Target mutation-driven drug discovery

In the era of precision medicine, genetics is becoming a key factor for the choice of therapies. For decades, it has been intuitively thought that variability in drug response relies, at least partly, on gene variants that may affect the pharmacokinetics and pharmacodynamics of drugs. The most considerable evidences regard polymorphisms in genes encoding enzymes and carriers involved in the ADME processes. Nowadays, more attention should be paid to genetic variations in drug targets, especially in cancer and rare diseases

Increased sodium channel use-dependent inhibition by a new potent analogue of tocainide greatly enhances in vivo antimyotonic activity

Although the sodium channel blocker, mexiletine, is the first choice drug in myotonia, some myotonic patients remain unsatisfied due to contraindications, lack of tolerability, or incomplete response. More therapeutic options are thus needed for myotonic patients, which require clinical trials based on solid preclinical data. In previous structure-activity relationship studies, we identified two newly-synthesized derivatives of tocainide, To040 and To042, with greatly enhanced potency and use-dependent behavior in inhibiting sodium currents in frog skeletal muscle fibers. The current study was performed to verify their potential as antimyotonic agents. Patch-clamp experiments show that both compounds, especially To042, are greatly more potent and use-dependent blockers of human skeletal muscle hNav1.4 channels compared to tocainide and mexiletine. Reduced effects on F1586C hNav1.4 mutant suggest that the compounds bind to the local anesthetic receptor, but that the increased hindrance and lipophilia of the N-substituent may further strengthen drug-receptor interaction and use-dependence. Compared to mexiletine, To042 was 120 times more potent to block hNav1.4 channels in a myotonia-like cellular condition and 100 times more potent to improve muscle stiffness in vivo in a previously-validated rat model of myotonia. To explore toxicological profile, To042 was tested on hERG potassium currents, motor coordination using rotarod, and C2C12 cell line for cytotoxicity. All these experiments suggest a satisfactory therapeutic index for To042. This study shows that, owing to a huge use-dependent block of sodium channels, To042 is a promising candidate drug for myotonia and possibly other membrane excitability disorders, warranting further preclinical and human studies

Soil biodegradation of nutrients enriched cellulose- and chitosan-derived mulching films for sustainable horticulture

In 2019, global plastics production reached 370 million tons, of which 58 million tons were in Europe[1]. If the plastic use in agriculture accounts for 2% of the global production[2], more than 7 million tons of plastic were used in 2019 in the agricultural sector. Mulch films represent the major source of plastic contamination in agricultural soils[3]. The agricultural surface area covered by plastic films in Europe is four times larger than that covered by greenhouses and six times that of low tunnel hoops. Over the past decades, biodegradable biopolymer mulching films (BPMFs) have been developed to reduce soil pollution by non-biodegradable plastic debris[4] and to expand the circular bioeconomy[5]. In Europe, since 1999, low density polyethylene mulches (LDPMs) have to be dismissed after their use to remove source of pollutants that can reach up to 200 kg ha-1[6] and decline soil quality, crop growth, and yield[7]. BPMFs are a sustainable alternative to conventional LDPMs. Unlike LDPMs, BPMFs, at the end of their lifetime, are tilled into soil where they are expected to be biodegraded by soil microorganisms[8]. Moreover, BPMFs show an estimated saving of about 500 kg of CO2 equivalent per hectare in comparison with LDPMs. Conversely, the impact of LDPMs in intensive horticulture could result higher than weed control by herbicides as by life cycle assessment (LCA)[9]. BPMFs can be obtained by thermo-plasticizing, solvent casting and spraying processes by using renewable and biodegradable raw materials such as starch, cellulose, chitosan, alginate, glucomannan[10] and glycerin as plasticizer[11]. Cellulose and chitosan, being the two most abundant natural biopolymers on Earth, have been proposed as the best candidates for BPMFs production. Unfortunately, the high tendency for intra- and intermolecular hydrogen bonding confers undesirable mechanical properties. The addition of plasticizer as well as fillers overcome this problem[12] modifying mechanical and functional properties of the materials. To sum up, biopolymer blending is an effective strategy to reuse cellulose and chitosan-containing by-products and develop materials with novel mechanical characteristics[13]. Moreover, the functional properties of these materials can be tuned by doping them with suitable compounds[14]. Based on what stated above, and considering that soil fertility, crop growth and yield, are generally N and P limited, the core idea of this project is the preparation of N- and P-enriched BPMFs for soil mulching, in order to slowly release soluble nutrients into soils upon their biodegradation. The latter aspect is of great importance because a proper C:N:P ratio can lead to an increase of soil-dwelling organisms thus contributing to nutrient cycling in the soil-plant system, soil C sequestration and biological fertility status[15]. Moreover, repeated additions of BPMFs over long term can increase the amount of nutrients, thus reducing the use of external inputs (e.g. synthetic fertilizers) within a circular economy perspective. The specific aim of the proposed research are: i) to set up a method for the preparation of suitable BPMFs enriched with N and P; ii) to characterize novel BPMFs and evaluate their structure, degradation kinetics, and isotopic composition iii) to assess the impact of the innovative BPMFs on soil nutrient cycling and crop growth and yield; iv) to evaluate the effect of the innovative BPMFs on soil prokaryotes and micro-arthropods communities; v) to speed-up the biodegradation of the innovative BPMFs by spraying them at the end of their lifecycle with selected microorganisms and by adding the recipient soil with earthworms; vi) to evaluate the innovative BPMFs using the LCA methodology and to investigate its role within the circular economy. Bibliography [1] Plastic Europe, 2020. Website https://www.plasticseurope.org/it/resources/publications/4312-plastics-facts-2020 accessed on 05.01.2021 [2] Vox, G., Loisi, R.V., Blanco, I., Mugnozza, G.S., & Schettini, E. (2016). Agriculture and Agricultural Science Procedia, 8, 583-591. [3] Wenqing, H., Enke, L., Qin, L., Shuang, L., Turner, N., C. & Changrong, Y. 2014. World Agriculture, 4, 3236. [4] Sanchez-Hernandez J.C., Capowiez Y. & Ro K.S., 2020. ACS Sustainable Chemistry & Engineering, 8, 4292-4316. [5] Karan, H., Funk, C., Grabert, M., Oey, M., & Hankamer, B., 2019. Trends in Plant Science, 24, 237-249. [6] Razza, F., Guerrini, S., & Impallari, F.M., 2019. Acta Horticulturae, 1252, 77-84. [7] Hou, L., Xi, J., Chen, X., Li, X., Ma, W., Lu, J., Xu J. & Lin, Y. B, 2019. Journal of Hazardous Materials, 378, 120774. [8] Kyrikou, I., & Briassoulis, D., 2007. Journal of Polymers and the Environment, 15, 125–150 [9] Tasca, A. L., Nessi, S., & Rigamonti, L., 2017. Journal of Cleaner Production, 140, 725-741 [10] Santagata, G., Malinconico, M., Immirzi, B., Schettini, E., Scarascia Mugnozza, G., & Vox, G., 2014. Acta Horticulturae 1037(1037), 921-928. [11] D’Avino, L., Rizzuto, G., Guerrini, S., Sciaccaluga, M., Pagnotta, E., & Lazzeri, L. (2015). Industrial Crops and Products, 75, 29-35. [12] Chen, P., Xie, F., Tang, F., & McNally, T. (2021). Influence of plasticiser type and nanoclay on the properties of chitosan-based materials. European Polymer Journal, 144, 110225. [13] Bajpai, A.K., Shukla, S. K., Bhanu, S., & Kankane, S., 2008. Progress in Polymer Science, 33(11), 1088-1118 [14] Sohaimy, M.I.H.A., & Isa, M.I.N.M. et al., 2020. Polymers. 12, 2487 [15] Cleveland, C.C., & Liptzin, D., 2007. Biogeochemistry 85, 235–25

Novel mutations of TCOF1 gene in European patients with treacher Collins syndrome

Background: Treacher Collins syndrome (TCS) is one of the most severe autosomal dominant congenital disorders of craniofacial development and shows variable phenotypic expression. TCS is extremely rare, occurring with an incidence of 1 in 50.000 live births. The TCS distinguishing characteristics are represented by down slanting palpebral fissures, coloboma of the eyelid, micrognathia, microtia and other deformity of the ears, hypoplastic zygomatic arches, and macrostomia. Conductive hearing loss and cleft palate are often present. TCS results from mutations in the TCOF1 gene located on chromosome 5, which encodes a serine/alanine-rich nucleolar phosphoprotein called Treacle. However, alterations in the TCOF1 gene have been implicated in only 81-93% of TCS cases. Methods: In this study, the entire coding regions of the TCOF1 gene, including newly described exons 6A and 16A, were sequenced in 46 unrelated subjects suspected of TCS clinical indication. Results: Fifteen mutations were reported, including twelve novel and three already described in 14 sporadic patients and in 3 familial cases. Moreover, seven novel polymorphisms were also described. Most of the mutations characterised were microdeletions spanning one or more nucleotides, in addition to an insertion of one nucleotide in exon 18 and a stop mutation. The deletions and the insertion described cause a premature termination of translation, resulting in a truncated protein. Conclusion: This study confirms that almost all the TCOF1 pathogenic mutations fall in the coding region and lead to an aberrant protein

Guidelines for the use and interpretation of diagnostic methods in adult food allergy

Food allergy has an increasing prevalence in the general population and in Italy concerns 8 % of people with allergies. The spectrum of its clinical manifestations ranges from mild symptoms up to potentially fatal anaphylactic shock. A number of patients can be diagnosed easily by the use of first- and second-level procedures (history, skin tests and allergen specific IgE). Patients with complex presentation, such as multiple sensitizations and pollen-food syndromes, frequently require a third-level approach including molecular diagnostics, which enables the design of a component-resolved sensitization profile for each patient. The use of such techniques involves specialists' and experts' skills on the issue to appropriately meet the diagnostic and therapeutic needs of patients. Particularly, educational programs for allergists on the use and interpretation of molecular diagnostics are needed

Effect of RNS60 in amyotrophic lateral sclerosis: a phase II multicentre, randomized, double-blind, placebo-controlled trial

Background and purpose Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease with limited treatment options. RNS60 is an immunomodulatory and neuroprotective investigational product that has shown efficacy in animal models of ALS and other neurodegenerative diseases. Its administration has been safe and well tolerated in ALS subjects in previous early phase trials. Methods This was a phase II, multicentre, randomized, double-blind, placebo-controlled, parallel-group trial. Participants diagnosed with definite, probable or probable laboratory-supported ALS were assigned to receive RNS60 or placebo administered for 24 weeks intravenously (375 ml) once a week and via nebulization (4 ml/day) on non-infusion days, followed by an additional 24 weeks off-treatment. The primary objective was to measure the effects of RNS60 treatment on selected biomarkers of inflammation and neurodegeneration in peripheral blood. Secondary objectives were to measure the effect of RNS60 on functional impairment (ALS Functional Rating Scale-Revised), a measure of self-sufficiency, respiratory function (forced vital capacity, FVC), quality of life (ALS Assessment Questionnaire-40, ALSAQ-40) and survival. Tolerability and safety were assessed. Results Seventy-four participants were assigned to RNS60 and 73 to placebo. Assessed biomarkers did not differ between arms. The mean rate of decline in FVC and the eating and drinking domain of ALSAQ-40 was slower in the RNS60 arm (FVC, difference 0.41 per week, standard error 0.16, p = 0.0101; ALSAQ-40, difference -0.19 per week, standard error 0.10, p = 0.0319). Adverse events were similar in the two arms. In a post hoc analysis, neurofilament light chain increased over time in bulbar onset placebo participants whilst remaining stable in those treated with RNS60. Conclusions The positive effects of RNS60 on selected measures of respiratory and bulbar function warrant further investigation

Clinical Features, Cardiovascular Risk Profile, and Therapeutic Trajectories of Patients with Type 2 Diabetes Candidate for Oral Semaglutide Therapy in the Italian Specialist Care

Introduction: This study aimed to address therapeutic inertia in the management of type 2 diabetes (T2D) by investigating the potential of early treatment with oral semaglutide. Methods: A cross-sectional survey was conducted between October 2021 and April 2022 among specialists treating individuals with T2D. A scientific committee designed a data collection form covering demographics, cardiovascular risk, glucose control metrics, ongoing therapies, and physician judgments on treatment appropriateness. Participants completed anonymous patient questionnaires reflecting routine clinical encounters. The preferred therapeutic regimen for each patient was also identified. Results: The analysis was conducted on 4449 patients initiating oral semaglutide. The population had a relatively short disease duration (42%  60% of patients, and more often than sitagliptin or empagliflozin. Conclusion: The study supports the potential of early implementation of oral semaglutide as a strategy to overcome therapeutic inertia and enhance T2D management

Genome-wide Analyses Identify KIF5A as a Novel ALS Gene

To identify novel genes associated with ALS, we undertook two lines of investigation. We carried out a genome-wide association study comparing 20,806 ALS cases and 59,804 controls. Independently, we performed a rare variant burden analysis comparing 1,138 index familial ALS cases and 19,494 controls. Through both approaches, we identified kinesin family member 5A (KIF5A) as a novel gene associated with ALS. Interestingly, mutations predominantly in the N-terminal motor domain of KIF5A are causative for two neurodegenerative diseases: hereditary spastic paraplegia (SPG10) and Charcot-Marie-Tooth type 2 (CMT2). In contrast, ALS-associated mutations are primarily located at the C-terminal cargo-binding tail domain and patients harboring loss-of-function mutations displayed an extended survival relative to typical ALS cases. Taken together, these results broaden the phenotype spectrum resulting from mutations in KIF5A and strengthen the role of cytoskeletal defects in the pathogenesis of ALS.Peer reviewe

Pre-Proceedings of the 4th European Workshop on "Modeling Autonomous Agents in a Multi-Agent World"

The purpose of this Workshop is to stimulate exchange and discussion of research in the field of multi-agent systems. MAAMAW is the European forum for DAI studies. While classical DAI research was mainly concerned with distributed problem solving and task allocation in view of a common goal, MAAMAW emphasizes the problems arising when several autonomous agents, endowed with their own goals, knowledge, and abilities, share a common environment and pursue either shared or competing goals. MAAMAW is therefore interested both in classical DAI problems (coordination, communication, cooperation, negotiation, etc.) and in theories of intention and action, or, more generally, in the architecture of the autonomous agent. Multi-agent models are a new area of research in rapid growth, of relevant interest both for AI and for social and management sciences. Both novel theoretical and computational approaches to multi-agent topics are encouraged. A 60 people international workshop to discuss your ideas or exhibit your systems