High-throughput screening in multicellular spheroids for target discovery in the tumor microenvironment

Area of Science:

• Oncology
• Biotechnology
• Drug Discovery

Background:

• Conventional 2D cell cultures inadequately model the complex tumor microenvironment (TME), leading to discrepancies between in vitro and in vivo results.
• This limitation contributes to the high failure rate in anti-cancer drug development.
• Three-dimensional multicellular tumor spheroids (MTS) offer a more accurate in vitro model, particularly for avascularized tumors.

Purpose of the Study:

• To review the application of MTS in anti-cancer drug discovery.
• To highlight high-throughput screening (HTS) compatible assays for MTS.
• To focus on target discovery within the TME using MTS-based assays.

Main Methods:

• Utilizing arrayed MTS in microtiter plates for HTS compatibility.
• Implementing multiplexed readouts to maximize data acquisition from MTS screens.
• Employing high-content screening to identify microenvironmental dependencies.

Main Results:

• MTS, when arrayed in microtiter plates, are compatible with HTS but are more resource-intensive than 2D cultures.
• High-content screening of MTS provides unique insights into microenvironmental dependencies, a key advantage over 2D screening.
• Multiplexed assays are crucial for maximizing information yield from MTS screens.

Conclusions:

• MTS represent a significant advancement for anti-cancer drug discovery and target identification within the TME.
• High-content screening of MTS offers superior value compared to 2D cultures by revealing microenvironmental factors.
• Further validation of MTS screening in animal models is essential for broader industry adoption.