Elsevier

Matrix Biology

Volume 83, October 2019, Pages 26-47
Matrix Biology

Tenascin-C increases lung metastasis by impacting blood vessel invasions

https://doi.org/10.1016/j.matbio.2019.07.001Get rights and content
Under a Creative Commons license
open access

Highlights

  • Host tenascin-C (TNC) encapsulates blood vessel invasions, BVI, promoting metastasis.

  • TNC increases endothelialization, tumor cell survival, platelet attraction and plasticity.

  • Through TGFbeta signaling TNC induces EMT and enhances cell migration and survival.

  • Unlike lymphatic invasions BVI in human cancer are endothelialized and express TNC.

Abstract

Metastasis is a major cause of death in cancer patients. The extracellular matrix molecule tenascin-C is a known promoter of metastasis, however the underlying mechanisms are not well understood. To further analyze the impact of tenascin-C on cancer progression we generated MMTV-NeuNT mice that develop spontaneous mammary tumors, on a tenascin-C knockout background. We also developed a syngeneic orthotopic model in which tumor cells derived from a MMTV-NeuNT tumor. Tumor cells were transfected with control shRNA or with shRNA to knockdown tenascin-C expression and, were grafted into the mammary gland of immune competent, wildtype or tenascin-C knockout mice. We show that stromal-derived tenascin-C increases metastasis by reducing apoptosis and inducing the cellular plasticity of cancer cells located in pulmonary blood vessels invasions (BVI), before extravasation. We characterized BVI as organized structures of tightly packed aggregates of proliferating tumor cells with epithelial characteristics, surrounded by Fsp1+ cells, internally located platelets and, a luminal monolayer of endothelial cells. We found extracellular matrix, in particular, tenascin-C, between the stromal cells and the tumor cell cluster. In mice lacking stromal-derived tenascin-C, the organization of pulmonary BVI was significantly affected, revealing novel functions of host-derived tenascin-C in supporting the integrity of the endothelial cell coat, increasing platelet abundance, tumor cell survival, epithelial plasticity, thereby promoting overall lung metastasis. Many effects of tenascin-C observed in BVI including enhancement of cellular plasticity, survival and migration, could be explained by activation of TGF-β signaling. Finally, in several human cancers, we also observed BVI to be surrounded by an endothelial monolayer and to express tenascin-C. Expression of tenascin-C is specific to BVI and is not observed in lymphatic vascular invasions frequent in breast cancer, which lack an endothelial lining. Given that BVI have prognostic significance for many tumor types, such as shorter cancer patient survival, increased metastasis, vessel occlusion, and organ failure, our data revealing a novel mechanism by which stromal tenascin-C promotes metastasis in human cancer, may have potential for diagnosis and therapy.

Abbreviations

BVI
blood vessel invasions
CD31
cluster of differentiation 31
CD41
cluster of differentiation 41, integrin alpha chain 2b
Cl. Cas-3
cleaved caspase-3
CK8/18
cytokeratin 8/18
DAPI
4′,6-diamidino-2-phenylindole
DMEM
Dulbecco's Modified Eagle's Medium
ELISA
Enzyme-linked immunosorbent assay
EPC
endothelial progenitor cells
EMT
epithelial-to-mesenchymal transition
ERK
extracellular signal-regulated kinases
ECM
extracellular matrix
FN
fibronectin
Fsp1
fibroblast-specific protein 1
GAPDH
Glyceraldehyde 3-phosphate dehydrogenase
GW788388
4-(4-[3-(Pyridin-2-yl)-1H-pyrazol-4-yl]pyridin-2-yl)-N-(tetrahydro-2Hpyran-4-yl) benzamide
HCC
hepatocellular carcinoma
HE
Hematoxylin and eosin
IDC NST
invasive ductal carcinoma, no special type
KD
knock down
KO
knock out
LM
laminin
LVI
lymph vessel invasions
MaCa
mammary carcinomas
Mmp9
Matrix metallopeptidase 9
MMTV-NeuNT
mouse mammary tumor virus driven NeuNT (activated rat ErbB2 homologue) transformed
NOS
not otherwise specified
NT193
MMTV-NeuNT breast tumor derived cell line
Pai-1
plasminogen activator inhibitor-1
PBS
phosphate-buffered saline
PDAC
pancreatic ductal adenocarcinomas
PNET
pancreatic neuroendocrine tumors
PyMT
polyoma middle T antigen
RAM1
reduced Arbuscular Mycorrhization 1
RCC
renal cell carcinoma
Sh
short hairpin
Slug
snail family zinc finger 2
Snail
snail family zinc finger 1
STS
staurosporine
TGF-β
transforming growth factor β
TME
tumor microenvironment
TNBC
triple negative breast cancer
TNC
tenascin-C
VEGFA
vascular endothelial growth factor A
WT
wild type
Zeb1
zinc finger E-box-binding homeobox 1
α-SMA
alpha-smooth muscle actin

Keywords

Blood vessel invasions
Circulating tumor cells
Tumor emboli
Tenascin-C
Cellular plasticity
TGF-β signaling
Lung metastasis
Endothelialization
Fsp1+ cells
Endothelial cells

Cited by (0)

1

Current address: 1. Tongji Cancer Research Institute, Tongji Hospital, Tongji Medical College in Huazhong University of Science and Technology, Wuhan, Hubei, China. 2. Department of Gastrointestinal Surgery, Tongji Hospital, Tongji Medical College in Huazhong University of Science and Technology, Wuhan, Hubei, China.

2

Equal contribution.