Lysyl hydroxylase 2 induces a collagen cross-link switch in tumor stroma
Yulong Chen, … , Mitsuo Yamauchi, Jonathan M. Kurie
Yulong Chen, … , Mitsuo Yamauchi, Jonathan M. Kurie
Published February 9, 2015
Citation Information: J Clin Invest. 2015;125(3):1147-1162. https://doi.org/10.1172/JCI74725.
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Lysyl hydroxylase 2 induces a collagen cross-link switch in tumor stroma

Abstract

Epithelial tumor metastasis is preceded by an accumulation of collagen cross-links that heighten stromal stiffness and stimulate the invasive properties of tumor cells. However, the biochemical nature of collagen cross-links in cancer is still unclear. Here, we postulated that epithelial tumorigenesis is accompanied by changes in the biochemical type of collagen cross-links. Utilizing resected human lung cancer tissues and a p21CIP1/WAF1-deficient, K-rasG12D-expressing murine metastatic lung cancer model, we showed that, relative to normal lung tissues, tumor stroma contains higher levels of hydroxylysine aldehyde–derived collagen cross-links (HLCCs) and lower levels of lysine aldehyde–derived cross-links (LCCs), which are the predominant types of collagen cross-links in skeletal tissues and soft tissues, respectively. Gain- and loss-of-function studies in tumor cells showed that lysyl hydroxylase 2 (LH2), which hydroxylates telopeptidyl lysine residues on collagen, shifted the tumor stroma toward a high-HLCC, low-LCC state, increased tumor stiffness, and enhanced tumor cell invasion and metastasis. Together, our data indicate that LH2 enhances the metastatic properties of tumor cells and functions as a regulatory switch that controls the relative abundance of biochemically distinct types of collagen cross-links in the tumor stroma.

Authors

Yulong Chen, Masahiko Terajima, Yanan Yang, Li Sun, Young-Ho Ahn, Daniela Pankova, Daniel S. Puperi, Takeshi Watanabe, Min P. Kim, Shanda H. Blackmon, Jaime Rodriguez, Hui Liu, Carmen Behrens, Ignacio I. Wistuba, Rosalba Minelli, Kenneth L. Scott, Johannah Sanchez-Adams, Farshid Guilak, Debananda Pati, Nishan Thilaganathan, Alan R. Burns, Chad J. Creighton, Elisabeth D. Martinez, Tomasz Zal, K. Jane Grande-Allen, Mitsuo Yamauchi, Jonathan M. Kurie

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Figure 5

LH2 expression is upregulated by STAT3 and paracrine signals from CAFs.

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LH2 expression is upregulated by STAT3 and paracrine signals from CAFs. ...
(A) HIF-1α levels in cell lysates determined by ELISA. Data from a single experiment expressed as mean ± SD of triplicate samples. (B and C) Immunoblot analysis of KC (B) and human (C) lung cancer cell lines. Actin was used as loading control. (D) Correlation of LH2 mRNA levels in Supplemental Figure 11A with pSTAT3 (Y705) levels determined densitometrically from C. P and r2 values, linear regression analysis. (E) Immunoblotting (gels) and Q-PCR analysis (bar graphs) of data from a single experiment in which human lung cancer cells were untreated (0) or treated with a Janus kinase inhibitor, P6, for 48 hours. Normalized Q-PCR values are expressed as mean of triplicate samples. (F) Immunoblotting (gels) and Q-PCR analysis (bar graph) of a single experiment in which A549 cells were stably transfected with 1 of 2 STAT3 shRNAs (SHF and SHG) or control shRNA (FF2). Q-PCR results (mean values of triplicate samples) are expressed relative to control transfectants, which were set at 100%. Actin was used as a loading control for immunoblot. (G) Immunoblotting (gels) and Q-PCR analysis (bar graphs) of A549 cells were monocultured (–) or co-cultured with human cancer-associated fibroblasts (hCAF) (+) for 24 hours in Boyden chambers. Q-PCR results are expressed as mean values of triplicate samples. Actin was used as a loading control for immunoblot. P values, 2-tailed Student’s t test unless otherwise specified.