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3. A Novel Microfluidic Platform for Personalized Anticancer Drug Screening Through Image Analysis.

Author

Lipreri, Maria Veronica, Totaro, Marilina Tamara, Boos, Julia Alicia, Basile, Maria Sofia, Baldini, Nicola, and Avnet, Sofia

Subjects
MICROFLUIDIC devices, MEDICAL screening, CHONDROSARCOMA, ANTINEOPLASTIC agents, INDIVIDUALIZED medicine
Abstract

The advancement of personalized treatments in oncology has garnered increasing attention, particularly for rare and aggressive cancer with low survival rates like the bone tumors osteosarcoma and chondrosarcoma. This study introduces a novel PDMS–agarose microfluidic device tailored for generating patient-derived tumor spheroids and serving as a reliable tool for personalized drug screening. Using this platform in tandem with a custom imaging index, we evaluated the impact of the anticancer agent doxorubicin on spheroids from both tumor types. The device produces 20 spheroids, each around 300 µm in diameter, within a 24 h timeframe, facilitating assessments of characteristics and reproducibility. Following spheroid generation, we measured patient-derived spheroid diameters in bright-field images, calcein AM-positive areas/volume, and the binary fraction area, a metric analyzing fluorescence intensity. By employing a specially developed equation that combines viability signal extension and intensity, we observed a substantial decrease in spheroid viability of around 75% for both sarcomas at the highest dosage (10 µM). Osteosarcoma spheroids exhibited greater sensitivity to doxorubicin than chondrosarcoma spheroids within 48 h. This approach provides a reliable in vitro model for aggressive sarcomas, representing a personalized approach for drug screening that could lead to more effective cancer treatments tailored to individual patients, despite some implementation challenges. [ABSTRACT FROM AUTHOR]

Published
2024
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13. Sarcoma treatment in the era of molecular medicine

Author

Grünewald, Thomas GP, Alonso, Marta, Avnet, Sofia, Banito, Ana, Burdach, Stefan, Cidre‐Aranaz, Florencia, Di Pompo, Gemma, Distel, Martin, Dorado‐Garcia, Heathcliff, Garcia‐Castro, Javier, González‐González, Laura, Grigoriadis, Agamemnon E, Kasan, Merve, Koelsche, Christian, Krumbholz, Manuela, Lecanda, Fernando, Lemma, Silvia, Longo, Dario L, Madrigal‐Esquivel, Claudia, Morales‐Molina, Álvaro, Musa, Julian, Ohmura, Shunya, Ory, Benjamin, Pereira‐Silva, Miguel, Perut, Francesca, Rodriguez, Rene, Seeling, Carolin, Al Shaaili, Nada, Shaabani, Shabnam, Shiavone, Kristina, Sinha, Snehadri, Tomazou, Eleni M, Trautmann, Marcel, Vela, Maria, Versleijen‐Jonkers, Yvonne MH, Visgauss, Julia, Zalacain, Marta, Schober, Sebastian J, Lissat, Andrej, English, William R, Baldini, Nicola, and Heymann, Dominique

Published
2020
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15. Data from MET Overexpression Turns Human Primary Osteoblasts into Osteosarcomas

Author

Patanè, Salvatore, primary, Avnet, Sofia, primary, Coltella, Nadia, primary, Costa, Barbara, primary, Sponza, Simone, primary, Olivero, Martina, primary, Vigna, Elisa, primary, Naldini, Luigi, primary, Baldini, Nicola, primary, Ferracini, Riccardo, primary, Corso, Simona, primary, Giordano, Silvia, primary, Comoglio, Paolo M., primary, and Di Renzo, Maria Flavia, primary

Published
2023
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16. Supplementary Figure S3 from MET Overexpression Turns Human Primary Osteoblasts into Osteosarcomas

Author

Patanè, Salvatore, primary, Avnet, Sofia, primary, Coltella, Nadia, primary, Costa, Barbara, primary, Sponza, Simone, primary, Olivero, Martina, primary, Vigna, Elisa, primary, Naldini, Luigi, primary, Baldini, Nicola, primary, Ferracini, Riccardo, primary, Corso, Simona, primary, Giordano, Silvia, primary, Comoglio, Paolo M., primary, and Di Renzo, Maria Flavia, primary

Published
2023
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17. Supplementary Figures S4 & S5 from MET Overexpression Turns Human Primary Osteoblasts into Osteosarcomas

Author

Patanè, Salvatore, primary, Avnet, Sofia, primary, Coltella, Nadia, primary, Costa, Barbara, primary, Sponza, Simone, primary, Olivero, Martina, primary, Vigna, Elisa, primary, Naldini, Luigi, primary, Baldini, Nicola, primary, Ferracini, Riccardo, primary, Corso, Simona, primary, Giordano, Silvia, primary, Comoglio, Paolo M., primary, and Di Renzo, Maria Flavia, primary

Published
2023
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19. Behavioral Voluntary and Social Bioassays Enabling Identification of Complex and Sex-Dependent Pain-(-Related) Phenotypes in Rats with Bone Cancer

Author

Segelcke, Daniel, primary, Linnemann, Jan, additional, Pradier, Bruno, additional, Kronenberg, Daniel, additional, Stange, Richard, additional, Richter, S. Helene, additional, Görlich, Dennis, additional, Baldini, Nicola, additional, Di Pompo, Gemma, additional, Verri, Waldiceu A., additional, Avnet, Sofia, additional, and Pogatzki-Zahn, Esther M., additional

Published
2023
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23. IDH mutations in G2-3 conventional central bone chondrosarcoma: a mono institutional experience

Author

Setola, Elisabetta, primary, Benini, Stefania, additional, Righi, Alberto, additional, Gamberi, Gabriella, additional, Carretta, Elisa, additional, Ferrari, Cristina, additional, Avnet, Sofia, additional, Palmerini, Emanuela, additional, Magagnoli, Giovanna, additional, Gambarotti, Marco, additional, Lollini, Pier Luigi, additional, Cesari, Marilena, additional, Cocchi, Stefania, additional, Paioli, Anna, additional, Longhi, Alessandra, additional, Scotlandi, Katia, additional, Laginestra, Maria Antonella, additional, Donati, Davide Maria, additional, Baldini, Nicola, additional, and Ibrahim, Toni, additional

Published
2022
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28. Behavioural Voluntary and Social Bioassays Enabling Identification of Complex and Sex-Dependent Pain- (-Related) Phenotypes in Rats with Bone Cancer

Author

Segelcke, Daniel, primary, Linnemann, Jan, additional, Pradier, Bruno, additional, Kronenberg, Daniel, additional, Stange, Richard, additional, Richter, Helene, additional, Görlich, Dennis, additional, Baldini, Nicola, additional, Di Pompo, Gemma, additional, Verri, Waldiceu A., additional, Avnet, Sofia, additional, and Pogatzki-Zahn, Esther, additional

Published
2022
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35. Incorporation/Enrichment of 3D Bioprinted Constructs by Biomimetic Nanoparticles: Tuning Printability and Cell Behavior in Bone Models.

Author

Fischetti, Tiziana, Borciani, Giorgia, Avnet, Sofia, Rubini, Katia, Baldini, Nicola, Graziani, Gabriela, and Boanini, Elisa

Subjects
BIOPRINTING, CELL culture, DRUG discovery, BONE cells, CALCIUM phosphate, NANOPARTICLES
Abstract

Reproducing in vitro a model of the bone microenvironment is a current need. Preclinical in vitro screening, drug discovery, as well as pathophysiology studies may benefit from in vitro three-dimensional (3D) bone models, which permit high-throughput screening, low costs, and high reproducibility, overcoming the limitations of the conventional two-dimensional cell cultures. In order to obtain these models, 3D bioprinting offers new perspectives by allowing a combination of advanced techniques and inks. In this context, we propose the use of hydroxyapatite nanoparticles, assimilated to the mineral component of bone, as a route to tune the printability and the characteristics of the scaffold and to guide cell behavior. To this aim, both stoichiometric and Sr-substituted hydroxyapatite nanocrystals are used, so as to obtain different particle shapes and solubility. Our findings show that the nanoparticles have the desired shape and composition and that they can be embedded in the inks without loss of cell viability. Both Sr-containing and stoichiometric hydroxyapatite crystals permit enhancing the printing fidelity of the scaffolds in a particle-dependent fashion and control the swelling behavior and ion release of the scaffolds. Once Saos-2 cells are encapsulated in the scaffolds, high cell viability is detected until late time points, with a good cellular distribution throughout the material. We also show that even minor modifications in the hydroxyapatite particle characteristics result in a significantly different behavior of the scaffolds. This indicates that the use of calcium phosphate nanocrystals and structural ion-substitution is a promising approach to tune the behavior of 3D bioprinted constructs. [ABSTRACT FROM AUTHOR]

Published
2023
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44. The Release of Inflammatory Mediators from Acid-Stimulated Mesenchymal Stromal Cells Favours Tumour Invasiveness and Metastasis in Osteosarcoma

Author

Avnet, Sofia, primary, Lemma, Silvia, additional, Cortini, Margherita, additional, Di Pompo, Gemma, additional, Perut, Francesca, additional, Lipreri, Maria Veronica, additional, Roncuzzi, Laura, additional, Columbaro, Marta, additional, Errani, Costantino, additional, Longhi, Alessandra, additional, Zini, Nicoletta, additional, Heymann, Dominique, additional, Dominici, Massimo, additional, Grisendi, Giulia, additional, Golinelli, Giulia, additional, Consolino, Lorena, additional, Longo, Dario Livio, additional, Nanni, Cristina, additional, Righi, Alberto, additional, and Baldini, Nicola, additional

Published
2021
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47. Acid microenvironment promotes cell survival of human bone sarcoma through the activation of cIAP proteins and NF-κB pathway

Author

Avnet, Sofia, Chano, Tokuhiro, Massa, Annamaria, Bonuccelli, Gloria, Lemma, Silvia, FALZETTI, LUIGI, Grisendi, Giulia, Dominici, Massimo, Baldini, Nicola, Avnet, Sofia, Chano, Tokuhiro, Massa, Annamaria, Bonuccelli, Gloria, Lemma, Silvia, Falzetti, Luigi, Grisendi, Giulia, Dominici, Massimo, and Baldini, Nicola

Subjects
Bone sarcoma, BIRC/TRAF pathway, acid tumor microenvironment, cell survival, Original Article
Abstract

Extracellular acidification is a very common cause of stress in tumor microenvironment and of Darwinian pressure. In acid areas of the tumor, most cancer cells are-albeit slowly proliferating-more resistant to cell death than those in well-perfused regions. Tumor acidosis can directly regulate the expression of pro-survival proteins since a low extracellular pH activates the caspase-dependent cell death machinery. This mechanism has never been explored in bone sarcomas. We cultured osteosarcoma and Ewing sarcoma cells under low pH (pH 6.5), and we performed deep-sequencing and protein analysis. Both in in vitro and in vivo models, acidification activity enhanced tumor cells survival. However, we did not observe any change in ERK1 phosphorylation. On the contrary, both at the mRNA and protein level, we found a significant induction of TRAF adaptor proteins and of cIAP proteins (BIRC2 and/or BIRC3). As a consequence, the downstream nuclear transcription factor kappa B (NF-κB) survival pathway was increased. Furthermore, the treatment with the cIAP inhibitor LCL161 reverted the protection from apoptosis under low pH. In vitro results were confirmed both in Ewing sarcoma xenograft and in osteosarcoma patients, since the analysis of tumor tissues demonstrated that the levels of expression of TRAF1 or NF-κB1 significantly correlate with the level of expression of the vacuolar ATPase (V-ATPase), the most important proton pump in eukaryotes. Moreover, in the tissue sections of xenograft model, the nuclear translocation of RelB, a key subunit of the NF-κB transcriptional complex, localized in the tumor region that also corresponded to the acid microenvironment associated with the highest levels of expression of LAMP2 and V-ATPase, in the internal area of the tumor, as revealed by immunohistochemistry. Our data confirm that tumor acid microenvironment activates a stress-regulated switch to promote cell survival of bone sarcoma, and support the hypothesis that this mechanism is mediated by the recruitment of TRAF/cIAP complexes. Altogether, these results suggest that TRAF/cIAP can be considered as a target for anti-cancer therapies.

Published
2019

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