Oncogenomic Approaches in Exploring Gain of Function of Mutant p53

Cancer is caused by the spatial and temporal accumulation of alterations in the genome of a given cell. This leads to the deregulation of key signalling pathways that play a pivotal role in the control of cell proliferation and cell fate. The p53 tumor suppressor gene is the most frequent target in genetic alterations in human cancers. The primary selective advantage of such mutations is the elimination of cellular wild type p53 activity. In addition, many evidences in vitro and in vivo have demonstrated that at least certain mutant forms of p53 may possess a gain of function, whereby they contribute positively to cancer progression. The fine mapping and deciphering of specific cancer phenotypes is taking advantage of molecular-profiling studies based on genome-wide approaches. Currently, high-throughput methods such as array-based comparative genomic hybridization (CGH array), single nucleotide polymorphism array (SNP array), expression arrays and ChIP-on-chip arrays are available to study mutant p53-associated alterations in human cancers. Here we will mainly focus on the integration of the results raised through oncogenomic platforms that aim to shed light on the molecular mechanisms underlying mutant p53 gain of function activities and to provide useful information on the molecular stratification of tumor patients

Double role of HMTA in ZnO nanorods grown by chemical bath deposition

ZnO nanorods (NRs) grown by chemical bath deposition (CBD) are among the most promising semiconducting nanostructures currently investigated for a variety of applications. Still, contrasting experimental results appear in the literature on the microscopic mechanisms leading to high aspect ratio and vertically aligned ZnO NRs. Here, we report on CBD of ZnO NRs using Zn nitrate salt and hexamethylenetetramine (HMTA), evidencing a double role of HMTA in the NRs growth mechanism. Beyond the well-established pH buffering activity, HMTA is shown to introduce a strong steric hindrance effect, biasing growth along the c-axis and ensuring the vertical arrangement. This twofold function of HMTA should be taken into account for avoiding detrimental phenomena such as merging or suppression of NRs, which occur at low HMTA concentration

Project “Biodiversity MARE Tricase”: A Species Inventory of the Coastal Area of Southeastern Salento (Ionian Sea, Italy)

Biodiversity is a broad concept that encompasses the diversity of nature, from the genetic to the habitat scale, and ensures the proper functioning of ecosystems. The Mediterranean Sea, one of the world’s most biodiverse marine basins, faces major threats, such as overexploitation of resources, pollution and climate change. Here we provide the first multi-taxa inventory of marine organisms and coastal terrestrial flora recorded in southeastern Salento (Ionian Sea, Italy), realized during the project “Biodiversity MARE Tricase”, which provided the first baseline of species living in the area. Sampling was carried out by SCUBA and free diving, fishing gears, and citizen science from 0 to 70 m. Overall, 697 taxa were found between March 2016 and October 2017, 94% of which were identified to the species level. Of these, 19 taxa represented new records for the Ionian Sea (36 additional new records had been reported in previous publications on specific groups, namely Porifera and Mollusca Heterobranchia), and two findings represented the easternmost records in the Mediterranean Sea (Helicosalpa virgula and Lampea pancerina). For eight other taxa, our findings represented the only locality in the Ionian Sea, besides the Straits of Messina. In addition to the species list, phenological events (e.g., blooms, presence of reproductive traits and behaviour) were also reported, with a focus on gelatinous plankton. Our results reveal that even for a relatively well-known area, current biodiversity knowledge may still be limited, and targeted investigations are needed to fill the gaps. Further research is needed to understand the distribution and temporal trends of Mediterranean biodiversity and to provide baseline data to identify ongoing and future changes

RESCUE OF HIPPO CO-ACTIVATOR YAP1 TRIGGERS DNA DAMAGE-INDUCED APOPTOSIS IN HEMATOLOGICAL CANCERS

Oncogene–induced DNA damage elicits genomic instability in epithelial cancer cells, but apoptosis is blocked through inactivation of the tumor suppressor p53. In hematological cancers, the relevance of ongoing DNA damage and mechanisms by which apoptosis is suppressed are largely unknown. We found pervasive DNA damage in hematologic malignancies including multiple myeloma, lymphoma and leukemia, which leads to activation of a p53–independent, pro-apoptotic network centered on nuclear relocalization of ABL1 kinase. Although nuclear ABL1 triggers cell death through its interaction with the Hippo pathway co–activator YAP1 in normal cells, we show that low YAP1 levels prevent nuclear ABL1–induced apoptosis in these hematologic malignancies. YAP1 is under the control of a serine–threonine kinase, STK4. Importantly, genetic inactivation of STK4 restores YAP1 levels, triggering cell death in vitro and in vivo. Our data therefore identify a novel synthetic–lethal strategy to selectively target cancer cells presenting with endogenous DNA damage and low YAP1 levels

TP63 and TP73 in cancer, an unresolved “family” puzzle of complexity, redundancy and hierarchy

AbstractTP53 belongs to a small gene family that includes, in mammals, two additional paralogs, TP63 and TP73. The p63 and p73 proteins are structurally and functionally similar to p53 and their activity as transcription factors is regulated by a wide repertoire of shared and unique post-translational modifications and interactions with regulatory cofactors. p63 and p73 have important functions in embryonic development and differentiation but are also involved in tumor suppression. The biology of p63 and p73 is complex since both TP63 and TP73 genes are transcribed into a variety of different isoforms that give rise to proteins with antagonistic properties, the TA-isoforms that act as tumor-suppressors and DN-isoforms that behave as proto-oncogenes. The p53 family as a whole behaves as a signaling “network” that integrates developmental, metabolic and stress signals to control cell metabolism, differentiation, longevity, proliferation and death. Despite the progress of our knowledge, the unresolved puzzle of complexity, redundancy and hierarchy in the p53 family continues to represent a formidable challenge

Association of kidney disease measures with risk of renal function worsening in patients with type 1 diabetes

Background: Albuminuria has been classically considered a marker of kidney damage progression in diabetic patients and it is routinely assessed to monitor kidney function. However, the role of a mild GFR reduction on the development of stage 653 CKD has been less explored in type 1 diabetes mellitus (T1DM) patients. Aim of the present study was to evaluate the prognostic role of kidney disease measures, namely albuminuria and reduced GFR, on the development of stage 653 CKD in a large cohort of patients affected by T1DM. Methods: A total of 4284 patients affected by T1DM followed-up at 76 diabetes centers participating to the Italian Association of Clinical Diabetologists (Associazione Medici Diabetologi, AMD) initiative constitutes the study population. Urinary albumin excretion (ACR) and estimated GFR (eGFR) were retrieved and analyzed. The incidence of stage 653 CKD (eGFR < 60 mL/min/1.73 m2) or eGFR reduction > 30% from baseline was evaluated. Results: The mean estimated GFR was 98 \ub1 17 mL/min/1.73m2 and the proportion of patients with albuminuria was 15.3% (n = 654) at baseline. About 8% (n = 337) of patients developed one of the two renal endpoints during the 4-year follow-up period. Age, albuminuria (micro or macro) and baseline eGFR < 90 ml/min/m2 were independent risk factors for stage 653 CKD and renal function worsening. When compared to patients with eGFR > 90 ml/min/1.73m2 and normoalbuminuria, those with albuminuria at baseline had a 1.69 greater risk of reaching stage 3 CKD, while patients with mild eGFR reduction (i.e. eGFR between 90 and 60 mL/min/1.73 m2) show a 3.81 greater risk that rose to 8.24 for those patients with albuminuria and mild eGFR reduction at baseline. Conclusions: Albuminuria and eGFR reduction represent independent risk factors for incident stage 653 CKD in T1DM patients. The simultaneous occurrence of reduced eGFR and albuminuria have a synergistic effect on renal function worsening

Effects of marine heatwaves on temperate sponge physiology and reproduction

Climate change is causing not only a gradual rise in global temperatures but also an increase in the frequency and severity of climate extremes. Marine Heat Waves (MHWs) are events of extreme sea surface temperature that can range from a few days to years, affecting small areas to entire oceans, and their frequency and intensity are expected to increase in the future. Sponges are considered the most ancient metazoans populating global oceans and make an important contribution to biodiversity and ecosystem functioning in shallow, mesophotic and deep ecosystems due to the multiple ecological functions they provide. The occurrence of MHWs has often been associated with sponge mass mortality and bleaching events in tropical, temperate and polar benthic ecosystems, but the biological effects and adaptive responses of sponges to these conditions are largely unknown. Crella incrustans is a poecilosclerid sponge commonly distributed in subtidal habitats of temperate Pacific regions and it is increasingly being considered a model organism for ecophysiological studies. The main goal of this thesis is to assess the biological impacts of MHWs on temperate sponge physiology and reproduction and to assess possible adaptive strategies to overcome these extreme climate events with these specific aims: 1) characterise reproductive period and modality of C. incrustans, including the description of larval ontogeny, release, settlement and metamorphosis; 2) investigate the impacts of near-future extreme temperatures on C. incrustans morphology, physiology and reproductive success; 3) investigate the occurrence of carryover effects of MHW conditions on the microbiome and developmental performance in C. incrustans; 4) to investigate the function of symbiotic bacteria that are vertically transmitted to larvae under MHW conditions in C. incrustans. In my first chapter, I characterised the reproductive modality and the early life stages of C. incrustans. Through histological analyses, I described the gametogenesis and larval ontogeny and using in vivo observations, I characterised larval release, settlement and metamorphosis. C. incrustans presented spermatocytes, oocytes and embryos in the sponge mesohyl during the Austral summer. Non-tufted parenchymella larvae were also released in laboratory conditions during the Austral summer. I found that 94.3% of larvae settled within two days and metamorphosed within a week. My main finding was that C. incrustans is a simultaneous hermaphrodite with asynchronous gametogenesis that releases larvae during the Austral summer, which is consistent with the majority of other poecilosclerid sponges. Importantly, in this chapter, I showed that C. incrustans is a suitable species for ecophysiological studies involving sponge reproduction and early life stages. In my second chapter, I investigated the impact of mean and extreme temperatures predicted to occur within the next 40 years, on the physiology, morphology and recruitment success of C. incrustans. In particular, I experimentally exposed adult sponges to future predicted average sea temperature (+ 2.5° C, according to the SSP3-7.0 scenario) and then simulated a MHW (16 days duration and a thermal peak at 22°C). The main finding was that under predicted average temperatures, C. incrustans did not show any morphological or physiological modifications. However, when exposed to the MHW conditions there was a significant increase in sponge respiration rate, significant weight loss with the occurrence of tissue regression and around 50% mortality. MHW conditions also caused a significantly shorter period of recruitment, a lower recruitment rate and a higher mortality rate of sponge settlers. Interestingly, sponges that survived the simulated MHW showed respiration rates like controls two weeks after the thermal peak, indicating some resilience to MHWs. These results indicate that MHW conditions have a much stronger impact on C. incrustans physiology, morphology and recruitment than future mean temperatures but also that this sponge species is likely to persist under future climate scenarios. In my third chapter, I investigated the occurrence of carryover effects of heatwave conditions on the sponge microbiome and post-settlement developmental features. I experimentally tested the effects of heatwave conditions (10 days at 21°C) on adult and larval microbiomes of C. incrustans. I also tested the effects of parental exposure to heatwave conditions on subsequent settler mortality, growth rate and metamorphosis duration under prolonged heatwave conditions (30 days at 21°C). My main finding was that the microbial community of adult sponges changed significantly after ten days at 21°C, with a significant decrease in symbiotic bacteria (i.e., Arenicellales) and a significant increase in stress-associated bacteria (e.g., Flavobacteriales and Clostridiales). Sponge larvae derived from control sponges were mainly characterised by a few bacterial taxa also abundant in adults (i.e., Arenicellales), confirming the occurrence of vertical transmission. The microbial community of sponge larvae derived from MHW-exposed sponges showed similar levels of Arenicellales to control larvae and a significant increase in endosymbiotic Verrucomicrobia. Settlers derived from heatwave-exposed sponges had better developmental performance, including a greater growth rate and longer metamorphosis period compared to settlers derived from control sponges and exposed to heatwave conditions. These results show the occurrence of MHW-induced carryover effects across life-stages in C. incrustans and suggest the occurrence of selective vertical transmission under MHW conditions as a possible adaptive strategy to overcome extreme thermal events. In my fourth data chapter, I investigated the function of vertically transmitted bacteria, the Arenicellales, under heatwave conditions in C. incrustans. To characterise the genetic potential of vertically transferred bacteria under MHW conditions, I enriched bacterial cells and performed shotgun sequencing of genomic DNA in C. incrustans. My main finding was that the metagenomic assembled genome (MAG) classified as Arenicellales belongs to a new family, genus and species, herein named Candidatus Larvaebacter crellansis. Predicted metabolic pathways revealed that this MAG has the genetic capacity to utilise sponge-derived molecules such as amino acids, aromatic compounds and osmolytes as a source of nutrients. However, it also has the genetic potential to produce molecules required by the sponge host, such as essential vitamins, including cobalamin, which is an important cofactor of several metabolic reactions and to enhance thermal tolerance in unicellular algae. Ca. Larvaebacter crellansis has the genetic potential for the production of terpenes, known for being antifoulants and feeding deterrents in sponges. This bacterium also has the potential for the production of catechol, known to be important for substrate adhesion by marine invertebrates. Considering the great potential of metabolic interactions between this bacterial species and the sponge host, my study supports the occurrence of selective vertical transmission of biologically relevant symbionts under heatwave conditions in C. incrustans. Overall, my thesis shows that MHWs can profoundly affect temperate sponge physiology and reproductive success. Despite that, temperate sponges may show great phenotypic plasticity in response to these extreme temperatures, such as morphological modifications, with the occurrence of tissue regression and selective vertical transmission of biologically relevant bacteria in C. incrustans. In conclusion, despite the detrimental effects of MHWs on temperate sponges, some species are likely to persist under future climate conditions.</p

Inclusive innovation and the role of partnerships: The case of Semi di Libertà

Purpose – This chapter aims to understand how partnerships and networks can aid the development and growth of organizations whose goal is to foster social inclusion along the agro-food supply chain, with particular reference to the social entrepreneurship sector. Methodology/approach – The chapter draws on a framework proposed by Newth and Woods (2014) to identify the main drivers of resistance to the development of social entrepreneurship. The empirical evidence is based on a single case, involving an Italian social enterprise, Semi di Liberta`, which produces high-quality artisan beer. Case material included an analysis of organization documents and interviews with key actors. Findings – The case study shows how Semi di Liberta` faced different types of resistance, related to formal and informal institutions and market drivers, and leveraged partnerships with other actors in the ecosystem. Some of these partnerships were planned a priori to overcome specific problems (e.g., the Prison’s Authority, Mastri Birrai). Other partnerships were developed “by chance” (e.g., “peer” associations) but turned out to be particularly important to deal with the above resistances. Research limitations/implications – The case study methodology prevents the authors from generalizing too far past the obtained results. However, key elements from the case, such as the relevance of “spontaneous” partnerships and those with “peer” organizations, could be taken into account for similar initiatives in different contexts. Originality/value – Recent literature has highlighted the relevance of partnerships in scaling social enterprises but has not explored the dynamics whereby these partnerships are created and developed. This chapter provides some preliminary evidence of how partnerships can be used to overcome the resistance limiting the growth of social entrepreneurship and the sustainability of socially inclusive initiatives