil Hetzel Institute for Translational Health Research, The Queen Elizabeth Hospital, Adelaide, SA, AustraliaDiscipline of Pharmacology, School of Biomedicine, The University of Adelaide, Adelaide, SA, AustraliaDiscipline of Surgery, Adelaide Medical School, The University of Adelaide, Adelaide, SA, AustraliaThe Basil Hetzel Institute for Translational Health Research, The Queen Elizabeth Hospital, Adelaide, SA, AustraliaDepartment of Surgery-Otolaryngology Head and Neck Surgery, Central Adelaide Local Health Network, Adelaide, SA, AustraliaBackgroundDysregulated inflammation is important in the pathogenesis of many diseases including cancer, allergy, and autoimmunity. Macrophage activation and polarisation are commonly involved in the initiation, maintenance and resolution of inflammation. Perhexiline (PHX), an antianginal drug, has been suggested to modulate macrophage function, but the molecular effects of PHX on macrophages are unknown. In this study we investigated the effect of PHX treatment on macrophage activation and polarization and reveal the underlying proteomic changes induced.MethodsWe used an established protocol to differentiate human THP-1 monocytes into M1 or M2 macrophages involving three distinct, sequential stages (priming, rest, and differentiation). We examined the effect of PHX treatment at each stage on the polarization into either M1 or M2 macrophages using flow cytometry, quantitative polymerase chain reaction (qPCR) and enzyme linked immunosorbent assay (ELISA). Quantitative changes in the proteome were investigated using data independent acquisition mass spectrometry (DIA MS).ResultsPHX treatment promoted M1 macrophage polarization, including increased STAT1 and CCL2 expression and IL-1β secretion. This effect occurred when PHX was added at the differentiation stage of the M1 cultures. Proteomic profiling of PHX treated M1 cultures identified changes in metabolic (fatty acid metabolism, cholesterol homeostasis and oxidative phosphorylation) and immune signalling (Receptor Tyrosine Kinase, Rho GTPase and interferon) pathways.ConclusionThis is the first study to report on the action of PHX on THP-1 macrophage polarization and the associated changes in the proteome of these cells.https://www.frontiersin.org/articles/10.3389/fimmu.2023.1054588/fullM1 macrolphageperhexilinequantitative proteomicsTHP-1 derived macrophagesmacrophage polarisation |
| spellingShingle | Bimala Dhakal Bimala Dhakal Celine Man Ying Li Celine Man Ying Li Mahnaz Ramezanpour Mahnaz Ramezanpour Mahnaz Ramezanpour Ghais Houtak Ghais Houtak Ghais Houtak Runhao Li Runhao Li Runhao Li George Bouras George Bouras George Bouras Alex Collela Nusha Chegeni Tim Kennion Chataway Paul Drew Paul Drew Benedetta C. Sallustio Benedetta C. Sallustio Sarah Vreugde Sarah Vreugde Sarah Vreugde Eric Smith Eric Smith Eric Smith Guy Maddern Guy Maddern Giovanni Licari Giovanni Licari Kevin Fenix Kevin Fenix Kevin Fenix Proteomic characterisation of perhexiline treatment on THP-1 M1 macrophage differentiation Frontiers in Immunology M1 macrolphage perhexiline quantitative proteomics THP-1 derived macrophages macrophage polarisation |
| title | Proteomic characterisation of perhexiline treatment on THP-1 M1 macrophage differentiation |
| title_full | Proteomic characterisation of perhexiline treatment on THP-1 M1 macrophage differentiation |
| title_fullStr | Proteomic characterisation of perhexiline treatment on THP-1 M1 macrophage differentiation |
| title_full_unstemmed | Proteomic characterisation of perhexiline treatment on THP-1 M1 macrophage differentiation |
| title_short | Proteomic characterisation of perhexiline treatment on THP-1 M1 macrophage differentiation |
| title_sort | proteomic characterisation of perhexiline treatment on thp 1 m1 macrophage differentiation |
| topic | M1 macrolphage perhexiline quantitative proteomics THP-1 derived macrophages macrophage polarisation |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1054588/full |
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