CEMIP ( HYBID , KIAA1199 ): structure, function and expression in health and disease

CEMIP (cell migration-inducing protein), also known as KIAA1199 or HYBID, is a protein involved in the depolymerisation of hyaluronic acid (HA), a major glycosaminoglycan component of the extracellular matrix. CEMIP was originally described in patients affected by nonsyndromic hearing loss and has subsequently been shown to play a key role in tumour initiation and progression, as well as arthritis, atherosclerosis and idiopathic pulmonary fibrosis. Despite the vast literature associating CEMIP with these diseases, its biology remains elusive. The present review article summarises all the major scientific evidence regarding its structure, function, role and expression, and attempts to cast light on a protein that modulates EMT, fibrosis and tissue inflammation, an unmet key aspect in several inflammatory disease conditions

Utilisation and effect of dried apple peels in preparation of a new dry sausage prototype

The purpose of this study was to develop a new recipe to obtain a special innovative pork sausage, to develop the manufacturing process, technological stages, and nutritional analysis of the final product. We obtained this product starting from a classic recipe (of pork sausage), it was added by adding of dry apple peels The apples are fruits with a rich content of ascorbic acid, and biominerals, and also have a high content of antioxidants. Our work has focused on evaluation of the sensory characteristics, biochemical, and nutritional characteristics a type of sausage obtained by adding of dry apple peels (sausage DAP). Sensory examination was performed by analyzing the appearance, texture and taste. The main physico-chemical features observed in the sample of sausage (simple sausage and sausages prepared with added dry apple peels) were: the content of humidity (%), ash (%), soduim chloride (%), protein (%), fat (%), carbohydrates (%) and energetic value (kcal/100g). Following the research that have been undertaken in this work, the obtained product (sausage obtained by adding of dry apple peels – sausages DAP) can be included in the category of secure products of consuming. This prototype can be considered a food variant due to its high nutritious properties and to its distinguished taste too

Stable isotope labeling and ultra-high-resolution NanoSIMS imaging reveal alpha-synuclein-induced changes in neuronal metabolism in vivo

In Parkinson’s disease, pathogenic factors such as the intraneuronal accumulation of the protein α-synuclein affect key metabolic processes. New approaches are required to understand how metabolic dysregulations cause degeneration of vulnerable subtypes of neurons in the brain. Here, we apply correlative electron microscopy and NanoSIMS isotopic imaging to map and quantify 13C enrichments in dopaminergic neurons at the subcellular level after pulse-chase administration of 13C-labeled glucose. To model a condition leading to neurodegeneration in Parkinson’s disease, human α-synuclein was unilaterally overexpressed in the substantia nigra of one brain hemisphere in rats. When comparing neurons overexpressing α-synuclein to those located in the control hemisphere, the carbon anabolism and turnover rates revealed metabolic anomalies in specific neuronal compartments and organelles. Overexpression of α-synuclein enhanced the overall carbon turnover in nigral neurons, despite a lower relative incorporation of carbon inside the nucleus. Furthermore, mitochondria and Golgi apparatus showed metabolic defects consistent with the effects of α-synuclein on inter-organellar communication. By revealing changes in the kinetics of carbon anabolism and turnover at the subcellular level, this approach can be used to explore how neurodegeneration unfolds in specific subpopulations of neurons

Exploring CEMIP axis to identify new therapeutic strategies in glomerular diseases

Glomerular diseases, also called glomerulonephritides (GNs), refer to all those conditions that affect glomeruli, the filtering units of the kidney. If left untreated or not managed effectively, progressive GNs may result in a range of complications, including chronic kidney disease and end-stage renal disease. Some pathophysiological aspects of GNs are treatable, with several novel mode of action (MoA) drugs reaching the market in recent years, but a strong unmet medical need persists and identification of novel potential GN therapeutic targets is still an active field of research in nephrology. In the present thesis work, I present evidence of the relevance of a newly identified hyaluronic acid degrading protein named Cell migration-inducing and hyaluronan-binding protein (CEMIP). CEMIP is specifically upregulated upon activation of the collagen receptor Discoidin Domain Receptor 1 (DDR1), an accepted molecular target shown to have a role in GN initiation and progression. Analysis of patient biopsies and CEMIP deletion experiment in a mouse model of GN suggested that the DDR1/CEMIP axis could play a role in renal pathologies characterized by a crescentic phase. This role may be mediated by CEMIP hyaluronidase activity, which generates low molecular weight hyaluronic acid (LMW-HA) fragments, known to be signaling molecules in inflammation. To date, very little scientific knowledge has been reported on the activity, mechanism and mode of action of these CEMIP-generated fragments. Our current grasp of CEMIP biology is dominated by the vast body of literature that has investigated CEMIP expression and/or deletion in cancer models such as in the context of tumor proliferation and invasion. After preliminary results assessed the role of CEMIP in GN, in the present work I sought to study its molecular MoA. Mutagenesis experiments confirmed the position of the key amino acid residues used by CEMIP to bind selectively to hyaluronic acid and identified, for the first time, the ones belonging to CEMIP postulated catalytic site. Additional experiments shed light on the role and importance of other domains of CEMIP as well as on the catalytic function of intracellular and secreted CEMIP. This molecular knowledge was used to design, synthesize and evaluate, using newly developed cellular assays, small molecule inhibitors on CEMIP hyaluronidase activity. A parallel effort was conducted to solve CEMIP structure by cryo-EM and one of its domains (the GG domain) by X-ray crystallography. All those efforts will hopefully result in the creation of CEMIP inhibitors active in vivo. The availability of such tool compounds will be then tested in preclinical model of GNs and cancer

Stable isotope labeling and ultra-high-resolution NanoSIMS imaging reveal alpha-synuclein-induced changes in neuronal metabolism in vivo

Abstract In Parkinson’s disease, pathogenic factors such as the intraneuronal accumulation of the protein α-synuclein affect key metabolic processes. New approaches are required to understand how metabolic dysregulations cause degeneration of vulnerable subtypes of neurons in the brain. Here, we apply correlative electron microscopy and NanoSIMS isotopic imaging to map and quantify 13C enrichments in dopaminergic neurons at the subcellular level after pulse-chase administration of 13C-labeled glucose. To model a condition leading to neurodegeneration in Parkinson’s disease, human α-synuclein was unilaterally overexpressed in the substantia nigra of one brain hemisphere in rats. When comparing neurons overexpressing α-synuclein to those located in the control hemisphere, the carbon anabolism and turnover rates revealed metabolic anomalies in specific neuronal compartments and organelles. Overexpression of α-synuclein enhanced the overall carbon turnover in nigral neurons, despite a lower relative incorporation of carbon inside the nucleus. Furthermore, mitochondria and Golgi apparatus showed metabolic defects consistent with the effects of α-synuclein on inter-organellar communication. By revealing changes in the kinetics of carbon anabolism and turnover at the subcellular level, this approach can be used to explore how neurodegeneration unfolds in specific subpopulations of neurons

Aerobic Fitness protects from Atherosclerotic Cardiovascular Risk Paralympic Athletes with a Locomotor Impairment

Aim: This study, carried out on Paralympic athletes (PA) with a locomotor impairment (LI), was aimed at: 1. assessing the prevalence of atherosclerotic cardiovascular disease (ACVD) risk factors (RF) in PA with either a spinal cord injury (PA-SCI) or other (different from SCI) LI (PA-OLI); 2. evaluating the hypothesis that aerobic fitness (oxygen uptake peak - VO2peak) was inversely related to ACVD RF. Methods: A total of 135 male PA (72 PA-SCI, 28 PA with lower limb amputation, 12 PA with a cerebral palsy/brain injury, 7 PA with poliomyelitis, 9 PA with other neurological disorders and 7 PA with other orthopedic disorders) were screened through anthropometric and blood pressure (BP) measurements, laboratory blood tests and graded cardiopulmonary maximal exercise test, to estimate both an ACVD-RF score and VO2peak. The ACVD-RF score was assessed summing 1 point for each of the following RF: obesity –OB- (BMI≥30 or waist circumference ≥102 cm), hypertension –HT- (systolic BP ≥ 140 mm Hg and diastolic BP ≥ 90 mm Hg), dyslipidemia -DL- (total Cholesterol -C- ≥200 mg·dl-1 or LDL-C ≥130 mg·dl-1 or HDL-

Allele-specific gene editing prevents deafness in a model of dominant progressive hearing loss

Since most dominant human mutations are single nucleotide substitutions(1,2), we explored gene editing strategies to disrupt dominant mutations efficiently and selectively without affecting wild-type alleles. However, single nucleotide discrimination can be difficult to achieve(3) because commonly used endonucleases, such as Streptococcus pyogenes Cas9 (SpCas9), can tolerate up to seven mismatches between guide RNA (gRNA) and target DNA. Furthermore, the protospacer-adjacent motif (PAM) in some Cas9 enzymes can tolerate mismatches with the target DNA(3,4). To circumvent these limitations, we screened 14 Cas9/gRNA combinations for specific and efficient disruption of a nucleotide substitution that causes the dominant progressive hearing loss, DFNA36. As a model for DFNA36, we used Beethoven mice(5), which harbor a point mutation in Tmc1, a gene required for hearing that encodes a pore-forming subunit of mechanosensory transduction channels in inner-ear hair cells(6). We identified a PAM variant of Staphylococcus aureus Cas9 (SaCas9-KKH) that selectively and efficiently disrupted the mutant allele, but not the wild-type Tmc1/ TMC1 allele, in Beethoven mice and in a DFNA36 human cell line. Adeno-associated virus (AAV)-mediated SaCas9-KKH delivery prevented deafness in Beethoven mice up to one year post injection. Analysis of current ClinVar entries revealed that similar to 21% of dominant human mutations could be targeted using a similar approach