The Cardiovascular Continuum describes a sequence of events from cardiovascular risk factors to end‐stage heart disease. It includes conventional pathologies affecting cardiovascular functions such ...as hypertension, atherosclerosis or thrombosis and was traditionally considered from the metabolic point of view. This Cardiovascular Continuum, originally described by Dzau and Braunwald, was extended by O'Rourke to consider also the crucial role played by degradation of elastic fibers, occurring during aging, in the appearance of vascular stiffness, another deleterious risk factor of the continuum. However, the involvement of the elastin degradation products, named elastin‐derived peptides, to the Cardiovascular Continuum progression has not been considered before. Data from our laboratory and others clearly showed that these bioactive peptides are central regulators of this continuum, thereby amplifying appearance and evolution of cardiovascular risk factors such as diabetes or hypertension, of vascular alterations such as atherothrombosis and calcification, but also nonalcoholic fatty liver disease and nonalcoholic steatohepatitis. The Elastin Receptor Complex has been shown to be a crucial actor in these processes. We propose here the participation of these elastin‐derived peptides and of the Elastin Receptor Complex in these events, and introduce a revisited Cardiovascular Continuum based on their involvement, for which elastin‐based pharmacological strategies could have a strong impact in the future.
During aging and/or related processes an intense elastin remodeling is observed, which leads to the release of bioactive elastin‐derived peptides (EDP). These peptides drive both the development of cardiovascular risk factors and pathologies that profoundly influence the course of the cardiovascular continuum. In this review, we summarize the study of the biochemistry and biology of EDP, which leads to the definition of innovative pharmacological strategies.
The incorporation of unnatural amino acids (uAAs) into protein‐based polymers has emerged as a powerful methodology to expand their chemical repertoire. Recently, we demonstrated that incorporating ...uAAs into two temperature‐responsive protein‐based polymers—namely resilin‐ and elastin‐like polypeptides (RLPs and ELPs, respectively)—can alter their properties. In this study, we incorporated aromatic uAAs into the protein sequence of RLP–ELP diblocks to yield new and diverse assemblies from a single DNA template. Specifically, we show that incorporating aromatic uAAs can modulate the phase‐transition behaviors and self‐assembly of the diblocks into various morphologies, including spherical and cylindrical micelles and single‐ and double‐layered vesicles, with some constructs also demonstrating a temperature‐responsive shape‐shifting behavior. Next, we evaluated the ability of the RLP–ELP assemblies to encapsulate a chemotherapeutic drug, doxorubicin, and show how the identity of the incorporated uAAs and the morphology of the nanostructure affect the encapsulation efficiency. Taken together, our findings demonstrate that the multi‐site incorporation of uAAs into temperature‐responsive, amphiphilic protein‐based diblock copolymers is a promising approach for the functionalization and tuning of self‐assembled nanostructures.
Abstract Hyaluronic acid (HA) is a major component of cartilage extracellular matrix and is an attractive material for use as 3D injectable matrices for cartilage regeneration. While previous studies ...have shown the promise of HA-based hydrogels to support cell-based cartilage formation, varying HA concentration generally led to simultaneous changes in both biochemical cues and stiffness. How cells respond to the change of biochemical content of HA remains largely unknown. Here we report an adaptable elastin-like protein-hyaluronic acid (ELP-HA) hydrogel platform using dynamic covalent chemistry, which allows variation of HA concentration without affecting matrix stiffness. ELP-HA hydrogels were created through dynamic hydrazone bonds via the reaction between hydrazine-modified ELP (ELP-HYD) and aldehyde-modified HA (HA-ALD). By tuning the stoichiometric ratio of aldehyde groups to hydrazine groups while maintaining ELP-HYD concentration constant, hydrogels with variable HA concentration (1.5%, 3%, or 5%) (w/v) were fabricated with comparable stiffness. To evaluate the effects of HA concentration on cell-based cartilage regeneration, chondrocytes were encapsulated within ELP-HA hydrogels with varying HA concentration. Increasing HA concentration led to a dose-dependent increase in cartilage-marker gene expression and enhanced sGAG deposition while minimizing undesirable fibrocartilage phenotype. The use of adaptable protein hydrogels formed via dynamic covalent chemistry may be broadly applicable as 3D scaffolds with decoupled niche properties to guide other desirable cell fates and tissue repair.
A major goal in materials science is to develop bioinspired functional materials based on the precise control of molecular building blocks across length scales. Here we report a protein-mediated ...mineralization process that takes advantage of disorder-order interplay using elastin-like recombinamers to program organic-inorganic interactions into hierarchically ordered mineralized structures. The materials comprise elongated apatite nanocrystals that are aligned and organized into microscopic prisms, which grow together into spherulite-like structures hundreds of micrometers in diameter that come together to fill macroscopic areas. The structures can be grown over large uneven surfaces and native tissues as acid-resistant membranes or coatings with tuneable hierarchy, stiffness, and hardness. Our study represents a potential strategy for complex materials design that may open opportunities for hard tissue repair and provide insights into the role of molecular disorder in human physiology and pathology.
Diseases bring about the need for interventions that pinpoint each specific aspect of the illness. Commonly, remission of a complex disease is accomplished by mixing treatments, medications, and ...therapeutics together in a fashion where they may negatively interact with each other or never arrive at the diseased site as a systemic heterogeneous mixture. Chronic wounds display intricacy as they are very localized and have their own environment where tissue deconstruction due to high levels of numerous proteases outweighs normal tissue reconstruction. This idea leads to the necessity of a protein that contains low diffusivity rates for localized treatment, strength against high concentrations of proteolytic species that lead to degradation of short chain peptides, while encompassing broad inhibitory effects against multiple proteases. Elastin‐like peptides are an attractive, thermoresponsive, protein‐based drug delivery partner as they contain low diffusivity and serve as a stable architecture for short chain peptide fusion. In this project, a novel elastin‐like peptide‐based protein has been created to target the inhibition of both human neutrophil elastase and matrix metalloprotease‐2. As a biologic, this is unique as it is a protein with specific biological activities against multiple proteases, ultimately displaying the potential to mix and match differing biologically active peptides within one amino acid sequence.
A new, bifunctional recombinant protein was expressed as the fusion product of human elastin‐like polypeptide (HELP) and the bilirubin‐binding protein UnaG. The engineered product displays both the ...HELP‐specific property of forming a functional hydrogel matrix and the UnaG‐specific capacity of emitting green fluorescence upon ligand binding. The new fusion protein has been proven to be effective at detecting bilirubin in complex environments with high background noise. A cell culture model of the stress response, consisting of bilirubin released in the cell culture medium, was set up to assess the bilirubin‐sensing properties of the functional matrix obtained by cross‐linking the HELP moiety. Our engineered protein allowed us to monitor cell induction by the release of bilirubin in the culture medium on a nanomolar scale. This study shows that elastin‐like protein fusion represents a versatile platform for the development of novel and commercially viable analytical and biosensing devices.
A new, bi‐functional protein was obtained by fusing the Human Elastin‐like Polypeptide (HELP) and the UnaG bilirubin‐specific binding protein. In this study, the Authors show that this new expression product formed a solid‐state 3D matrix that retained the property of binding bilirubin with high affinity even in complex multicomponent biological mixtures. Fluorescence interfering signals were removed by simply changing the soaking medium of the matrix. Nanomolar bilirubin concentrations in microwells could be detected by using a standard multiplate fluorimeter.However, if the Editor decided that it has to be shortened, we agree with that proposed in this version.
Elastin-like polypeptides (ELPs) constitute a genetically engineered class of ‘protein polymers’ derived from human tropoelastin. They exhibit a reversible phase separation whereby samples remain ...soluble below a transition temperature (Tt) but form amorphous coacervates above Tt. Their phase behavior has many possible applications in purification, sensing, activation, and nanoassembly. As humanized polypeptides, they are non-immunogenic, substrates for proteolytic biodegradation, and can be decorated with pharmacologically active peptides, proteins, and small molecules. Recombinant synthesis additionally allows precise control over ELP architecture and molecular weight, resulting in protein polymers with uniform physicochemical properties suited to the design of multifunctional biologics. As such, ELPs have been employed for various uses including as anti-cancer agents, ocular drug delivery vehicles, and protein trafficking modulators. This review aims to offer the reader a catalogue of ELPs, their various applications, and potential for commercialization across a broad spectrum of fields.
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Proteins are essential throughout the biological and biomedical sciences and the purification strategies of proteins of interest have advanced over centuries. Elastin-like polypeptides (ELPs) are ...compound polymers that have recently been highlighted for their sharp and reversible phase transition property when heated above their lower critical solution temperature (LCST). ELPs preserve this behavior when fused to a protein, and as a result providing a simple method to isolate a recombinant ELP fusion protein from cell contaminants by taking the solution through the soluble and insoluble phase of the ELP fusion protein, a technique designated as the inverse transition cycle (ITC). ITC is considered an inexpensive and efficient way of purifying recombinant ELP fusion proteins. In addition, ELPs render recombinant fusion protein more stability and a longer clear time in blood stream, which give ELPs a lot of valuable applications in the biotechnological and pharmaceutical industry. This article reviews the modernizations of ELPs and briefly highlights on the possible use of technologies such as the automatic piston discharge (APD) centrifuges to improve the efficiency of the ITC in the pharmaceutical industry to obtain benefits.
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