DESTABILIZE THE GLIOMA CELL NICHE THROUGH HSP90-GRP78 NETWORK TARGETING

INTRODUCTION: Gliomas are the most common and lethal malignant tumor of the central nervous system representing 1.3% of all human cancers. Glioblastoma (GBM, WHO grade IV) is the most common astrocytic-derived brain tumor in adults, more frequent in older patients (mean age 55 years). They are characterized by a highly aggressive behavior and poor prognosis. Survival rates for gliomas are poor with a median survival that is not exceeding 14 months. Over the last years, the absence of adequate therapeutic options has led the scientific community to seek different molecular targets and bio-markers able to allow a more adequate prognostic and therapeutic stratification of patients. In fact, the absence of adequate therapeutic options makes GBM a fatal disease. A feature of gliomas is the presence of tumor necrotic foci surrounded by severely hypoxic pseudopalisading cells. This unfavorable microenvironment elicits in tumor cells compensatory pathways as autophagy and the unfolded-protein response (UPR). They are two interconnected pathways exploited by tumor cells to cope with low nutrient conditions, hypoxia or other microenvironment-generated stress. Moreover, evidences showed that tumor hypoxia fuel cell motility and preserve the so-called cancer stem cells niche (CSCs). Heat shock proteins (HSPs) are highly conserved proteins which are expressed in response to various forms of stress, in response to environmental challenges, and in specific stages of development and differentiation. Some HSPs are also expressed under non stress conditions and act as molecular chaperones. Thus, HSPs fulfill a variety of biologic functions, including the regulation of synthesis, folding, assembly, and degradation of different proteins. HSPs are constitutively expressed in human carcinomas or cancer cell lines of various histogenetic origins. The endoplasmic reticulum chaperone GRP78 is aberrantly overexpressed by human cancers and preliminary reports showed that targeting of GRP78 in glioma animal model can delay cancer growth. The high expression of HSPs in malignant glioma indicates a possible role of these proteins in resistance to cancer chemotherapy. They in fact, protect cells from many cell-damaging agents or conditions of environmental stress. AIM OF THE STUDY: Assess whether the HSP90-GRP78 axe expression in perinecrotic tumor areas and/or pseudo-palisading cells contributes in maintaining a pro-survival environment and whether it is regulated by hypoxic stimuli, plays roles in glioma cancer stem cell maintenance and in glioma cell motility. Define the contribution of molecular chaperones in glioma modulation of HSPGRP78 axe by specific drugs to provide targets for novel therapies useful to improve prognosis of patients affected by brain tumors. MATERIALS AND METHODS: Clinical series. A series of patients\u2019 (n\ub0 123) recruited at IRCCS Ca\u2019 Granda between 2013-2016, with correlation of molecular profiles to clinicopathological characteristic were collected. From each patient the total RNA was isolated with Tizol\uae reagent (Thermo Fisher Scientific). For gene expression quantification Human TaqManAssays (Applied Biosystems) and reagents were used. Glioma cell lines. LN229, SW1088 and T98G were from ATCC and maintained as suggested by manufacturer. U251-HRE cell were a generous gift from Dr. Ottobrini. Gene knock-out experiments were carried out as transient transfection of short interfering RNA (Sigma Aldrich) using Lipofectamine3000 as transfection reagent (LifeTechnologies). Organotypic tissue cultures. Organotypic glioma tissue were generated through vibratome (VT1200 Leica Microsystems) serial cutting of fresh tumors, and slices were cultured on dedicated supports from Millipore, were cultured with or without the drugs: Temozolomide and/or Gamitrinibs. RESULTS: The expression of GRP78 and TRAP1 are increased in high-grade compared to lower grade or normal brain parenchyma and, high protein levels correlated to poor prognosis. Moreover, GRP78 appears to be enriched in perinecrotic areas of high-grade human gliomas where GRP78 overexpression correlates with HIF-1\u3b1 expression. GRP78 targeting by siRNA or Gamitrinibs decreased the side population of LN229 cells as well as tumor cell motility. The hypoxia regulator HIF1\u3b1 was decreased in TRAP1 knock-out cells respect to control. Lastly, an increase in apoptosis gene was evidenced after treatment with drugs. Finally, Gamitrinibs treatment of glioma organotypic cultures significantly affected tumor cell viability respect to Temozolomide. CONCLUSIONS: These data suggest that GRP78 is a poor prognostic marker for high grades gliomas and HSPs are enriched in peri-necrotic tumor areas sustaining a pro-survival environment. Moreover, these data suggest that GRP78 overexpression in brain tumors could sustain the stem cell population in stress conditions such as hypoxia. This study confirms the potential role of GRP78 as a molecular target in the treatment of malignant glioma

Reconceptualising Personas Across Cultures: Archetypes, Stereotypes & Collective Personas in Pastoral Namibia

The paucity of projects where persona is the research foci and a lack of consensus on this artefact keep many reticent about its purpose and value. Besides crafting personas is expected to differ across cultures, which contrasts the advancements in Western theory with studies and progress in other sites. We postulate User-Created Personas reveal specific characteristics of situated contexts by allowing laypeople to design persona artefacts in their own terms. Hence analysing four persona sessions with an ethnic group in pastoral Namibia –ovaHerero– brought up a set of fundamental questions around the persona artefact regarding stereotypes, archetypes, and collective persona representations: (1) to what extent user depictions are stereotypical or archetypal? If stereotypes prime (2) to what degree are current personas a useful method to represent end-users in technology design? And, (3) how can we ultimately read accounts not conforming to mainstream individual persona descriptions but to collectives

In vitro faecal fermentation of Tritordeum breads and its effect on the human gut health

Spontaneous fermentation of Tritordeum flour enhances the nutritional potential of this hybrid cereal. However, the effect of consumption of Tritordeum sourdough bread (SDB) on gut health remains to be elucidated. This study investigated the effect of in vitro digestion and faecal fermentation of SDB compared to that of traditional baker's yeast (BYB) Tritordeum bread. After 24-h anaerobic faecal fermentation, both SDB and BYB (1% w/v) induced an increase in the relative abundances of Bifidobacterium, Megasphaera, Mitsuokella, and Phascolarctobacterium genera compared to baseline, while concentrations of acetate and butyrate were significantly higher at 24 h for SDB compared to those for BYB. Integrity of intestinal epithelium, as assessed through in vitro trans-epithelial electrical resistance (TEER) assay, was slightly increased after incubation with SDB fermentation supernatants, but not after incubation with BYB fermentation supernatants. The SDB stimulated in vitro mucosal immune response by inducing early secretion of inflammatory cytokines, IL-6 and TNF-α, followed by downregulation of the inflammatory trigger through induction of anti-inflammatory IL-10 expression. Overall, our findings suggest that Tritordeum sourdough can modulate gut microbiota fermentation activity and positively impact the gut health

Pten alterations and their role in cancer management: Are we making headway on precision medicine?

Alterations in the tumor suppressor phosphatase and tensin homolog (PTEN) occur in a substantial proportion of solid tumors. These events drive tumorigenesis and tumor progression. Given its central role as a downregulator of the phosphoinositide 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) pathway, PTEN is deeply involved in cell growth, proliferation, and survival. This gene is also implicated in the modulation of the DNA damage response and in tumor immune microenvironment modeling. Despite the actionability of PTEN alterations, their role as biomarkers remains controversial in clinical practice. To date, there is still a substantial lack of validated guidelines and/or recommendations for PTEN testing. Here, we provide an update on the current state of knowledge on biologic and genetic alterations of PTEN across the most frequent solid tumors, as well as on their actual and/or possible clinical applications. We focus on possible tailored schemes for cancer patients\u2019 clinical management, including risk assessment, diagnosis, prognostication, and treatment

New onset of loss of smell or taste in household contacts of home-isolated SARS-CoV-2-positive subjects

Purpose: To estimate the prevalence of smell or taste impairment in household contacts of mildly symptomatic home-isolated SARS-CoV-2-positive patients. Methods: Cross-sectional study based on ad hoc questions. Results: Of 214 mildly symptomatic COVID-19 patients managed at home under self-isolation, 179 reported to have at least one household contact, with the total number of no study participants contacts being 296. Among 175 household contacts not tested for SARS-CoV-2 infection, 67 (38.3%) had SARS-CoV-2 compatible symptoms, 39 (22.3%) had loss of smell or taste with 7 (4.0%) having loss of smell or taste in the absence of other symptoms. The prevalence of smell or taste impairment was 1.5% in patients tested negative compared to 63.0% of those tested positive for SARS-CoV-2 (p < 0.001). Conclusion: Smell or taste impairment are quite common in not-tested household contacts of mildly symptomatic home-isolated SARS-CoV-2-positive patients. This should be taken into account when estimating the burden of loss of sense of smell and taste during COVID-19 pandemic, and further highlights the value of loss of sense of smell and taste as a marker of infection

Cytosolic phosphorylated EGFR is predictive of recurrence in early stage penile cancer patients: A retropective study

Background: Penile cancer (PC) is a rare tumor, and therapeutic options are limited for this disease, with an overall 5-year overall survival around 65-70%. Adjuvant therapy is not recommended for patients with N0-1 disease, despite up to 60% of these patients will die within 5 years from diagnosis. Methods: Medical records of all patients who underwent radical surgery at University Federico II of Naples and at National Tumor Institute "Pascale" of Naples for early squamous cell carcinoma of the penis from January, 2000 to December, 2011 were retrieved. Paraffin wax embedded tissue specimens were retrieved from the pathology archives of the participating Institutions for all patients. Expression of p-EGFR, EGFR and positivity to HPV were evaluated along with other histological variables of interest. Demographic data of eligible patients were retrieved along with clinical characteristics such as type of surgical operation, time of follow up, time of recurrence, overall survival. A multivariable model was constructed using a forward stepwise selection procedure. Results: Thirty eligible patients were identified. All patients were positive for EGFR by immunohistochemistry, while 13 and 16 were respectively positive for nuclear and cytosolic p-EGFR. No EGFR amplification was detected by FISH. Eight patients were positive for high-risk HPV by ISH. On univariable analysis, corpora cavernosa infiltration (OR 7.8; 95% CI = 0,8 to 75,6; P = 0,039) and positivity for cytosolic p-EGFR (OR 7.6; 95% CI = 1.49 to 50; P = 0.009) were predictive for recurrence, while only positivity for cytosolic p-EGFR (HR = 9.0; 95% CI 1.0-100; P = 0,0116) was prognostic for poor survival. Conclusion: It is of primary importance to identify patients with N0-1 disease who are at increased risk of recurrence, as they do not normally receive any adjuvant therapy. Expression of p-EGFR was found in this series to be strongly related to increase risk of recurrence and shorter overall survival. This finding is consistent with the role of p-EGFR in other solid malignancies. Integration of p-EGFR with classic prognostic factors and other histology markers should be pursued to establish optimal adjuvant therapy for N0-1 PC patients

Current-Driven Conformational Changes, Charging and Negative Differential Resistance in Molecular Wires

We introduce a theoretical approach based on scattering theory and total energy methods that treats transport non-linearities, conformational changes and charging effects in molecular wires in a unified way. We apply this approach to molecular wires consisting of chain molecules with different electronic and structural properties bonded to metal contacts. We show that non-linear transport in all of these systems can be understood in terms of a single physical mechanism and predict that negative differential resistance at high bias should be a generic property of such molecular wires.Comment: 9 pages, 4 figure