Biology subjects

Gentili, C.

Publications and source records attributed to Gentili, C..

9 recordsLinked to original sources

bioRxiv2026

PTHrP drives aggressive traits in colorectal cancer cells: Implications of tumor-stromal cells

In the tumor microenvironment (TME), dynamic interactions between cells and soluble factors promote tumor progression. We previously demonstrated that parathyroid hormone-related peptide (PTHrP), a TME-associated cytokine, enhances the aggressive phenotype of HCT116 colorectal cancer (CRC) cells, and that conditioned medium from PTHrP-treated HMEC-1 endothelial stromal cells (CM) induces epithelial-to-mesenchymal transition (EMT) in CRC cells. Here, Western blot analysis showed that CM modulates Met receptor expression and activation and promotes cancer stem cell (CSC) traits in HCT116 cells. Since PTHrP induces CPT-11 chemoresistance through Met signaling, we investigated the involvement of the CM-Met axis in this process. Viability assays revealed that CM increases cell number and confers CPT11 resistance through Met activation. Transforming growth factor beta 1 (TGF{beta}1), upregulated in PTHrP-treated HMEC-1 cells, was evaluated as a potential mediator. Its neutralization reversed the CM-induced increase in cell number but did not affect chemoresistance. In silico analyses revealed differences between CRC and normal tissues related to TGF{beta}1 signaling and Met activation, along with positive correlations among the analyzed markers. Immunohistochemical observation of human samples is consistent with our previous findings. Overall, these findings support a role for PTHrP in promoting CRC aggressiveness through coordinated effects on tumor and stromal compartments

cancer biology↗

bioRxiv2026

In Silico study of clinical implication of markers associated with PTHrP regulatory mechanisms and linked to angiogenesis and EMT program of colorectal cancer

Parathyroid hormone-related peptide (PTHrP), encoded by PTHLH, has been implicated in tumor progression through its involvement in epithelial-mesenchymal transition (EMT), angiogenesis, and tumor cell migration. Previous experimental studies suggest that PTHrP may promote these processes in colorectal cancer (CRC), partly through the modulation of factors such as secreted protein acidic and rich in cysteine (SPARC) and vascular endothelial growth factor (VEGFA). These events play a key role in the acquisition of an aggressive phenotype in our experimental models. In this study, we performed an integrative in silico analysis of multiple transcriptomic datasets to investigate the potential role of PTHLH in CRC. Differential expression analysis identified a set of consistently dysregulated genes across independent datasets. Functional enrichment and network analyses revealed that PTHLH expression is associated with biological processes related to extracellular matrix remodeling, EMT, and angiogenesis. Correlation analyses showed a positive association between PTHLH and SPARC expression, while network-based approaches suggested a potential functional connection with VEGFA. To assess the clinical relevance of these findings, survival analysis was performed using publicly available datasets. High expression levels of PTHLH, SPARC, and VEGFA were significantly associated with reduced overall survival in patients. Notably, a combined gene signature based on these three factors demonstrated a stronger prognostic effect than individual genes, indicating enhanced predictive value. These findings suggest that PTHrP is associated with molecular pathways involved in tumor progression and, together with SPARC and VEGF, may contribute to a coordinated regulatory axis with prognostic relevance in CRC, warranting further experimental validation.

cancer biology↗

bioRxiv2025

Therapeutic efficacy of hiMSC-derived Extracellular Vesicles from Serum-containing and Xeno-free media for osteoarthritis treatment

BackgroundExtracellular vesicles derived from human induced mesenchymal stromal cells (hiEVs) constitute a promising cell-free therapeutic option for osteoarthritis. To allow translation to the clinic we evaluated the therapeutic effects of hiEVs for osteoarthritis treatment. Specifically, we assessed the efficacy of hiEVs collected from serum-containing and serum-free, PurStem (PS), media in an OA mice model. MethodshiEVs were administered with or without hydrogel via intra-articular (i.a.) injection in a destabilization of the medial meniscus (DMM) mouse model. Fluorescence imaging was used to monitor the retention of IR780-labeled hiEVs in the joint cavity. The therapeutic effects were evaluated by analyzing damage scores as well as catabolic and anabolic markers, including Mmp13 and Col2 expression, in joint tissues. ResultsFluorescence imaging confirmed that hiEVs remained localized at the injection site without systemic migration. HiEVs demonstrated significant protective effects against joint tissue degeneration in the DMM mouse OA model, as evidenced by reduced damage scores, decreased Mmp13 expression, and increased anabolic processes (Col2 expression). The hydrogel alone also exerted beneficial therapeutic effects, including reduced damage scores, increased Col2 expression, and reduced Mmp13 levels; however, these effects were notably smaller than those achieved with hiEV treatment while it was independent of the medium used for hiEV collection. ConclusionsTogether, our findings demonstrate that hiEVs from xeno-free conditions effectively prevent cartilage degradation and promote its repair. This paves the way for future clinical translation of hiEV-based therapies as a safe, scalable, and effective approach to treat osteoarthritis.

developmental biology↗

bioRxiv2024

Impaired functional brain-heart interplay sustains emotion dysregulation in depressed individuals

Depression is a leading worldwide cause of mental disorders and disability, strongly affecting emotional processing and regulation. Its dysfunctional psycho-physiological dynamics may be part of the a nervous-system-wise symptomatology, impacting not only patients psyche but also significantly influencing functional cardiovascular dynamics. Therefore, depression serves as an exemplary pathological manifestation of the dysfunctional interaction between the central and autonomic nervous systems. While recent literature has been developing specific techniques to quantify such interactions, often referred to as functional Brain-Heart Interplay (BHI), the quantitative role of BHI dynamics in depression is largely unknown. This study aims to experimentally unveil BHI patterns specific to emotional regulation and processing in subjects exhibiting depressive symptoms compared to healthy controls. Results were gathered from a cohort of 72 individuals and indicate that depressive symptoms are associated with a continuous efferent central-to-peripheral hyperactivity and an afferent peripheral-to-central hypoactivity. This hypoactivity appears to be specific to negative emotional processing. This study offers novel insights into the systemic investigation of the neuro-physiological bases of depression.

neuroscience↗

bioRxiv2021

Cardiac sympathovagal activity initiates a functional brain-body response to emotional processing

A century-long debate on bodily states and emotions persists. While the involvement of bodily activity in emotion physiology is widely recognized, the specificity and causal role of such activity related to brain dynamics has not yet been demonstrated. We hypothesize that the peripheral neural monitoring and control of cardiovascular activity prompts and sustains brain dynamics during an emotional experience, so these afferent inputs are processed by the brain by triggering a concurrent efferent information transfer to the body. To this end, we investigated the functional brain-heart interplay under emotion elicitation in publicly available data from 62 healthy participants using a computational model based on synthetic data generation of EEG and ECG signals. Our findings show that sympathovagal activity plays a leading and causal role in initiating the emotional response, in which ascending modulations from vagal activity precede neural dynamics and correlate to the reported level of arousal. The subsequent dynamic interplay observed between the central and autonomic nervous systems sustains emotional processing. These findings should be particularly revealing for the psychophysiology and neuroscience of emotions. SignificanceWe investigate the temporal dynamics of brain and cardiac activities in healthy subjects who underwent an emotional elicitation through videos. We demonstrate that, within the first few seconds, emotional stimuli modulate the heart activity, which in turn stimulate an emotion-specific cortical response in the brain. Then, the conscious emotional experience is sustained by a bidirectional brain-heart interplay and information exchange. Moreover, the perceived intensity of an emotional stimulus is predicted by the intensity of neural control regulating the heart activity. These findings may constitute the fundamental knowledge linking neurophysiology and psychiatric disorders, including the link between depressive symptoms and cardiovascular disorders.

neuroscience↗

bioRxiv2020

Brain activity during facial processing in autism spectrum disorder: an activation likelihood estimation (ALE) meta-analysis of neuroimaging studies

BackgroundThough aberrant face processing is a hallmark of autistic spectrum disorder (ASD), findings on accompanying brain activity are divergent. Therefore, we conducted an activation likelihood estimation (ALE) meta-analysis of studies examining brain activity during face processing. MethodsWe searched PubMed and PsycINFO using combinations of terms as fMRI, Autism Spectrum Disorder, Face Perception. Eligible studies reported on DSM-diagnosed ASD patients, compared to controls (HC), using face stimuli presented in fMRI and reporting whole-brain analysis coordinates. We compared two approaches: "convergence of differences" (primary analysis) using study-level coordinates from ASD vs. HC contrasts, and "differences in convergence" (secondary) pooling coordinates within each group separately, and contrasting the resultant ALE-maps. ResultsThirty-five studies (655 ASD and 668 HC) were included. Primary analysis identified a cluster in amygdala/parahippocampus where HC showed greater convergence of activation. Secondary analysis yielded no significant results. ConclusionsResults suggest that ASD dysfunction in face processing relies on structures involved in emotional processing rather than perception. We also demonstrate that the two ALE methodologies lead to divergent results.

neuroscience↗

bioRxiv2019

The Human Exonuclease-1 Interactome And Phosphorylation Sites

Error-free repair of DNA double-strand break is orchestrated by homologous recombination (HR) pathways and requires the concerted action of several factors. Among these, EXO1 and DNA2/BLM execute extensive resection of DNA ends to produce 3-overhangs, which are key intermediates for downstream steps of HR. To help shedding light on regulatory aspects of DNA repair pathways in which EXO1 participates, we set out to identify proteins interacting with EXO1. Affinity purification of EXO1 followed by Orbitrap mass spectrometry led to the identification of novel partners that are involved in RNA processing or that are the causative agents of rare X-linked disorders. Depletion of a selected subset of EXO1 interacting proteins led to reduction of the DNA damage response. Among those, we examined the RRP5-homologue and NF{kappa}B-interacting protein PDCD11/ALG-4, which has roles in apoptosis and is a putative driver gene in cutaneous T-cell lymphoma. We provide evidence that depletion of PDCD11 decreased the formation of {gamma}H2AX foci and the phosphorylation of DNA damage response signaling intermediates in response to camptothecin or bleomycin, resulting in increased cellular resistance to DNA damage. Furthermore, extensive coverage of EXO1 sequence (>85%) by mass spectrometry allowed conducting an in-depth analysis of its phosphorylation sites, with the identification of 26 residues that are differentially modified in untreated conditions or upon induction of DNA damage.\n\nAs a whole, these results provide the basis for future in-depth studies on novel roles of EXO1 in genome stability and indicate targets for pharmacological inhibition of pathways of cancer development.\n\nHIGHLIGHTS O_LIProteome-wide analysis of Exonuclease-1 (EXO1) interacting proteins revealed novel partners involved in RNA processing or that are the causative agents of rare X-linked disorders.\nC_LIO_LIWe provide evidence for a role of PDCD11 in the DNA Damage Response.\nC_LIO_LIWe conducted a comprehensive identification of EXO1 phosphorylation sites.\nC_LI

cell biology↗

bioRxiv2018

Aurora A depletion reveals centrosome-independent polarization mechanism in C. elegans

How living systems break symmetry in an organized manner is an important question in biology. In C. elegans zygotes, symmetry breaking normally occurs in the vicinity of centrosomes, resulting in anterior-directed cortical flows and establishment of a single posterior PAR-2 domain. Here, we report that zygotes depleted of the Aurora A kinase AIR-1 or of centrosomes establish two posterior domains, one at each pole. Using transgenic animals and microfabricated triangular chambers, we establish that such bipolarity occurs in a PAR-2- and curvature-dependent manner. Furthermore, we develop an integrated physical model of symmetry breaking, establishing that local PAR-dependent weakening of the actin cortex, together with mutual inhibition of anterior and posterior PAR proteins, provides a mechanism for self-organized PAR polarization without functional centrosomes in C. elegans.\n\nOne Sentence SummaryWe uncover a novel centrosome-independent mechanism of polarization in C. elegans zygotes

cell biology↗

bioRxiv2018

Pharmacophore-guided discovery of CDC25 inhibitors causing cell cycle arrest and cell death

CDC25 phosphatases have a key role in cell cycle transitions and are important targets for cancer therapy. Here, we set out to discover novel CDC25 inhibitors. Using a combination of computational approaches we defined a minimal common pharmacophore in established CDC25 inhibitors and performed a virtual screening of a proprietary library. Taking advantage of the availability of crystal structures for CDC25A and CDC25B and using a molecular docking strategy, we carried out hit expansion/optimization. Enzymatic assays revealed that naphthoquinone scaffolds were the most promising CDC25 inhibitors among selected hits. At the molecular level, the compounds acted through a mixed-type mechanism of inhibition of phosphatase activity, involving reversible oxidation of cysteine residues. In 2D cell cultures, the compounds caused arrest of the cell cycle at the G1/S or at the G2/M transition. Mitotic markers analysis and time-lapse microscopy confirmed that CDK1 activity was impaired and that mitotic arrest was followed by death. Finally, studies on 3D organoids derived from intestinal crypt stem cells of Apc/K-Ras mice revealed that the compounds caused arrest of proliferation.

cancer biology↗