Carcinomas contain a matrix metalloproteinase-resistant isoform of type I collagen exerting selective support to invasion.

Collagen fibers affect metastasis in two opposing ways, by supporting invasive cells but also by generating a barrier to invasion. We hypothesized that these functions might be performed by different isoforms of type I collagen. Carcinomas are reported to contain alpha1(I)(3) homotrimers, a type I collagen isoform normally not present in healthy tissues, but the role of the homotrimers in cancer pathophysiology is unclear. In this study, we found that these homotrimers were resistant to all collagenolytic matrix metalloproteinases (MMP). MMPs are massively produced and used by cancer cells and cancer-associated fibroblasts for degrading stromal collagen at the leading edge of tumor invasion. The MMP-resistant homotrimers were produced by all invasive cancer cell lines tested, both in culture and in tumor xenografts, but they were not produced by cancer-associated fibroblasts, thereby comprising a specialized fraction of tumor collagen. We observed the homotrimer fibers to be resistant to pericellular degradation, even upon stimulation of the cells with proinflammatory cytokines. Furthermore, we confirmed an enhanced proliferation and migration of invasive cancer cells on the surface of homotrimeric versus normal (heterotrimeric) type I collagen fibers. In summary, our findings suggest that invasive cancer cells may use homotrimers for building MMP-resistant invasion paths, supporting local proliferation and directed migration of the cells whereas surrounding normal stromal collagens are cleaved. Because the homotrimers are universally secreted by cancer cells and deposited as insoluble, MMP-resistant fibers, they offer an appealing target for cancer diagnostics and therapy.

[1]  S. Krane,et al.  Severely impaired wound healing in the collagenase-resistant mouse. , 2003, The Journal of investigative dermatology.

[2]  D. Eyre,et al.  Collagen polymorphism in extracellular matrix of human osteosarcoma. , 1982, Journal of the National Cancer Institute.

[3]  S. Yamagata,et al.  FBJ virus-induced osteosarcoma contains type I, type I trimer, type III as well as type V collagens. , 1984, Journal of biochemistry.

[4]  R. Hegerl.  Three-dimensional Reconstruction from Projections in Electron Microscopy , 1989 .

[5]  Z. Werb,et al.  New functions for the matrix metalloproteinases in cancer progression , 2002, Nature Reviews Cancer.

[6]  C. Luparello,et al.  Collagen biosynthesis by a breast carcinoma cell strain and biopsy fragments of the primary tumour. , 1988, Cell biology international reports.

[7]  R. Jaenisch,et al.  A targeted mutation at the known collagenase cleavage site in mouse type I collagen impairs tissue remodeling , 1995, The Journal of cell biology.

[8]  Michael P. Sheetz,et al.  Differential Matrix Rigidity Response in Breast Cancer Cell Lines Correlates with the Tissue Tropism , 2009, PloS one.

[9]  S. Krane,et al.  Osteocyte and osteoblast apoptosis and excessive bone deposition accompany failure of collagenase cleavage of collagen. , 2000, The Journal of clinical investigation.

[10]  Frank H. Ruddle,et al.  Procollagen and collagen produced by a teratocarcinoma-derived cell line, TSD4: Evidence for a new molecular form of collagen , 1977, Cell.

[11]  Raghu Kalluri,et al.  Fibroblasts in cancer , 2006, Nature Reviews Cancer.

[12]  E. Jaffee,et al.  Ectopic expression of vascular cell adhesion molecule-1 as a new mechanism for tumor immune evasion. , 2007, Cancer research.

[13]  H. Birkedal‐Hansen,et al.  Matrix Metalloproteinases , 2008, Current protocols in cell biology.

[14]  J. Ward,et al.  MT1-MMP-Deficient Mice Develop Dwarfism, Osteopenia, Arthritis, and Connective Tissue Disease due to Inadequate Collagen Turnover , 1999, Cell.

[15]  Y. DeClerck,et al.  Differential collagen biosynthesis by human neuroblastoma cell variants. , 1987, Cancer research.

[16]  Gerhard Christofori,et al.  EMT, the cytoskeleton, and cancer cell invasion , 2009, Cancer and Metastasis Reviews.

[17]  Jason C. Ho,et al.  The Integrin-Extracellular Matrix Axis in Pancreatic Cancer , 2007, Pancreas.

[18]  P. Fratzl,et al.  Collagen from the osteogenesis imperfecta mouse model (oim) shows reduced resistance against tensile stress. , 1997, The Journal of clinical investigation.

[19]  M. Stack,et al.  Membrane type 1-matrix metalloproteinase: substrate diversity in pericellular proteolysis. , 2008, Seminars in cell & developmental biology.

[20]  R. Hoover,et al.  Collagen polymorphism in cultured rat kidney mesangial cells. , 1987, Laboratory investigation; a journal of technical methods and pathology.

[21]  Gillian Murphy,et al.  Structure and function of matrix metalloproteinases and TIMPs. , 2006, Cardiovascular research.

[22]  R. Page,et al.  Action of mammalian collagenases on type I trimer collagen. , 1984, Collagen and related research.

[23]  E. Marsilio,et al.  Absence of procollagen alpha 2(I) mRNA in chemically transformed rat liver epithelial cells. , 1984, The Journal of biological chemistry.

[24]  I. Damjanov,et al.  Synthesis of type I homotrimer collagen molecules by cultured human lung adenocarcinoma cells. , 1988, The American journal of pathology.

[25]  D. Rifkin,et al.  Biology and biochemistry of proteinases in tumor invasion. , 1993, Physiological reviews.

[26]  Joan C. Marini,et al.  Structural Heterogeneity of Type I Collagen Triple Helix and Its Role in Osteogenesis Imperfecta* , 2008, Journal of Biological Chemistry.

[27]  R. Puvanakrishnan,et al.  Purification and characterization of collagens from rat fibrosarcoma induced by 3-methylcholanthrene , 1993, Molecular and Cellular Biochemistry.

[28]  D. Dinakarpandian,et al.  Collagenase unwinds triple‐helical collagen prior to peptide bond hydrolysis , 2004, The EMBO journal.

[29]  G. Balian,et al.  Two cell lines from bone marrow that differ in terms of collagen synthesis, osteogenic characteristics, and matrix mineralization. , 1993, The Journal of bone and joint surgery. American volume.

[30]  Wolfgang Losert,et al.  Segregation of type I collagen homo- and heterotrimers in fibrils. , 2008, Journal of molecular biology.

[31]  R. Garbelli,et al.  Absence of regular alpha2(I) collagen chains in colon carcinoma biopsy fragments. , 1998, Carcinogenesis.

[32]  Sanjay Kumar,et al.  The mechanical rigidity of the extracellular matrix regulates the structure, motility, and proliferation of glioma cells. , 2009, Cancer research.

[33]  Scott A. Guelcher,et al.  Extracellular Matrix Rigidity Promotes Invadopodia Activity , 2008, Current Biology.

[34]  Barbara Smith,et al.  DNA hypermethylation near the transcription start site of collagen alpha2(I) gene occurs in both cancer cell lines and primary colorectal cancers. , 2003, Cancer research.

[35]  S. Leikin,et al.  Changes in thermal stability and microunfolding pattern of collagen helix resulting from the loss of alpha2(I) chain in osteogenesis imperfecta murine. , 2003, Journal of molecular biology.

[36]  L. Reneker,et al.  Type I collagen glomerulopathy: postnatal collagen deposition follows glomerular maturation. , 2007, Kidney international.

[37]  M T Davisson,et al.  Defective pro alpha 2(I) collagen synthesis in a recessive mutation in mice: a model of human osteogenesis imperfecta. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[38]  B. Leheup,et al.  Extracellular matrix biosynthesis by cultured fetal rat lung epithelial cells. I. Characterization of the clone and the major genetic types of collagen produced. , 1989, Laboratory investigation; a journal of technical methods and pathology.

[39]  Cynthia A. Reinhart-King,et al.  Tensional homeostasis and the malignant phenotype. , 2005, Cancer cell.

[40]  P. Byers,et al.  Rare autosomal recessive cardiac valvular form of Ehlers-Danlos syndrome results from mutations in the COL1A2 gene that activate the nonsense-mediated RNA decay pathway. , 2004, American journal of human genetics.

[41]  E. J. Miller,et al.  Analysis of changes in collagen biosynthesis that occur when chick chondrocytes are grown in 5-bromo-2'-deoxyuridine. , 1975, Proceedings of the National Academy of Sciences of the United States of America.

[42]  L. Liotta,et al.  Tumor invasion and metastasis: an imbalance of positive and negative regulation. , 1991, Cancer research.

[43]  J. Quigley,et al.  Matrix metalloproteinases and tumor metastasis , 2006, Cancer and Metastasis Reviews.

[44]  C. Luparello,et al.  A comparison of spreading and motility behaviour of 8701-BC breast carcinoma cells on type I, I-trimer and type V collagen substrata. Evidence for a permissive effect of type I-trimer collagen on cell locomotion. , 1991, Journal of cell science.

[45]  L. Moro,et al.  Identification of collagen alpha1(I) trimer and normal type I collagen in a polyoma virus-induced mouse tumor. , 1977, Archives of biochemistry and biophysics.

[46]  S. Leikin,et al.  Molecular mechanism of alpha 1(I)-osteogenesis imperfecta/Ehlers-Danlos syndrome: unfolding of an N-anchor domain at the N-terminal end of the type I collagen triple helix. , 2006, The Journal of biological chemistry.

[47]  B. Smith,et al.  Characterization of collagen synthesized by normal and chemically transformed rat liver epithelial cell lines. , 1980, Biochemistry.