Ciro Isidoro | Università degli Studi del Piemonte Orientale "Amedeo Avogadro" (original) (raw)

Papers by Ciro Isidoro

Journal of cancer prevention, Mar 29, 2024

Disease Markers, 2010

The lysosomal protease Cathepsin D (CD) has been implicated in the homeostasis of lymphatic tissu... more The lysosomal protease Cathepsin D (CD) has been implicated in the homeostasis of lymphatic tissues. We investigated whether the level of CD expression influences the progression and the clinical outcome in Non-Hodgkin's Lymphomas (NHLs). The expression of CD was assessed by immunohistochemistry and immunofluorescence in biopsies of Diffuse Large B Cell Lymphomas (DLBCL, 35 cases), Follicular Lymphomas (FL, 9 cases of grade I-II plus 14 cases of grade IIIB), Chronic Lymphocytic Leukaemias (CLL, 17 cases) and Peripheral T-cell Lymphomas (PTCL, 5 cases). CD staining showed a cytoplasmic punctate pattern compatible with its lysosomal localization. Based on the level of CD expression and the proportion of positive cells, lymphomas were classified as 'low expressing' (< 20% of tumor cells) or 'highly expressing' (20% of tumor cells). Lymphomas highly expressing CD were associated with a worse stage (III-IV) at diagnosis (31/34 cases; p = 0.002) and with a poor clinical outcome (i.e., partial remission and death; 28/34 cases; p = 0.03). In the subgroup of aggressive/high grade of malignancy lymphomas (i.e., DLBCL, FL IIIB and PTCL), the Kaplan-Meier curve revealed a very low cumulative overall survival probability (∼20% at 5 year) for patients bearing a NHL with > 40% CD-positive cells compared to that of patients bearing a NHL with < 20% CD-positive cells (∼70% at 5 year). This correlation was statistically significant (log-rank test, p = 0.01). In Cox multivariate analysis CD failed to be a prognosticator independent of pathologic stage, though the hazard ratio confirmed the association of low expression with a better survival probability. These data indicate that the presence of a high percentage of CD-positive tumor cells negatively reflects on the progression of NHLs.

Biomedicine & Pharmacotherapy, Mar 1, 2021

Keloids are characterized by increased deposition of fibrous tissue in the skin and subcutaneous ... more Keloids are characterized by increased deposition of fibrous tissue in the skin and subcutaneous tissue following an abnormal wound healing process. Although keloid etiology is yet to be fully understood, fibroblasts are known to be key players in its development. Here we analyze the antifibrotic mechanisms of Halofuginone (HF), a drug reportedly able to inhibit the TGF-β1-Smad3 pathway and to attenuate collagen synthesis, in an in-vitro keloid model using patient-derived Keloid Fibroblasts (KFs) isolated from fibrotic tissue collected during the "Scar Wars" clinical study (NCT NCT03312166). TGF-β1 was used as a pro-fibrotic agent to stimulate fibroblasts response under HF treatment. The fibrotic related properties of KFs, including survival, migration, proliferation, myofibroblasts conversion, ECM synthesis and remodeling, were investigated in 2D and 3D cultures. HF at 50 nM concentration impaired KFs proliferation, and decreased TGF-β1-induced expression of α-SMA and type I procollagen production. HF treatment also reduced KFs migration, prevented matrix contraction and increased the metallo-proteases/inhibitors (MMP/TIMP) ratio. Overall, HF elicits an anti-fibrotic contrasting the TGF-β1 stimulation of KFs, thus supporting its therapeutic use for keloid prevention and management.

Gli Autori presentano i risultati di un'attivita di ricerca e sviluppo finalizzata alla defin... more Gli Autori presentano i risultati di un'attivita di ricerca e sviluppo finalizzata alla definizione di un approccio innovativo alla progettazione ergonomica dei sistemi produttivi di beni e servizi (industria e terziario, settore pubblico e privato). L'attivita si e svolta nell'ambito del progetto "Modelli di gestione dell'ergonomia per la sicurezza sui luoghi di lavoro" finanziato dalla Regione Piemonte ed e stata orientata all'impostazione di un sistema informativo e formativo di riferimento in ambito territoriale. Nucleo fondante del progetto e stato l'assunto, fatto proprio dai decisori regionali, che l'Ergonomia nei luoghi di lavoro non rappresenta unicamente uno strumento per tutelare la sicurezza ed il benessere dei lavoratori, ma anche un valido strumento di innovazione e progresso dei sistemi produttivi. I principali risultati del lavoro svolto nella prima fase del progetto (conclusasi nel febbraio 2010) sono stati resi disponibili in un sito internet pubblico (www.ergonomia.corep.it). Il lavoro di gruppo ha portato, tra l'altro, alla definizione di una check-list di controllo in fase progettuale e alla realizzazione di un prototipo di strumento per l'analisi comparata delle piu diffuse metodologie di valutazione ergonomica (Toolkit)

European Journal of Neuroscience, Mar 1, 2008

The intracellular assembly of tau aggregates is a pathological hallmark shared by Alzheimer's dis... more The intracellular assembly of tau aggregates is a pathological hallmark shared by Alzheimer's disease and other neurodegenerative disorders known collectively as tauopathies. To model how tau fibrillogenesis evolves in tauopathies, we previously established transfectant M1C cultures from human neuroblastoma BE(2)-M17D cells that inducibly express human tau. In the present study, these cells were used to determine the role of the autophagic-lysosomal system in the degradation and aggregation of wild-type tau. Tau induction for 5 days led to the accumulation of tau with nominal assembly of tau aggregates within cells. When the lysosomotropic agent, chloroquine (CQ), was added following the termination of tau induction, tau clearance was delayed. Decreased tau truncation and increased levels of intact tau were observed. When present during tau induction, CQ led to tau accumulation and promoted the formation of sarkosyl-insoluble aggregates containing both truncated and full-length tau. CQ treatment significantly decreased the activities of cathepsins D, B and L, and the inhibition of cathepsins B and L mimicked the effect of CQ and increased tau levels in cells. Additionally, exposure of cells to the autophagy inhibitor, 3-methyladenine, led to tau accumulation and aggregation. These results suggest that the autophagic-lysosomal system plays a role in the clearance of tau, and that dysfunction of this system results in the formation of tau oligomers and insoluble aggregates.

Journal of Traditional and Complementary Medicine, Jul 1, 2020

The antiviral and the coronavirus-host protein pathways inhibiting properties of herbs and natura... more The antiviral and the coronavirus-host protein pathways inhibiting properties of herbs and natural compounds-Additional weapons in the fight against the COVID-19 pandemic?,

Collection of Czechoslovak Chemical Communications, Oct 16, 2014

ABSTRACT NaGdF4 nanoparticles (NPs) coated with organic ligands are known to provide efficient “p... more ABSTRACT NaGdF4 nanoparticles (NPs) coated with organic ligands are known to provide efficient “positive” contrast in magnetic resonance imaging (MRI). Strongly chelating ligands, such as ethylenediaminetetraacetic acid (EDTA) and d,l-1,2-diaminopropionic-N,N,N′,N′-tetraacetic acid (EDTACOOH), which is an EDTA derivative with a functionalisable carboxylic group, were used to coat NaGdF4. The carboxylic group was used to insert a polyethylene glycol (PEG) functionality (to give EDTAPEG) to favour better suspension and stealth and/or a fluorescent dye to obtain dual optical/MRI probes. The relaxometric behaviour of these NPs was investigated as a function of the magnetic field strength, and a significant contribution from water molecules hydrogen bonded to the organic coating was evidenced. The chemical stability of the NPs was evaluated both in a physiological medium and in the presence of a strong chelating agent. Finally, the dye-functionalised NPs were tested in ovarian carcinoma cells as dual optical and MRI probes.

Journal of Neurology, Neurosurgery & Psychiatry, 2016

Methods Hearts from two different HD murine models, the transgenic N171-82Q and the knock-in zQ17... more Methods Hearts from two different HD murine models, the transgenic N171-82Q and the knock-in zQ175, were studied by standard histological and biochemical techniques and by echocardiography. Results Disease in HD mouse models is associated with smaller gross heart mass. When tracked over time, hearts in disease animals initially gain mass appropriately, but fail to maintain the same rate of mass increase as WT littermates after symptom onset. Interestingly, evidence at the functional, tissue, and cellular levels suggests lack of heart failure. Analysis of the mTORC1 pathway revealed decreased mTORC1 activity in the hearts of HD animals that is not the result of decreased axis activation or increased energy stress. Finally, systemic treatment with an AAV9-transduced anti-HTT miRNA gene restored many molecular abnormalities observed in HD animal hearts. Conclusions These studies suggest that mutant HTT is not sufficient to cause heart failure. Additionally, mTORC1 activity may be impaired by cardiac expression of mutant HTT through as yet unknown cellular mechanisms, and may contribute to decreased heart mass. These findings have implications for the ability of the heart to respond to stress, and may help explain the mortality from heart disease in HD patients.

British Journal of Haematology, Mar 1, 2006

The cellular Fas-associated death domain-like interleukin-1bconverting enzyme (FLICE) inhibitory ... more The cellular Fas-associated death domain-like interleukin-1bconverting enzyme (FLICE) inhibitory protein (cFLIP), negatively modulates the caspase 8-dependent apoptotic cascade triggered by stimulation of death receptors, such as the tumour necrosis factor receptor (TNFR)-1 and Fas, belonging to the TNFR superfamily (Bodmer et al, 2002). Two isoforms of cFLIP, termed cFLIP S (short) and cFLIP L (long), have been described so far (Krueger et al, 2001; Thome & Tschopp, 2001). Both these isoforms bear two death effector domains (DED) downstream of the N-terminus that allow for a specific interaction with the adapter molecule Fas-associated death domain (FADD), thus preventing procaspase 8 recruitment and formation of the death-inducing signalling complex (DISC; Kischkel et al, 1995). cFLIPs and cFLIP L act, however, in different ways: while the former completely inhibits procaspase 8 activation at DISC level, the latter allows generation of the caspase 8 p43/p41 intermediates but not the processing to the active p18/p10 caspase 8 fragments (Krueger et al, 2001; Micheau et al, 2002). Upon stimulation of Fas, the cFLIP L recruited at the FADD is cleaved and a 43 kDa fragment, formed at the N-terminus, remains bound to the DISC (Scaffidi et al, 1999; Mathas et al, 2004). Disruption of death factors-mediated apoptotic cell death is believed to play a major role in the pathogenesis and tumour progression of lymphomas. Considering the regulatory function of cFLIP in this pathway, this molecule is suspected to be implicated in the carcinogenesis of lymphoid tissues. Consistently, strong cytoplasmic staining for cFLIP has been reported in tissues from classic Hodgkin lymphomas (

Modern Pathology, Jul 1, 2010

The expression of beclin-1, an oncosuppressor monoallelically deleted in 460% epithelial cancers,... more The expression of beclin-1, an oncosuppressor monoallelically deleted in 460% epithelial cancers, has been shown to be developmentally regulated in T and B lymphocytes. By interacting with either bcl-2 or class III phosphatidyl-inositol-3-phosphate kinase, beclin-1 regulates apoptosis and autophagy, two processes crucial for lymphatic tissue homeostasis. We analyzed the potential link between beclin-1-mediated autophagy and the malignant behaviour of lymphomas. The tissue expression of beclin-1 was analyzed in a large series of non-Hodgkin lymphomas and correlated with patient's clinical outcome. By immunofluorescence, beclin-1 staining showed faintly detectable and diffusely distributed in the cytoplasm (regarded as negative) or confined to the perinuclear region as large and brilliant puncta suggestive of macro-aggregate reactivity (regarded as positive). The positive expression of beclin-1 well correlated with the presence of LC3-positive autophagic vacuoles and was inversely correlated with the expression of bcl-2. Non-Hodgkin lymphomas in which X20% of tumour cells expressed high level of beclin-1 aggregates were associated with a complete (57%) or partial (35%) remission. The 5-year overall survival probability, calculated by the Kaplan-Meier method, was 92% and 42% in beclin-1expressing non-Hodgkin lymphomas with X20% and o20% positive cells, respectively (log-rank test, Po0.000.1). In Cox multivariate analysis, the level of beclin-1 expression, adjusted for patient's age and pathologic stage, revealed to be significantly correlated with patient's survival (Po0.0001). This is the first demonstration of the involvement of beclin-1 and autophagy in the clinical behaviour of non-Hodgkin lymphomas. The present data are compatible with the hypothesis that non-Hodgkin lymphomas with upregulated autophagy are more responsive to chemotherapy and indicate that beclin-1 could be a valuable independent prognostic factor in this heterogeneous group of tumours.

International Journal of Cancer, Nov 28, 2001

Neuroblastoma is the most common type of cancer in infants. In children this tumor is particularl... more Neuroblastoma is the most common type of cancer in infants. In children this tumor is particularly aggressive; despite various new therapeutic approaches, it is associated with poor prognosis. Given the importance of endosomal-lysosomal proteolysis in cellular metabolism, we hypothesized that inhibition of lysosomal protease would impact negatively on neuroblastoma cell survival. Treatment with E-64 or CA074Me (2 specific inhibitors of cathepsin B) or with pepstatin A (a specific inhibitor of cathepsin D) was cytotoxic for 2 neuroblastoma cell lines having different degrees of malignancy. Cell death was associated with condensation and fragmentation of chromatin and externalization of plasma membrane phosphatidylserine, 2 hallmarks of apoptosis. Concomitant inhibition of the caspase cascade protected neuroblastoma cells from cathepsin inhibitor-induced cytotoxicity. These data indicate that prolonged inhibition of the lysosomal proteolytic pathway is incompatible with cell survival, leading to apoptosis of neuroblastoma cells, and that the cathepsin-mediated and caspase-mediated proteolytic systems are connected and cooperate in the regulation of such an event. Since modern antitumor chemotherapy is aimed at restoring the normal rate of apoptosis in neoplastic tissues, the demonstration that endosomal-lysosomal cathepsins are involved in this process may constitute a basis for novel strategies that include cathepsin inhibitors in the therapeutic regimen.

Free Radical Biology and Medicine, May 1, 2007

Hydrogen peroxide, the major oxidoradical species in the central nervous system, has been involve... more Hydrogen peroxide, the major oxidoradical species in the central nervous system, has been involved in neuronal cell death and associated neurodegenerative diseases. In this study, we have investigated the involvement of the lysosomal pathway in the cytotoxic mechanism of hydrogen peroxide in human neuroblastoma cells. Alteration of lysosomal and mitochondrial membrane integrity was shown to be an early event in the lethal cascade triggered by oxidative stress. Desferrioxamine (DFO), an iron chelator that abolishes the formation of reactive oxygen species within lysosomes, prevented lysosome leakage, mitochondrial permeabilization and caspase-dependent apoptosis in hydrogen peroxide-treated cells. Inhibition of cathepsin D, not of cathepsin B, as well as small-interference RNA-mediated silencing of the cathepsin D gene prevented hydrogen peroxide-induced injury of mitochondria, caspase activation, and TUNEL-positive cell death. Cathepsin D activity was shown indispensable for translocation of Bax onto mitochondrial membrane associated with oxidative stress. DFO abolished both the cytosolic relocation of Cathepsin D and the mitochondrial relocation of Bax in hydrogen peroxide-treated cells. siRNA-mediated down-regulation of Bax expression protected the cells from oxidoradical injury. The present study identifies the lysosome as the primary target and the axis cathepsin D-Bax as the effective pathway of hydrogen peroxide lethal activity in neuroblastoma cells.

Carcinogenesis, Nov 28, 2007

In human colorectal DLD1 cancer cells, the dietary bioflavonoid resveratrol (RV) rapidly induced ... more In human colorectal DLD1 cancer cells, the dietary bioflavonoid resveratrol (RV) rapidly induced autophagy. This effect was reversible (on removal of the drug) and was associated with increased expression and cytosolic redistribution of the proteins Beclin1 and LC3 II. Supplementing the cells with asparagine (Asn) abrogated the Beclin-dependent autophagy. When applied acutely (2 h), RV was not toxic; however, reiterate chronic (48 h) exposure to RV eventually led to annexin V-and terminal deoxinucleotidyl transferase-mediated dUTP-biotin nick end labelingpositive cell death. This toxic effect was autophagy dependent, as it was prevented either by Asn, by expressing a dominant-negative lipid kinase-deficient class III phosphoinositide 3-phosphate kinase, or by RNA interference knockdown of Beclin1. Lamp2b silencing abolished the fusion of autophagosomes with lysosomes and preserved cell viability despite the ongoing formation of autophagosomes in cells chronically exposed to RV. The pan-caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone inhibited RV-induced cell death, but not autophagy. These results uncover a novel pathway of RV cytotoxicity in which autophagy plays a dual role: (i) at first, it acts as a prosurvival stress response and (ii) at a later time, it switches to a caspase-dependent apoptosis pathway. The present data also indicate that genetic or epigenetic inactivation of autophagy proteins in cancer cells may confer resistance to RV-mediated killing.

Carcinogenesis, Nov 27, 2006

In human colorectal cancer cells, the polyphenol resveratrol (RV) activated the caspase-dependent... more In human colorectal cancer cells, the polyphenol resveratrol (RV) activated the caspase-dependent intrinsic pathway of apoptosis. This effect was not mediated via estrogen receptors. Pepstatin A, an inhibitor of lysosomal cathepsin D (CD), not (2S,3S)-transepoxysuccinyl-L-leucylamido-3-methylbutane ethyl ester, an inhibitor of cathepsins B and L, prevented RV cytotoxicity. Similar protection was attained by small interference RNA-mediated knockdown of CD protein expression. RV promoted the accumulation of mature CD, induced lysosome leakage and increased cytosolic immunoreactivity of CD. Inhibition of CD or its posttranscriptional down-regulation precluded Bax oligomerization, permeabilization of mitochondrial membrane, cytosolic translocation of cytochrome c, caspase 3 activation and terminal deoxinucleotidyl transferase-mediated dUTP-biotin nick end labeling positivity occurring in RV-treated cells. The present study identifies the lysosome as a novel target of RV activity and demonstrates a hierarchy of the proteolytic pathways involved in its cytotoxic mechanism in which the lysosomal CD acts upstream of the cytosolic caspase activation. Our data indicate that metabolic, pharmacologic or genetic conditions affecting CD expression and/or activity could reflect on the sensitivity of cancer cells to RV.

Endocrinology, Aug 1, 2008

Cathepsin D (CD), a lysosomal aspartic protease present in mammary tissue and milk in various mol... more Cathepsin D (CD), a lysosomal aspartic protease present in mammary tissue and milk in various molecular forms, is also found in the incubation medium of mammary acini in molecular forms that are proteolytically active on prolactin at a physiological pH. Because prolactin controls the vesicular traffic in mammary cells, we studied, in vivo and in vitro, its effects on the polarized transport and secretion of various forms of CD in the rat mammary gland. CD accumulated in vesicles not involved in endocytosis in the basal region of cells. Prolactin increased this accumulation and the release of endosomal active single-chain CD at the basal side of acini. The CD-mediated proteolysis of prolactin, leading to the antiangiogenic 16-kDa form, at a physiological pH, was observed only in conditioned medium but not milk. These data support the novel concept that an active molecular form of CD, secreted at the basal side of the mammary epithelium, participates in processing blood-borne prolactin outside the cell, this polarized secretion being controlled by prolactin itself.

Bioscience Reports, Apr 1, 2013

CD (cathepsin D) is a ubiquitous lysosomal hydrolase involved in a variety of pathophysiological ... more CD (cathepsin D) is a ubiquitous lysosomal hydrolase involved in a variety of pathophysiological functions, including protein turnover, activation of pro-hormones, cell death and embryo development. CD-mediated proteolysis plays a pivotal role in tissue and organ homoeostasis. Altered expression and compartmentalization of CD have been observed in diseased muscle fibres. Whether CD is actively involved in muscle development, homoeostasis and dystrophy remains to be demonstrated. Zebrafish (Danio rerio) is emerging as a valuable 'in vivo' vertebrate model for muscular degeneration and congenital myopathies. In this work, we report on the perturbance of the somitic musculature development in zebrafish larvae caused by MPO (morpholino)-mediated silencing of CD in oocytes at the time of fertilization. Restoring CD expression, using an MPO-non-matching mutated mRNA, partially rescued the normal phenotype, confirming the indispensable role of CD in the correct development and integrity of the somitic musculature. This is the first report showing a congenital myopathy caused by CD deficiency in a vertebrate experimental animal model.

DOAJ (DOAJ: Directory of Open Access Journals), 2010

Journal of cancer prevention, Mar 29, 2024

Disease Markers, 2010

Biomedicine & Pharmacotherapy, Mar 1, 2021

European Journal of Neuroscience, Mar 1, 2008

Journal of Traditional and Complementary Medicine, Jul 1, 2020

Collection of Czechoslovak Chemical Communications, Oct 16, 2014

Journal of Neurology, Neurosurgery & Psychiatry, 2016

British Journal of Haematology, Mar 1, 2006

Modern Pathology, Jul 1, 2010

International Journal of Cancer, Nov 28, 2001

Free Radical Biology and Medicine, May 1, 2007

Carcinogenesis, Nov 28, 2007

Carcinogenesis, Nov 27, 2006

Endocrinology, Aug 1, 2008

Bioscience Reports, Apr 1, 2013

DOAJ (DOAJ: Directory of Open Access Journals), 2010

New paradigms in Cancer Science, 2025

The conference aims to discuss the latest advances in Cancer Sciences and establish collaborative... more The conference aims to discuss the latest advances in Cancer Sciences and establish collaborative networking. We invite senior scientists to join and share their knowledge with colleagues and young researchers. We encourage young researchers to take this opportunity to present their data and benefit from advice and constructive criticism in an open context.