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

Area of Science:

• Oncology
• Biomedical Engineering
• Drug Development

Background:

• Standard preclinical oncology models fail to fully recapitulate disease biology, leading to high clinical trial attrition rates.
• Three-dimensional (3D) culture systems offer enhanced preclinical model relevance but lack robust efficacy evaluation assays.
• Current limitations in cellular assays hinder the widespread adoption of 3D culture systems in drug development.

Purpose of the Study:

• To assess the compatibility of the lactate dehydrogenase (LDH) assay in 3D spheroid cultures.
• To develop a standardized protocol for applying the LDH assay to ex vivo cultures, considering growth dynamics.
• To establish a reliable readout method for evaluating therapeutic efficacy in 3D tumor models.

Main Methods:

• Assessed lactate dehydrogenase (LDH) assay compatibility in 3D spheroid cultures against other commercial assays.
• Developed a standardized protocol for ex vivo LDH assay application, incorporating culture growth dynamics.
• Validated the LDH assay for longitudinal monitoring and endpoint assessment in xenospheres and patient-derived explants.

Main Results:

• The LDH assay is compatible with 3D spheroid cultures and provides reliable efficacy data.
• Accounting for growth rates and background LDH levels ensures assay suitability.
• The LDH assay is non-destructive and independent of reagent penetration or material manipulation.

Conclusions:

• The LDH assay, with standardized protocols, is a suitable methodology for monitoring therapeutic efficacy in 3D tumor models.
• This non-destructive method enhances the utility of 3D cultures in preclinical and co-clinical drug development.
• Reliable readout methods are crucial for advancing the use of complex 3D tumor models in drug discovery.