Targeting CD47 in Anaplastic Thyroid Carcinoma Enhances Tumor Phagocytosis by Macrophages and Is a Promising Therapeutic Strategy.

Background: Anaplastic thyroid carcinoma (ATC) is one of the most aggressive human cancers, with a median survival of only three to six months. Standard treatment options and even targeted therapies have so far failed to improve long-term overall survival. Thus, novel treatment modalities for ATC, such as immunotherapy, are urgently needed. CD47 is a "don't eat me" signal, which prevents cancer cells from phagocytosis by binding to signal regulatory protein alpha on macrophages. So far, the role of macrophages and the CD47-signal regulatory protein alpha signaling axis in ATC is not well understood. Methods: This study analyzed 19 primary human ATCs for macrophage markers, CD47 expression, and immune checkpoints by immunohistochemistry. ATC cell lines and a fresh ATC sample were assessed by flow cytometry for CD47 expression and macrophage infiltration, respectively. CD47 was blocked in phagocytosis assays of co-cultured macrophages and ATC cell lines. Anti-CD47 antibody treatment was administered to ATC cell line xenotransplanted immunocompromised mice, as well as to tamoxifen-induced ATC double-transgenic mice. Results: Human ATC samples were heavily infiltrated by CD68- and CD163-expressing tumor-associated macrophages (TAMs), and expressed CD47 and calreticulin, the dominant pro-phagocytic molecule. In addition, ATC tissues expressed the immune checkpoint molecules programmed cell death 1 and programmed death ligand 1. Blocking CD47 promoted the phagocytosis of ATC cell lines by macrophages in vitro. Anti-CD47 antibody treatment of ATC xenotransplanted mice increased the frequency of TAMs, enhanced the expression of macrophage activation markers, augmented tumor cell phagocytosis, and suppressed tumor growth. In double-transgenic ATC mice, CD47 was expressed on tumor cells, and blocking CD47 increased TAM frequencies. Conclusions: Targeting CD47 or CD47 in combination with programmed cell death 1 may potentially improve the outcomes of ATC patients and may represent a valuable addition to the current standard of care

Metrics for short-term coastal characterization, protection and planning decisions of Sentina Natural Reserve, Italy

Geomorphological and sedimentological surveys of the emerged and submerged beach-dune system are fundamental for a successful management and protection strategies for coastal planning and development. In particular, these surveys can reveal if coastal areas are affected by erosion, pollution and loss of habitats under the seasonal anthropic pressure related to tourism, leisure and professional fishing, urbanisation and/or other activities impacting the coastal marine resources. In the present study we discuss the results of the multidisciplinary monitoring activities carried out within the Sentina Natural Reserve (Municipality of San Benedetto del Tronto, Adriatic side of Central Italy, at the southeastern end of the Marche Region) proposing an exportable methodological approach. Due to the absence of buildings, the study area has not been considered in the coastal protection plan by regional and local authorities and, as a consequence, it is currently exposed to severe coastal erosion, the rate of which has been more precisely determined during the present study. This monitoring testifies that most of the seaward surface of the beach disappeared resulting in a general set back of the whole beach environment. In the last decades, several restoration strategies have been adopted to protect and restore the dunes and the back dunes habitats and the municipality also carried out an emergency action to nourish the beach, including the use of sand dredged from the nearby city harbor. With this sediment management approach, a tradeoff between safety of navigation of harbor inlet and habitat conservation of Natural Reserve have been reached, since beach nourishment can reduce coastal vulnerability and risk, even though its sustainability in the long term is still debated

Genetic basis of congenital upper limb anomalies: analysis of 487 cases of a specialized clinic

Background: Specific data regarding the frequencies of the congenital upper limb anomalies (CULA) according to their aetiology are hardly available due to the heterogeneity across classification systems. In this study, we aim at defining the aetiology of CULA of the patients attending Modena University Hospital’s Congenital Hand Malformations multidisciplinary clinic in the years 2004-2012. Methods: Medical records of 487 patients were retrospectively reviewed. On the basis of clinical, anamnestic and genetic data the CULA were distributed into two main groups: (1) non-mendelian aetiology, including prenatal exposure, somatic mutations and amniotic bands and (2) mendelian aetiology including single gene and genomic/chromosomal diseases. CULA were further grouped according to the embryological damage (formation, separation and growth defects) and to the involved axis (radial, ulnar, central). Results: A mendelian aetiology was diagnosed in 199 patients (40.9%), whereas the remaining 288 cases (59.1%) were described as non-mendelian. The involvement of the lower limbs, the presence of malformations in other organs and facial dysmorphisms were significantly more represented in the mendelian cases. The formation defects were significantly more frequent in the non-mendelian group (p<0.001), whereas the frequency of separation defects was higher in the mendelian cases (p=0.0025). Patients with non-mendelian aetiologies showed a significantly higher frequency of central defects (p=0.0031). Conclusion: The two aetiologies differ in terms of patient’s clinical features, morphology defect and axis involvement. These data may be helpful in the diagnostic workup for the indication to perform genetic testing and for the recurrence risk assessment

Membrane lipids are key-modulators of the endocannabinoid-hydrolase FAAH

Lipid composition is expected to play an important role in modulating membrane enzyme activity, in particular if the substrates are themselves lipid molecules. A paradigmatic case is fatty acid amide hydrolase (FAAH), a critical enzyme in terminating the endocannabinoid signalling and an important therapeutic target. Here, using a combined experimental and computational approach, we show that membrane lipids modulate structure, subcellular localization and activity of FAAH. We report that FAAH dimer is stabilized by the lipid bilayer and shows higher membrane binding affinity and enzymatic activity within membranes containing both cholesterol and the natural FAAH substrate, anandamide (AEA). Additionally, colocalization of cholesterol, AEA, and FAAH in mouse neuroblastoma cells suggests a mechanism through which cholesterol increases the substrate accessibility of FAAH

The endocannabinoid system shows different alterations in in vitro and in vivo models of Huntington’s disease

Introduction: In this investigation we analyzed the main components of the so-called “endocannabinoid system” (ECS) in R6/2 mice, a widely used model of Huntington’s disease (HD). Methods: We measured the endogenous content of anandamide (AEA) and 2-arachidonoylglycerol (2-AG), of their biosynthetic (NAPE-PLD and DAGL, respectively) and hydrolytic enzymes (FAAH and MAGL, respectively), and of their target receptors (CB1, CB2 and TRPV1) in the brain of wild-type and R6/2 mice of different ages. In addition, we measured FAAH activity in lymphocytes of R6/2 mice, in order to evaluate whether central ECS alterations were mirrored by peripheral cells. Results: In 12-week-old R6/2 mice we found a reduction of NAPE-PLD and DAGL activity, and of CB binding, as well as an increase in 2-AG content when compared to wild-type littermates, without any other change in ECS elements. Our analysis was extended to HD43 cells, an inducible cellular model of HD derived from rat ST14A cells. In both induced and non-induced conditions we demonstrated a fully functional ECS, and we showed that HD43 cells replicate the decrease in FAAH activity (half of that measured in ST14A cells) already observed in human brain and lymphocytes of HD patients. Conclusions: Overall, our data suggest that ECS is differently affected in mouse and human HD, and that HD43 cells are suitable for high throughput screening of FAAH-oriented drugs affecting HD progression. [...

as manuscript BJ20130960 THIS IS NOT THE VERSION OF RECORD -see Biochemical Journal Immediate Publication

Abstract Lipid composition is expected to play an important role in modulating membrane enzyme activity, in particular if the substrates are themselves lipid molecules. A paradigmatic case is fatty acid amide hydrolase (FAAH), a critical enzyme in terminating the endocannabinoid signalling and an important therapeutic target. Here, using a combined experimental and computational approach, we show that membrane lipids modulate structure, subcellular localization and activity of FAAH. We report that FAAH dimer is stabilized by the lipid bilayer and shows higher membrane binding affinity and enzymatic activity within membranes containing both cholesterol and the natural FAAH substrate, anandamide (AEA). Additionally, colocalization of cholesterol, AEA, and FAAH in mouse neuroblastoma cells suggests a mechanism through which cholesterol increases the substrate accessibility of FAAH. Keywords: cholesterol/endocannabinoids/FAAH/membrane Summary statement The dimeric structure of FAAH is stabilized by the membrane. Membrane binding affinity of FAAH is relevant for subcellular localization. Free cholesterol within the membrane increases the activity of the enzyme. Molecular dynamic studies suggest that cholesterol increases substrate accessibility. INTRODUCTION Fatty acid amide hydrolase (FAAH) is a membrane-bound enzyme that is responsible for the intracellular hydrolysis of the bioactive lipid anandamide (N-arachidonoylethanolamine, AEA) and other congeners known as endocannabinoids (eCBs) Membrane lipid composition affects eCB uptake and signalling, and accumulated evidence demonstrates that cholesterol is a key determinant of this regulation With the aim of dissecting the requirements for the catalytic activity and those for the enzyme interaction with membranes, we analysed by small angle X-ray scattering (SAXS) and fluorescence resonance energy transfer (FRET) the conformational changes induced by lipid bilayers on FAAH. Both using synthetic or reconstructed lipid vesicles from different cell compartments (i.e., plasma membrane (PM) or endoplasmic reticulum (ER)), we demonstrated a key-role of membrane lipids in stabilizing a dimeric form of FAAH with cholesterol and AEA, that both modulate its enzymatic activity within the membrane. Furthermore, molecular dynamics simulations supported a novel mechanism by which cholesterol may help to open the membrane port of FAA