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Loss of one Tgfbr2 allele in fibroblasts promotes metastasis in MMTV: polyoma middle T transgenic and transplant mouse models of mammary tumor progression

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Abstract

Accumulation of fibroblasts is a phenomenon that significantly correlates with formation of aggressive cancers. While studies have shown that the TGF-β signaling pathway is an important regulator of fibroblast activation, the functional contribution of TGF-β signaling in fibroblasts during multi-step tumor progression remains largely unclear. In previous studies, we used a sub-renal capsule transplantation model to demonstrate that homozygous knockout of the Tgfbr2 gene (Tgbr2FspKO) enhanced mammary tumor growth and metastasis. Here, we show for the first time a significant role for loss of one Tgfbr2 allele during multi-step mammary tumor progression. Heterozygous deletion of Tgfbr2 in stromal cells in MMTV–PyVmT transgenic mice (PyVmT/Tgfbr2hetFspKO mice) resulted in earlier tumor formation and increased stromal cell accumulation. In contrast to previous studies of Tgbr2FspKO fibroblasts, Tgfbr2hetFspKO fibroblasts did not significantly increase tumor growth, but enhanced lung metastasis in PyVmT transgenic mice and in co-transplantation studies with PyVmT mammary carcinoma cells. Furthermore, Tgfbr2hetFspKO fibroblasts enhanced mammary carcinoma cell invasiveness associated with expression of inflammatory cytokines including CXCL12 and CCL2. Analyses of Tgbr2FspKO and Tgfbr2hetFspKO fibroblasts revealed differences in the expression of factors associated with metastatic spread, indicating potential differences in the mechanism of action between homozygous and heterozygous deletion of Tgfbr2 in stromal cells. In summary, these studies demonstrate for the first time that loss of one Tgfbr2 allele in fibroblasts enhances mammary metastases in a multi-step model of tumor progression, and demonstrate the importance of clarifying the functional contribution of genetic alterations in stromal cells in breast cancer progression.

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Abbreviations

CCL2:

Chemokine (C–C motif) ligand 2

CXCL12:

Chemokine (C–X–C motif) ligand 12

Fsp1:

Fibroblast specific protein 1

HGF:

Hepatocyte growth factor

MMTV:

Mouse mammary tumor virus promoter

PyVmT:

Polyoma virus middle T

α-sma:

Alpha smooth muscle actin

TGF-β:

Transforming growth factor beta

Tgfbr2 :

TGF-beta type II receptor gene

Tgfbr2FspKO :

Homozygous deletion of Tgfbr2 in stromal cells

Tgfbr2hetFspKO :

Heterozygous deletion of Tgfbr2 in stromal cells

TUNEL:

Terminal deoxynucleotidyl transferase dUTP nick end labeling

VWF8:

Von Willebrand factor 8

WT:

Wildtype

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Acknowledgments

We thank Stacey Hembruff for technical assistance, Fang Fan, M.D (University of Kansas Medical Center), Osama Tawfik, M.D (University of Kansas Medical Center), Roy Jensen, M.D (University of Kansas Medical Center), and Rebecca Cook, Ph.D (Vanderbilt University, Nashville, TN) for scientific discussion. This work was supported by the following grants: CA102162 and CA85492 (National Cancer Institute DHHS), and the TJ Martell Foundation to HL Moses, CA127357-01A2 (National Cancer Institute DHHS) and University of Kansas Endowment to N Cheng.

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Authors and Affiliations

  1. Department of Pathology and Laboratory Medicine, University of Kansas Cancer Center, University of Kansas Medical Center, Lied 3001, 3901 Rainbow Blvd, Kansas City, KS, 66160, USA

    Wei Bin Fang, Iman Jokar & Nikki Cheng

  2. Department of Cancer Biology, Vanderbilt-Ingram Cancer Center, Nashville, TN, 33232, USA

    Anna Chytil & Harold L. Moses

  3. Department of Pathology, Vanderbilt University Medical Center, Nashville, TN, 33232, USA

    Ty Abel

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  1. Wei Bin Fang
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Correspondence to Nikki Cheng.

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Fang, W.B., Jokar, I., Chytil, A. et al. Loss of one Tgfbr2 allele in fibroblasts promotes metastasis in MMTV: polyoma middle T transgenic and transplant mouse models of mammary tumor progression. Clin Exp Metastasis 28, 351–366 (2011). https://doi.org/10.1007/s10585-011-9373-0

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