Background: Child maltreatment (CM) covers various forms of physical, emotional, and sexual abuse and neglect. Although the scientific literature has extensively documented that exposure to sexual abuse and/or neglect during childhood can cause long-term harm to an individual's overall well-being, the psycho-biological impact of these specific forms of CM requires further exploration. This pilot study tested the hypothesis that experiencing childhood neglect and sexual abuse are associated with psychological alterations as well as biological alterations, namely blood gene expression changes.Methods: This study encompasses a group of volunteer university students, who completed a battery of questionnaires to evaluate the presence of neglect and sexual abuse experience during childhood (CTQ-SF) and psychological distress (SCL-90-R; BDI-II). Both subsets were compared with control groups. Peripheral blood mononuclear cells were collected from all groups to extract RNA and perform genome wide expression analyses.Results: Neglected and sexually abused individuals showed evidence of biological alterations. Through a genome wide transcriptomic analysis, combined with multivariate nomogram analysis, we identified two groups of 5 genes, the changes in expression of each group identified a subject who experienced either neglect or sexual abuse, with a probability of 95%. Among the first group of genes, the expression of ARMC1 correlated significantly with depressive scores in neglected individuals. Among the second group of genes, the expression of ABTB1 correlated significantly with general psychological distress in sexually abused individuals.Conclusion: These results support that childhood neglect and sexual abuse are associated with gene expression changes and psychological outcomes, underscoring the importance of refining the diagnostic process with more objective screening and assessment tools.

As nanocarriers of a new generation, biomimetic nanovesicles are an emerging class of therapeutic tools whose surface is integrated or fabricated with biomaterials capable of mimicking the biological features and functions of native cells. Thanks to this, biomimetic nanovesicles, in particular, those made by plasma membrane moieties, possess greatly improved biocompatibility, high target specificity, a long retention time, and minimal undesired immune responses. For these reasons, a multitude of progenitor cells including cancer ones were employed as templates to generate biomimetic or membrane-camouflaged nanovesicles hosting different therapeutic compounds. In this contribution, different membrane-derived biomimetic vesicles (M-NVs) were generated by osmotic lysis or plasma membrane isolation approaches from normal and cancer cell lines and assayed against in vitro models of human glioblastoma. M-NVs were compared in their cellular internalization degrees of DNA and proteins, morphologically and molecularly characterized, expressing an extracellular membrane-associated marker. Then, Rose Bengal (RB), a photoactivable drug characterized by a relatively low cellular uptake, was incorporated into nascent glioblastoma-derived M-NVs and finally administered to homotypic receiving cells, showing an increased degree of internalization as well as induced cytotoxic effects, even in the absence of photodynamic direct stimulation. Similar results were also obtained assaying lyophilized M-NVs loaded with RB. In conclusion, M-NVs generated by cell membranes effectively deliver several cargoes, including therapeutic molecules, maintain functionality after lyophilization, and show significant internalization effects, making them a promising strategy for therapeutic applications against human glioblastoma cells.

Nanoscale extracellular vesicles (EVs), consisting of exomers, exosomes and microvesicles/ectosomes, have been extensively investigated in the last 20 years, although their biological role is still something of a mystery. EVs are involved in the transfer of lipids, nucleic acids and proteins from donor to recipient cells or distant organs as well as regulating cell-cell communication and signaling. Thus, EVs are important in intercellular communication and this is not limited to sister cells, but may also mediate the crosstalk between different cell types even over long distances. EVs play crucial functions in both cellular homeostasis and the pathogenesis of diseases, and since their contents reflect the status of the donor cell, they represent an additional valuable source of information for characterizing complex biological processes. Recent advances in isolation and analytical methods have led to substantial improvements in both characterizing and engineering EVs, leading to their use either as novel biomarkers for disease diagnosis/prognosis or even as novel therapies. Due to their capacity to carry biomolecules, various EV-based therapeutic applications have been devised for several pathological conditions, including eye diseases. In the eye, EVs have been detected in the retina, aqueous humor, vitreous body and also in tears. Experiences with other forms of intraocular drug applications have opened new ways to use EVs in the treatment of retinal diseases. We here provide a comprehensive summary of the main in vitro, in vivo, and ex vivo literaturebased studies on EVs' role in ocular physiological and pathological conditions. We have focused on age-related macular degeneration, diabetic retinopathy, glaucoma, which are common eye diseases leading to permanent blindness, if not treated properly. In addition, the putative use of EVs in retinitis pigmentosa and other retinopathies is discussed. Finally, we have reviewed the potential of EVs as therapeutic tools and/or biomarkers in the above-mentioned retinal disorders. Evidence emerging from experimental disease models and human material strongly suggests future diagnostic and/or therapeutic exploitation of these biological agents in various ocular disorders with a good possibility to improve the patient's quality of life.

Study question Do cumulus cells (CCs), during mouse germinal vesicle (GV)-to-metaphase II (MII) transition, produce miRNAs with a role in the acquisition of the oocyte developmental competence? Summary answer We identified a group of 74 miRNAs with a known role in key processes of oocytes maturation and developmental competence. What is known already We recently developed a co-culture platform whereby CC-free, denuded, GV oocytes (DOs) were matured to MII upon a feeder layer (FL) of CCs derived either from developmentally competent (FL-SN-CCs) or incompetent (FL-NSN-CCs) oocytes. When cultured upon FL-SN-CCs, DOs displayed a developmental competence equal to that showed by control cumulus-enclosed oocytes (COCs). Instead, when cultured upon FL-NSN-CCs, all DOs arrested at the 2-cell stage. We found that CCs released into the medium large amounts of extracellular vesicles (EVs), whose content analysis by NGS led to the identification of candidate miRNAs. Study design, size, duration Five-eight-week-old CD1 mouse females (Charles River) were injected with 10 I.U. Pregnant Mare Serum Gonadotropin (PMSG). After 48 hr, the ovaries were isolated and their surface punctured with a thin sterile needle to collect fully-grown antral oocytes which were separated from the surrounding CCs. Eight-ten DOs were cultured upon FL-SN-CCs or FL-NSN-CCs for 15 hr in 50 µl exosome-free alpha-MEM Glutamax medium for EVs production. Experiments were performed in triplicates. Participants/materials, setting, methods EVs were collected and processed for EVs characterisation by Amnis-Imaging-flow-cytometer and transmission electron microscopy (TEM), or for RNA extraction (miRNeasy Micro Kit, Qiagen), libraries production (Small RNA-Seq Library Prep kit, Lexogen) and NGS (Illumina Genome Analyzer and the NextSeq 500/550 High Output v2.5). To identify miRNA’s target genes, we used “mirPath v.3” server of the DIANA tools website (https://dianalab.e-ce.uth.gr/html/mirpathv3/index.php?r=mirpath). Then, STRING and KEGG were used to highlight the pathways in which these genes are involved. Main results and the role of chance Imaging flow cytometry recorded an amount of released EVs in both FL-SN-CCs or FL-NSN-CCs in the mean range of 5-5.9 x 10(8) particles/ml (p > 0.05), with >96% positive to CD9 exosome surface marker. TEM analysis showed an EVs size in diameter comprised between 18.5-54.9 nm. By comparing FL-SN-CCs vs. FL-NSN-CCs, NGS of EVs’ miRNAs content displayed 74 differentially expressed miRNAs, 43 up- and 31 down-regulated. Of them, 7 resulted involved in the regulation of 71 target genes with a known function in meiosis resumption (N = 24), follicle growth (N = 23), fertilisation (N = 1) and in the acquisition of the oocyte’s developmental competence (N = 23). Limitations, reasons for caution We used an in vitro platform to mimic the cumulus-oocyte-complex environment and, thus, the artificial set-up may differ from that of the natural condition. Also, our hypothesis on the role of these miRNAs, needs to be further confirmed by functional assays (e.g., miRNA inactivation). Wider implications of the findings Overall, our results indicate CC-derived miRNAs as candidates of the CC-to-oocyte bidirectional communication and suggest a group of miRNAs as potential regulative molecules in the acquisition of the oocyte developmental competence. Trial registration number NOT APPLICABLE

The role of cumulus cells (CCs) in the acquisition of oocyte developmental competence is not yet fully understood. In a previous study, we matured cumulus-denuded fully-grown mouse oocytes to metaphase II (MII) on a feeder layer of CCs (FL-CCs) isolated from developmentally competent (FL-SN-CCs) or incompetent (FL-NSN-CCs) SN (surrounded nucleolus) or NSN (not surrounding nucleolus) oocytes, respectively. We observed that oocytes cultured on the former could develop into blastocysts, while those matured on the latter arrested at the 2-cell stage. To investigate the CC factors contributing to oocyte developmental competence, here we focused on the CCs' release into the medium of extracellular vesicles (EVs) and on their miRNA content. We found that, during the 15-h transition to MII, both FL-SN-CCs and FL-NSN-CCs release EVs that can be detected, by confocal microscopy, inside the zona pellucida (ZP) or the ooplasm. The majority of EVs are <200 nm in size, which is compatible with their ability to cross the ZP. Next-generation sequencing of the miRNome of FL-SN-CC versus FL-NSN-CC EVs highlighted 74 differentially expressed miRNAs, with 43 up- and 31 down-regulated. Although most of these miRNAs do not have known roles in the ovary, in silico functional analysis showed that seven of these miRNAs regulate 71 target genes with specific roles in meiosis resumption (N = 24), follicle growth (N = 23), fertilization (N = 1), and the acquisition of oocyte developmental competence (N = 23). Overall, our results indicate CC EVs as emerging candidates of the CC-to-oocyte communication axis and uncover a group of miRNAs as potential regulatory factors.

Do cumulus cells (CCs), during mouse germinal vesicle (GV)-to-metaphase II (MII) transition, produce miRNAs with a role in the acquisition of the oocyte developmental competence? We identified a group of 74 miRNAs with a known role in key processes of oocytes maturation and developmental competence. We recently developed a co-culture platform whereby CC-free, denuded, GV oocytes (DOs) were matured to MII upon a feeder layer (FL) of CCs derived either from developmentally competent (FL-SN-CCs) or incompetent (FL-NSN-CCs) oocytes. When cultured upon FL-SN-CCs, DOs displayed a developmental competence equal to that showed by control cumulus-enclosed oocytes (COCs). Instead, when cultured upon FL-NSN-CCs, all DOs arrested at the 2-cell stage. We found that CCs released into the medium large amounts of extracellular vesicles (EVs), whose content analysis by NGS led to the identification of candidate miRNAs. Five-eight-week-old CD1 mouse females (Charles River) were injected with 10 I.U. Pregnant Mare Serum Gonadotropin (PMSG). After 48 hr, the ovaries were isolated and their surface punctured with a thin sterile needle to collect fully-grown antral oocytes which were separated from the surrounding CCs. Eight-ten DOs were cultured upon FL-SN-CCs or FL-NSN-CCs for 15 hr in 50 µl exosome-free alpha-MEM Glutamax medium for EVs production. Experiments were performed in triplicates. EVs were collected and processed for EVs characterisation by Amnis-Imaging-flow-cytometer and transmission electron microscopy (TEM), or for RNA extraction (miRNeasy Micro Kit, Qiagen), libraries production (Small RNA-Seq Library Prep kit, Lexogen) and NGS (Illumina Genome Analyzer and the NextSeq 500/550 High Output v2.5). To identify miRNA’s target genes, we used “mirPath v.3” server of the DIANA tools website (https://dianalab.e-ce.uth.gr/html/mirpathv3/index.php?r=mirpath). Then, STRING and KEGG were used to highlight the pathways in which these genes are involved. Imaging flow cytometry recorded an amount of released EVs in both FL-SN-CCs or FL-NSN-CCs in the mean range of 5-5.9 x 10(8) particles/ml (p > 0.05), with >96% positive to CD9 exosome surface marker. TEM analysis showed an EVs size in diameter comprised between 18.5-54.9 nm. By comparing FL-SN-CCs vs. FL-NSN-CCs, NGS of EVs’ miRNAs content displayed 74 differentially expressed miRNAs, 43 up- and 31 down-regulated. Of them, 7 resulted involved in the regulation of 71 target genes with a known function in meiosis resumption (N = 24), follicle growth (N = 23), fertilisation (N = 1) and in the acquisition of the oocyte’s developmental competence (N = 23). We used an in vitro platform to mimic the cumulus-oocyte-complex environment and, thus, the artificial set-up may differ from that of the natural condition. Also, our hypothesis on the role of these miRNAs, needs to be further confirmed by functional assays (e.g., miRNA inactivation). Overall, our results indicate CC-derived miRNAs as candidates of the CC-to-oocyte bidirectional communication and suggest a group of miRNAs as potential regulative molecules in the acquisition of the oocyte developmental competence. NOT APPLICABLE

Aldehyde dehydrogenases of the subfamily 1A (ALDH1A) are enzymes necessary for the oxidation of all-trans or 9-cis retinal to retinoic acid (RA). Retinoic acid and its derivatives are important for normal development and maintenance of epithelia, reproduction, memory, and immune function in adults. Moreover, in recent years, it has been demonstrated that ALDH1A members are also expressed and functional in several human cancers where their role is not limited to the synthesis of RA. Here, we review the current knowledge about ALDH1A3, one of the 1A isoforms, in cancers with an emphasis on two of the deadliest tumors that affect humans: glioblastoma multiforme and mesothelioma. In both tumors, ALDH1A3 is considered a negative prognostic factor, and its level correlates with excessive proliferation, chemoresistance, and invasiveness. We also review the recent attempts to develop both ALDH1A3-selective inhibitors for cancer therapy and ALDH1A3-specific fluorescent substrates for fluorescence-guided tumor resection.

Berberine (BBR) is known for its antitumor activity and photosensitizer properties in anti-cancer photodynamic therapy (PDT), and it has previously been favorably assayed against glioblastoma multiforme (GBM)-derived cells. In this work, two BBR hydrophobic salts, dodecyl sulfate (S) and laurate (L), have been encapsulated in PLGA-based nanoparticles (NPs), chitosan-coated by the addition of chitosan oleate in the preparation. NPs were also further functionalized with folic acid. All the BBR-loaded NPs were efficiently internalized into T98G GBM established cells, and internalization increased in the presence of folic acid. However, the highest mitochondrial co-localization percentages were obtained with BBR-S NPs without folic acid content. In the T98G cells, BBR-S NPs appeared to be the most efficient in inducing cytotoxicity events and were therefore selected to assess the effect of photodynamic stimulation (PDT). As a result, PDT potentiated the viability reduction for the BBR-S NPs at all the studied concentrations, and a roughly 50% reduction of viability was obtained. No significant cytotoxic effect on normal rat primary astrocytes was observed. In GBM cells, a significant increase in early and late apoptotic events was scored by BBR NPs, with a further increase following the PDT scheme. Furthermore, a significantly increased depolarization of mitochondria was highlighted following BBR-S NPs' internalization and mostly after PDT stimulation, compared to untreated and PDT-only treated cells. In conclusion, these results highlighted the efficacy of the BBR-NPs-based strategy coupled with photoactivation approaches to induce favorable cytotoxic effects in GBM cells.

Celiac disease (CD) is a chronic and systemic autoimmune disorder that affects preferentially the small intestine of individuals with a genetic predisposition. CD is promoted by the ingestion of gluten, a storage protein contained in the endosperm of the seeds of wheat, barley, rye, and related cereals. Once in the gastrointestinal (GI) tract, gluten is enzymatically digested with the consequent release of immunomodulatory and cytotoxic peptides, i.e., 33mer and p31-43. In the late 1970s a new group of biologically active peptides, called gluten exorphins (GEs), was discovered and characterized. In particular, these short peptides showed a morphine-like activity and high affinity for the δ-opioid receptor (DOR). The relevance of GEs in the pathogenesis of CD is still unknown. Recently, it has been proposed that GEs could contribute to asymptomatic CD, which is characterized by the absence of symptoms that are typical of this disorder. In the present work, GEs cellular and molecular effects were in vitro investigated in SUP-T1 and Caco-2 cells, also comparing viability effects with human normal primary lymphocytes. As a result, GEs treatments increased tumor cell proliferation by cell cycle and Cyclins activation as well as by induction of mitogenic and pro-survival pathways. Finally, a computational model of GEs interaction with DOR is provided. Altogether, the results might suggest a possible role of GEs in CD pathogenesis and on its associated cancer comorbidities.

Dimethyl fumarate (DMF) is a well-characterized molecule that exhibits immuno-modulatory, anti-inflammatory, and antioxidant properties and that is currently approved for the treatment of psoriasis and multiple sclerosis. Due to its Nrf2-dependent and independent mechanisms of action, DMF has a therapeutic potential much broader than expected. In this comprehensive review, we discuss the state-of-the-art and future perspectives regarding the potential repurposing of DMF in the context of chronic inflammatory diseases of the intestine, such as inflammatory bowel disorders (i.e., Crohn's disease and ulcerative colitis) and celiac disease. DMF's mechanisms of action, as well as an exhaustive analysis of the in vitro/in vivo evidence of its beneficial effects on the intestine and the gut microbiota, together with observational studies on multiple sclerosis patients, are here reported. Based on the collected evidence, we highlight the new potential applications of this molecule in the context of inflammatory and immune-mediated intestinal diseases.

Enteric glial cells (EGCs), the major components of the enteric nervous system (ENS), are implicated in the maintenance of gut homeostasis, thereby leading to severe pathological conditions when impaired. However, due to technical difficulties associated with EGCs isolation and cell culture maintenance that results in a lack of valuable in vitro models, their roles in physiological and pathological contexts have been poorly investigated so far. To this aim, we developed for the first time, a human immortalized EGC line (referred as ClK clone) through a validated lentiviral transgene protocol. As a result, ClK phenotypic glial features were confirmed by morphological and molecular evaluations, also providing the consensus karyotype and finely mapping the chromosomal rearrangements as well as HLA-related genotypes. Lastly, we investigated the ATP- and acetylcholine, serotonin and glutamate neurotransmitters mediated intracellular Ca2+ signaling activation and the response of EGCs markers (GFAP, SOX10, S100β, PLP1, and CCL2) upon inflammatory stimuli, further confirming the glial nature of the analyzed cells. Overall, this contribution provided a novel potential in vitro tool to finely characterize the EGCs behavior under physiological and pathological conditions in humans.

Cystic formation in human primary brain tumors is a relatively rare event whose incidence varies widely according to the histotype of the tumor. Composition of the cystic fluid has mostly been characterized in samples collected at the time of tumor resection and no indications of the evolution of cystic content are available. We characterized the evolution of the proteome of cystic fluid using a bottom-up proteomic approach on sequential samples obtained from secretory meningioma (SM), cystic schwannoma (CS) and cystic high-grade glioma (CG). We identified 1008 different proteins; 74 of these proteins were found at least once in the cystic fluid of all tumors. The most abundant proteins common to all tumors studied derived from plasma, with the exception of prostaglandin D2 synthase, which is a marker of cerebrospinal fluid origin. Overall, the protein composition of cystic fluid obtained at different times from the same tumor remained stable. After the identification of differentially expressed proteins (DEPs) and the protein-protein interaction network analysis, we identified the presence of tumor-specific pathways that may help to characterize tumor-host interactions. Our results suggest that plasma proteins leaking from local blood-brain barrier disruption are important contributors to cyst fluid formation, but cerebrospinal fluid (CSF) and the tumor itself also contribute to the cystic fluid proteome and, in some cases, as with immunoglobulin G, shows tumor-specific variations that cannot be simply explained by differences in vessel permeability or blood contamination.

A long-standing practice in the treatment of cancer is that of hitting hard with the maximum tolerated dose to eradicate tumors. This continuous therapy, however, selects for resistant cells, leading to the failure of the treatment. A different type of treatment strategy, adaptive therapy, has recently been shown to have a degree of success in both preclinical xenograft experiments and clinical trials. Adaptive therapy is used to maintain a tumor's volume by exploiting the competition between drug-sensitive and drug-resistant cells with minimum effective drug doses or timed drug holidays. To further understand the role of competition in the outcomes of adaptive therapy, we developed a 2D on-lattice agent-based model. Our simulations show that the superiority of the adaptive strategy over continuous therapy depends on the local competition shaped by the spatial distribution of resistant cells. Intratumor competition can also be affected by fibroblasts, which produce microenvironmental factors that promote cancer cell growth. To this end, we simulated the impact of different fibroblast distributions on treatment outcomes. As a proof of principle, we focused on five types of distribution of fibroblasts characterized by different locations, shapes, and orientations of the fibroblast region with respect to the resistant cells. Our simulation shows that the spatial architecture of fibroblasts modulates tumor progression in both continuous and adaptive therapy. Finally, as a proof of concept, we simulated the outcomes of adaptive therapy of a virtual patient with four metastatic sites composed of different spatial distributions of fibroblasts and drug-resistant cell populations. Our simulation highlights the importance of undetected metastatic lesions on adaptive therapy outcomes.

Photodynamic therapy (PDT) is a promising anticancer strategy based on the light energy stimulation of photosensitizers (PS) molecules within a malignant cell. Among a multitude of recently challenged PS, Rose bengal (RB) has been already reported as an inducer of cytotoxicity in different tumor cells. However, RB displays a low penetration capability across cell membranes. We have therefore developed a short-term amino acids starvation protocol that significantly increases RB uptake in human astrocytoma cells compared to normal rat astrocytes. Following induced starvation uptake, RB is released outside cells by the exocytosis of extracellular vesicles (EVs). Thus, we have introduced a specific pharmacological treatment, based on the GW4869 exosomes inhibitor, to interfere with RB extracellular release. These combined treatments allow significantly reduced nanomolar amounts of administered RB and a decrease in the time interval required for PDT stimulation. The overall conditions affected astrocytoma viability through the activation of apoptotic pathways. In conclusion, we have developed for the first time a combined scheme to simultaneously increase the RB uptake in human astrocytoma cells, reduce the extracellular release of the drug by EVs, and improve the effectiveness of PDT-based treatments. Importantly, this strategy might be a valuable approach to efficiently deliver other PS or chemotherapeutic drugs in tumor cells.

Extracellular vesicles (EVs), comprising large microvesicles (MVs) and exosomes (EXs), play a key role in intercellular communication, both in physiological and in a wide variety of pathological conditions. However, the education of EV target cells has so far mainly been investigated as a function of EX cargo, while few studies have focused on the characterization of EV surface membrane molecules and the mechanisms that mediate the addressability of specific EVs to different cell types and tissues. Identifying these mechanisms will help fulfill the diagnostic, prognostic, and therapeutic promises fueled by our growing knowledge of EVs. In this review, we first discuss published studies on the presumed EV "delivery code" and on the combinations of the hypothesized EV surface membrane "sender" and "recipient" molecules that may mediate EV targeting in intercellular communication. Then we briefly review the main experimental approaches and techniques, and the bioinformatic tools that can be used to identify and characterize the structure and functional role of EV surface membrane molecules. In the final part, we present innovative techniques and directions for future research that would improve and deepen our understandings of EV-cell targeting.

Photodynamic therapy (PDT) has recently attracted interest as an innovative and adjuvant treatment for different cancers including malignant gliomas. Among these, Glioblastoma (GBM) is the most prevalent neoplasm in the central nervous system. Despite conventional therapeutic approaches that include surgical removal, radiation, and chemotherapy, GBM is characterized by an extremely poor prognosis and a high rate of recurrence. PDT is a physical process that induces tumor cell death through the genesis and accumulation of reactive oxygen species (ROS) produced by light energy interaction with a photosensitizing agent. In this contribution, we explored the potentiality of the plant alkaloid berberine (BBR) as a photosensitizing and cytotoxic agent coupled with a PDT scheme using a blue light source in human established astrocytoma cell lines. Our data mainly indicated for the combined BBR-PDT scheme a potent activation of the apoptosis pathway, through a massive ROS production, a great extent of mitochondria depolarization, and the sub-sequent activation of caspases. Altogether, these results demonstrated that BBR is an efficient photosensitizer agent and that its association with PDT may be a potential anticancer strategy for high malignant gliomas.

Glioblastoma multiforme (GBM) is the most lethal primary brain tumor in adults, with an average survival time of about one year from initial diagnosis. In the attempt to overcome the complexity and drawbacks associated with in vivo GBM models, together with the need of developing systems dedicated to screen new potential drugs, considerable efforts have been devoted to the implementation of reliable and affordable in vitro GBM models. Recent findings on GBM molecular features, revealing a high heterogeneity between GBM cells and also between other non-tumor cells belonging to the tumoral niche, have stressed the limitations of the classical 2D cell culture systems. Recently, several novel and innovative 3D cell cultures models for GBM have been proposed and implemented. In this review, we first describe the different populations and their functional role of GBM and niche non-tumor cells that could be used in 3D models. An overview of the current available 3D in vitro systems for modeling GBM, together with their major weaknesses and strengths, is presented. Lastly, we discuss the impact of groundbreaking technologies, such as bioprinting and multi-omics single cell analysis, on the future implementation of 3D in vitro GBM models.

Since invasive bladder cancer (BC) can progress to life threatening metastases, understanding the molecular mechanisms underlying BC invasion is crucial for potentially decreasing the mortality of this disease. Herein, it is discovered that autophagy-related gene 7 (ATG7) is remarkably overexpressed in human invasive BC tissues. The knockdown of ATG7 in human BC cells dramatically inhibits cancer cell invasion, revealing that ATG7 is a key player in regulating BC invasion. Mechanistic studies indicate that MIR190A is responsible for ATG7 mRNA stability and protein overexpression by directly binding to ATG7 mRNA 3'-UTR. Furthermore, ATG7-mediated autophagy promotes HNRNPD (ARE/poly(U)-binding/degradation factor 1) protein degradation, and in turn reduces HNRNPD interaction with ARHGDIB mRNA, resulting in the elevation of ARHGDIB mRNA stability, and subsequently leading to BC cell invasion. The identification of the MIR190A/ATG7 autophagic mechanism regulation of HNRNPD/ARHGDIB expression provides an important insight into understanding the nature of BC invasion and suggests that autophagy may represent a potential therapeutic strategy for the treatment of human BC patients.