Engineered red blood cells (RBCs) appear to be a promising method for therapeutic drug and protein delivery. With a number of agents in clinical trials (e.g., dexamethasone 21-phosphate in ataxia telangiectasia, asparaginase in pancreatic cancer/acute lymphoblastic leukemia, thymidine phosphorylase in mitochondrial neurogastrointestinal encephalomyopathy, RTX-134 in phenylketonuria, etc.), this leading article summarizes the ongoing efforts in developing these agents, focuses on the clinical progress, and provides a brief background into engineered RBCs and the different ways in which they can be exploited for therapeutic/diagnostic purposes. References to available data on safety, efficacy, and tolerability are reported. Due to the continuous progress in this field, the information is updated as of January 2020 from databases, websites, and press releases of the involved companies and information that is in the public domain.

Background: The current COVID-19 pandemic has demonstrated the enormous importance of maintaining good hygienic conditions in everyday indoor environments for the prevention of infectious diseases. This includes sanitization methods capable of significantly reducing the microbial load in the air and on surfaces. However, in line with the ecological transition, alternative systems for environmental sanitization with reduced environmental impact are urgently needed. The photocatalytic reaction using UV-C light-emitting diode (UV-C LED) lamps with short wavelengths, especially in the range of 200–280 nanometers (nm), can significantly reduce the microbial load, safeguarding the environment thanks to reduced energy consumption. The objective of this review is to describe the latest innovations in the use of UV-C LED technology in the sanitization of indoor environments, reporting the fundamental principles on which its activity relies. Methods: Two databases (PubMed, Web of Science), were searched, following PRISMA guidelines. Results: A total of 1348 publications were identified, of which 379 were assessed in detail and, of these, 16 were included in the review. Conclusions: This literature review highlighted that UV-C LEDs irradiation represents a valid, eco-sustainable sanitization method that could be exploited as an alternative to chemical compounds to contain indoor microbiological pollution in living and working environments.

Improving indoor air quality present in environments where people live is important to protect human health. This particularly applies to public transportation, where air quality may affect the health and safety of passengers, workers and staff. To provide better air quality, many buildings and transports are provided with heating, ventilation and air conditioning (HVAC) systems, which are always equipped with filters to retain the particulate present in the airflow, but they lack continuous air sanitization systems. In this study, a new UV-C LED and ionizer-based continuous sanitation air (CSA) system to be installed in a train HVAC was developed (international patent: N.PCT/IB2021/054194) and its sanitation efficacy against various microbial species (bacteria and fungi) was assessed. The device proved to be very effective at the microbial killing of aerodispersed microorganisms, both in its experimental configuration (ISO 15714:2019) and in a train setting. The installation of this CSA system on public transportation appears to be a promising solution to guarantee high microbiological air quality with a very low environmental impact due to its eco-friendly components.

The beneficial effects that physical activity gives to our bodies are well known, but they could be compromised by reduced indoor air quality (IAQ) in confined sports environments. Within the latter, indoor air often may host high levels of biological pollutants (bacteria, viruses and moulds). The use of UV-C light-emitting diode (UV-C LED) technology can significantly reduce the airborne microbial load, giving many advantages both on a hygienic and ecological level thanks to their reduced environmental impact. In this study, a new eco-friendly technology, named ‘CSA System technology’ (international patent: US20220268458A1), was integrated into an innovative continuous sanitation air (CSA) device. A prototype called CSA System S14000 was installed in a university sport pressostatic structure, and its sanitation efficacy and efficiency were assessed. The prototype demonstrated high efficacy in eliminating airborne microorganisms, both in efficacy tests (inactivation rate around 99.92 ± 0.07%, log reduction of 3.22 ± 0.39) and in on-site environmental tests (inactivation rate around 70.7 ± 3.6% and 72.4 ± 3.3% for airborne bacteria and moulds, respectively). Based on the results, the installation of the tested device in indoor sports environments appears to be a promising solution to guarantee efficient and healthy sanitation for athletes during sporting activity.

Here we report, for the first time, the engineering of human red blood cells (RBCs) with an entire metabolic pathway as a potential strategy to treat patients with guanidinoacetate methyltransferase (GAMT) deficiency, capable of reducing the high toxic levels of guanidinoacetate acid (GAA) and restoring proper creatine levels in blood and tissues. We first produced a recombinant form of native human GAMT without any tags to encapsulate into RBCs. Due to the poor solubility and stability features of the recombinant enzyme, both bioinformatics studies and extensive optimization work were performed to select a mutant GAMT enzyme, where only four critical residues were replaced, as a lead candidate. However, GAMT-loaded RBCs were ineffective in GAA consumption and creatine production because of the limiting intra-erythrocytic S-adenosyl methionine (SAM) content unable to support GAMT activity. Therefore, a recombinant form of human methionine adenosyl transferase (MAT) was developed. RBCs co-entrapped with both GAMT and MAT enzymes performed, in vitro, as a competent cellular bioreactor to remove GAA and produce creatine, fueled by physiological concentrations of methionine and the ATP generated by glycolysis. Our results highlight that metabolic engineering of RBCs is possible and represents proof of concept for the design of novel therapeutic approaches.

Most of the ATM variants associated with Ataxia Telangiectasia are still classified as variants with uncertain significance. Ataxia Telangiectasia is a multisystemic disorder characterized by “typical” and “atypical” phenotypes, with early-onset and severe symptoms or with late-onset and mild symptoms, respectively. Here we classified the c.7157C > A ATM variant found in homozygosity in two brothers of Lebanese ethnicity. The brothers presented with an atypical phenotype, showing less than 50% of the positive criteria considered for classification. We performed several in silico analyses to predict the effect of c.7157C > A at the DNA, mRNA and protein levels, revealing that the alteration causes a missense substitution in a highly conserved alpha helix in the FAT domain. 3D structural analyses suggested that the variant might be pathogenic due to either loss of activity or to a structural damage affecting protein stability. Our subsequentin vitrostudies showed that the second hypothesis is the most likely, as indicated by the reduced protein abundance found in the cells carrying the variant. Moreover, two different functional assays showed that the mutant protein partially retains its kinase activity. Finally, we investigated thein vitroeffect of Dexamethasone showing that the drug is able to increase both protein abundance and activity. In conclusion, our results suggest that the c.7157C > A variant is pathogenic, although it causes an atypical phenotype, and that dexamethasone could be therapeutically effective on this and possibly other missense ATM variants.