Organotypic 3D decellularized matrix tumor spheroids for high-throughput drug screening

resumo

Decellularized extracellular matrix (dECM) is emerging as a valuable tool for generating 3D in vitro tumor models that better recapitulate tumor-stroma interactions. However, the development of dECM-3D heterotypic microtumors exhibiting a controlled morphology is yet to be materialized. Precisely controlling microtumors morphologic features is key to avoid an inaccurate evaluation of therapeutics performance during preclinical screening. To address this, herein we employed ultra-low adhesion surfaces for bioengineering organotypic 3D metastatic breast cancer-fibroblast models enriched with dECM microfibrillar fragments, as a bottom-up strategy to include major matrix components and their associated biomolecular cues during the early stages of 3D microtissue spheroids assembly, simulating pre-existing ECM presence in the in vivo setting. This biomimetic approach enabled the self-assembly of dECM-3D tumor-stroma spheroids with tunable size and reproducible morphology. Along time, dECM enriched and stroma-rich microtumors exhibited necrotic core formation, secretion of key biomarkers and higher cancer-cell specific resistance to different chemotherapeutics in comparison to standard spheroids. Exometabolomics profiling of dECM-Spheroid in vitro models further identified important breast cancer metabolic features including glucose/pyruvate consumption and lactate excretion, which suggest an intense glycolytic activity, recapitulating major hallmarks of the native microenvironment. Such organotypic dECM-enriched microtumors overcome the morphologic variability generally associated with cell-laden dECM models, while providing a scalable testing platform that can be foreseeable leveraged for highthroughput screening of candidate therapeutics.

palavras-chave

BREAST-CANCER PATIENTS; EXTRACELLULAR-MATRIX; IN-VITRO; ADIPOSE-TISSUE; LUNG-CANCER; CELLS; METABOLISM; BIOMATERIALS; GENERATION; RESISTANCE

categoria

Engineering, Biomedical; Materials Science, Biomaterials

autores

Ferreira, LP; Gaspar, VM; Mendes, L; Duarte, IF; Mano, JF

Grupos

G5 - Materiais Biomiméticos, Biológicos e Vivos

Projectos

CICECO - Aveiro Institute of Materials (UIDB/50011/2020)

CICECO - Aveiro Institute of Materials (UIDP/50011/2020)

Produção de Plataformas Microcapsulares 3D+ para Avaliação Laboratorial em Larga Escala de Novas Terapias Combinatórias para o Cancro do Pâncreas (PANGEIA)

Rede Nacional de Ressonância Magnética Nuclear (PTNMR)

agradecimentos

This work was developed within the scope of the project CICECO-Aveiro Institute of Materials, UIDB/50011/2020 & UIDP/50011/2020, financed by national funds through the FCT/MEC and when appropriate co-financed by FEDER under the PT2020 Partnership Agreement. This work was also supported by the Programa Operacional Competitividade e Internacionalizacao (POCI), in the component FEDER, and by national funds (OE) through FCT/MCTES, in the scope of the project PANGEIA (PTDC/BTM-SAL/30503/2017). Vitor Gaspar acknowledges funding in the form of a Junior Researcher Contract under the scope of the project PANGEIA (PTDC/BTM-SAL/30503/2017). The authors acknowledge the financial support by the Portuguese Foundation for Science and Technology (FCT) through an individual Doctoral Grant (SFRH/BD/141718/2018, Luis Ferreira). Confocal imaging acquisition was performed in the LiM Facility of iBiMED, a node of PPBI (Portuguese Platform of BioImaging): POCI-01-0145-FEDER-022122. The NMR spectrometer is part of the National NMR Network (PTNMR), partially supported by Infrastructure Project No 022161 (co-financed by FEDER through COMPETE 2020, POCI and PORL and FCT through PIDDAC).