Biology subjects

Huerta-Lopez, C.

Publications and source records attributed to Huerta-Lopez, C..

3 recordsLinked to original sources

bioRxiv2025

Tuning viscoelasticity of dynamic covalent hydrogels for human tissue modeling

The development of three-dimensional (3D) in vitro tissue culture models is critical for biomedical research. Hydrogel-based systems have become a preferred scaffold for 3D models, as they have tunable viscoelastic properties, which are well-known to influence cell morphology and function. In particular, reversible hydrogel crosslinks formed through dynamic covalent chemistry (DCC) can introduce viscoelastic behavior including stress relaxation. However, traditional strategies to increase stress-relaxation rates in DCC gels rely on faster bond kinetics, resulting in faster erosion rates that prevent their use for long-term 3D culture. As an alternative strategy, we explore the use of molecular parameters (specifically molecular weight and degree of functionalization) to independently control the stiffness and stress relaxation behavior while preventing rapid erosion. As demonstration, we develop and validate a modified theoretical model of gel viscoelasticity applied to a two-component DCC gel composed of modified hyaluronic acid and elastin-like protein. Finally, we utilize this tunable gel platform to explore the impact of scaffold viscoelasticity on encapsulated human neural progenitor cells. In summary, this work expands the molecular design space of DCC hydrogels to achieve tunable viscoelastic properties for 3D in vitro models.

bioengineering↗

bioRxiv2025

Titin cleavage in living cardiomyocytes induces sarcomere disassembly but does not trigger cell proliferation

AimsAdult mammalian hearts have limited regenerative capacity due to the inability of cardiomyocytes to proliferate, a major clinical hurdle in contemporary cardiology. The presence of highly organized, contractile sarcomeres has long been considered an impediment for cardiomyocyte division. Indeed, sarcomere disassembly is a crucial step to complete the cell cycle in the few situations where cardiomyocytes have been observed to proliferate. However, whether sarcomere disassembly can per se trigger cell cycle re-entry remains unknown, a possibility that we have tested here. Methods and resultsWe have engineered a system to induce sarcomere disassembly in living murine cardiomyocytes based on the specific cleavage of the structural protein titin by tobacco etch virus protease (TEVp). Although isolated neonatal cardiomyocytes with disassembled sarcomeres remain viable and retain low-amplitude contractile activity, our results show no evidence of increased cardiomyocyte proliferation in targeted cells, as indicated by analyses of markers of DNA synthesis and cytokinesis. We obtain equivalent results when titin is cleaved in the adult myocardium in vivo. ConclusionThe removal of sarcomere structural barriers is necessary, but not sufficient, for cardiomyocyte proliferation, which implies that additional factors are required for cardiomyocytes to undergo cell division. Translational perspectiveThere is a clinical need to identify therapeutic strategies that promote cardiac regeneration through the proliferation of cardiomyocytes that survive an injury to the heart, for instance after myocardial infarction. Based on the observation that cardiomyocytes require sarcomere disassembly for proliferation, we have examined if the sole disassembly of sarcomeres is enough to promote cell division in cardiomyocytes. Our work demonstrates a strategy to induce specific sarcomere disassembly, which, however does not result in increased proliferative capacity of cardiomyocytes. These results imply that additional factors need to be considered to promote cardiomyocyte proliferation by facilitating sarcomere disassembly.

cell biology↗

bioRxiv2022

Cell response to extracellular matrix energy dissipation outweighs rigidity sensing

The mechanical properties of the extracellular matrix (ECM) determine cell differentiation, proliferation and migration through mechanoresponsive proteins including YAP. However, how different mechanical signals cooperate, synergize or compete to steer cell behavior remains poorly understood. Here, we have examined competition between the two major ECM mechanical cues, i.e. rigidity, which activates cell mechanosensing, and viscous energy dissipation, which reduces stiffness blunting cell mechanotransduction. To trigger competition, we have engineered protein hydrogels allowing concomitant modulation of stiffness and viscosity by mechanisms characteristic of native ECM. Culturing cells on these hydrogels, we have found that substrate energy dissipation attenuates YAP mechanosensing prevailing over stiffness cues. Hampered YAP activation on more dissipative substrates correlates with faster actin flow and smaller focal adhesions. Mechanistically, inhibition of actomyosin contractility reverses the outcome of the competition between rigidity and energy dissipation. Our results highlight the dominating contribution of substrate viscosity to the biology of the cell.

bioengineering↗