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@ARTICLE{Zucchini:294418,
author = {C. Zucchini and M. C. Manara and C. Cristalli and M.
Carrabotta and S. Greco and R. S. Pinca and C. Ferrari and
L. Landuzzi and M. Pasello and P.-L. Lollini and M.
Gambarotti and D. M. Donati and K. Scotlandi},
title = {{ROCK}2 deprivation leads to the inhibition of tumor growth
and metastatic potential in osteosarcoma cells through the
modulation of {YAP} activity.},
journal = {Journal of experimental $\&$ clinical cancer research},
volume = {38},
number = {1},
issn = {0392-9078},
address = {London},
publisher = {BioMed Central},
reportid = {DKFZ-2024-02247},
pages = {503},
year = {2019},
abstract = {The treatment of metastatic osteosarcoma (OS) remains a
challenge for oncologists, and novel therapeutic strategies
are urgently needed. An understanding of the pathways that
regulate OS dissemination is required for the design of
novel treatment approaches. We recently identified
Rho-associated coiled-coil containing protein kinase 2
(ROCK2) as a crucial driver of OS cell migration. In this
study, we explored the impact of ROCK2 disruption on the
metastatic capabilities of OS cells and analyzed its
functional relationship with Yes-associated protein-1 (YAP),
the main transcriptional mediator of mechanotransduction
signaling.The effects of ROCK2 depletion on metastasis were
studied in NOD Scid gamma (NSG) mice injected with U-2OS
cells in which ROCK2 expression had been stably silenced.
Functional studies were performed in vitro in human U-2OS
cells and in three novel cell lines derived from
patient-derived xenografts (PDXs) by using standard methods
to evaluate malignancy parameters and signaling
transduction. The nuclear immunostaining of YAP and the
evaluation of its downstream targets Cysteine Rich
Angiogenic Inducer 6, Connective Tissue Growth Factor and
Cyclin D1 by quantitative PCR were performed to analyze YAP
activity. The effect of the expression and activity of ROCK2
and YAP on tumor progression was analyzed in 175 OS primary
tumors.The silencing of ROCK2 markedly reduced tumor growth
and completely abolished the metastatic ability of U-2OS
cells. The depletion of ROCK2, either by pharmacological
inhibition or silencing, induced a dose- and time-dependent
reduction in the nuclear expression and transcriptional
activity of YAP. The nuclear expression of YAP was observed
in 80/175 $(46\%)$ tumor samples and was significantly
correlated with worse patient prognosis and a higher
likelihood of metastasis and death. The use of verteporfin,
a molecule that specifically inhibits the TEAD-YAP
association, remarkably impaired the growth and migration of
OS cells in vitro. Moreover to inhibiting YAP activity, our
findings indicate that verteporfin also affects the ROCK2
protein and its functions.We describe the functional
connection between ROCK2 and YAP in the regulation of OS
cell migration and metastasis formation. These data provide
support for the use of verteporfin as a possible therapeutic
option to prevent OS cell dissemination.},
keywords = {Adaptor Proteins, Signal Transducing: genetics / Adaptor
Proteins, Signal Transducing: metabolism / Adolescent /
Animals / Bone Neoplasms: genetics / Bone Neoplasms:
metabolism / Bone Neoplasms: pathology / Bone Neoplasms:
therapy / Cell Line, Tumor / Cell Movement / Cell
Proliferation / Child / Disease Models, Animal / Female /
Heterografts / Humans / Immunohistochemistry / Kaplan-Meier
Estimate / Mice / Osteosarcoma: genetics / Osteosarcoma:
metabolism / Osteosarcoma: pathology / Osteosarcoma: therapy
/ Prognosis / Protein Binding / RNA Interference /
Transcription Factors: genetics / Transcription Factors:
metabolism / Verteporfin: pharmacology / YAP-Signaling
Proteins / rho-Associated Kinases: genetics / rho-Associated
Kinases: metabolism / Metastasis (Other) / Osteosarcoma
(Other) / ROCK2 (Other) / Verteporfin (Other) / YAP (Other)
/ Adaptor Proteins, Signal Transducing (NLM Chemicals) /
Transcription Factors (NLM Chemicals) / YAP-Signaling
Proteins (NLM Chemicals) / YAP1 protein, human (NLM
Chemicals) / Verteporfin (NLM Chemicals) / ROCK2 protein,
human (NLM Chemicals) / rho-Associated Kinases (NLM
Chemicals)},
ddc = {610},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:31878963},
pmc = {pmc:PMC6933701},
doi = {10.1186/s13046-019-1506-3},
url = {https://inrepo02.dkfz.de/record/294418},
}