Mantarray 3D Engineered Muscle Tissue Platform Demonstrates Clinically-relevant Disease Stratification Of An In Vitro Human Duchenne Muscular Dystrophy Model

Background & Aim Accurate in vitro modeling of healthy and disease conditions is crucial for assessing the safety and efficacy of novel therapeutics before clinical trials. For muscle diseases, direct evaluation of muscle contraction is a reliable indicator of overall tissue function, however, this often necessitates expensive and low-throughput non-human in vivo models. Animal models may not accurately predict human responses to compound exposure. Conversely, 3D engineered heart tissues (EHTs) from human induced pluripotent stem cells (iPSCs) offer promising opportunities for in vitro disease modeling. However, generating functional, reproducible, and predictive 3D human tissue models is challenging for many investigators. Methods, Results & Conclusion Here, we have developed a turnkey platform for facile fabrication of 3D muscle organoids that enables label-free, push-button measurements of contractility. The platform ensures highly reproducible tissues from virtually any cell source. Furthermore, it allows for customized stimulation protocols to individual constructs, while simultaneously measuring contractility across 24 tissues in parallel. This approach enables the stratification of healthy and diseased muscle phenotypes and facilitates compound safety and efficacy screening for evaluation of a drug's effect on contractile output.We present a 3D model of Duchenne muscular dystrophy (DMD) that utilizes EHTs formed from an isogenic pair of healthy and diseased cells. DMD tissues display functional deficits consistent with in vivo disease indicators, including reduced contractile force and slowed relaxation kinetics, though beat rate remains unchanged. EHTs remain functional over months in culture and provide a large experimental window to not only study therapeutic effect, but also disease phenotypes that may present at later stages of development and maturity. We show both acute and chronic effects of compounds in EHTs, including a drug (BMS-986094) that failed clinical trials due to unanticipated cardiotoxicity.These data demonstrate a first-and-only commercial platform that integrates individual, well-based control of electrical stimulation across a 24-well plate of tissues to pace muscle constructs, model force-frequency, and enable exercise or damage protocols. Stimulation is coupled with automated assessment of 3D muscle contraction, providing an inclusive, medium-throughput platform for studying muscle-related disorders, advancing therapeutic discovery using a modality-agnostic biosystem.