Inhibitors of Apoptosis Proteins (IAPs) as Targets for Anti-Cancer Treatment

Smac mimetics (SMs) constitute a class of compounds that target the inhibitor of apoptosis proteins (IAPs) and enhance the cytotoxic activity of several drugs. In our work, we built and characterized a library of about 140 SMs and focused on SM83 due to its high affinity for the targets, cytotoxic activity and good pharmacokinetic profile. In vivo, SM83 reduced in monotherapy the primary tumor growth of two triple negative breast cancer xenografts. Furthermore, SM83 treatment, alone or in combinations with TRAIL-armed CD34+cell, resulted in the reduction of spontaneous lung metastasis formation. Mechanistically, by depleting cIAP1, SM83 affects the expression of the tumor genes and inhibits the metastasis-promoting gene Snai2, thus preventing cancer cell motility. Moreover, we tested SM83 as a standalone in ascites cancer models and described an in vivo anti-tumor effect against cancer cell lines that are intrinsically resistant to SM treatment in vitro. In the in vivo settings, SM83, in fact, triggered an inflammation event of the host, characterized by macrophage secretion of TNF, IL-1β and interferon-γ (IFNγ), and rapidly killed floating tumor cells within the ascites by a non-apoptotic mechanism. Of note, SM83 treatment caused the massive accumulation of neutrophils within the ascites and tumor nodules, which, however, was not responsible for cancer cell killing. Finally, we described the capability of SM83 to enhance the cytotoxic activity of camptothecin especially in human epithelial cells expressing oncogenic KRAS. The increased sensitivity of these premalignant cells is caused by an ERK2-dependent up-regulation of NOXA. Of note, oncogenic KRAS fails to sensitize a panel of isogenic cancer cell lines with wild type and mutated KRAS, and we demonstrated that this unresponsiveness could be reverted by concomitant inhibition of AKT. Therefore, our work suggests that the activation of AKT is capable of counterbalancing the potential pro-death stimulus triggered by oncogenic KRAS

Social impact assessment of wind power generation. An innovative method for decision making processes

This paper explores the social impact for population in the energy sector combining LCA and SIA (social impact assessment). As case study, a new 66 MW wind power plant under development in the countryside of Southern Sardinia has been considered. The innovative method, based on the analysis of the context, aims to empirically analyze some selected sustainability indicators. The proposed method starts from a detailed analysis of the wind power project, with particular reference to the plant site characteristics, technical features of the wind farm, opinions of the stakeholders, environmental and social impacts and expected economic benefits. The acquired data are validated with a Severity statistical method that identifies the KPIs. The indicators are classified into general categories of damage Human life, Safety guarantee, Social resources, Public participation and analyzed through a combined SIA-LCA method to identify indicators damage weights. This work shows the importance of putting together indicators already explored in the environmental field such as Human health, Ecosystem quality, Resource, Climate Change and as social indicators Renewable Energy with Noise, Visual Impact, Shadow Flichers, the perceptions of the local community

Life Cycle Assessment of an Integrated PV-ACAES System

The aim of this paper is to evaluate the overall life cycle environmental impact of an adiabatic compressed air energy storage (ACAES) system, which is designed to achieve the best match between the power production of a photovoltaic (PV) power plant and the power demand from the final user. The electrical energy demand of a small town, with a maximum power load of about 10 MW, is considered a case study. The ACAES system is designed with a compressor-rated power of about 10 MW and charging and discharging times of 10 and 24 h, respectively. Different sizes of the PV plant, ranging from 20 to 40 MWp, and two different solutions for the compressed air storage, an underground cavern, and a gas pipeline, are analyzed. The aim of this analysis is to compare the impacts on human health, ecosystem quality, climate change, and resource consumption of the PV power generation plant and the integrated PV-ACAES system with those of a reference scenario in which the end user demand is met entirely by the grid. The best results in terms of a reduction in environmental impact in comparison to the reference scenario are obtained for a small PV plant (20 MW) without the ACAES section, with reductions of about 85–95% depending on the category of impact. The integration of the ACAES system improves energy self-consumption but worsens the environmental impact, especially for air storage in gas pipelines. The best configuration in terms of environmental impact is based on a 30 MW PV plant integrated with an ACAES section using an underground cavern for air storage and allows for improvements in the energy self-consumption of between 38% and 61%, with a reduction in the environmental impact compared to the reference scenario of about 80–91% depending on the impact categor

Life Cycle Analysis of a Hydrogen Valley with multiple end-users

This paper aims to evaluate the environmental impact along the overall life cycle of the various components of a Hydrogen Valley with multiple end-users fed by green hydrogen. As case study, a hydrogen valley including a MW-scale electrolyser powered by different percentages of energy supplied by a wind farm and/or a photovoltaic plant, and an H2 storage section is considered. The H2 produced is used to feed a fleet of fuel cell electric vehicles and a stationary fuel cell, while the residue H2 is injected in a natural gas pipeline considering a maximum safety limit of 5%vol. When the safety limit is reached, the H2 overproduction can be used to produce biomethane through a biological hydrogen methanation process. With the aim of analysing the actual contribution of these hydrogen-based ecosystems towards more sustainable energy systems, a Life Cycle Analysis of the hydrogen valley is carried out. The results show that the final use of hydrogen for fuel cell electric vehicles produces the most valuable environmental benefits. Moreover, Hydrogen Valley solutions integrated with photovoltaic plants allows to maximize the use of H2 in fuel cell electric vehicles and therefore are the most valuable choice from an environmental point of view

Antibody-mediated blockade of JMJD6 interaction with collagen I exerts antifibrotic and antimetastatic activities

JMJD6 is known to localize in the nucleus, exerting histone arginine demethylase and lysyl hydroxylase activities. A novel localization of JMJD6 in the extracellular matrix, resulting from its secretion as a soluble protein, was unveiled by a new anti-JMJD6 mAb called P4E11, which was developed to identify new targets in the stroma. Recombinant JMJD6 binds with collagen type I (Coll-I), and distinct JMJD6 peptides interfere with collagen fibrillogenesis, collagen-fibronectin interaction, and adhesion of human tumor cells to the collagen substrate. P4E11 and collagen binding to JMJD6 are mutually exclusive because the amino acid sequences of JMJD6 necessary for the interaction with Coll-I are part of the conformational epitope recognized by P4E11. In mice injected with mouse 4T1 breast carcinoma cells, treatment with P4E11 reduced fibrosis at the primary tumor and prevented lung metastases. Reduction of fibrosis has also been documented in human breast and ovarian tumors (MDA-MB-231 and IGROV1, respectively) xenotransplanted into immunodeficient mice treated with P4E11. In summary, this study uncovers a new localization and function for JMJD6 that is most likely independent from its canonical enzymatic activities, and demonstrates that JMJD6 can functionally interact with Coll-I. P4E11 mAb, inhibiting JMJD6/Coll-I interaction, represents a new opportunity to target fibrotic and tumor diseases

Targeting COPZ1 non-oncogene addiction counteracts the viability of thyroid tumor cells

Thyroid carcinoma is generally associated with good prognosis, but no effective treatments are currently available for aggressive forms not cured by standard therapy. To find novel therapeutic targets for this tumor type, we had previously performed a siRNA-based functional screening to identify genes essential for sustaining the oncogenic phenotype of thyroid tumor cells, but not required to the same extent for the viability of normal cells (non-oncogene addiction paradigm). Among those, we found the coatomer protein complex ζ1 (COPZ1) gene, which is involved in intracellular traffic, autophagy and lipid homeostasis. In this paper, we investigated the mechanisms through which COPZ1 depletion leads to thyroid tumor cell death. We showed that siRNA-mediated COPZ1 depletion causes abortive autophagy, endoplasmic reticulum stress, unfolded protein response and apoptosis. Interestingly, we observed that mouse tumor xenografts, locally treated with siRNA targeting COPZ1, showed a significant reduction of tumor growth. On the whole, we demonstrated for the first time the crucial role of COPZ1 in the viability of thyroid tumor cells, suggesting that it may be considered an attractive target for novel therapeutic approaches for thyroid cancer

Mitochondrial permeabilization engages NF-κB-dependent anti-tumour activity under caspase deficiency

Apoptosis represents a key anti-cancer therapeutic effector mechanism. During apoptosis, mitochondrial outer membrane permeabilization (MOMP) typically kills cells even in the absence of caspase activity. Caspase activity can also have a variety of unwanted consequences that include DNA damage. We therefore investigated whether MOMP-induced caspase-independent cell death (CICD) might be a better way to kill cancer cells. We find that cells undergoing CICD display potent pro-inflammatory effects relative to apoptosis. Underlying this, MOMP was found to stimulate NF-κB activity through the downregulation of inhibitor of apoptosis proteins. Strikingly, engagement of CICD displays potent anti-tumorigenic effects, often promoting complete tumour regression in a manner dependent on intact immunity. Our data demonstrate that by activating NF-κB, MOMP can exert additional signalling functions besides triggering cell death. Moreover, they support a rationale for engaging caspase-independent cell death in cell-killing anti-cancer therapies

EGFR-targeted TRAIL and a Smac mimetic synergize to overcome apoptosis resistance in KRAS mutant colorectal cancer cells

TRAIL is a death receptor ligand that induces cell death preferentially in tumor cells. Recombinant soluble TRAIL, however, performs poorly as an anti-cancer therapeutic because oligomerization is required for potent biological activity. We previously generated a diabody format of tumor-targeted TRAIL termed DbαEGFR-scTRAIL, comprising single-stranded TRAIL molecules (scTRAIL) and the variable domains of a humanized variant of the EGFR blocking antibody Cetuximab. Here we define the bioactivity of DbαEGFR-scTRAIL with regard to both EGFR inhibition and TRAIL receptor activation in 3D cultures of Caco-2 colorectal cancer cells, which express wild-type K-Ras. Compared with conventional 2D cultures, Caco-2 cells displayed strongly enhanced sensitivity toward DbαEGFR-scTRAIL in these 3D cultures. We show that the antibody moiety of DbαEGFR-scTRAIL not only efficiently competed with ligand-induced EGFR function, but also determined the apoptotic response by specifically directing DbαEGFR-scTRAIL to EGFR-positive cells. To address how aberrantly activated K-Ras, which leads to Cetuximab resistance, affects DbαEGFR-scTRAIL sensitivity, we generated stable Caco-2tet cells inducibly expressing oncogenic K-RasG12V. In the presence of doxycycline, these cells showed increased resistance to DbαEGFR-scTRAIL, associated with the elevated expression of the anti-apoptotic proteins cIAP2, Bcl-xL and FlipS. Co-treatment of cells with the Smac mimetic SM83 restored the DbαEGFR-scTRAIL-induced apoptotic response. Importantly, this synergy between DbαEGFR-scTRAIL and SM83 also translated to 3D cultures of oncogenic K-Ras expressing HCT-116 and LoVo colorectal cancer cells. Our findings thus support the notion that DbαEGFR-scTRAIL therapy in combination with apoptosis-sensitizing agents may be promising for the treatment of EGFR-positive colorectal cancers, independently of their KRAS status