Intracellular delivery of therapeutic proteins. New advancements and future directions

Achieving the full potential of therapeutic proteins to access and target intracellular receptors will have enormous benefits in advancing human health and fighting disease. Existing strategies for intracellular protein delivery, such as chemical modification and nanocarrier-based protein delivery approaches, have shown promise but with limited efficiency and safety concerns. The development of more effective and versatile delivery tools is crucial for the safe and effective use of protein drugs. Nanosystems that can trigger endocytosis and endosomal disruption, or directly deliver proteins into the cytosol, are essential for successful therapeutic effects. This article aims to provide a brief overview of the current methods for intracellular protein delivery to mammalian cells, highlighting current challenges, new developments, and future research opportunities

Selective Protein Conjugation of Poly(glycerol monomethacrylate) and Poly(polyethylene glycol methacrylate) with Tunable Topology via Reductive Amination with Multifunctional ATRP Initiators for Activity Preservation

In this study, we compare poly(glycerol monomethacrylate) (PGMA) of different chain lengths and architectures (linear and two-arm) with poly(poly(ethylene glycol) methyl ether methacrylate) (PPEGMA) as an alternative polymer platform for the synthesis of a new generation of protein-polymer conjugates. Mono-and two-arm functional atom-transfer radical polymerization (ATRP) initiators were designed and selectively attached to lysozyme at the N-terminus via reductive amination. Site-specific, grafting from activator regenerated by electron transfer (ARGET) ATRP was carried out in phosphate buffer, and the reaction parameters were optimized to obtain polymer conjugates with predetermined molar mass and topology. The activity preservation under proteolytic and high-temperature conditions showed a clear dependence on the structure of the repeating unit and on the macromolecular architecture. These results highlighted the potential of PGMA as a poly(ethylene glycol) (PEG) alternative for the half-life extension of biotherapeutics. Moreover, this synthetic approach may inspire the design of a new class of protein-polymer conjugates through an optimal combination of macromolecular composition and topology

Synthesis and Characterization of Thermally and Chemically Gelling Injectable Hydrogels for Tissue Engineering

Novel, injectable hydrogels were developed that solidify through a dual-gelation, physical and chemical, mechanism upon preparation and elevation of temperature to 37°C. A thermogelling, poly(N-isopropylacrylamide)-based macromer with pendant epoxy rings and a hydrolyticallydegradable polyamidoamine-based diamine crosslinker were synthesized, characterized, and combined to produce in situ forming hydrogel constructs. Network formation through the epoxyamine reaction was shown to be rapid and facile, and the progressive incorporation of the hydrophilic polyamidoamine crosslinker into the hydrogel was shown to mitigate the often problematic tendency of thermogelling materials to undergo significant post-formation gel syneresis. The results suggest that this novel class of injectable hydrogels may be attractive substrates for tissue engineering applications due to the synthetic versatility of the component materials and beneficial hydrogel gelation kinetics and stability

A multi-element psychosocial intervention for early psychosis (GET UP PIANO TRIAL) conducted in a catchment area of 10 million inhabitants: study protocol for a pragmatic cluster randomized controlled trial

Multi-element interventions for first-episode psychosis (FEP) are promising, but have mostly been conducted in non-epidemiologically representative samples, thereby raising the risk of underestimating the complexities involved in treating FEP in 'real-world' services