A three-dimensional bioprinted model to evaluate the effect of stiffness on neuroblastoma cell cluster dynamics and behavior

Monferrer, Ezequiel; Martín-Vañó, Susana; Carretero, Aitor; García-Lizarribar, Andrea; Burgos‐Panadero, Rebeca; Navarro, Samuel; Samitier, J.; Noguera, Rosa

doi:10.1038/s41598-020-62986-w

Cited by 44 publications

(50 citation statements)

References 60 publications

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“…The adaptation process was faster in VN-KO tumor cells but less manageable than in hydrogels. Furthermore, cells grown in rigid 3D hydrogels showed faster selection for SCAs and less presence of intratumoral heterogeneity than those cells with long and soft 3D growth, which could re ect a more e cient adaptation to niche [39] as occurs in aggressive patients tumors with rigid ECMs [58].…”

Section: Discussionmentioning

confidence: 96%

“…To get closer to con rming this hypothesis, we selected two representative NB cell lines characterized by genomic alterations typically observed in HR-NB (MNA in SK-N-BE(2) [42] and ALK mutation in SH-SY5Y) [47]. Both cell lines were grown separately in hydrogels with ECM rigidity similar to the ECM of human HR-NB and xenograft tumors [39]. To make the 3D models even more biomimetic, malignant neuroblasts were cocultured with 10% Schwann cells in some cases, since even in poor stroma NB a low proportion can be found accompanying the undifferentiated or poorly differentiated neuroblasts [51].…”

Section: Discussionmentioning

confidence: 99%

“…Composite hydrogels were synthetized using gelatin (denatured collagen) and alginate as previously described [39,40]. Polymer solutions were prepared to obtain nal concentrations of 5% w/v GelMA and 0, 0.5, 1, 1.5 and 2% of AlgMA.…”

Section: Preparation Of Gelatin-based Hydrogelsmentioning

confidence: 99%

“…We recently published a study using 3D bioprinted hydrogels composed of methacrylated gelatin (GelMA) and increasing concentrations of methacrylated alginate (AlgMA) to recreate different ECM stiffness. We described a pattern of aggressive behavior in NB cells cultivated for long times in rigid hydrogels [39]; however, this study did not analyze how scaffolding stiffness and growing time affected genomic heterogeneity. In this work the described models (xenograft VN-KO mice and 3D bioprinted hydrogels) are used to discern how tumor genomics patterns and clonal heterogeneity in two NB cell lines are affected by the VN host status, and tumor microenvironment stiffness and also to extrapolate the results in primary human HR-NB.…”

Section: Introductionmentioning

confidence: 99%

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Impact of extracellular matrix properties on neuroblastoma genomic heterogeneity

López-Carrasco¹,

Martín-Vañó²,

Burgos‐Panadero³

et al. 2020

Preprint

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Background Increased tissue stiffness is a common feature of malignant solid tumors, often associated with metastasis and poor patient outcomes. Vitronectin, as an extracellular matrix anchorage glycoprotein related to a stiff matrix, is present in a particularly increased quantity and specific distribution in high-risk neuroblastoma. Furthermore, as cells can sense and transform the proprieties of the extracellular matrix into chemical signals through mechanotransduction, genotypic changes related to stiffness are possible. Methods We have applied high density SNPa and NGS techniques to in vivo and in vitro models (orthotropic xenograft vitronectin knock-out mice and 3D bioprinted hydrogels with different stiffness) using two representative neuroblastoma cell lines (the MYCN amplified SK-N-BE(2) and the ALK mutated SH-SY5Y), to discern how tumor genomics patterns and clonal heterogeneity of both cell lines are affected. Results We describe a remarkable subclonal selection of some genomic aberrations in SK-N-BE(2) cells grown in knock-out vitronectin xenograft mice that also emerged when cultured for long times in stiff hydrogels. Specially, we detected an enlarged subclonal cell population with chromosome 9 aberrations in both models. Similar abnormalities were found in human high-risk neuroblastoma with MYCN amplification. Genomics of the SH-SY5Y cell line remained stable when cultured in both models. Conclusions Focus on heterogeneous intratumor segmental chromosome aberrations and mutations, as a mirror image of tumor microenvironment, is a vital area of future research.

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Section: Discussionmentioning

confidence: 96%

Section: Discussionmentioning

confidence: 99%

Section: Preparation Of Gelatin-based Hydrogelsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Impact of extracellular matrix properties on neuroblastoma genomic heterogeneity

López-Carrasco¹,

Martín-Vañó²,

Burgos‐Panadero³

et al. 2020

Preprint

Self Cite

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“…3D cultures are emerging as good models for ECM-related biological studies, also reducing use of animal models. These scaffolds can reproduce complex ECM features absent from traditional, 2D or monolayer cultures, and allow us to add degrees of complexity to delve deeper into the physiopathological tumor microenvironment [37,38]. We recently published a study using 3D bioprinted hydrogels composed of methacrylated gelatin (GelMA) and increasing concentrations of methacrylated alginate (AlgMA) to recreate different ECM stiffness.…”

Section: Introductionmentioning

confidence: 99%

Impact of extracellular matrix stiffness on genomic heterogeneity in MYCN-amplified neuroblastoma cell line

López-Carrasco

Martín-Vañó

Burgos‐Panadero

et al. 2020

Preprint

Self Cite

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Background: Increased tissue stiffness is a common feature of malignant solid tumors, often associated with metastasis and poor patient outcomes. Vitronectin, as an extracellular matrix anchorage glycoprotein related to a stiff matrix, is present in a particularly increased quantity and specific distribution in high-risk neuroblastoma. Furthermore, as cells can sense and transform the proprieties of the extracellular matrix into chemical signals through mechanotransduction, genotypic changes related to stiffness are possible.Methods: We applied high density SNPa and NGS techniques to in vivo and in vitro models (orthotropic xenograft vitronectin knock-out mice and 3D bioprinted hydrogels with different stiffness) using two representative neuroblastoma cell lines (the MYCN-amplified SK-N-BE(2) and the ALK -mutated SH-SY5Y), to discern how tumor genomics patterns and clonal heterogeneity of the two cell lines are affected. Results: We describe a remarkable subclonal selection of genomic aberrations in SK-N-BE(2) cells grown in knock-out vitronectin xenograft mice that also emerged when cultured for long times in stiff hydrogels. In particular, we detected an enlarged subclonal cell population with chromosome 9 aberrations in both models. Similar abnormalities were found in human high-risk neuroblastoma with MYCN amplification. The genomics of the SH-SY5Y cell line remained stable when cultured in both models. Conclusions: Focus on heterogeneous intratumor segmental chromosome aberrations and mutations, as a mirror image of tumor microenvironment, is a vital area of future research.

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Bioprinted Constructs that Mimic the Ossification Center Microenvironment for Targeted Innervation in Bone Regeneration

Miao

Liu

et al. 2021

Adv Funct Materials

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Although great progress has been made in engineered bone tissues, delayed or ineffective bone regeneration remains an issue due to the lack of neural network reconstruction in their design. Therefore, an engineered bone tissue construct that mimics the ossification center microenvironment to promote innervation is proposed. Based on this, the NGF@Lap constructs are constructed through bioprinting technology, which can release nerve growth factor (NGF) for a long time and simulate the ossification center's microenvironment with high expression NGF. In vitro, NGF@Lap‐GA can promote axonal extension. Meanwhile, the NGF and Laponite from the constructs can respectively promote the expression and secretion of calcitonin gene‐related peptide (CGRP) in sensory neurons. Further, the constructs show a CGRP‐dependent osteogenic and inhibition of adipogenesis, which is mainly regulated by AMP‐activated protein kinase‐peroxisome proliferator activated receptor pathway. In vivo, the constructs increased neurovascular network density in the tissue surrounding the implant, promoted bone marrow mesenchymal stem cells osteogenic differentiation, and effectively improved bone regeneration in the cranial defect model. In conclusion, the novel tissue‐engineered bone simulates the ossification center microenvironment, promotes innervation, and has promising potential for future application in bone regeneration.

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A three-dimensional bioprinted model to evaluate the effect of stiffness on neuroblastoma cell cluster dynamics and behavior

Cited by 44 publications

References 60 publications

Impact of extracellular matrix properties on neuroblastoma genomic heterogeneity

Impact of extracellular matrix properties on neuroblastoma genomic heterogeneity

Impact of extracellular matrix stiffness on genomic heterogeneity in MYCN-amplified neuroblastoma cell line

Bioprinted Constructs that Mimic the Ossification Center Microenvironment for Targeted Innervation in Bone Regeneration

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