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Osteosarcoma cells exhibit functional interactions with stromal cells, fostering a lung microenvironment conducive to the establishment of metastatic tumor cells

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Abstract

Background

Osteosarcoma (OS) stands out as the most common bone tumor, with approximately 20% of the patients receiving a diagnosis of metastatic OS at their initial assessment. A significant challenge lies in the frequent existence of undetected metastases during the initial diagnosis. Mesenchymal stem cells (MSCs) possess unique abilities that facilitate tumor growth, and their interaction with OS cells is crucial for metastatic spread.

Methods and results

We demonstrated that, in vitro, MSCs exhibited a heightened migration response toward the secretome of non-metastatic OS cells. When challenged to a secretome derived from lungs preloaded with OS cells, MSCs exhibited greater migration toward lungs colonized with metastatic OS cells. Moreover, in vivo, MSCs displayed preferential migratory and homing behavior toward lungs colonized by metastatic OS cells. Metastatic OS cells, in turn, demonstrated an increased migratory response to the MSCs’ secretome. This behavior was associated with heightened cathepsin D (CTSD) expression and the release of active metalloproteinase 2 (MMP2) by metastatic OS cells.

Conclusions

Our assessment focused on two complementary tumor capabilities crucial to metastatic spread, emphasizing the significance of inherent cell features. The findings underscore the pivotal role of signaling integration within the niche, with a complex interplay of migratory responses among established OS cells in the lungs, prometastatic OS cells in the primary tumor, and circulating MSCs. Pulmonary metastases continue to be a significant factor contributing to OS mortality. Understanding these mechanisms and identifying differentially expressed genes is essential for pinpointing markers and targets to manage metastatic spread and improve outcomes for patients with OS.

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Data availability

All relevant data supporting the findings of this study are available within the paper and it’s Supporting Information files.

Abbreviations

BM:

Bone marrow

CCL2:

C–C motif chemokine ligand 2

CCR2:

C–C motif chemokine receptor 2

CM:

Conditioned medium

CTSA:

Cathepsin A

CTSD:

Cathepsin D

CTSs:

Cathepsins

CXCR4:

C–X–C motif chemokine receptor 4

CXCL12:

C–X–C motif chemokine ligand 12

DAPI:

4′,6-Diamidino-2-phenylindole dihydrochloride

DiR:

1,1′-Dioctadecyl-3,3,3′,3′-tetramethylindotricarbocyanine iodide

DMEM:

Dulbecco’s modified eagle’s medium

DMEM/F12:

Dulbecco’s modified eagle’s medium/nutrient mixture F12

ECM:

Extracellular matrix

FBS:

Fetal bovine serum

FI:

Fluorescence intensity

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

GEO:

Gene expression omnibus

HMEC-1:

Human microvascular endothelial cells

IACUC:

Institutional Animal Care and Use Committee

IL-1:

Interleukin 1 alpha

i.v.:

Intravenously

MMPs:

Metalloproteinases

MMP2:

Metalloproteinase 2

MMP9:

Metalloproteinase 9

MSCs:

Mesenchymal stem/stromal cells

NIH:

National Institutes of Health

qPCR:

Real time polymerase chain reaction

PBS:

Phosphate buffer solution

PECAM:

Platelet–endothelial cell adhesion molecule 1

OS:

Osteosarcoma

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

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Funding

This work was supported in part by the Fund for Scientific and Technological Research (FONCyT) PICT No. 1974 and by Florencio Fiorini Foundation.

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Authors and Affiliations

  1. Remodeling Processes and cellular niches laboratory, Instituto de Medicina Traslacional e Ingeniería Biomédica (IMTIB)–CONICET-Hospital Italiano Buenos Aires (HIBA)-Instituto Universitario del Hospital Italiano (IUHI), 4240, C1199ACL, Potosí, CABA, Argentina

    Matías J. P. Valenzuela Alvarez, Luciana M. Gutierrez & Marcela F. Bolontrade

  2. IIMT–CONICET, Facultad de Ciencias Biomédicas, Universidad Austral, Av. Perón 1500, EPB1629AHJ, Pilar, Argentina

    Juan M. Bayo, María J. Cantero & Mariana G. Garcia

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  1. Matías J. P. Valenzuela Alvarez
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  2. Luciana M. Gutierrez
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  3. Juan M. Bayo
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  4. María J. Cantero
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  5. Mariana G. Garcia
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  6. Marcela F. Bolontrade
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Contributions

MVA and LMG performed experiments, analyzed data and designed figures. MVA also contributed to the writing of the manuscript and conceived experiments. MJC, MGG, and JB performed experiments; MGG contributed with reagents and analyzed data. MB conceived research and experiments, analyzed data and wrote manuscript. Authors discussed and commented the results on the manuscript.

Corresponding author

Correspondence to Marcela F. Bolontrade.

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All authors declare no competing financial interests.

Research involving animals right

The animal handling procedures in this study were conducted in accordance with the guidelines and regulations established by the Institutional Animal Care and Use Committee (IACUC) under the protocol number IACUC IMTIB 0001/19. Furthermore, all procedures complied with relevant national and international standards for the ethical use of animals in scientific research. Study design and execution followed the principles outlined in the Guide for the Care and Use of Laboratory Animals published by the National Institutes of Health (NIH).

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Valenzuela Alvarez, M.J.P., Gutierrez, L.M., Bayo, J.M. et al. Osteosarcoma cells exhibit functional interactions with stromal cells, fostering a lung microenvironment conducive to the establishment of metastatic tumor cells. Mol Biol Rep 51, 467 (2024). https://doi.org/10.1007/s11033-024-09315-w

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