MMP Mediated Degradation of Type IV Collagen Alpha 1 and Alpha 3 Chains Reflects Basement Membrane Remodeling in Experimental and Clinical Fibrosis – Validation of Two Novel Biomarker Assays
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{"title"=>"MMP mediated degradation of type IV collagen alpha 1 and alpha 3 chains reflects basement membrane remodeling in experimental and clinical fibrosis - Validation of two novel biomarker assays", "type"=>"journal", "authors"=>[{"first_name"=>"Jannie Marie", "last_name"=>"Sand", "scopus_author_id"=>"55365925700"}, {"first_name"=>"Lise", "last_name"=>"Larsen", "scopus_author_id"=>"57197149480"}, {"first_name"=>"Cory", "last_name"=>"Hogaboam", "scopus_author_id"=>"7005896872"}, {"first_name"=>"Fernando", "last_name"=>"Martinez", "scopus_author_id"=>"35374549400"}, {"first_name"=>"Mei Lan", "last_name"=>"Han", "scopus_author_id"=>"12786822400"}, {"first_name"=>"Martin Roøssel", "last_name"=>"Larsen", "scopus_author_id"=>"7201730952"}, {"first_name"=>"Arkadiusz", "last_name"=>"Nawrocki", "scopus_author_id"=>"7004449640"}, {"first_name"=>"Qinlong", "last_name"=>"Zheng", "scopus_author_id"=>"36342144400"}, {"first_name"=>"Morten Asser", "last_name"=>"Karsdal", "scopus_author_id"=>"6602506854"}, {"first_name"=>"Diana Julie", "last_name"=>"Leeming", "scopus_author_id"=>"12798086700"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"sgr"=>"84893320758", "doi"=>"10.1371/journal.pone.0084934", "pui"=>"372285519", "pmid"=>"24376856", "scopus"=>"2-s2.0-84893320758", "issn"=>"19326203", "isbn"=>"1932-6203"}, "id"=>"85afdddc-24e7-3a01-8d8c-65d34074cd7f", "abstract"=>"OBJECTIVES: Fibrosis is characterized by excessive tissue remodeling resulting from altered expression of various growth factors, cytokines and proteases. We hypothesized that matrix metalloproteinase (MMP) mediated degradation of type IV collagen, a main component of the basement membrane, will release peptide fragments (neo-epitopes) into the circulation. Here we present the development of two competitive enzyme-linked immunosorbent assays (ELISAs) for assessing the levels of specific fragments of type IV collagen α1 (C4M12a1) and α3 (C4M12a3) chains in serum as indicators of fibrosis.\\n\\nMETHODS: Fragments of type IV collagen cleaved in vitro by MMP-12 were identified by mass spectrometry, and two were chosen for ELISA development due to their unique sequences. The assays were evaluated using samples from a carbon tetrachloride (CCl₄) rat model of liver fibrosis and from patients with idiopathic pulmonary fibrosis (IPF) or chronic obstructive pulmonary disease (COPD).\\n\\nRESULTS: Two technically robust ELISAs were produced using neo-epitope specific monoclonal antibodies. Mean serum C4M12a1 levels were significantly elevated in CCl₄-treated rats compared with controls in weeks 12, 16, and 20, with a maximum increase of 102% at week 16 (p < 0.0001). Further, C4M12a1 levels correlated with the total collagen content of the liver in CCl₄-treated rats (r = 0.43, p = 0.003). Mean serum C4M12a3 levels were significantly elevated in patients with mild, moderate, and severe IPF, and COPD relative to healthy controls, with a maximum increase of 321% in COPD (p < 0.0001).\\n\\nCONCLUSIONS: Two assays measuring C4M12a1 and C4M12a3 enabled quantification of MMP mediated degradation of type IV collagen in serum. C4M12a1 was elevated in a pre-clinical model of liver fibrosis, and C4M12a3 was elevated in IPF and COPD patients. This suggests the use of these assays to investigate pathological remodeling of the basement membrane in different organs. However, validations in larger clinical settings are needed.", "link"=>"http://www.mendeley.com/research/mmp-mediated-degradation-type-iv-collagen-alpha-1-alpha-3-chains-reflects-basement-membrane-remodeli", "reader_count"=>72, "reader_count_by_academic_status"=>{"Unspecified"=>3, "Professor > Associate Professor"=>2, "Librarian"=>1, "Researcher"=>14, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>19, "Student > Postgraduate"=>2, "Student > Master"=>12, "Other"=>5, "Student > Bachelor"=>5, "Lecturer"=>1, "Lecturer > Senior Lecturer"=>1, "Professor"=>5}, "reader_count_by_user_role"=>{"Unspecified"=>3, "Professor > Associate Professor"=>2, "Librarian"=>1, "Researcher"=>14, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>19, "Student > Postgraduate"=>2, "Student > Master"=>12, "Other"=>5, "Student > Bachelor"=>5, "Lecturer"=>1, "Lecturer > Senior Lecturer"=>1, "Professor"=>5}, "reader_count_by_subject_area"=>{"Unspecified"=>7, "Engineering"=>3, "Biochemistry, Genetics and Molecular Biology"=>14, "Materials Science"=>1, "Agricultural and Biological Sciences"=>19, "Medicine and Dentistry"=>17, "Arts and Humanities"=>1, "Business, Management and Accounting"=>1, "Pharmacology, Toxicology and Pharmaceutical Science"=>4, "Sports and Recreations"=>1, "Chemistry"=>2, "Immunology and Microbiology"=>2}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>3}, "Materials Science"=>{"Materials Science"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>17}, "Chemistry"=>{"Chemistry"=>2}, "Sports and Recreations"=>{"Sports and Recreations"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>2}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>19}, "Business, Management and Accounting"=>{"Business, Management and Accounting"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>14}, "Unspecified"=>{"Unspecified"=>7}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>4}, "Arts and Humanities"=>{"Arts and Humanities"=>1}}, "reader_count_by_country"=>{"Denmark"=>3, "Italy"=>1}, "group_count"=>4}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1326051"], "description"=>"<p>Reactivity to the standard peptide (A: ILGHVPGMLL; B: PGDIVFRKGP), the elongated peptide (A: <u>E</u>ILGHVPGMLL; B: <u>P</u>PGDIVFRKGP) and a nonsense peptide (A + B: PGPPGIVIGT) was tested for the assays C4M12a1 (A) and C4M12a3 (B). The background signal from the system was tested using a nonsense coating peptide (A+B: biotin-K-PGPPGIVIGT). Signals are shown as optical density (OD) at 450 nm, subtracting the background at 650 nm, as a function of standard peptide concentration.</p>", "links"=>[], "tags"=>[], "article_id"=>884672, "categories"=>["Biological Sciences"], "users"=>["Jannie Marie Sand", "Lise Larsen", "Cory Hogaboam", "Fernando Martinez", "MeiLan Han", "Martin Røssel Larsen", "Arkadiusz Nawrocki", "Qinlong Zheng", "Morten Asser Karsdal", "Diana Julie Leeming"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0084934.g003", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Assay_specificity_/884672", "title"=>"Assay specificity.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-12-23 02:46:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/1326053"], "description"=>"<p>C4M12a1 (A and C) and C4M12a3 (B and D) marker levels were assessed in samples of purified type IV collagen cleaved <i>in </i><i>vitro</i> by MMP-12, pepsin, and MMP-12 in combination with pepsin (A and B), as well as MMP-2 and MMP-9 (C and D). Data were normalized by subtraction of intact type IV collagen background.</p>", "links"=>[], "tags"=>["cleaved", "iv"], "article_id"=>884674, "categories"=>["Biological Sciences"], "users"=>["Jannie Marie Sand", "Lise Larsen", "Cory Hogaboam", "Fernando Martinez", "MeiLan Han", "Martin Røssel Larsen", "Arkadiusz Nawrocki", "Qinlong Zheng", "Morten Asser Karsdal", "Diana Julie Leeming"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0084934.g004", "stats"=>{"downloads"=>0, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Reactivity_to_cleaved_type_IV_collagen_/884674", "title"=>"Reactivity to cleaved type IV collagen.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-12-23 02:46:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/1326054"], "description"=>"<p><i>A</i>: Serum C4M12a1 was assessed in control rats and carbon tetrachloride (CCl<sub>4</sub>)-treated rats at termination at week 8, 12, 16, and 20. Results are shown as mean ± SEM. <i>B</i>: Serum C4M12a1 levels of CCl<sub>4</sub>-treated rats were stratified into quartiles according to total collagen content of the liver as determined by Sirius red staining. Results are shown as mean ± SEM. <i>C+D</i>: Correlation of total collagen content of the liver (Sirius red) with serum C4M12a1 levels for controls (C) and CCl<sub>4</sub>-treated rats (D). Spearman r: Spearman correlation coefficient. Significance levels: *: p < 0.05, **: p < 0.01, ***: p < 0.001, ****: p < 0.0001. </p>", "links"=>[], "tags"=>["c4m12a1", "carbon", "tetrachloride-treated"], "article_id"=>884675, "categories"=>["Biological Sciences"], "users"=>["Jannie Marie Sand", "Lise Larsen", "Cory Hogaboam", "Fernando Martinez", "MeiLan Han", "Martin Røssel Larsen", "Arkadiusz Nawrocki", "Qinlong Zheng", "Morten Asser Karsdal", "Diana Julie Leeming"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0084934.g005", "stats"=>{"downloads"=>2, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Serum_C4M12a1_levels_in_carbon_tetrachloride_treated_rats_/884675", "title"=>"Serum C4M12a1 levels in carbon tetrachloride-treated rats.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-12-23 02:46:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/1326055"], "description"=>"<p>Serum C4M12a3 was assessed in healthy controls (n = 23) and patients with mild IPF (n = 10), moderate IPF (n = 10), severe IPF (n = 10), and mild COPD (n = 10). Results are presented as geometric mean ± SEM. Significance levels: *: p< 0.05, **: p < 0.01, ***: p < 0.001, ****: p < 0.0001. </p>", "links"=>[], "tags"=>["c4m12a3", "ipf", "copd"], "article_id"=>884676, "categories"=>["Biological Sciences"], "users"=>["Jannie Marie Sand", "Lise Larsen", "Cory Hogaboam", "Fernando Martinez", "MeiLan Han", "Martin Røssel Larsen", "Arkadiusz Nawrocki", "Qinlong Zheng", "Morten Asser Karsdal", "Diana Julie Leeming"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0084934.g006", "stats"=>{"downloads"=>2, "page_views"=>30, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Serum_C4M12a3_levels_in_IPF_and_COPD_patients_/884676", "title"=>"Serum C4M12a3 levels in IPF and COPD patients.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-12-23 02:46:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/1326048"], "description"=>"<p>The sequence of the type IV collagen α1 chain (left) and α3 chain (right) were aligned for the species human, rat, and mouse. Bold types indicate the target sequence, while dots indicate the MMP-12 cleavage site recognized by the monoclonal antibody employed in the respective assay (left: C4M12a1, right: C4M12a3).</p>", "links"=>[], "tags"=>["iv", "collagen"], "article_id"=>884669, "categories"=>["Biological Sciences"], "users"=>["Jannie Marie Sand", "Lise Larsen", "Cory Hogaboam", "Fernando Martinez", "MeiLan Han", "Martin Røssel Larsen", "Arkadiusz Nawrocki", "Qinlong Zheng", "Morten Asser Karsdal", "Diana Julie Leeming"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0084934.g001", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Alignment_of_type_IV_collagen_945_1_and_945_3_chains_/884669", "title"=>"Alignment of type IV collagen α1 and α3 chains.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-12-23 02:46:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/1326050"], "description"=>"<p>Typical standard curves and native reactivity against human, rat, and mouse material diluted to 1:8 as indicated for the assays C4M12a1 (A) and C4M12a3 (B). The reactivity of the C4M12a3 ELISA to the rat sequence (PGDTVFQPGP) was also tested (B). Signals are shown as optical density (OD) at 450 nm, subtracting the background at 650 nm, as a function of standard peptide concentration. UD: undiluted sample.</p>", "links"=>[], "tags"=>["curves"], "article_id"=>884671, "categories"=>["Biological Sciences"], "users"=>["Jannie Marie Sand", "Lise Larsen", "Cory Hogaboam", "Fernando Martinez", "MeiLan Han", "Martin Røssel Larsen", "Arkadiusz Nawrocki", "Qinlong Zheng", "Morten Asser Karsdal", "Diana Julie Leeming"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0084934.g002", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Standard_curves_and_native_reactivity_/884671", "title"=>"Standard curves and native reactivity.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-12-23 02:46:28"}

PMC Usage Stats | Further Information

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Relative Metric

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