Oncolytic virotherapy for anaplastic and poorly differentiated thyroid cancer: a promise or a clinical reality?

Oncolytic viruses (OVs) selectively infect and lyse cancer cells. A direct lytic effect of OVs has been theorized in the initial studies; however, the antineoplastic effect of OVs is also due to the induction of an immune response against cancer cells. Anaplastic thyroid cancer is one of the most aggressive human malignancies with a short survival time of about 6–12 months from the diagnosis. The lack of effective therapies has prompted to investigate the efficacy of OVs in anaplastic thyroid carcinoma. Different OVs have been tested in preclinical studies, either as single agents or in combinatorial treatments. In this review, the results of these studies are summarized and future perspective discussed

Stainless steel crown vs bulk fill composites for the restoration of primary molars post‐pulpectomy: 1‐year survival and acceptance results of a randomized clinical trial

Background A key factor for the success and longevity of the endodontic treatment is sealing of the cavity after restorative treatment. Aim The aim of this randomised clinical trial was to evaluate the 1-year survival of endodontic treatment in primary molars restored with stainless steel crowns (SSCs) and bulk fill composite resin (BF). As a secondary outcome, the acceptance of both children and parents was evaluated. Design Ninety-one 3- to 8-year-old children with at least one primary molar requiring endodontic treatment were selected. Participants were randomized to SSC or BF and evaluated after 1, 3, 6, and 12 months. An acceptance questionnaire was completed immediately after the treatment. The primary outcome was the endodontic treatment success, evaluated in the intention-to-treat (ITT) population using the Kaplan-Meier and non-inferiority Cox regression analyses, with a non-inferiority limit of 15%. Sensitivity analysis between the success rates after 1 year was performed using Miettinen-Nurminen's method. The Mann-Whitney test was used to compare the treatment acceptance (α = 5%). Results The survival rate after 1 year was BF = 75% and SSC = 88% (HR = 1.41; 90% CI 0.57-3.43). ITT analysis showed a success rate of BF = 86.7% and SSC = 82.6% (RR = 0.95; 0.78-1.16). The non-inferiority hypothesis between the survival of endodontic treatment could not be proved in both analyses (P > .05). The overall acceptance scores did not differ between the restorative groups (P > .05). Conclusion This study failed to show non-inferiority of BF compared with the SSC. The materials were well accepted by both children and their parents

PARP inhibition as new approach to increase the oncolytic activity of the adenoviral mutant dl922-947 against Anaplastic Thyroid Carcinoma

Anaplastic Thyroid Carcinoma (ATC) is one of the most aggressive tumor characterized by an average survival time of 3-6 months after diagnosis. Multimodality therapy, which includes surgical debulking, external radiation therapy and chemotherapy, has failed to show any improvements in survival. Therefore, novel therapies with different mechanisms of action are required. Oncolytic viruses (OVs) represent a novel therapeutic tool for the treatment of aggressive tumors. OVs as single agents have demonstrated limited efficacy in a clinical setting, thus highlighting the need of novel combinatorial therapies (i.e. OVs plus a rationally selected drug) with a potential great impact on clinical use. In this study, we showed that the oncolytic adenovirus dl922-947 induces massive S-phase entry in ATC cell lines, followed by an extensive DNA damage, quantified by γH2AX staining. Moreover, we demonstrated that dl922-947 triggered a DNA damage response, characterized by the activation of the checkpoint kinases ATM and Chk1, and at the same time impaired the ability of the DNA repair, by affecting MRN complex protein levels. The virus-induced single-strand DNA breaks (SSBs) activate the Poly-ADP-Ribose Polymerase (PARP) to signal SSBs to the enzymatic machinery involved in their repair. Here, we demonstrate that the pharmacological inhibition of PARP increases dl922-947 cytotoxicity against Anaplastic Thyroid Carcinoma both in vitro and in vivo. We also show that PARP inhibitor AZD2281 synergizes with dl922-947 increasing viral replication and accelerating cell death pathways. Furthermore, we showed that the virus as single treatment induced an apoptotic-like cell death, as indicated by the presence of Caspase 3 activation, PARP cleavage, Annexin V positivity and Cytochrome C release from mitochondria, although lacking the morphological features of apoptosis. In this regard we also proved that dl922-947 acts on the pathway involved in membrane blebbing by inhibiting Myosin Light Chain 2 (MLC2) phosphorylation. Using a pan caspase inhibitor, both caspase-dependent and -independent cell death pathways are found activated upon dl922-947 infection. Our data clearly suggest that the combination of the oncolytic adenovirus dl922-947 with PARP inhibitor could represent a novel and effective therapeutic approach for the treatment of Anaplastic Thyroid Carcinoma

PARP inhibitor olaparib increases the oncolytic activity of dl922-947 in in vitro and in vivo model of anaplastic thyroid carcinoma

PARP inhibitors are mostly effective as anticancer drugs in association with DNA damaging agents. We have previously shown that the oncolytic adenovirus dl922-947 induces extensive DNA damage, therefore we hypothesized a synergistic antitumoral effect of the PARP inhibitor olaparib in association with dl922-947. Anaplastic thyroid carcinoma was chosen as model since it is a particularly aggressive tumor and, because of its localized growth, it is suitable for intratumoral treatment with oncolytic viruses. Here, we show that dl922-947 infection induces PARP activation, and we confirm in vitro and in vivo that PARP inhibition increases dl922-947 replication and oncolytic activity. In vitro, the combination with olaparib exacerbates the appearance of cell death markers, such as Annexin V positivity, caspase 3 cleavage, cytochrome C release and propidium iodide permeability. In vivo, we also observed a better viral distribution upon PARP inhibition. Changes in CD31 levels suggest a direct effect of olaparib on tumor vascularization and on the viral distribution within the tumor mass. The observation that PARP inhibition enhances the effects of dl922-947 is highly promising not only for the treatment of anaplastic thyroid carcinoma but, in general, for the treatment of other tumors that could benefit from the use of oncolytic viruses

Oncolytic Virus Therapy Alters the Secretome of Targeted Glioblastoma Cells

Oncolytic virus (OV) therapy, which is being tested in clinical trials for glioblastoma, targets cancer cells, while triggering immune cells. Yet OV sensitivity varies from patient to patient. As OV therapy is regarded as an anti-tumor vaccine, by making OV-infected cancer cells secrete immunogenic proteins, linking these proteins to transcriptome would provide a measuring tool to predict their sensitivity. A set of six patient-derived glioblastoma cells treated ex-vivo with herpes simplex virus type 1 (HSV1) modeled a clinical setting of OV infection. The cellular transcriptome and secreted proteome (separated into extracellular vesicles (EV) and EV-depleted fractions) were analyzed by gene microarray and mass-spectroscopy, respectively. Data validation and in silico analysis measured and correlated the secretome content with the response to infection and patient survival. Glioblastoma cells reacted to the OV infection in a seemingly dissimilar fashion, but their transcriptomes changed in the same direction. Therefore, the upregulation of transcripts encoding for secreted proteins implies a common thread in the response of cancer cells to infection. Indeed, the OV-driven secretome is linked to the immune response. While these proteins have distinct membership in either EV or EV-depleted fractions, it is their co-secretion that augments the immune response and associates with favorable patient outcomes

Predicted sea-level changes and evolutionary estimates for age of isolation in Central Mediterranean insular lizards

Rates of biological evolution on islands are often presumed to exceed rates on the mainland. We tested this postulation by computing the evolutionary rate of head shape in Italian wall lizard Podarcis siculus, occurring on four islands off the coast of Southern Italy. We calculated the evolutionary rate using a phylogenetic tree whose node ages were derived from Lambeck et al. predicted ages of geographic isolation of the islands. Such ages are based on a relative sea-level change model for the late Pleistocene–Holocene. Through a likelihood optimization procedure, our method allows computing, besides the evolutionary rate, biological estimates of the ages of insular populations, with this indirectly testing Lambeck et al.’s model estimates. We found that the rate of evolution in Podarcis head shapes on islands is not statistically different from the mainland rate, although insular lizards have distinctive head shapes. Overall, the insular phenotype took 1–4000 years to arise (differing among islands). The estimated ages of insular populations are lower than Lambeck et al.’s estimates and fall in the 5- to 6-ka interval

Glucose impairs tamoxifen responsiveness modulating connective tissue growth factor in breast cancer cells

Type 2 diabetes and obesity are negative prognostic factors in patients with breast cancer (BC). We found that sensitivity to tamoxifen was reduced by 2-fold by 25 mM glucose (High Glucose; HG) compared to 5.5 mM glucose (Low Glucose; LG) in MCF7 BC cells. Shifting from HG to LG ameliorated MCF7 cell responsiveness to tamoxifen. RNA-Sequencing of MCF7 BC cells revealed that cell cycle-related genes were mainly affected by glucose. Connective Tissue Growth Factor (CTGF) was identified as a glucose-induced modulator of cell sensitivity to tamoxifen. Co-culturing MCF7 cells with human adipocytes exposed to HG, enhanced CTGF mRNA levels and reduced tamoxifen responsiveness of BC cells. Inhibition of adipocyte-released IL8 reverted these effects. Interestingly, CTGF immuno-detection in bioptic specimens from women with estrogen receptor positive (ER+) BC correlated with hormone therapy resistance, distant metastases, reduced overall and disease-free survival. Thus, glucose affects tamoxifen responsiveness directly modulating CTGF in BC cells, and indirectly promoting IL8 release by adipocytes

Cytomegalovirus promotes murine glioblastoma growth via pericyte recruitment and angiogenesis.

Cytomegalovirus (CMV) has been implicated in glioblastoma (GBM); however, a mechanistic connection in vivo has not been established. The purpose of this study is to characterize the effects of murine CMV (MCMV) on GBM growth in murine models. Syngeneic GBM models were established in mice perinatally infected with MCMV. We found that tumor growth was markedly enhanced in MCMV+ mice, with a significant reduction in overall survival compared with that of controls (P \u3c 0.001). We observed increased angiogenesis and tumor blood flow in MCMV+ mice. MCMV reactivation was observed in intratumoral perivascular pericytes and tumor cells in mouse and human GBM specimens, and pericyte coverage of tumor vasculature was strikingly augmented in MCMV+ mice. We identified PDGF-D as a CMV-induced factor essential for pericyte recruitment, angiogenesis, and tumor growth. The antiviral drug cidofovir improved survival in MCMV+ mice, inhibiting MCMV reactivation, PDGF-D expression, pericyte recruitment, and tumor angiogenesis. These data show that MCMV potentiates GBM growth in vivo by increased pericyte recruitment and angiogenesis due to alterations in the secretome of CMV-infected cells. Our model provides evidence for a role of CMV in GBM growth and supports the application of antiviral approaches for GBM therapy