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Despite its essential functions, the impact of age-associated post-translational modifications on ECM-driven fibroblast activities such as proliferation, survival, fibroblast-to-myofibroblast transformation (FMT), and extracellular matrix production remains largely unknown. N\u03b5-carboxymethyl-lysine (CML) is one of the well-characterized advanced glycation end-products (AGEs) that can occur on lysine residues within ECM proteins through non-enzymatic glycation. In this study, we determined the accumulation and the effects of the CML-modified ECM (CML-ECM) on fibroblast activation. Immunostainings and immunoblot analysis demonstrated significant increases in CML-AGE content in idiopathic pulmonary fibrosis (IPF) compared to age-matched healthy lungs. Gene expression analysis and fibroblast activation assays collectively implicate the ECM as a negative regulator of fibroblast activation. Notably, the CML modification of the ECM resulted in a significant decrease in its anti-fibrotic effects including proliferation, FMT, apoptosis, and ECM production. Together, the results of this study revealed an unexplored pathological role played by the CML-ECM on fibroblast activation, which has wide implications in IPF and other fibrotic diseases.<\/jats:p>","DOI":"10.3390\/ijms242115811","type":"journal-article","created":{"date-parts":[[2023,10,31]],"date-time":"2023-10-31T11:16:27Z","timestamp":1698750987000},"page":"15811","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["N\u03b5-Carboxymethyl-Lysine Modification of Extracellular Matrix Proteins Augments Fibroblast Activation"],"prefix":"10.3390","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2809-5449","authenticated-orcid":false,"given":"Harshavardhana H.","family":"Ediga","sequence":"first","affiliation":[{"name":"Division of Pulmonary, Critical Care and Sleep Medicine, Department of Internal Medicine, University of Cincinnati, Cincinnati, OH 45267-0564, USA"},{"name":"Department of Biochemistry, ICMR-National Institute of Nutrition, Hyderabad 500007, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrick","family":"Hester","sequence":"additional","affiliation":[{"name":"Division of Pulmonary, Critical Care and Sleep Medicine, Department of Internal Medicine, University of Cincinnati, Cincinnati, OH 45267-0564, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adithi","family":"Yepuri","sequence":"additional","affiliation":[{"name":"Division of Pulmonary, Critical Care and Sleep Medicine, Department of Internal Medicine, University of Cincinnati, Cincinnati, OH 45267-0564, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4787-3944","authenticated-orcid":false,"given":"Geereddy Bhanuprakash","family":"Reddy","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, ICMR-National Institute of Nutrition, Hyderabad 500007, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6042-5321","authenticated-orcid":false,"given":"Satish K.","family":"Madala","sequence":"additional","affiliation":[{"name":"Division of Pulmonary, Critical Care and Sleep Medicine, Department of Internal Medicine, University of Cincinnati, Cincinnati, OH 45267-0564, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,10,31]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2031","DOI":"10.1007\/s00018-020-03693-7","article-title":"Emerging cellular and molecular determinants of idiopathic pulmonary fibrosis","volume":"78","author":"Phan","year":"2021","journal-title":"Cell. 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