The influence of metformin treatment on the circulating proteomeResearch in context
Summary: Background: Metformin is one of the most used drugs worldwide. Given the increasing use of proteomics in trials, bioresources, and clinics, it is crucial to understand the influence of metformin on the levels of the circulating proteome. Methods: We analysed a combined longitudinal proteom...
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| author | Ben Connolly Laura McCreight Roderick C. Slieker Khaled F. Bedair Louise Donnelly Juliette A. de Klerk Joline W.J. Beulens Petra J.M. Elders Göran Bergström Mun-Gwan Hong Robert W. Koivula Paul W. Franks Jochen M. Schwenk Anders Gummesson Ewan R. Pearson Leen M. ‘t Hart Moustafa Abdalla Jonathan Adam Jerzy Adamski Kofi Adragni Rosa Lundbye L. Allesøe Kristine H. Allin Anna A. Artati Manimozhiyan Arumugam Naeimeh Atabaki Pasdar Tania Baltauss Karina Banasik Anna Barnett Patrick Baum Jimmy D. Bell Susanna Bianzano Roberto Bizzotto Amelie Bonnefond Caroline Anna A. Brorsson Andrew A. Brown Søren Brunak Louise Cabrelli Robert Caiazzo Henna Cederberg Elizaveta Chabanova Marc Clos-Garcia Matilda Dale David Davtian Adem Y. Dawed Federico De Masi Nathalie de Preville Koen F. Dekkers Harshal A. Deshmukh Christiane Dings Avirup Dutta Beate Ehrhardt Line Engelbrechtsen Rebeca Eriksen Yong Fan Juan Fernandez Jorge Ferrer Hugo Fitipaldi Ian M. Forgie Annemette Forman Francesca Frau Philippe Froguel Gary Frost Johann Gassenhuber Giuseppe (Nick) N. Giordano Toni Giorgino Stephen Gough Harald Grallert Rolf Grempler Lenka Groeneveld Leif Groop Valborg Gudmundsdóttir Ramneek Gupta Mark Haid Torben Hansen Tue H. Hansen Andrew T. Hattersley Ragna Haussler Alison J. Heggie Anita M. Hennige Anita V. Hill Reinhard W. Holl Michelle Hudson Bernd Jablonka Ulrik Plesner Jacobsen Christopher Jennison Joachim Johansen Angus G. Jones Tugce Karaderi Jane Kaye Gwen Kennedy Maria Klintenberg Robert W. Koivula Tarja Kokkola Anitra D. Koopman Azra Kurbasic Teemu Kuulasmaa Markku Laakso Thorsten Lehr Heather Loftus Agnete Troen T. Lundgaard Liwei Lyu Anubha Mahajan Andrea Mari Gianluca Mazzoni Mark I. McCarthy Timothy J. McDonald Donna McEvoy Nicky McRobert Ian McVittie Miranda Mourby Petra Musholt Pascal Mutie Rachel Nice Claudia Nicolay Giel Nijpels Birgitte Nilsson Colin N. Palmer Francois Pattou Imre Pavo Helle K. Pedersen Oluf Pedersen Mandy H. Perry Hugo Pomares-Millan Cornelia P. Prehn Anna Ramisch Simon Rasmussen Violeta Raverdi Martin Ridderstråle Neil Robertson Marianne Rodriquez Hartmut Ruetten Femke Rutters Peter Sackett Nina Scherer Nisha Shah Sapna Sharma Iryna Sihinevich Nadja B. Sondertoft Hans-Henrik Staerfeldt Birgit Steckel-Hamann Harriet Teare Cecilia Engel E. Thomas Elizabeth Louise L. Thomas Melissa K. Thomas Henrik S. Thomsen Barbara Thorand Claire E. Thorne Joachim Tillner Konstantinos D. Tsirigos Andrea Tura Mathias Uhlen Sabine van Oort Jagadish Vangipurapu Helene Verkindt Henrik Vestergaard Ana Viñuela Josef K. Vogt Peter W. Wad Sackett Mark Walker Agata Wesolowska-Andersen Brandon Whitcher Margaret W. White Alexander Efanov Giuseppe N. Giordano Gerard A. Bouland Frédéric Burdet Iulian Dragan Andreas Festa Michael K. Hansen Dmitry Kuznetsov Florence Mehl Diana Marek Imre Pavo Kevin Duffin Samreen K. Syed Janice L. Shaw Over Cabrera Timothy J. Pullen Bernard Thorens Mark Ibberson Guy A. Rutter |
| author_facet | Ben Connolly Laura McCreight Roderick C. Slieker Khaled F. Bedair Louise Donnelly Juliette A. de Klerk Joline W.J. Beulens Petra J.M. Elders Göran Bergström Mun-Gwan Hong Robert W. Koivula Paul W. Franks Jochen M. Schwenk Anders Gummesson Ewan R. Pearson Leen M. ‘t Hart Moustafa Abdalla Jonathan Adam Jerzy Adamski Kofi Adragni Rosa Lundbye L. Allesøe Kristine H. Allin Anna A. Artati Manimozhiyan Arumugam Naeimeh Atabaki Pasdar Tania Baltauss Karina Banasik Anna Barnett Patrick Baum Jimmy D. Bell Susanna Bianzano Roberto Bizzotto Amelie Bonnefond Caroline Anna A. Brorsson Andrew A. Brown Søren Brunak Louise Cabrelli Robert Caiazzo Henna Cederberg Elizaveta Chabanova Marc Clos-Garcia Matilda Dale David Davtian Adem Y. Dawed Federico De Masi Nathalie de Preville Koen F. Dekkers Harshal A. Deshmukh Christiane Dings Avirup Dutta Beate Ehrhardt Line Engelbrechtsen Rebeca Eriksen Yong Fan Juan Fernandez Jorge Ferrer Hugo Fitipaldi Ian M. Forgie Annemette Forman Francesca Frau Philippe Froguel Gary Frost Johann Gassenhuber Giuseppe (Nick) N. Giordano Toni Giorgino Stephen Gough Harald Grallert Rolf Grempler Lenka Groeneveld Leif Groop Valborg Gudmundsdóttir Ramneek Gupta Mark Haid Torben Hansen Tue H. Hansen Andrew T. Hattersley Ragna Haussler Alison J. Heggie Anita M. Hennige Anita V. Hill Reinhard W. Holl Michelle Hudson Bernd Jablonka Ulrik Plesner Jacobsen Christopher Jennison Joachim Johansen Angus G. Jones Tugce Karaderi Jane Kaye Gwen Kennedy Maria Klintenberg Robert W. Koivula Tarja Kokkola Anitra D. Koopman Azra Kurbasic Teemu Kuulasmaa Markku Laakso Thorsten Lehr Heather Loftus Agnete Troen T. Lundgaard Liwei Lyu Anubha Mahajan Andrea Mari Gianluca Mazzoni Mark I. McCarthy Timothy J. McDonald Donna McEvoy Nicky McRobert Ian McVittie Miranda Mourby Petra Musholt Pascal Mutie Rachel Nice Claudia Nicolay Giel Nijpels Birgitte Nilsson Colin N. Palmer Francois Pattou Imre Pavo Helle K. Pedersen Oluf Pedersen Mandy H. Perry Hugo Pomares-Millan Cornelia P. Prehn Anna Ramisch Simon Rasmussen Violeta Raverdi Martin Ridderstråle Neil Robertson Marianne Rodriquez Hartmut Ruetten Femke Rutters Peter Sackett Nina Scherer Nisha Shah Sapna Sharma Iryna Sihinevich Nadja B. Sondertoft Hans-Henrik Staerfeldt Birgit Steckel-Hamann Harriet Teare Cecilia Engel E. Thomas Elizabeth Louise L. Thomas Melissa K. Thomas Henrik S. Thomsen Barbara Thorand Claire E. Thorne Joachim Tillner Konstantinos D. Tsirigos Andrea Tura Mathias Uhlen Sabine van Oort Jagadish Vangipurapu Helene Verkindt Henrik Vestergaard Ana Viñuela Josef K. Vogt Peter W. Wad Sackett Mark Walker Agata Wesolowska-Andersen Brandon Whitcher Margaret W. White Alexander Efanov Giuseppe N. Giordano Gerard A. Bouland Frédéric Burdet Iulian Dragan Andreas Festa Michael K. Hansen Dmitry Kuznetsov Florence Mehl Diana Marek Imre Pavo Kevin Duffin Samreen K. Syed Janice L. Shaw Over Cabrera Timothy J. Pullen Bernard Thorens Mark Ibberson Guy A. Rutter |
| author_sort | Ben Connolly |
| collection | DOAJ |
| description | Summary: Background: Metformin is one of the most used drugs worldwide. Given the increasing use of proteomics in trials, bioresources, and clinics, it is crucial to understand the influence of metformin on the levels of the circulating proteome. Methods: We analysed a combined longitudinal proteomics dataset from the IMPOCT, RAMP and S3WP-T2D clinical trials in 98 participants before and after metformin exposure. This discovery analysis contained 372 proteins measured by proximity extension assays (Olink). We followed up experiment–wise statistically significant findings in two cross-sectional cohorts of people with type 2 diabetes comparing metformin treated and untreated individuals: IMI-DIRECT (784 participants, 372 proteins, Olink) and IMI-RHAPSODY (1175 participants, 1195 proteins, SomaLogic). Findings: Overall, 23 protein analytes were robustly associated with exposure to metformin in the discovery and replication. This includes 11 protein-metformin associations that replicated in both replication sets and platforms (REG4, GDF15, REG1A, t-PA, TFF3, CDH5, CNTN1, OMD, NOTCH3, THBS4 and CD93), with the remaining 12 protein-metformin associations replicated using the Olink platform (EPCAM, SPINK1, SAA-4, COMP, ITGB2, ADGRG2, FAM3C, MERTK, COL1A1, HAOX1, VCAN, TIMD4) but not measured on the SomaLogic platform. Gene-set enrichment analysis revealed that the metformin exposure was associated with intestinal associated proteins. Interpretation: These data highlight the need to account for exposure to metformin, and potentially other drugs, in proteomic studies and where protein biomarkers are used for clinical care. Funding: Innovative Medicines Initiative Joint Undertaking 2, under grant agreement no. 115881 (RHAPSODY) and the Innovative Medicines Initiative Joint Undertaking under grant agreement no. 115317 (DIRECT), resources of which are composed of financial contribution from the European Union's Seventh Framework Programme (FP7/2007-2013) and EFPIA companies in kind contribution as well as the Swiss State Secretariat for Education Research' and Innovation (SERI), under contract no. 16.0097 (RHAPSODY). |
| format | Article |
| id | doaj-art-c5ab2db824d94938ae0c666a8ef1f5a6 |
| institution | Kabale University |
| issn | 2352-3964 |
| language | English |
| publishDate | 2025-08-01 |
| publisher | Elsevier |
| record_format | Article |
| series | EBioMedicine |
| spelling | doaj-art-c5ab2db824d94938ae0c666a8ef1f5a62025-08-20T03:36:35ZengElsevierEBioMedicine2352-39642025-08-0111810585910.1016/j.ebiom.2025.105859The influence of metformin treatment on the circulating proteomeResearch in contextBen Connolly0Laura McCreight1Roderick C. Slieker2Khaled F. Bedair3Louise Donnelly4Juliette A. de Klerk5Joline W.J. Beulens6Petra J.M. Elders7Göran Bergström8Mun-Gwan Hong9Robert W. Koivula10Paul W. Franks11Jochen M. Schwenk12Anders Gummesson13Ewan R. Pearson14Leen M. ‘t Hart15Moustafa AbdallaJonathan AdamJerzy AdamskiKofi AdragniRosa Lundbye L. AllesøeKristine H. AllinAnna A. ArtatiManimozhiyan ArumugamNaeimeh Atabaki PasdarTania BaltaussKarina BanasikAnna BarnettPatrick BaumJimmy D. BellSusanna BianzanoRoberto BizzottoAmelie BonnefondCaroline Anna A. BrorssonAndrew A. BrownSøren BrunakLouise CabrelliRobert CaiazzoHenna CederbergElizaveta ChabanovaMarc Clos-GarciaMatilda DaleDavid DavtianAdem Y. DawedFederico De MasiNathalie de PrevilleKoen F. DekkersHarshal A. DeshmukhChristiane DingsAvirup DuttaBeate EhrhardtLine EngelbrechtsenRebeca EriksenYong FanJuan FernandezJorge FerrerHugo FitipaldiIan M. ForgieAnnemette FormanFrancesca FrauPhilippe FroguelGary FrostJohann GassenhuberGiuseppe (Nick) N. GiordanoToni GiorginoStephen GoughHarald GrallertRolf GremplerLenka GroeneveldLeif GroopValborg GudmundsdóttirRamneek GuptaMark HaidTorben HansenTue H. HansenAndrew T. HattersleyRagna HausslerAlison J. HeggieAnita M. HennigeAnita V. HillReinhard W. HollMichelle HudsonBernd JablonkaUlrik Plesner JacobsenChristopher JennisonJoachim JohansenAngus G. JonesTugce KaraderiJane KayeGwen KennedyMaria KlintenbergRobert W. KoivulaTarja KokkolaAnitra D. KoopmanAzra KurbasicTeemu KuulasmaaMarkku LaaksoThorsten LehrHeather LoftusAgnete Troen T. LundgaardLiwei LyuAnubha MahajanAndrea MariGianluca MazzoniMark I. McCarthyTimothy J. McDonaldDonna McEvoyNicky McRobertIan McVittieMiranda MourbyPetra MusholtPascal MutieRachel NiceClaudia NicolayGiel NijpelsBirgitte NilssonColin N. PalmerFrancois PattouImre PavoHelle K. PedersenOluf PedersenMandy H. PerryHugo Pomares-MillanCornelia P. PrehnAnna RamischSimon RasmussenVioleta RaverdiMartin RidderstråleNeil RobertsonMarianne RodriquezHartmut RuettenFemke RuttersPeter SackettNina SchererNisha ShahSapna SharmaIryna SihinevichNadja B. SondertoftHans-Henrik StaerfeldtBirgit Steckel-HamannHarriet TeareCecilia Engel E. ThomasElizabeth Louise L. ThomasMelissa K. ThomasHenrik S. ThomsenBarbara ThorandClaire E. ThorneJoachim TillnerKonstantinos D. TsirigosAndrea TuraMathias UhlenSabine van OortJagadish VangipurapuHelene VerkindtHenrik VestergaardAna ViñuelaJosef K. VogtPeter W. Wad SackettMark WalkerAgata Wesolowska-AndersenBrandon WhitcherMargaret W. WhiteAlexander EfanovGiuseppe N. GiordanoGerard A. BoulandFrédéric BurdetIulian DraganAndreas FestaMichael K. HansenDmitry KuznetsovFlorence MehlDiana MarekImre PavoKevin DuffinSamreen K. SyedJanice L. ShawOver CabreraTimothy J. PullenBernard ThorensMark IbbersonGuy A. RutterDivision of Population Health & Genomics, School of Medicine, University of Dundee, UKDivision of Population Health & Genomics, School of Medicine, University of Dundee, UKDepartment of Cell and Chemical Biology, Leiden University Medical Center, Leiden, the NetherlandsDivision of Population Health & Genomics, School of Medicine, University of Dundee, UKDivision of Population Health & Genomics, School of Medicine, University of Dundee, UKDepartment of Cell and Chemical Biology, Leiden University Medical Center, Leiden, the Netherlands; Department of Internal Medicine (Nephrology), Leiden University Medical Center, Leiden, the NetherlandsDepartment of Epidemiology and Data Sciences, Amsterdam University Medical Center, Location Vrije Universiteit Amsterdam, Amsterdam, the Netherlands; Amsterdam Cardiovascular Sciences Research Institutes, Amsterdam Public Health, Amsterdam, the NetherlandsHealth Behaviors & Chronic Diseases Research Program & Personalised Medicine Research Program Amsterdam Public Health, Amsterdam, the Netherlands; Department of General Practice Medicine, Amsterdam UMC, Location Vrije Universiteit, Amsterdam, the NetherlandsRegion Västra Götaland, Sahlgrenska University Hospital, Department of Clinical Physiology, Gothenburg, Sweden; Department of Molecular and Clinical Medicine, Institute of Medicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, SwedenDepartment of Protein Science, SciLifeLab, KTH Royal Institute of Technology, Stockholm, SwedenOxford Centre for Diabetes, Endocrinology and Metabolism, Radcliffe Department of Medicine, University of Oxford, Oxford, UK; Genetic and Molecular Epidemiology Unit, Lund University Diabetes Centre, Department of Clinical Sciences, Lund University, Malmo, SwedenGenetic and Molecular Epidemiology Unit, Lund University Diabetes Centre, Department of Clinical Sciences, Lund University, Malmo, Sweden; Precision Health University Research Institute, Queen Mary University of London, London, UKDepartment of Protein Science, SciLifeLab, KTH Royal Institute of Technology, Stockholm, SwedenDepartment of Molecular and Clinical Medicine, Institute of Medicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden; Region Västra Götaland, Sahlgrenska University Hospital, Department of Clinical Genetics and Genomics, Gothenburg, SwedenDivision of Population Health & Genomics, School of Medicine, University of Dundee, UK; Corresponding author. Division of Population Health & Genomics, School of Medicine, University of Dundee, DD1 9SY.Department of Cell and Chemical Biology, Leiden University Medical Center, Leiden, the Netherlands; Department of Epidemiology and Data Sciences, Amsterdam University Medical Center, Location Vrije Universiteit Amsterdam, Amsterdam, the Netherlands; Health Behaviors & Chronic Diseases Research Program & Personalised Medicine Research Program Amsterdam Public Health, Amsterdam, the Netherlands; Department of Biomedical Data Sciences, Section Molecular Epidemiology, Leiden University Medical Center, Leiden, the Netherlands; Corresponding author. Leiden University Medical Center, Department of Cell and Chemical Biology, Albinusdreef 2, 2333ZA, Leiden, the Netherlands.Summary: Background: Metformin is one of the most used drugs worldwide. Given the increasing use of proteomics in trials, bioresources, and clinics, it is crucial to understand the influence of metformin on the levels of the circulating proteome. Methods: We analysed a combined longitudinal proteomics dataset from the IMPOCT, RAMP and S3WP-T2D clinical trials in 98 participants before and after metformin exposure. This discovery analysis contained 372 proteins measured by proximity extension assays (Olink). We followed up experiment–wise statistically significant findings in two cross-sectional cohorts of people with type 2 diabetes comparing metformin treated and untreated individuals: IMI-DIRECT (784 participants, 372 proteins, Olink) and IMI-RHAPSODY (1175 participants, 1195 proteins, SomaLogic). Findings: Overall, 23 protein analytes were robustly associated with exposure to metformin in the discovery and replication. This includes 11 protein-metformin associations that replicated in both replication sets and platforms (REG4, GDF15, REG1A, t-PA, TFF3, CDH5, CNTN1, OMD, NOTCH3, THBS4 and CD93), with the remaining 12 protein-metformin associations replicated using the Olink platform (EPCAM, SPINK1, SAA-4, COMP, ITGB2, ADGRG2, FAM3C, MERTK, COL1A1, HAOX1, VCAN, TIMD4) but not measured on the SomaLogic platform. Gene-set enrichment analysis revealed that the metformin exposure was associated with intestinal associated proteins. Interpretation: These data highlight the need to account for exposure to metformin, and potentially other drugs, in proteomic studies and where protein biomarkers are used for clinical care. Funding: Innovative Medicines Initiative Joint Undertaking 2, under grant agreement no. 115881 (RHAPSODY) and the Innovative Medicines Initiative Joint Undertaking under grant agreement no. 115317 (DIRECT), resources of which are composed of financial contribution from the European Union's Seventh Framework Programme (FP7/2007-2013) and EFPIA companies in kind contribution as well as the Swiss State Secretariat for Education Research' and Innovation (SERI), under contract no. 16.0097 (RHAPSODY).http://www.sciencedirect.com/science/article/pii/S2352396425003032MetforminProteomicsType 2 diabetesBiomarker |
| spellingShingle | Ben Connolly Laura McCreight Roderick C. Slieker Khaled F. Bedair Louise Donnelly Juliette A. de Klerk Joline W.J. Beulens Petra J.M. Elders Göran Bergström Mun-Gwan Hong Robert W. Koivula Paul W. Franks Jochen M. Schwenk Anders Gummesson Ewan R. Pearson Leen M. ‘t Hart Moustafa Abdalla Jonathan Adam Jerzy Adamski Kofi Adragni Rosa Lundbye L. Allesøe Kristine H. Allin Anna A. Artati Manimozhiyan Arumugam Naeimeh Atabaki Pasdar Tania Baltauss Karina Banasik Anna Barnett Patrick Baum Jimmy D. Bell Susanna Bianzano Roberto Bizzotto Amelie Bonnefond Caroline Anna A. Brorsson Andrew A. Brown Søren Brunak Louise Cabrelli Robert Caiazzo Henna Cederberg Elizaveta Chabanova Marc Clos-Garcia Matilda Dale David Davtian Adem Y. Dawed Federico De Masi Nathalie de Preville Koen F. Dekkers Harshal A. Deshmukh Christiane Dings Avirup Dutta Beate Ehrhardt Line Engelbrechtsen Rebeca Eriksen Yong Fan Juan Fernandez Jorge Ferrer Hugo Fitipaldi Ian M. Forgie Annemette Forman Francesca Frau Philippe Froguel Gary Frost Johann Gassenhuber Giuseppe (Nick) N. Giordano Toni Giorgino Stephen Gough Harald Grallert Rolf Grempler Lenka Groeneveld Leif Groop Valborg Gudmundsdóttir Ramneek Gupta Mark Haid Torben Hansen Tue H. Hansen Andrew T. Hattersley Ragna Haussler Alison J. Heggie Anita M. Hennige Anita V. Hill Reinhard W. Holl Michelle Hudson Bernd Jablonka Ulrik Plesner Jacobsen Christopher Jennison Joachim Johansen Angus G. Jones Tugce Karaderi Jane Kaye Gwen Kennedy Maria Klintenberg Robert W. Koivula Tarja Kokkola Anitra D. Koopman Azra Kurbasic Teemu Kuulasmaa Markku Laakso Thorsten Lehr Heather Loftus Agnete Troen T. Lundgaard Liwei Lyu Anubha Mahajan Andrea Mari Gianluca Mazzoni Mark I. McCarthy Timothy J. McDonald Donna McEvoy Nicky McRobert Ian McVittie Miranda Mourby Petra Musholt Pascal Mutie Rachel Nice Claudia Nicolay Giel Nijpels Birgitte Nilsson Colin N. Palmer Francois Pattou Imre Pavo Helle K. Pedersen Oluf Pedersen Mandy H. Perry Hugo Pomares-Millan Cornelia P. Prehn Anna Ramisch Simon Rasmussen Violeta Raverdi Martin Ridderstråle Neil Robertson Marianne Rodriquez Hartmut Ruetten Femke Rutters Peter Sackett Nina Scherer Nisha Shah Sapna Sharma Iryna Sihinevich Nadja B. Sondertoft Hans-Henrik Staerfeldt Birgit Steckel-Hamann Harriet Teare Cecilia Engel E. Thomas Elizabeth Louise L. Thomas Melissa K. Thomas Henrik S. Thomsen Barbara Thorand Claire E. Thorne Joachim Tillner Konstantinos D. Tsirigos Andrea Tura Mathias Uhlen Sabine van Oort Jagadish Vangipurapu Helene Verkindt Henrik Vestergaard Ana Viñuela Josef K. Vogt Peter W. Wad Sackett Mark Walker Agata Wesolowska-Andersen Brandon Whitcher Margaret W. White Alexander Efanov Giuseppe N. Giordano Gerard A. Bouland Frédéric Burdet Iulian Dragan Andreas Festa Michael K. Hansen Dmitry Kuznetsov Florence Mehl Diana Marek Imre Pavo Kevin Duffin Samreen K. Syed Janice L. Shaw Over Cabrera Timothy J. Pullen Bernard Thorens Mark Ibberson Guy A. Rutter The influence of metformin treatment on the circulating proteomeResearch in context EBioMedicine Metformin Proteomics Type 2 diabetes Biomarker |
| title | The influence of metformin treatment on the circulating proteomeResearch in context |
| title_full | The influence of metformin treatment on the circulating proteomeResearch in context |
| title_fullStr | The influence of metformin treatment on the circulating proteomeResearch in context |
| title_full_unstemmed | The influence of metformin treatment on the circulating proteomeResearch in context |
| title_short | The influence of metformin treatment on the circulating proteomeResearch in context |
| title_sort | influence of metformin treatment on the circulating proteomeresearch in context |
| topic | Metformin Proteomics Type 2 diabetes Biomarker |
| url | http://www.sciencedirect.com/science/article/pii/S2352396425003032 |
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