A synthetic mechanogenetic gene circuit for autonomous drug delivery in engineered tissues

Abstract

Mechanobiologic signals play critical roles in regulating cellular responses under both physiologic and pathologic conditions. Using a combination of synthetic biology and tissue engineering, we developed a mechanically-responsive bioartificial tissue that responds to mechanical loading to produce a pre-programmed therapeutic biologic drug. By deconstructing the signaling networks induced by activation of the mechanically-sensitive ion channel transient receptor potential vanilloid 4 (TRPV4), we created synthetic TRPV4-responsive genetic circuits in chondrocytes. These cells were then engineered into living tissues constructs that respond to mechanical compression to drive the production of the antiinflammatory biologic interleukin-1 receptor antagonist. Mechanical loading of these tissues constructs in the presence of the cytokine interleukin-1a protected constructs from inflammatory degradation. This "mechanogenetic" approach enables long-term autonomous delivery of therapeutic compounds that are driven by physiologically-relevant mechanical loading with cell-scale mechanical force resolution. The development of synthetic mechanogenetic gene circuits provides a novel approach for the autonomous regulation of cell-based drug delivery systems.

Urolithin A Protects Chondrocytes From Mechanical Overloading-Induced Injuries

“…Based on our results, these senescent hallmarks were observed in the excessive loading group, thus providing, for the first time, direct evidence of compressive overloading inducing chondrocyte senescence. Recently, the mechanically sensitive ion channel, transient receptor potential vanilloid 4 (TRPV4), was shown to mediate mechanical loading-induced inflammation, which could also participate in generating senescent phenotype (Nims et al, 2020). Of note, we did not observe a p16 level change in response to the mechanical stimuli.…”

A genome-engineered bioartificial implant for autoregulated anticytokine drug delivery

“…While protein-based drug delivery, including enzyme-activated systems, have been successful in a number of applications (23)(24)(25)(26)(27), such approaches are generally based on repeated injections or on biodegradable materials and thus cannot easily supply continuously biologic molecules for long-term delivery, unlike the rapid dynamic stimulus-response functions that living cells can provide. In previous studies, "designer" or "smart" cell approaches have been used to develop cell therapies for metabolic diseases such as diabetes but have been based on transient transfection or viral gene delivery, resulting in unpredictable gene insertion sites or copy numbers (4).…”

Transient Receptor Potential Vanilloid 4 as a Regulator of Induced Pluripotent Stem Cell Chondrogenesis

“…development. 46,47 The role of TRPV4 signaling in iPSC-derived chondrocytes mimics that seen in native articular chondrocytes; thus, iPSC chondrogenesis could provide a model system for studying the role of TRPV4 and TRPV4 channelopathies in cartilage development and pathogenesis. Controlled modulation of activity of TRPV4 evolves as a promising novel approach for optimizing stem cell differentiation in functional tissue engineering of cartilage replacements for joint repair.…”

“…Within a heterogeneous population of cells in a micromass, the enhancement of chondrogenesis appears to occur specifically in the GFP+, chondroprogenitor cell population. An improved understanding of the role of TRPV4 in chondrogenesis promises to provide new insights into the development of new therapeutic approaches for diseases of cartilage such as osteoarthritis or hereditary disorders that affect cartilage during development 46,47 . The role of TRPV4 signaling in iPSC‐derived chondrocytes mimics that seen in native articular chondrocytes; thus, iPSC chondrogenesis could provide a model system for studying the role of TRPV4 and TRPV4 channelopathies in cartilage development and pathogenesis.…”