115 research outputs found
The first-in-class alkylating deacetylase inhibitor molecule tinostamustine shows antitumor effects and is synergistic with radiotherapy in preclinical models of glioblastoma
Background: The use of alkylating agents such as temozolomide in association with radiotherapy (RT) is the
therapeutic standard of glioblastoma (GBM). This regimen modestly prolongs overall survival, also if, in light of
the still dismal prognosis, further improvements are desperately needed, especially in the patients with O6-
methylguanine-DNA-methyltransferase (MGMT) unmethylated tumors, in which the benefit of standard treatment
is less. Tinostamustine (EDO-S101) is a first-in-class alkylating deacetylase inhibitor (AK-DACi) molecule that fuses
the DNA damaging effect of bendamustine with the fully functional pan-histone deacetylase (HDAC) inhibitor,
vorinostat, in a completely new chemical entity.
Methods: Tinostamustine has been tested in models of GBM by using 13 GBM cell lines and seven patient-derived
GBM proliferating/stem cell lines in vitro. U87MG and U251MG (MGMT negative), as well as T98G (MGMT positive),
were subcutaneously injected in nude mice, whereas luciferase positive U251MG cells and patient-derived GBM stem
cell line (CSCs-5) were evaluated the orthotopic intra-brain in vivo experiments.
Results: We demonstrated that tinostamustine possesses stronger antiproliferative and pro-apoptotic effects than
those observed for vorinostat and bendamustine alone and similar to their combination and irrespective of MGMT
expression. In addition, we observed a stronger radio-sensitization of single treatment and temozolomide used as
control due to reduced expression and increased time of disappearance of ÎłH2AX indicative of reduced signal and
DNA repair. This was associated with higher caspase-3 activation and reduction of RT-mediated autophagy. In vivo,
tinostamustine increased time-to-progression (TTP) and this was additive/synergistic to RT. Tinostamustine had
significant therapeutic activity with suppression of tumor growth and prolongation of DFS (disease-free survival) and
OS (overall survival) in orthotopic intra-brain models that was superior to bendamustine, RT and temozolomide and
showing stronger radio sensitivity.
Conclusions: Our data suggest that tinostamustine deserves further investigation in patients with glioblastoma
The small molecule ephrin receptor inhibitor, GLPG1790, Reduces renewal capabilities of cancer stem cells, showing anti-tumour efficacy on preclinical glioblastoma models
Therapies against glioblastoma (GBM) show a high percentage of failure associated with the survival of glioma stem cells (GSCs) that repopulate treated tumours. Forced differentiation of GSCs is a promising new approach in cancer treatment. Erythropoietin-producing hepatocellular (Eph) receptors drive tumourigenicity and stemness in GBM. We tested GLPG1790, a first small molecule with inhibition activity versus inhibitor of various Eph receptor kinases, in preclinical GBM models using in vitro and in vivo assays. GLPG1790 rapidly and persistently inhibited Ephrin-A1-mediated phosphorylation of Tyr588 and Ser897, completely blocking EphA2 receptor signalling. Similarly, this compound blocks the ephrin B2-mediated EphA3 and EphB4 tyrosine phosphorylation. This resulted in anti-glioma effects. GLPG1790 down-modulated the expression of mesenchymal markers CD44, Sox2, nestin, octamer-binding transcription factor 3/4 (Oct3/4), Nanog, CD90, and CD105, and up-regulated that of glial fibrillary acidic protein (GFAP) and pro-neural/neuronal markers, ÎČIII tubulin, and neurofilaments. GLPG1790 reduced tumour growth in vivo. These effects were larger compared to radiation therapy (RT; U251 and T98G xenografts) and smaller than those of temozolomide (TMZ; U251 and U87MG cell models). By contrast, GLPG1790 showed effects that were higher than Radiotherapy (RT) and similar to Temozolomide (TMZ) in orthotopic U87MG and CSCs-5 models in terms of disease-free survival (DFS) and overall survival (OS). Further experiments were necessary to study possible interactions with radio- and chemotherapy. GLPG1790 demonstrated anti-tumor effects regulating both the differentiative status of Glioma Initiating Cells (GICs) and the quality of tumor microenvironment, translating into efficacy in aggressive GBM mouse models. Significant common molecular targets to radio and chemo therapy supported the combination use of GLPG1790 in ameliorative antiglioma therapy
CXCR1/2 pathways in paclitaxel-induced neuropathic pain
Chemotherapy-induced peripheral neuropathy (CIPN) is a type of neuropathic pain that represents a frequent and serious consequence of chemotherapy agents. Over the last years, significant progress has been achieved in elucidating the underlying pathogenesis of CIPN. The interference of taxanes with microtubule has been proposed as a mechanism that leads to altered axonal transport and to permanent neurological damages. The inflammatory process activated by chemotherapeutic agents has been considered as a potential trigger of nociceptive process in CIPN.In this study we investigated the effect of reparixin, an inhibitor of CXCR1/CXCR2, in suppressing the development of paclitaxel-induced nociception in rats. Moreover, reparixin activity in reversing the neurotoxic effects induced by paclitaxel or GRO/KC in F11 cells was also analyzed.Reparixin administered by continuous infusion ameliorated paclitaxel-induced mechanical and cold allodynia in rats. In F11 cells, reparixin was able to inhibit the increase of acetyladed α-tubulin induced both by paclitaxel and GRO/KC. The subsequent experiments were performed in order to dissect the signal transduction pathways under GRO/KC control, eventually modulated by paclitaxel and/or reparixin. To this aim we found that reparixin significantly counteracted p-FAK, p-JAK2/p-STAT3, and PI3K-p-cortactin activation induced either by paclitaxel or GRO/KC.Overall the present results have identified IL-8/CXCR1/2 pathway as a mechanism involved in paclitaxel-induced peripheral neuropathy. In particular, the obtained data suggest that the inhibition of CXCR1/2 combined with standard taxane therapy, in addition to potentiating the taxane anti-tumor activity can reduce chemotherapy-induced neurotoxicity, thus giving some insight for the development of novel treatments
Heavy metals in follicular fluid affect the ultrastructure of the human mature cumulus-oocyte complex
It is known that exposure to heavy metals such as lead (Pb) and cadmium (Cd) has several
adverse effects, particularly on the human reproductive system. Pb and Cd have been associated
with infertility in both men and women. In pregnant women, they have been associated with
spontaneous abortion, preterm birth, and impairment of the development of the fetus. Since these
heavy metals come from both natural and anthropogenic activities and their harmful effects have
been observed even at low levels of exposure, exposure to them remains a public health issue,
especially for the reproductive system. Given this, the present study aimed to investigate the
potential reproductive effects of Pb and Cd levels in the follicular fluid (FF) of infertile women and
non-smokers exposed to heavy metals for professional reasons or as a result of living in rural areas
near landfills and waste disposal areas in order to correlate the intrafollicular presence of these
metals with possible alterations in the ultrastructure of human cumulus-oocyte complexes (COCs),
which are probably responsible for infertility. Blood and FF metals were measured using atomic
Citation: Miglietta, S.; Cristiano, L.;
Battaglione, E.; Macchiarelli, G.;
Nottola, S.A.; De Marco, M.P.;
Costanzi, F.; Schimberni, M.;
Colacurci, N.; Caserta, D.; Familiari,
G. Heavy Metals in Follicular Fluid
Affect the Ultrastructure of the
Human Mature Cumulus-Oocyte
Complex. Cells 2023, 12, 2577.
https://doi.org/10.3390/cells12212577
Academic Editor: Elena Llano
Received: 11 October 2023
Revised: 30 October 2023
Accepted: 31 October 2023
Published: 5 November 2023
Copyright: © 2023 by the authors.
Licensee MDPI, Basel, Switzerland.
This article is an open access article
distributed under the terms and
conditions of the Creative Commons
Attribution (CC BY) license
(https://creativecommons.org/license
s/by/4.0/).
Cells 2023, 12, 2577 2 of 24
absorption spectrometry. COCs corresponding to each FF analyzed were subjected to ultrastructural
analyses using transmission electron microscopy. We demonstrated for the first time that
intrafollicular levels of Pb (0.66 ”g/dLâ0.85 ”g/dL) and Cd (0.26 ”g/Lâ0.41 ”g/L) could be associated
with morphological alterations of both the oocyte and cumulus cellsâ (CCs) ultrastructure. Since
blood Cd levels (0.54 ”g/Lâ1.87 ”g/L) were above the current reference values established by the
guidelines of the Agency for Toxic Substances and Disease Registry (ATSDR) and the Environmental
Protection Agency (EPA) (0.4 ”g/L), whereas blood Pb levels (1.28 ”g/dLâ3.98 ”g/dL) were below
the ATSDR reference values (â€5 ”g/dL), we believe that these alterations could be due especially to
Cd, even if we cannot exclude a possible additional effect of Pb. Our results highlighted that oocytes
were affected in maturation and quality, whereas CCs showed scarcely active steroidogenic
elements. Regressing CCs, with cytoplasmic alterations, were also numerous. According to Cdâs
endocrine-disrupting activity, the poor steroidogenic activity of CCs might correlate with delayed
oocyte cytoplasmic maturation. So, we conclude that levels of heavy metals in the blood and the FF
might negatively affect fertilization, embryo development, and pregnancy, compromising oocyte
competence in fertilization both directly and indirectly, impairing CC steroidogenic activity, and
inducing CC apoptosis
Adult IDH wild-type glioblastoma ultrastructural investigation suggests a possible correlation between morphological biomarkers and Ki-67 index
Glioblastoma is an aggressive brain tumor with an average life expectancy between 14 and 16 months after diagnosis. The Ki-67 labeling index (LI), a measure of cellular proliferation, is emerging as a prognostic marker in GBM. In this study, we investigated the ultrastructure of glioblastoma tissue from 9 patients with the same molecular profile (adult IDH wild-type glioblastoma, wild-type ATRX, and positive for TP53 expression, GFAP expression, and EGFR overexpression) to find possible ultrastructural features to be used as biomarkers and correlated with the only parameter that differs among our samples, the Ki-67 LI. Our main results were the visualization of the anatomical basis of astrocyte-endothelial cells crosstalk; the ultrastructural in situ imaging of clusters of hyperactivated microglia cells (MsEVs); the ultrastructural in situ imaging of microglia cells storing lipid vesicles (MsLVs); the ultrastructural in situ imaging of neoplastic cells mitophagy (NCsM). The statistical analysis of our data indicated that MsEVs and MsLVs correlate with the Ki-67 LI value. We can thus assume they are good candidates to be considered morphological biomarkers correlating to Ki-67 LI. The role of NCsM instead must be further evaluated. Our study findings demonstrate that by combining ultrastructural characteristics with molecular information, we can discover biomarkers that have the potential to enhance diagnostic precision, aid in treatment decision-making, identify targets for therapy, and enable personalized treatment plans tailored to each patient. However, further research with larger sample sizes is needed to validate these findings and fully utilize the potential of ultrastructural analysis in managing glioblastoma
Effects of simulated microgravity In vitro on human metaphase II oocytes: an electron microscopy-based study
The Gravity Force to which living beings are subjected on Earth rules the functionality
of most biological processes in many tissues. It has been reported that a situation of Microgravity
(such as that occurring in space) causes negative effects on living beings. Astronauts returning from
space shuttle missions or from the International Space Station have been diagnosed with various
health problems, such as bone demineralization, muscle atrophy, cardiovascular deconditioning, and
vestibular and sensory imbalance, including impaired visual acuity, altered metabolic and nutritional
status, and immune system dysregulation. Microgravity has profound effects also on reproductive
functions. Female astronauts, in fact, suppress their cycles during space travels, and effects at the
cellular level in the early embryo development and on female gamete maturation have also been
observed. The opportunities to use space flights to study the effects of gravity variations are limited
because of the high costs and lack of repeatability of the experiments. For these reasons, the use
of microgravity simulators for studying, at the cellular level, the effects, such as those, obtained
during/after a spatial trip, are developed to confirm that these models can be used in the study of
body responses under conditions different from those found in a unitary Gravity environment (1 g).
In view of this, this study aimed to investigate in vitro the effects of simulated microgravity on the
ultrastructural features of human metaphase II oocytes using a Random Positioning Machine (RPM).
We demonstrated for the first time, by Transmission Electron Microscopy analysis, that microgravity
might compromise oocyte quality by affecting not only the localization of mitochondria and cortical
granules due to a possible alteration of the cytoskeleton but also the function of mitochondria and
endoplasmic reticulum since in RPM oocytes we observed a switch in the morphology of smooth
endoplasmic reticulum (SER) and associated mitochondria from mitochondria-SER aggregates to
mitochondriaâvesicle complexes. We concluded that microgravity might negatively affect oocyte
quality by interfering in vitro with the normal sequence of morphodynamic events essential for
acquiring and maintaining a proper competence to fertilization in human oocyte
Age-dependent roles of peroxisomes in the hippocampus of a transgenic mouse model of Alzheimerâs disease
BACKGROUND: Alzheimer's Disease (AD) is a progressive neurodegenerative disease, especially affecting the hippocampus. Impairment of cognitive and memory functions is associated with amyloid beta-peptide-induced oxidative stress and alterations in lipid metabolism. In this scenario, the dual role of peroxisomes in producing and removing ROS, and their function in fatty acids beta-oxidation, may be critical. This work aims to investigating the possible involvement of peroxisomes in AD onset and progression, as studied in a transgenic mouse model, harboring the human Swedish familial AD mutation. We therefore characterized the peroxisomal population in the hippocampus, focusing on early, advanced, and late stages of the disease (3, 6, 9, 12, 18 months of age). Several peroxisome-related markers in transgenic and wild-type hippocampal formation were comparatively studied, by a combined molecular/immunohistochemical/ultrastructural approach.
RESULTS:
Our results demonstrate early and significant peroxisomal modifications in AD mice, compared to wild-type. Indeed, the peroxisomal membrane protein of 70 kDa and acyl-CoA oxidase 1 are induced at 3 months, possibly reflecting the need for efficient fatty acid beta-oxidation, as a compensatory response to mitochondrial dysfunction. The concomitant presence of oxidative damage markers and the altered expression of antioxidant enzymes argue for early oxidative stress in AD. During physiological and pathological brain aging, important changes in the expression of peroxisome-related proteins, also correlating with ongoing gliosis, occur in the hippocampus. These age- and genotype-based alterations, strongly dependent on the specific marker considered, indicate metabolic and/or numerical remodeling of peroxisomal population.
CONCLUSIONS:
Overall, our data support functional and biogenetic relationships linking peroxisomes to mitochondria and suggest peroxisomal proteins as biomarkers/therapeutic targets in pre-symptomatic AD
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