Modeling the Effect of TNF-α upon Drug-Induced Toxicity in Human, Tissue-Engineered Myobundles

Abstract

A number of significant muscle diseases, such as cachexia, sarcopenia, systemic chronic inflammation, along with inflammatory myopathies share TNF-α-dominated inflammation in their pathogenesis. In addition, inflammatory episodes may increase susceptibility to drug toxicity. To assess the effect of TNF-α-induced inflammation on drug responses, we engineered 3D, human skeletal myobundles, chronically exposed them to TNF-α during maturation, and measured the combined response of TNF-α and the chemotherapeutic doxorubicin on muscle function. First, the myobundle inflammatory environment was characterized by assessing the effects of TNF-α on 2D human skeletal muscle cultures and 3D human myobundles. High doses of TNF-α inhibited maturation in human 2D cultures and maturation and function in 3D myobundles. Then, a tetanus force dose-response curve was constructed to characterize doxorubicin's effects on function alone. The combination of TNF-α and 10nM doxorubicin exhibited a synergistic effect on both twitch and tetanus force production. Overall, the results demonstrated that inflammation of a 3D, human skeletal muscle inflammatory system alters the response to doxorubicin.

Modeling Patient-Specific Muscular Dystrophy Phenotypes and Therapeutic Responses in Reprogrammed Myotubes Engineered on Micromolded Gelatin Hydrogels

“…Of note, however, iDRMs require less time, cost, and expertise to generate compared to iPSC-derived myoblasts, which may be especially beneficial for generating patient-specific muscle tissues in time-sensitive or resource-limited settings. Extending culture time ( Santoso and McCain, 2021 ), integrating supporting cell types ( Juhas et al, 2018 ; Santosa et al, 2018 ; Santoso and McCain, 2021 ), providing electrical ( Nedachi et al, 2008 ; Chen et al, 2021 ) or mechanical stimulation ( Heher et al, 2015 ; Chang et al, 2016 ), or engineering 3-D tissues ( Madden et al, 2015 ; Uzel et al, 2016 ; Costantini et al, 2017 ; Davis et al, 2019 ; Ariyasinghe et al, 2020 ; Ebrahimi et al, 2021 ) or earlier exposure to AO could also help induce muscle maturation and proper localization of dystrophin and α-actinin.…”

A Pound of Flesh: What Cachexia Is and What It Is Not

“…Decreased capillary density is associated with poor prognoses of cachexia and is influenced by TNFα [ 167 ]. TNFα was also shown to inhibit myofiber maturation in tissue-engineered myobundles [ 168 ].…”

Neuromuscular disease modeling on a chip

“…Over the past two decades, approaches for engineering 3D muscle bundles have been advanced and refined. Several different types of culture chambers with anchor points have been fabricated ( Costantini et al, 2017a ; Smith et al, 2016 ), including Velcro and nylon frames ( Davis et al, 2017 , 2019 ; Madden et al, 2015 ; Rao et al, 2018 ; Smith et al, 2016 ; Zhang et al, 2018 ) and microfabricated chambers with pillars ( Osaki et al, 2018 , 2020 ; Uzel et al, 2016 ). Testing of multiple ECM solutions has also revealed that fibrin hydrogels are optimal for encapsulating myotubes due to their strength ( Hinds et al, 2011 ; Pollot et al, 2018 ), although these hydrogel compositions are not necessarily physiological.…”

“…Testing of multiple ECM solutions has also revealed that fibrin hydrogels are optimal for encapsulating myotubes due to their strength ( Hinds et al, 2011 ; Pollot et al, 2018 ), although these hydrogel compositions are not necessarily physiological. To improve assay capabilities, contractile forces have been measured in 3D muscle bundles with custom force transducers ( Davis et al, 2019 ; Madden et al, 2015 ; Rao et al, 2018 ) or by tracking the displacement of pillars ( Osaki et al, 2018 ; Uzel et al, 2016 ). Similar to 2D tissues, biophysical cues, such as mechanical stretch ( Powell et al, 2002 ) and optogenetic stimulation ( Mills et al, 2019 ), or addition of fibroblasts ( Dennis et al, 2001 ) have also been shown to mature 3D muscle bundles.…”