Spastin and alsin protein interactome analyses begin to reveal key canonical pathways and suggest novel druggable targets

Developing effective and long-term treatment strategies for rare and complex neurodegenerative diseases is challenging. One of the major roadblocks is the extensive heterogeneity among patients. This hinders understanding the underlying disease-causing mechanisms and building solutions that have implications for a broad spectrum of patients. One potential solution is to develop personalized medicine approaches based on strategies that target the most prevalent cellular events that are perturbed in patients. Especially in patients with a known genetic mutation, it may be possible to understand how these mutations contribute to problems that lead to neurodegeneration. Protein–protein interaction analyses offer great advantages for revealing how proteins interact, which cellular events are primarily involved in these interactions, and how they become affected when key genes are mutated in patients. This line of investigation also suggests novel druggable targets for patients with different mutations. Here, we focus on alsin and spastin, two proteins that are identified as “causative” for amyotrophic lateral sclerosis and hereditary spastic paraplegia, respectively, when mutated. Our review analyzes the protein interactome for alsin and spastin, the canonical pathways that are primarily important for each protein domain, as well as compounds that are either Food and Drug Administration–approved or are in active clinical trials concerning the affected cellular pathways. This line of research begins to pave the way for personalized medicine approaches that are desperately needed for rare neurodegenerative diseases that are complex and heterogeneous.

Yeast knockout library allows for efficient testing of genomic mutations for cell-free protein synthesis

Cell-free protein synthesis(CFPS)systems from crude lysates have benefitted from modifications to their enzyme composition.For example,functionally deleting enzymes in the source strain that are deleterious to CFPS can improve protein synthesis yields.However,making such modifications can take substantial time.As a proof-of-concept to accelerate prototyping capabilities,we assessed the feasibility of using the yeast knockout collection to identify negative effectors in a Saccharomyces cerevisiae CFPS platform.We analyzed extracts made from six deletion strains that targeted the single deletion of potentially negative effectors(e.g.,nucleases).We found a statistically significant increase in luciferase yields upon loss of function of GCN3,PEP4,PPT1,NGL3,and XRN1 with a maximum increase of over 6-fold as compared to the wild type.Our work has implications for yeast CFPS and for rapidly prototyping strains to enable cell-free synthetic biology applications.

Personalized composite scaffolds for accelerated cell-and growth factor-free craniofacial bone regeneration

Approaches to regenerating bone often rely on integrating biomaterials and biological signals in the form of cells or cytokines.However,from a translational point of view,these approaches are challenging due to the sourcing and quality of the biologic,unpredictable immune responses,complex regulatory paths,and high costs.We describe a simple manufacturing process and a material-centric 3D-printed composite scaffold system(CSS)that offers distinct advantages for clinical translation.The CSS comprises a 3D-printed porous polydiolcitrate-hydroxyapatite composite elastomer infused with a polydiolcitrate-graphene oxide hydrogel composite.Using a micro-continuous liquid interface production 3D printer,we fabricate a precise porous ceramic scaffold with 60 wt%hydroxyapatite resembling natural bone.The resulting scaffold integrates with a thermoresponsive hydrogel composite in situ to fit the defect,which is expected to enhance surface contact with surrounding tissue and facilitate biointegration.The antioxidative properties of citrate polymers prevent long-term inflammatory responses.The CSS stimulates osteogenesis in vitro and in vivo.Within 4 weeks in a calvarial critical-sized bone defect model,the CSS accelerated ECM deposition(8-fold)and mineralized osteoid(69-fold)compared to the untreated.Through spatial transcriptomics,we demonstrated the comprehensive biological processes of CSS for prompt osseointegration.Our material-centric approach delivers impressive osteogenic properties and streamlined manufacturing advantages,potentially expediting clinical application for bone reconstruction surgeries.

Peroxidase-Like Reactivity at Iron-Chelation Sites in a Mesoporous Synthetic Melanin

High catalytic activity and substrate specificity make enzymes a rich source of inspiration for catalyst development.Co-opting the advantages of natural materials while tuning them to a modified form and purpose,however,is not a straightforward synthetic task.Polymerization of L-3,4-dihydroxyphenylalanine(L-DOPA)results in amorphous polymer nanoparticles that are similar in many ways to natural eumelanin.Herein,the authors introduce mesoporosity and iron ion chelation to synthesize a variant of the L-DOPA polymer with high peroxidase-like activity.Our results indicate catalytic reaction with peroxide under mildly acidic conditions(pH 5.4 and 6)with a greater maximum reaction velocity(Vmax)than horseradish peroxidase(HRP)at optimal pH 3.5–4.5.Comparison between Fe(Ⅲ)and Fe(Ⅱ)loading indicates that either can be used as a starting point to trigger reactivity,though Fe(Ⅱ)loading leads to materials with twice the Vmax of the Fe(Ⅲ)-loaded sample.The lack of catalyst degradation despite the redox changes and presence of radical species is consistent with the robust nature and redox versatility of polydopamine-based materials and demonstrates strong potential as a versatile redox-catalysis platform.