Oncolytic Adenoviruses for Cancer Therapy

Many immuno-therapeutic strategies are currently being developed to fight cancer. In this scenario, oncolytic adenoviruses (Onc.Ads) have an interesting role for their peculiar tumor selectivity, safety, and transgene-delivery capability. The major strength of the Onc.Ads is the extraordinary immunogenicity that leads to a strong T-cell response, which, together with the possibility of the delivery of a therapeutic transgene, could be more effective than current strategies. In this review, we travel in the adenovirus (Ads) and Onc.Ads world, focusing on a variety of strategies that can enhance Onc.Ads antitumoral efficacy, passing through tumor microenvironment modulation. Onc.Ads-based therapeutic strategies constitute additional weapons in the fight against cancer and appear to potentiate conventional and immune checkpoint inhibitors (ICIs)-based therapies leading to a promising scenario.Peer reviewe

The Effect of Job–Education Vertical Mismatch on Wages Among Recent PhD Graduates: Evidence From an Instrumental Variable Analysis

Existing studies suggest that recent PhD graduates with a job vertically mismatched with their education tend to earn lower wages than their matched counterparts. However, by being based on cross-sectional ordinary least squares (OLS) estimates, these studies raise endogeneity concerns and can only be considered evidence of a correlation between vertical mismatch and wages. This paper improves this literature by applying a heteroskedasticity-based instrumental variable estimation approach to analyzing Italian PhD holders’ cross-sectional micro-data. Our analysis suggests that previous empirical studies have provided slightly upward estimates of the impact of vertical mismatch on wages. Nevertheless, our results show that the effect of overeducation on wages is sizeable. However, no wage effect is found for overskilling. The heterogeneity of these findings by field of study and gender are also inspected

The Effect of Job–Education Vertical Mismatch on Wages Among Recent PhD Graduates: Evidence From an Instrumental Variable Analysis

Existing studies suggest that recent PhD graduates with a job vertically mismatched with their education tend to earn lower wages than their matched counterparts. However, by being based on cross-sectional ordinary least squares (OLS) estimates, these studies raise endogeneity concerns and can only be considered evidence of a correlation between vertical mismatch and wages. This paper improves this literature by applying a heteroskedasticity-based instrumental variable estimation approach to analyzing Italian PhD holders’ cross-sectional micro-data. Our analysis suggests that previous empirical studies have provided slightly upward estimates of the impact of vertical mismatch on wages. Nevertheless, our results show that the efect of overeducation on wages is sizeable. However, no wage efect is found for overskilling. The heterogeneity of these fndings by feld of study and gender are also inspecte

Two-dimensional posture evaluation in Parkinson’s disease: effect of loads on the spinal angle during gait

Parkinson’s Disease patients present diminished coordination caused by neural degeneration. This leads to large motor difficulties during gait such as balance loss and pronounced forward inclination of the upper body. This work assessed the spinal sagittal plane angle alterations in two groups: six parkinsonian patients and six control healthy subjects. This parameter was analyzed during gait under three conditions: without external loads and with external loads applied either on the chest or on the lower back area. Results were statistically compared by means of t-test of paired samples in both groups. For patients, a significant effect was found when loads were applied on the chest. On the other hand, healthy subjects showed no significant differences in either case.Fil: Celoria, Paula. Instituto Tecnológico de Buenos Aires; ArgentinaFil: Nanni, Federico. Instituto Tecnológico de Buenos Aires; ArgentinaFil: Pastore, Flavia. Instituto Tecnológico de Buenos Aires; ArgentinaFil: Pulenta, Sebastian. Instituto Tecnológico de Buenos Aires; ArgentinaFil: Tajerian, Matias. Instituto Tecnológico de Buenos Aires; ArgentinaFil: Pantazis, Lucio José. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Tecnológico de Buenos Aires; ArgentinaFil: Moscoso Vásquez, Hilda Marcela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Tecnológico de Buenos Aires; ArgentinaFil: Cerquetti, Daniel. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; ArgentinaFil: Merello, Marcelo Jorge. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; ArgentinaFil: Risk, Marcelo. Instituto Tecnológico de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

179. Helper-Dependent Adenovirus-Mediated Gene Transfer of an LDL Receptor/Transferrin Chimeric Protein Reduces Aortic Atherosclerosis in LDL Receptor-Deficient Mice

Familial hypercholesterolemia (FH) is a well-characterized genetic hyperlipidemia due in most of the cases to mutations in the LDL receptor (LDLR) gene; FH is characterized by elevated concentration of plasma LDL cholesterol (LDL-C) with consequent deposition of LDL-C in tendons, skin and arteries. Statins can lower cholesterol levels but are not effective in all patients whose prognosis is still quite poor. In the past, we have developed safe and effective gene-therapy strategies for hepatocytes transduction and consequent expression of anti-atherogenic proteins using PEGylated helper-dependent adenoviral (HD-Ad) vectors. We have recently devised a therapeutic strategy for reducing LDL using a secreted protein that can potentially be expressed in non-hepatic tissues used as bioreactors. At this aim, we developed an HD-Ad vector for the expression of the soluble form of the extracellular portion of the human LDLR fused with transferrin (LDLR/Tf). We evaluated the efficacy of LDLR/Tf in cellular models such as CHOldla7 in which we restored the cell ability to uptake of labeled LDL; subsequently, we administered intravenously 1X10E11 vp/kg of the HD-Ad vector expressing LDLR/Tf in LDLR-deficient mice and demonstrated the efficacy of the above-mentioned vector in reducing total and LDL cholesterol levels; in addition, expression of LDLR/Tf significantly reduced aortic atherosclerotic lesions in treated LDLR-deficient mice compared to controls 1.78±0.48 vs. 5.38±0.54 sq.mm.). We therefore demonstrated the efficacy of serum secretion of LDLR/Tf in reducing aortic atherosclerosis in FH mice. These results will allow the evaluation of HD-Ad vector-mediated expression of LDLR/Tf in non-hepatic tissues using alternative routes of administration in order to develop safer gene transfer protocol more compatible with clinical applications

A new synthetic protein, TAT-RH, inhibits tumor growth through the regulation of NFκB activity

Based on its role in angiogenesis and apoptosis, the inhibition of NFkappaB activity is considered an effective treatment for cancer, hampered by the lack of selective and safe inhibitors. We recently demonstrated that the RH domain of GRK5 (GRK5-RH) inhibits NFkappaB, thus we evaluated its effects on cancer growth.The role of GRK5-RH on tumor growth was assessed in a human cancer cell line (KAT-4). RH overexpression was induced by adenovirus mediated gene transfer; alternatively we administered a synthetic protein reproducing the RH domain of GRK5 (TAT-RH), actively transported into the cells.In vitro, adenovirus mediated GRK5-RH overexpression (AdGRK5-NT) in human tumor cells (KAT-4) induces IkappaB accumulation and inhibits NFkappaB transcriptional activity leading to apoptotic events. In BALB/c nude mice harboring KAT-4 induced neoplasias, intra-tumor delivery of AdGRK5-NT reduces in a dose-dependent fashion tumor growth, with the highest doses completely inhibiting it. This phenomenon is paralleled by a decrease of NFkappaB activity, an increase of IkappaB levels and apoptotic events. To move towards a pharmacological setup, we synthesized the TAT-RH protein. In cultured KAT-4 cells, different dosages of TAT-RH reduced cell survival and increased apoptosis. In BALB/c mice, the anti-proliferative effects of TAT-RH appear to be dose-dependent and highest dose completely inhibits tumor growth.Our data suggest that GRK5-RH inhibition of NFkappaB is a novel and effective anti-tumoral strategy and TAT-RH could be an useful tool in the fighting of cancer

Comparative Analysis of Gene Expression Data Reveals Novel Targets of Senescence-Associated microRNAs

In the last decades, cellular senescence is viewed as a complex mechanism involved in different processes, ranging from tumor suppression to induction of age-related degenerative alterations. Senescence-inducing stimuli are myriad and, recently, we and others have demonstrated the role exerted by microRNAs in the induction and maintenance of senescence, by the identification of a subset of Senescence-Associated microRNAs (SAmiRs) up-regulated during replicative or stress-induced senescence and able to induce a premature senescent phenotype when over-expressed in human primary cells. With the intent to find novel direct targets of two specific SAmiRs, SAmiR-494 and -486-5p, and cellular pathways which they are involved in, we performed a comparative analysis of gene expression profiles available in literature to select genes down-regulated upon replicative senescence of human primary fibroblasts. Among them, we searched for SAmiR’s candidate targets by analyzing with different target prediction algorithms their 3’UTR for the presence of SAmiR-binding sites. The expression profiles of selected candidates have been validated on replicative and stress-induced senescence and the targeting of the 3’UTRs was assessed by luciferase assay. Results allowed us to identify Cell Division Cycle Associated 2 (CDCA2) and Inhibitor of DNA binding/differentiation type 4 (ID4) as novel targets of SAmiR-494 and SAmiR-486-5p, respectively. Furthermore, we demonstrated that the over-expression of CDCA2 in human primary fibroblasts was able to partially counteract etoposide-induced senescence by mitigating the activation of DNA Damage Response

Systems Biology Approaches for the Improvement of Oncolytic Virus-Based Immunotherapies

Oncolytic virus (OV)-based immunotherapy is mainly dependent on establishing an efficient cell-mediated antitumor immunity. OV-mediated antitumor immunity elicits a renewed antitumor reactivity, stimulating a T-cell response against tumor-associated antigens (TAAs) and recruiting natural killer cells within the tumor microenvironment (TME). Despite the fact that OVs are unspecific cancer vaccine platforms, to further enhance antitumor immunity, it is crucial to identify the potentially immunogenic T-cell restricted TAAs, the main key orchestrators in evoking a specific and durable cytotoxic T-cell response. Today, innovative approaches derived from systems biology are exploited to improve target discovery in several types of cancer and to identify the MHC-I and II restricted peptide repertoire recognized by T-cells. Using specific computation pipelines, it is possible to select the best tumor peptide candidates that can be efficiently vectorized and delivered by numerous OV-based platforms, in order to reinforce anticancer immune responses. Beyond the identification of TAAs, system biology can also support the engineering of OVs with improved oncotropism to reduce toxicity and maintain a sufficient portion of the wild-type virus virulence. Finally, these technologies can also pave the way towards a more rational design of armed OVs where a transgene of interest can be delivered to TME to develop an intratumoral gene therapy to enhance specific immune stimuli

Angiotensin receptor I stimulates osteoprogenitor proliferation through TGFβ-mediated signaling:AT1R SIGNALING IN OSTEOBLAST DIFFERENTIATION

Clinical studies of large human populations and pharmacological interventions in rodent models have recently suggested that anti-hypertensive drugs that target angiotensin II (Ang II) activity may also reduce loss of bone mineral density. Here, we identified in a genetic screening the Ang II type I receptor (AT1R) as a potential determinant of osteogenic differentiation and, implicitly, bone formation. Silencing of AT1R expression by RNA interference severely impaired the maturation of a multipotent mesenchymal cell line (W20-17) along the osteoblastic lineage. The same effect was also observed after the addition of the AT1R antagonist losartan but not the AT2R inhibitor PD123,319. Additional cell culture assays traced the time of greatest losartan action to the early stages of W20-17 differentiation, namely during cell proliferation. Indeed, addition of Ang II increased proliferation of differentiating W20-17 and primary mesenchymal stem cells and this stimulation was reversed by losartan treatment. Cells treated with losartan also displayed an appreciable decrease of activated (phosphorylated)-Smad2/3 proteins. Moreover, Ang II treatment elevated endogenous transforming growth factor β (TGFβ) expression considerably and in an AT1R-dependent manner. Finally, exogenous TGFβ was able to restore high proliferative activity to W20-17 cells that were treated with both Ang II and losartan. Collectively, these results suggest a novel mechanism of Ang II action in bone metabolism that is mediated by TGFβ and targets proliferation of osteoblast progenitors

Insulin stimulates fibroblast proliferation through calcium-calmodulin-dependent kinase II.

Insulin effects are mediated by multiple integrated signals generated by the insulin receptor. Fibroblasts, as most of mammalian cells, are a target of insulin action and are impor- tant actors in the vascular pathogenesis of hyperinsulinemia. A role for calcium-calmodulin-dependent kinases (CaMK) in insulin signaling has been proposed but has been under inves- tigated. We investigated the role of the CaMK isoform II in insulin signaling in human fibroblasts. A rapid and transient increase of intracellular calcium concentration was induced by insulin stimulation, followed by increase of CaMKII activity, via L type calcium channels. Concomitantly, insulin stimula- tion induced Raf-1 and ERK activation, followed by thymidine uptake. Inhibition of CaMKII abrogated the insulin-induced Raf-1 and ERK activation, resulting also in the inhibition of thymidine incorporation. These results demonstrate that in fibroblasts, insulin-activated CaMKII is necessary, together with Raf-1, for ERK activation and cell proliferation. This represents a novel mechanism in the control of insulin signals leading to fibroblast proliferation, as well as a putative site for pharmacological intervention