Application of LDH assay for therapeutic efficacy evaluation of ex vivo tumor models.

Autor: Cox MC; AbbVie, 1 North Waukegan Road, North Chicago, IL, 60064-6098, USA., Mendes R; IBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901, Oeiras, Portugal.; Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Avenida da República, 2780-157, Oeiras, Portugal., Silva F; CEDOC-FCM-NOVA, Centro de Estudos de Doenças Crónicas da Faculdade de Ciências Médicas, Universidade Nova de Lisboa, R. Câmara Pestana 6, 1150-078, Lisbon, Portugal., Mendes TF; IBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901, Oeiras, Portugal.; Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Avenida da República, 2780-157, Oeiras, Portugal., Zelaya-Lazo A; AbbVie, 1 North Waukegan Road, North Chicago, IL, 60064-6098, USA., Halwachs K; AbbVie, 1 North Waukegan Road, North Chicago, IL, 60064-6098, USA., Purkal JJ; AbbVie, 1 North Waukegan Road, North Chicago, IL, 60064-6098, USA., Isidro IA; IBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901, Oeiras, Portugal.; Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Avenida da República, 2780-157, Oeiras, Portugal., Félix A; CEDOC-FCM-NOVA, Centro de Estudos de Doenças Crónicas da Faculdade de Ciências Médicas, Universidade Nova de Lisboa, R. Câmara Pestana 6, 1150-078, Lisbon, Portugal.; IPOLFG, Instituto Português de Oncologia de Lisboa Francisco Gentil, R. Prof. Lima Basto, 1099-023, Lisbon, Portugal., Boghaert ER; AbbVie, 1 North Waukegan Road, North Chicago, IL, 60064-6098, USA., Brito C; IBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901, Oeiras, Portugal. anabrito@ibet.pt.; Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Avenida da República, 2780-157, Oeiras, Portugal. anabrito@ibet.pt.; The Discoveries Centre for Regenerative and Precision Medicine, Lisbon Campus, Av. da República, 2780-157, Oeiras, Portugal. anabrito@ibet.pt.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2021 Sep 17; Vol. 11 (1), pp. 18571. Date of Electronic Publication: 2021 Sep 17.
DOI: 10.1038/s41598-021-97894-0
Abstrakt: The current standard preclinical oncology models are not able to fully recapitulate therapeutic targets and clinically relevant disease biology, evidenced by the 90% attrition rate of new therapies in clinical trials. Three-dimensional (3D) culture systems have the potential to enhance the relevance of preclinical models. However, the limitations of currently available cellular assays to accurately evaluate therapeutic efficacy in these models are hindering their widespread adoption. We assessed the compatibility of the lactate dehydrogenase (LDH) assay in 3D spheroid cultures against other commercially available readout methods. We developed a standardized protocol to apply the LDH assay to ex vivo cultures, considering the impact of culture growth dynamics. We show that accounting for growth rates and background release levels of LDH are sufficient to make the LDH assay a suitable methodology for longitudinal monitoring and endpoint assessment of therapeutic efficacy in both cell line-derived xenografts (xenospheres) and patient-derived explant cultures. This method has the added value of being non-destructive and not dependent on reagent penetration or manipulation of the parent material. The establishment of reliable readout methods for complex 3D culture systems will further the utility of these tumor models in preclinical and co-clinical drug development studies.
(© 2021. The Author(s).)
Databáze: MEDLINE
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