Publications by authors named "Adelaide Greco"

50 Publications

The Use of Platelet-Rich Plasma for Treatment of Tenodesmic Lesions in Horses: A Systematic Review and Meta-Analysis of Clinical and Experimental Data.

Animals (Basel) 2021 Mar 12;11(3). Epub 2021 Mar 12.

Department of Veterinary Medicine and Animal Production, University of Napoli "Federico II", Via Federico Delpino 1, 80137 Napoli, Italy.

The use of platelet-rich plasma (PRP) to enhance tenodesmic lesion healing has been questioned over the years. The aim of this study was to evaluate current literature to establish the effectiveness of PRP for treating tenodesmic lesions through a systematic review, in accordance with the PRISMA guidelines, and a meta-analysis. Studies comparing PRP with placebo or other treatments for horses with tenodesmic injuries or evaluated PRP effect on tendon and ligament explants were included. Outcomes were clinical, ultrasound, histologic, molecular evaluation, and adverse effects. Two authors independently extracted data and assessed each study's risk of bias. Treatment effects were evaluated using risk ratios for dichotomous data, together with 95% CI. Data were pooled using the random-effects model. The quality of the evidence for each outcome was assessed using GRADE criteria. Twenty-four trials met inclusion criteria for systematic review, while fifteen studies were included in the meta-analysis. Results showed no significant differences in the outcomes between PRP and control groups. Finally, there is no definitive evidence that PRP enhances tendons and ligaments healing. Therefore, there is a need for more controlled trials to draw a firmer conclusion about the efficacy of PRP as a treatment for tenodesmic lesions in the horse.
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http://dx.doi.org/10.3390/ani11030793DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7998797PMC
March 2021

Ultrasonographic measurement of kidney-to-aorta parameters in Whippets.

Vet Radiol Ultrasound 2021 Feb 7. Epub 2021 Feb 7.

Interdepartmental Center of Veterinary Radiology, University of Napoli "Federico II", Napoli, Italy.

In a previous study, an ultrasonographic method to assess kidney size in dogs as a ratio of kidney length to aortic luminal diameter (KL/AoD ratio) was proposed. The main limitation of this method was the wide range of normal values (5.5-9.1), which resulted in poor sensitivity and specificity. The aim of this prospective, observational, reference interval study was to determine whether the KL/AoD normal cut-off values in a single breed (Whippets) would have a narrower range than the previously reported normal reference ranges. The influence of sex, age, weight, and side on kidney length (KL) and of sex, age, weight, and scanning plane (longitudinal vs transversal) on aortic luminal diameter (AoD) were also investigated. Thirty-six clinically healthy Whippets (16 males, 20 females) without ultrasonographic renal lesions were included in this study. The 95% confidence interval of mean KL/AoD was found to be narrower than the previously reported range (ie, 6.3-6.9 versus 5.5-9.1). This was considered to be especially notable in that the KL in this breed exhibits marked sexual dimorphism. The KL/AoD ratio did not differ between right versus left sides or male versus female sexes in Whippets (P > .05). Findings from the current study provided KL/AoD ratio normal reference range cut-off values for future use in Whippets and supported the use of breed-specific KL/AoD ratio values for characterizing abnormal renal size in other canine breeds.
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http://dx.doi.org/10.1111/vru.12958DOI Listing
February 2021

Gene Dosage Affects Thyroid Cancer Histology and Differentiation In Vivo.

Int J Mol Sci 2020 Dec 22;22(1). Epub 2020 Dec 22.

Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, via Pansini 5, 80131 Naples, Italy.

The transcription factor Forkhead box E1 () is a key player in thyroid development and function and has been identified by genome-wide association studies as a susceptibility gene for papillary thyroid cancer. Several cancer-associated polymorphisms fall into gene regulatory regions and are likely to affect expression levels. However, the possibility that changes in expression modulate thyroid cancer development has not been investigated. Here, we describe the effects of gene dosage reduction on cancer phenotype in vivo. Mice heterozygous for null allele () were crossed with a -inducible cancer model to develop thyroid cancer in either a or genetic background. In mice, cancer histological features are quite similar to that of human high-grade papillary thyroid carcinomas, while cancers developed with reduced gene dosage maintain morphological features resembling less malignant thyroid cancers, showing reduced proliferation index and increased apoptosis as well. Such cancers, however, appear severely undifferentiated, indicating that levels affect thyroid differentiation during neoplastic transformation. These results show that dosage exerts pleiotropic effects on thyroid cancer phenotype by affecting histology and regulating key markers of tumor differentiation and progression, thus suggesting the possibility that could behave as lineage-specific oncogene in follicular cell-derived thyroid cancer.
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http://dx.doi.org/10.3390/ijms22010025DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7792778PMC
December 2020

Ultrasound-guided percutaneous antegrade pyelography for suspected ureteral obstruction in 6 pet guinea pigs ().

Vet Q 2020 Dec;40(1):198-204

Interdepartmental Centre of Veterinary Radiology, University of Naples Federico II, Naples, Italy.

Objectives: To describe the feasibility and safety of ultrasound-guided percutaneous antegrade pyelography (US-PAP) in pet guinea pigs () with suspected ureteral obstruction.

Materials And Methods: Six adult pet guinea pigs (4 females and 2 males, all intact) were evaluated for suspected ureteral obstruction. The mean weight of the guinea pigs was 0.8 ± 0.25 kg (range 0.4-1.1 kg), and mean age was 4.07 ± 1.63 years (range 2-7 years). All animals were free from comorbid diseases, had clinical signs of urologic disease and were referred based on either strong clinical suspicion of, or diagnostic imaging of ureteral obstruction. Data on signalment and clinical examination findings, response to anaesthesia and imaging findings were recorded.

Results: Partial ureteral obstruction was confirmed in all guinea pigs but one, in which a complete ureteral obstruction occurred. Uroliths were in both ureters of 5 cases and in both the left renal pelvis and ureters in 1 case. All guinea pigs showed a normal appetite and regular defaecation within 2 h following the procedure. No intraoperative or immediate postoperative complications were encountered after the procedure. The only complication was contrast medium leakages in the subcapsular perinephric, retroperitoneal and, in one case, peritoneal space, which caused no overt clinical consequences afterwards. In one male patient, mobilisation of the ureteral calculus occurred and the urolith was found in the urinary bladder on the radiograph taken after contrast medium injection.

Clinical Significance: The US-PAP technique is a useful, safe and easy-to-perform diagnostic tool in guinea pigs with hydronephrosis and hydroureter.
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http://dx.doi.org/10.1080/01652176.2020.1803512DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7476506PMC
December 2020

New CXCR4 Antagonist Peptide R (Pep R) Improves Standard Therapy in Colorectal Cancer.

Cancers (Basel) 2020 Jul 18;12(7). Epub 2020 Jul 18.

Research Department, Microenvironment Molecular Targets, Istituto Nazionale Tumori IRCCS "Fondazione G. Pascale", 80131 Napoli, Italy.

The chemokine receptor CXCR4 is overexpressed and functional in colorectal cancer. To investigate the role of CXCR4 antagonism in potentiating colon cancer standard therapy, the new peptide CXCR4 antagonist Peptide R (Pep R) was employed. Human colon cancer HCT116 xenograft-bearing mice were treated with chemotherapeutic agents (CT) 5-Fluorouracil (5FU) and oxaliplatin (OX) or 5FU and radio chemotherapy (RT-CT) in the presence of Pep R. After two weeks, CT plus Pep R reduced by 4-fold the relative tumor volume (RTV) as compared to 2- and 1.6-fold reductions induced, respectively, by CT and Pep R. In vitro Pep R addition to CT/RT-CT impaired HCT116 cell growth and further reduced HCT116 and HT29 clonal capability. Thus, the hypothesis that Pep R could target the epithelial mesenchyme transition (EMT) process was evaluated. While CT decreased ECAD and increased ZEB-1 and CD90 expression, the addition of Pep R restored the pretreatment expression. In HCT116 and HT29 cells, CT/RT-CT induced a population of CD133+CXCR4+ cells, supposedly a stem-resistant cancer cell population, while Pep R reduced it. Taken together, the results showed that targeting CXCR4 ameliorates the effect of treatment in colon cancer through inhibition of cell growth and reversal of EMT treatment-induced markers, supporting further clinical studies.
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http://dx.doi.org/10.3390/cancers12071952DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7409147PMC
July 2020

Junctional adhesion molecule-A is down-regulated in anaplastic thyroid carcinomas and reduces cancer cell aggressiveness by modulating p53 and GSK3 α/β pathways.

Mol Carcinog 2019 07 4;58(7):1181-1193. Epub 2019 Mar 4.

IRCCS SDN, Napoli, Italy.

Junctional adhesion molecule A (JAM-A) is a transmembrane protein that contributes to different biological process, including the epithelial to mesenchymal transition (EMT). Through an EMT profiler array, we explored the molecular players associated with human thyroid cancer progression and identified JAM-A as one of the genes mostly deregulated. The quantitative real-time polymerase chain reaction and immunohistochemistry analyses showed that downregulation of JAM-A occurred in anaplastic thyroid carcinoma (ATC) compared with normal thyroid (NT) and papillary thyroid carcinoma (PTC) tissues and correlated with extrathyroid infiltration, tumor size, and ATC histotype. In ATC cell lines, JAM-A restoration suppressed malignant hallmarks of transformation including cell proliferation, motility, and transendothelial migration. Accordingly, knockdown of JAM-A enhanced thyroid cancer cell proliferation and motility in PTC cells. Through the proteome profiler human phospho-kinase array, we demonstrated that higher expression of JAM-A was associated with a significant increased level of phosphorylation of p53 and GSK3 α/β proteins. In conclusion, our findings highlight a novel role of JAM-A in thyroid cancer progression and suggest that JAM-A restoration could have potential clinical relevance in thyroid cancer treatment.
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http://dx.doi.org/10.1002/mc.23001DOI Listing
July 2019

Helper-dependent adenovirus-mediated gene transfer of a secreted LDL receptor/transferrin chimeric protein reduces aortic atherosclerosis in LDL receptor-deficient mice.

Gene Ther 2019 04 30;26(3-4):121-130. Epub 2019 Jan 30.

CEINGE-Biotecnologie Avanzate, Napoli, Italy.

Familial hypercholesterolemia (FH) is a genetic hyperlipidemia characterized by elevated concentrations of plasma LDL cholesterol. Statins are not always effective for the treatment of FH patients; unresponsive patients have poor prognosis and rely on LDL apheresis. In the past, we developed safe and effective gene therapy strategies for the expression of anti-atherogenic proteins using PEGylated helper-dependent adenoviral (HD-Ad) vectors. We recently developed a HD-Ad vector for the expression of the soluble form of the extracellular portion of the human LDL receptor (LDLR) fused with a rabbit transferrin dimer (LDLR-TF). We evaluated the efficacy of the LDLR-TF chimeric protein  in CHOLDLA7, a cell line lacking LDLR expression, restoring the ability to uptake LDL. Subsequently, we administered intravenously 1 × 10E13 vp/kg of this vector in LDLR-deficient mice and observed amelioration of lipid profile and reduction of aortic atherosclerosis. Finally, we studied LDL distribution after HD-Ad vector-mediated expression of LDLR-TF in LDLR-deficient mice and found LDL accumulation in liver, and in heart and intestine. These results support the possibility of lowering LDL-C levels and reducing aortic atherosclerosis using a secreted therapeutic transgene; the present strategy potentially can be modified and adapted to non-systemic gene transfer with expression of the secreted chimeric protein in muscle or other tissues. Intramuscular or local administration strategies could improve the safety profile of this strategy and facilitate applicability.
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http://dx.doi.org/10.1038/s41434-019-0061-zDOI Listing
April 2019

Preclinical Imaging for the Study of Mouse Models of Thyroid Cancer.

Int J Mol Sci 2017 Dec 16;18(12). Epub 2017 Dec 16.

Istituto di Biostrutture e Bioimmagini, Consiglio Nazionale delle Ricerche-IBB, CNR, 80145 Napoli, Italy.

Thyroid cancer, which represents the most common tumors among endocrine malignancies, comprises a wide range of neoplasms with different clinical aggressiveness. One of the most important challenges in research is to identify mouse models that most closely resemble human pathology; other goals include finding a way to detect markers of disease that common to humans and mice and to identify the most appropriate and least invasive therapeutic strategies for specific tumor types. Preclinical thyroid imaging includes a wide range of techniques that allow for morphological and functional characterization of thyroid disease as well as targeting and in most cases, this imaging allows quantitative analysis of the molecular pattern of the thyroid cancer. The aim of this review paper is to provide an overview of all of the imaging techniques used to date both for diagnosis and theranostic purposes in mouse models of thyroid cancer.
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http://dx.doi.org/10.3390/ijms18122731DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5751332PMC
December 2017

Multimodal imaging for a theranostic approach in a murine model of B-cell lymphoma with engineered nanoparticles.

Nanomedicine 2018 02 22;14(2):483-491. Epub 2017 Nov 22.

Dipartimento di Scienze Biomediche Avanzate, Università degli Studi di Napoli Federico II, Napoli, Italia; IBB, CNR, Napoli, Italia; CEINGE Biotecnologie Avanzate, s.c.ar.l., Napoli, Italia. Electronic address:

Nanoparticles (NPs) are a promising tool for in vivo multimodality imaging and theranostic applications. Hyaluronic acid (HA)-based NPs have numerous active groups that make them ideal as tumor-targeted carriers. The B-lymphoma neoplastic cells express on their surfaces a clone-specific immunoglobulin receptor (Ig-BCR). The peptide A20-36 (pA20-36) selectively binds to the Ig-BCR of A20 lymphoma cells. In this work, we demonstrated the ability of core-shell chitosan-HA-NPs decorated with pA20-36 to specifically target A20 cells and reduce the tumor burden in a murine xenograft model. We monitored tumor growth using high-frequency ultrasonography and demonstrated targeting specificity and kinetics of the NPs via in vivo fluorescent reflectance imaging. This result was also confirmed by ex vivo magnetic resonance imaging and confocal microscopy. In conclusion, we demonstrated the ability of NPs loaded with fluorescent and paramagnetic tracers to act as multimodal imaging contrast agents and hence as a non-toxic, highly specific theranostic system.
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http://dx.doi.org/10.1016/j.nano.2017.11.016DOI Listing
February 2018

Mouse models of neurodegenerative disease: preclinical imaging and neurovascular component.

Brain Imaging Behav 2018 Aug;12(4):1160-1196

IBB, CNR, Via T. De Amicis 95, 80145, Napoli, Italy.

Neurodegenerative diseases represent great challenges for basic science and clinical medicine because of their prevalence, pathologies, lack of mechanism-based treatments, and impacts on individuals. Translational research might contribute to the study of neurodegenerative diseases. The mouse has become a key model for studying disease mechanisms that might recapitulate in part some aspects of the corresponding human diseases. Neurodegenerative disorders are very complicated and multifactorial. This has to be taken in account when testing drugs. Most of the drugs screening in mice are very difficult to be interpretated and often useless. Mouse models could be condiderated a 'pathway models', rather than as models for the whole complicated construct that makes a human disease. Non-invasive in vivo imaging in mice has gained increasing interest in preclinical research in the last years thanks to the availability of high-resolution single-photon emission computed tomography (SPECT), positron emission tomography (PET), high field Magnetic resonance, Optical Imaging scanners and of highly specific contrast agents. Behavioral test are useful tool to characterize different animal models of neurodegenerative pathology. Furthermore, many authors have observed vascular pathological features associated to the different neurodegenerative disorders. Aim of this review is to focus on the different existing animal models of neurodegenerative disorders, describe behavioral tests and preclinical imaging techniques used for diagnose and describe the vascular pathological features associated to these diseases.
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http://dx.doi.org/10.1007/s11682-017-9770-3DOI Listing
August 2018

Inhibition of Bone Marrow-Derived Mesenchymal Stem Cells Homing Towards Triple-Negative Breast Cancer Microenvironment Using an Anti-PDGFRβ Aptamer.

Theranostics 2017 22;7(14):3595-3607. Epub 2017 Aug 22.

Istituto di Biostrutture e Bioimmagini-CNR, Naples, Italy.

Bone marrow-derived mesenchymal stem cells (BM-MSCs) are shown to participate in tumor progression by establishing a favorable tumor microenvironment (TME) that promote metastasis through a cytokine networks. However, the mechanism of homing and recruitment of BM-MSCs into tumors and their potential role in malignant tissue progression is poorly understood and controversial. Here we show that BM-MSCs increase aggressiveness of triple-negative breast cancer (TNBC) cell lines evaluated as capability to migrate, invade and acquire stemness markers. Importantly, we demonstrate that the treatment of BM-MSCs with a nuclease-resistant RNA aptamer against platelet-derived growth factor receptor β (PDGFRβ) causes the inhibition of receptor-dependent signaling pathways thus drastically hampering BM-MSC recruitment towards TNBC cell lines and BM-MSCs trans-differentiation into carcinoma-associated fibroblast (CAF)-like cells. Moreover, molecular imaging analysis demonstrated the aptamer ability to prevent BM-MSCs homing to TNBC xenografts. Collectively, our results indicate the anti-PDGFRβ aptamer as a novel therapeutic tool to interfere with BM-MSCs attraction to TNBC providing the rationale to further explore the aptamer in more complex pre-clinical settings.
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http://dx.doi.org/10.7150/thno.18974DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5596446PMC
July 2018

The chemokine scavenging receptor D6/ACKR2 is a target of miR-146a in thyroid cancer.

Genes Cancer 2017 May;8(5-6):577-588

Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Federico II University, Naples, Italy.

We have previously shown that miR-146a, a NF-κB-regulated microRNA, is strongly expressed in human specimens and cell lines derived from anaplastic thyroid carcinomas (ATC) where it mediates some of the NF-κB pro-tumorigenic functions. By using a bioinformatic analysis, we identified the chemokine scavenger receptor D6/ ACKR2 as a target of miR146a in human ATC. We found that the expression of D6/ ACKR2 was up-regulated in miR-146a-null ATC cell lines and that the 3' UTR of D6/ ACKR2 mRNA was able to inhibit its expression in parental, but not in miR-146a-null ATC cells. Since human specimens from primary ATC showed a low expression of D6/ ACKR2 compared to normal thyroid tissues, we analyzed the effects of D6/ACKR2 over-expression in ATC cells. Different chemokines added to the conditioned medium of D6/ACKR2 over-expressing ATC cells partially failed to drive monocyte migration, and tumors derived from the injection of the same cells in nude mice showed a decreased number of infiltrating macrophages. Taken together, these results indicate that ATC cells down-regulate D6/ACKR2 expression through miR-146a activity to sustain leukocyte trafficking inside tumor microenvironment and shed light on a novel mechanism by which NF-κB indirectly inhibits the expression and the function of anti-tumorigenic gene in thyroid cancer.
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http://dx.doi.org/10.18632/genesandcancer.141DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5511891PMC
May 2017

A novel CXCR4-targeted near-infrared (NIR) fluorescent probe (Peptide R-NIR750) specifically detects CXCR4 expressing tumors.

Sci Rep 2017 05 31;7(1):2554. Epub 2017 May 31.

Functional Genomics, Istituto Nazionale per lo Studio e la Cura dei Tumori, Fondazione "G. Pascale"-IRCCS, Napoli, Italy.

C-X-C chemokine receptor 4 (CXCR4) is over-expressed in multiple human cancers and correlates with tumor aggressiveness, poor prognosis and increased risk for distant metastases. Imaging agents for CXCR4 are thus highly desirable. We developed a novel CXCR4-targeted near-infrared (NIR) fluorescent probe (Peptide R-NIR750) conjugating the new developed CXCR4 peptidic antagonist Peptide R with the NIR fluorescent dye VivoTag-S750. Specific CXCR4 binding was obtained in cells overexpressing human CXCR4 (B16-hCXCR4 and human melanoma cells PES43), but not in CXCR4 low expressing cells (FB-1). Ex vivo evaluation demonstrated that PepR-NIR750 specifically detects B16-hCXCR4-derived subcutaneous tumors and lung metastases. Fluorescence Molecular Tomography (FMT) in vivo imaging was performed on mice carrying subcutaneous CHO and CHO-CXCR4 tumors. PepR-NIR750 accumulates only in CXCR4-positive expressing subcutaneous tumors. Additionally, an intense NIR fluorescence signal was detected in PES43-derived lung metastases of nude mice injected with PepR-NIR750 versus mice injected with VivoTag-S750. With a therapeutic intent, mice bearing PES43-derived lung metastases were treated with Peptide R. A the dramatic reduction in PES43-derived lung metastases was detected through a decrease of the PepR-NIR750 signal. PepR-NIR750 is a specific probe for non-invasive detection of human high CXCR4-expressing tumors and metastatic lesion and thus a valuable tool for cancer molecular imaging.
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http://dx.doi.org/10.1038/s41598-017-02818-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5451476PMC
May 2017

Novel Approach for Evaluation of Protective Role against Liver Damage in Immunocompromised Murine Model.

Front Microbiol 2016 7;7:1750. Epub 2016 Nov 7.

Department of Molecular Medicine and Medical Biotechnology, Federico II University Medical SchoolNaples, Italy; CEINGE-Advanced BiotechnologiesNaples, Italy.

is a gram-negative facultative intracellular bacterium and is the causative agent of cat-scratch disease. Our previous data have established that colonization is able to prevent damages through the polysaccharide A (PSA) in an experimental murine model. In order to determine whether the PSA is essential for the protection against pathogenic effects of in immunocompromised hosts, SCID mice were co-infected with wild type or its mutant ΔPSA and the effects of infection on murine tissues have been observed by High-Frequency Ultrasound (HFUS), histopathological examination, and Transmission Electron Microscopy (TEM). For the first time, echostructure, hepatic lobes length, vascular alterations, and indirect signs of hepatic dysfunctions, routinely used as signs of disease in humans, have been analyzed in an immunocompromised murine model. Our findings showed echostructural alterations in all infected mice compared with the Phosphate Buffer Solution (PBS) control group; further, those infected with and co-infected with ΔPSA presented the major echostructural alterations. Half of the mice infected with and all those co-infected with ΔPSA have showed an altered hepatic echogenicity compared with the renal cortex. The echogenicity score of co-infected mice with ΔPSA differed significantly compared with the PBS control group (p < 0.05). Moreover the inflammation score of the histopathological evaluation was fairly concordant with ultrasound findings. Ultrastructural analysis performed by TEM revealed no significant alterations in liver samples of SCID mice infected with wild type while those infected with ΔPSA showed the presence of collagen around the main vessels compared with the PBS control group. The liver samples of mice infected with showed macro-areas rich in collagen, stellate cells, and histiocytic cells. Interestingly, our data demonstrated that immunocompromised SCID mice infected with and co-infected with ΔPSA showed the most severe morpho-structural liver damage. In addition, these results suggests that the HFUS together with histopathological evaluation could be considered good imaging approach to evaluate hepatic alterations.
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http://dx.doi.org/10.3389/fmicb.2016.01750DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5097911PMC
November 2016

Advances in molecular preclinical therapy mediated by imaging.

Q J Nucl Med Mol Imaging 2017 Mar 18;61(1):76-94. Epub 2016 Nov 18.

Department of BioMolecular Sciences, National Center for Natural Products Research, University of Mississippi, Jackson, MS, USA -

Several advances have been made toward understanding the biology of cancer and most of them are due to robust genetic studies that led to the scientific recognition that although many patients have the same type of cancer their tumors may have harbored different molecular alterations. Personalized therapy and the development of advanced techniques of preclinical imaging and new murine models of disease are emerging concepts that are allowing mapping of disease markers in vivo and in some cases also receptor targeted therapy. Aim of this review is to illustrate some emerging models of disease that allow patient tumor implantation in mice for subsequent drug testing and advanced approaches for therapy mediated by preclinical imaging. In particular we discuss targeted therapy mediated by high frequency ultrasound and magnetic resonance, two emerging techniques in molecular preclinical therapy.
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http://dx.doi.org/10.23736/S1824-4785.16.02944-7DOI Listing
March 2017

Advances in multimodal molecular imaging.

Q J Nucl Med Mol Imaging 2017 Mar 18;61(1):19-32. Epub 2016 Nov 18.

Institute of Biostructures and Bioimaging, National Research Council, Naples, Italy -

Preclinical molecular imaging is an emerging field. Improving the ability of scientists to study the molecular basis of human pathology in animals is of the utmost importance for future advances in all fields of human medicine. Moreover, the possibility of developing new imaging techniques or of implementing old ones adapted to the clinic is a significant area. Cardiology, neurology, immunology and oncology have all been studied with preclinical molecular imaging. The functional techniques of photoacoustic imaging (PAI), fluorescence molecular tomography (FMT), positron emission tomography (PET), and single photon emission computed tomography (SPECT) in association with each other or with the anatomic reference provided by computed tomography (CT) as well as with anatomic and functional information provided by magnetic resonance (MR) have all been proficiently applied to animal models of human disease. All the above-mentioned imaging techniques have shown their ability to explore the molecular mechanisms involved in animal models of disease. The clinical translatability of most of the techniques motivates the ongoing study of their possible fields of application. The ability to combine two or more techniques allows obtaining as much information as possible on the molecular processes involved in pathologies, reducing the number of animals necessary in each experiment. Merging molecular probes compatible with various imaging technique will further expand the capability to achieve the best results.
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http://dx.doi.org/10.23736/S1824-4785.16.02943-5DOI Listing
March 2017

Original Research: Feasibility and safety of two surgical techniques for the development of an animal model of jugular vein occlusion.

Exp Biol Med (Maywood) 2017 Jan 5;242(1):22-28. Epub 2016 Jul 5.

IBB, CNR, Napoli 80145, Italy.

To date, no studies have explored the effect of abnormal cerebral venous circulation on brain disorders, whereas many studies have investigated neurodegenerative brain anomalies associated with arterial diseases. The aim of our study was to demonstrate the feasibility of different surgical techniques to induce venous obstruction of cerebral brain drainage. Six C57/black mice underwent bilateral occlusion of the external jugular vein (group EJV), six underwent bilateral occlusion of the internal jugular vein (group IJV), and six underwent bilateral occlusion of both the EJV and the IJV (group EJV/IJV). Within each group, the interruption of blood flow was obtained via monopolar electro-coagulation (ME) in three mice and via surgical ligation (SL) in the remaining three mice. A "sham group" of two mice was used as the control. High-frequency ultrasound (HFUS) was used to detect the absence of blood flow in the examined vessel. The ME procedure led to successful results in two of nine (22%) mice, one in the EJV group, one in the EJV/IJV group, and zero in the IJV group, and 4 of 18 (22%) mice when considering individual veins (i.e., total number of EJVs and IJVs occluded). The SL procedure was successful in two of three (67%) mice in the EJV group, in three of three (100%) mice in the IJV and in three of four (75%) mice in the EJV/IJV group. Therefore, the overall success rate was 8/10 (80%) when considering mice, and 20/26 (77%) when considering individual veins. The monopolar electro-coagulation method exhibited a high mortality due to cardiorespiratory arrest, while the results of the bilateral surgical ligation of EJVs and IJVs show that it is technically feasible and safe.
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http://dx.doi.org/10.1177/1535370216657446DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5206976PMC
January 2017

Intravenous immune globulin suppresses angiogenesis in mice and humans.

Signal Transduct Target Ther 2016;1. Epub 2016 Jan 28.

Department of Ophthalmology and Visual Sciences, University of Kentucky, Lexington, KY, USA; Department of Physiology, University of Kentucky, Lexington, KY, USA.

Human intravenous immune globulin (IVIg), a purified IgG fraction composed of ~ 60% IgG1 and obtained from the pooled plasma of thousands of donors, is clinically used for a wide range of diseases. The biological actions of IVIg are incompletely understood and have been attributed both to the polyclonal antibodies therein and also to their IgG (IgG) Fc regions. Recently, we demonstrated that multiple therapeutic human IgG1 antibodies suppress angiogenesis in a target-independent manner via FcγRI, a high-affinity receptor for IgG1. Here we show that IVIg possesses similar anti-angiogenic activity and inhibited blood vessel growth in five different mouse models of prevalent human diseases, namely, neovascular age-related macular degeneration, corneal neovascularization, colorectal cancer, fibrosarcoma and peripheral arterial ischemic disease. Angioinhibition was mediated by the Fc region of IVIg, required FcγRI and had similar potency in transgenic mice expressing human FcγRs. Finally, IVIg therapy administered to humans for the treatment of inflammatory or autoimmune diseases reduced kidney and muscle blood vessel densities. These data place IVIg, an agent approved by the US Food and Drug Administration, as a novel angioinhibitory drug in doses that are currently administered in the clinical setting. In addition, they raise the possibility of an unintended effect of IVIg on blood vessels.
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http://dx.doi.org/10.1038/sigtrans.2015.2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4768485PMC
February 2021

Human IgG1 antibodies suppress angiogenesis in a target-independent manner.

Signal Transduct Target Ther 2016;1. Epub 2016 Jan 28.

Division of Immunology, Department of Pathology, University of Cambridge, Cambridge, UK.

Aberrant angiogenesis is implicated in diseases affecting nearly 10% of the world's population. The most widely used anti-angiogenic drug is bevacizumab, a humanized IgG1 monoclonal antibody that targets human VEGFA. Although bevacizumab does not recognize mouse Vegfa, it inhibits angiogenesis in mice. Here we show bevacizumab suppressed angiogenesis in three mouse models not via Vegfa blockade but rather Fc-mediated signaling through FcγRI (CD64) and c-Cbl, impairing macrophage migration. Other approved humanized or human IgG1 antibodies without mouse targets (adalimumab, alemtuzumab, ofatumumab, omalizumab, palivizumab and tocilizumab), mouse IgG2a, and overexpression of human IgG1-Fc or mouse IgG2a-Fc, also inhibited angiogenesis in wild-type and FcγR humanized mice. This anti-angiogenic effect was abolished by Fcgr1 ablation or knockdown, Fc cleavage, IgG-Fc inhibition, disruption of Fc-FcγR interaction, or elimination of FcRγ-initated signaling. Furthermore, bevacizumab's Fc region potentiated its anti-angiogenic activity in humanized VEGFA mice. Finally, mice deficient in FcγRI exhibited increased developmental and pathological angiogenesis. These findings reveal an unexpected anti-angiogenic function for FcγRI and a potentially concerning off-target effect of hIgG1 therapies.
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http://dx.doi.org/10.1038/sigtrans.2015.1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4763941PMC
February 2021

High-Frequency Ultrasound-Guided Injection for the Generation of a Novel Orthotopic Mouse Model of Human Thyroid Carcinoma.

Thyroid 2016 Apr 3;26(4):552-8. Epub 2016 Mar 3.

4 IRCCS SDN , Naples, Italy .

Background: Thyroid carcinoma is the most common endocrine malignancy and has an increasing incidence. High-frequency ultrasound (HFUS) has a spatial resolution of 30 μm, which is a property that has been exploited for thyroid visualization and analysis in mice. The aim of this study was to generate a novel orthotopic mouse model of human follicular thyroid carcinoma (FTC) using an HFUS-guided injection system.

Methods: Twenty Balb/C nude mice were injected in the right lobe of the thyroid with 2 × 10(6) FTC-133 cells using the microinjection HFUS-guided system, and 20 mice, used as a control, underwent surgical orthotopic implantation of 2 × 10(6) FTC-133 cells in the right lobe of the thyroid. All mice underwent HFUS imaging two weeks after cell injection; HFUS examinations and tumor volume (TV) measurements were repeated weekly. Micro-computed tomography was performed at different time points to determine whether lung metastasis had occurred. TVs were compared between the two models (surgical vs. HFUS-guided) using the Mann-Whitney U-test, and the Mantel-Cox log-rank test was applied to evaluate the death hazard. Hematoxylin and eosin analysis of formalin-fixed, paraffin-embedded mouse tissue was performed to validate the in vivo imaging results.

Results: Of the HFUS-guided injected mice, 9/18 survived up to 40 days after the injection of tumor cells. Mice injected surgically had 100% mortality at day 29. Of 38 mice, 29 (14/18 HFUS, 15/20 surgical) showed metastasis in the salivary glands and lymph nodes, and 13 (10/18 HFUS, 3/20 surgical) also showed metastasis in the lungs, which was confirmed by histological analysis. In the surgical group, there was an evident, frequent (12/20 mice) involvement of the contralateral lobe of the thyroid, whereas this feature was only detected in 1/18 mice in the HFUS group. Statistical analysis showed the same pattern of growth in the two groups, and a significant hazard in the mice in the surgical group (p = 0.03).

Conclusions: This study demonstrated the technical feasibility of an HFUS-guided orthotopic mouse model of FTC. The HFUS-guided orthotopic model is easily reproducible and allows prolonged monitoring of the disease because the animals showed an increased survival rate.
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http://dx.doi.org/10.1089/thy.2015.0511DOI Listing
April 2016

miR-340 predicts glioblastoma survival and modulates key cancer hallmarks through down-regulation of NRAS.

Oncotarget 2016 Apr;7(15):19531-47

Department of Molecular Medicine and Medical Biotechnology, "Federico II" University of Naples, Naples, Italy.

Glioblastoma is the most common primary brain tumor in adults; with a survival rate of 12 months from diagnosis. However, a small subgroup of patients, termed long-term survivors (LTS), has a survival rate longer then 12-14 months. There is thus increasing interest in the identification of molecular signatures predicting glioblastoma prognosis and in how to improve the therapeutic approach. Here, we report miR-340 as prognostic tumor-suppressor microRNA for glioblastoma. We analyzed microRNA expression in > 500 glioblastoma patients and found that although miR-340 is strongly down-regulated in glioblastoma overall, it is up-regulated in LTS patients compared to short-term survivors (STS). Indeed, miR-340 expression predicted better prognosis in glioblastoma patients. Coherently, overexpression of miR-340 in glioblastoma cells was found to produce a tumor-suppressive activity. We identified NRAS mRNA as a critical, direct target of miR-340: in fact, miR-340 negatively influenced multiple aspects of glioblastoma tumorigenesis by down-regulating NRAS and downstream AKT and ERK pathways. Thus, we demonstrate that expression of miR-340 in glioblastoma is responsible for a strong tumor-suppressive effect in LTS patients by down-regulating NRAS. miR-340 may thus represent a novel marker for glioblastoma diagnosis and prognosis, and may be developed into a tool to improve treatment of glioblastoma.
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http://dx.doi.org/10.18632/oncotarget.6968DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4991399PMC
April 2016

Measurement of [123I]FP-CIT binding to the dopamine transporter (DAT) in healthy mouse striatum using dedicated small animal SPECT imaging: feasibility, optimization and validation.

Q J Nucl Med Mol Imaging 2018 Mar 1;62(1):112-117. Epub 2015 Sep 1.

IRCCS, SDN, Naples, Italy.

Background: In-vivo imaging of dopamine transporter (DAT), a reliable marker of degeneration of nigrostriatal dopaminergic innervation, has gained increasing interest in preclinical neurodegenerative research for studying disease mechanisms and testing new therapeutic strategies. We assessed the feasibility and the reliability of in vivo and ex vivo quantification of Methyl (3S,4S,5R)-8-(3-fluoropropyl)-3-(4-iodophenyl)-8-azabicyclo[3.2.1]octane-4-carboxylate ([123I]FP-CIT) binding to striatal DAT sites in mouse brain.

Methods: Dedicated small animal single-photon emission computed tomography (SPECT) images of [123I]FP-CIT binding were obtained in 3 groups of healthy mice: untreated (N.=6), pre-treated with lugol solution (N.=4), and pre-treated with selective dopamine transporter uptake inhibitor GBR12909 (N.=4). Ex-vivo autoradiography studies were performed at the end of SPECT studies with phosphor image system in 4 out of the 6 untreated mice and in all mice pre-treated with lugol. Regions of interest were defined over the striatum. The specific binding (SB) was calculated using the cerebral cortex as reference region.

Results: SPECT images in untreated mice showed high [123I]FP-CIT uptake in the striatum and extracerebral regions. Lugol pretreatment improved striatal images quality decreasing salivary and thyroid glands uptake. SB was higher (P<0.0001) in mice pre-treated with lugol (5.97±0.60) than in untreated mice (2.25±0.28). Autoradiography showed similar SB findings in untreated (2.27±0.33) and lugol-treated (4.27±0.57) mice (P<0.0001). In-vivo striatal 123I-FP-CIT SB and ex-vivo striatal 123I-FP-CIT SB were significantly correlated (r=0.87; P<0.0001). SPECT competition studies showed a significant (P<0.0001) reduction of [123I]FP-CIT SB in the striatum after GBR12909.

Conclusions: We demonstrated the feasibility of [123I]FP-CIT imaging of the normal mouse brain using small-animal SPECT without pinhole collimators. The reliability of quantitative measurement of striatal [123I]FP-CIT SB is supported by competition studies showing measurable inhibition of uptake induced by GBR12909 and by the strong correlation between in vivo and ex vivo striatal [123I]FP-CIT SB. Our data also demonstrate that pre-treatment with lugol might improve striatal [123I]FP-CIT SB in mice.
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http://dx.doi.org/10.23736/S1824-4785.17.02813-8DOI Listing
March 2018

High-Frequency Ultrasound for the Study of Early Mouse Embryonic Cardiovascular System.

Reprod Sci 2015 Dec 2;22(12):1649-55. Epub 2015 Jul 2.

Istituto di Biostrutture e Bioimmagini, Consiglio Nazionale delle Ricerche, Napoli, Italy.

An accurate diagnosis of congenital heart defects during fetal development is critical for interventional planning. Mice can be used to generate animal models with heart defects, and high-frequency ultrasound (HFUS) imaging enables in utero imaging of live mouse embryos. A wide range of physiological measurements is possible using Doppler-HFUS imaging; limitations of any single measurement warrant a multiparameter approach to characterize cardiovascular function. Doppler-HFUS was used to explore the embryonic (heart, aorta) and extraembryonic (umbilical blood flow) circulatory systems to create a database in normal mouse embryos between 9.5 and 16.5 days of gestation. Multivariate analyses were performed to explore correlations between gestational age and embryo echocardiographic parameters. Heart rate and peak velocity in the aorta were positively correlated with gestational time, whereas cardiac cycle length, isovolumetric relaxation time, myocardial performance index, and arterial deceleration time of the umbilical cord were negatively correlated with it. Doppler-HFUS facilitated detailed characterization of the embryonic mouse circulation and represents a useful tool for investigation of the early mouse embryonic cardiovascular system.
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http://dx.doi.org/10.1177/1933719115594017DOI Listing
December 2015

Head and Neck Veins of the Mouse. A Magnetic Resonance, Micro Computed Tomography and High Frequency Color Doppler Ultrasound Study.

PLoS One 2015 11;10(6):e0129912. Epub 2015 Jun 11.

IRCCS SDN, Naples, Italy.

To characterize the anatomy of the venous outflow of the mouse brain using different imaging techniques. Ten C57/black male mice (age range: 7-8 weeks) were imaged with high-frequency Ultrasound, Magnetic Resonance Angiography and ex-vivo Microcomputed tomography of the head and neck. Under general anesthesia, Ultrasound of neck veins was performed with a 20 MHz transducer; head and neck Magnetic Resonance Angiography data were collected on 9.4 T or 7 T scanners, and ex-vivo Microcomputed tomography angiography was obtained by filling the vessels with a radiopaque inert silicone rubber compound. All procedures were approved by the local ethical committee. The dorsal intracranial venous system is quite similar in mice and humans. Instead, the mouse Internal Jugular Veins are tiny vessels receiving the sigmoid sinuses and tributaries from cerebellum, occipital lobe and midbrain, while the majority of the cerebral blood, i.e. from the olfactory bulbs and fronto-parietal lobes, is apparently drained through skull base connections into the External Jugular Vein. Three main intra-extracranial anastomoses, absent in humans, are: 1) the petrosquamous sinus, draining into the posterior facial vein, 2) the veins of the olfactory bulb, draining into the superficial temporal vein through a foramen of the frontal bone 3) the cavernous sinus, draining in the External Jugular Vein through a foramen of the sphenoid bone. The anatomical structure of the mouse cranial venous outflow as depicted by Ultrasound, Microcomputed tomography and Magnetic Resonance Angiography is different from humans, with multiple connections between intra- and extra-cranial veins.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0129912PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4466257PMC
April 2016

In vivo imaging and characterization of [(18)F]DPA-714, a potential new TSPO ligand, in mouse brain and peripheral tissues using small-animal PET.

Nucl Med Biol 2015 Mar 6;42(3):309-16. Epub 2014 Dec 6.

Institute of Biostructure and Bioimaging, CNR, Naples, Italy. Electronic address:

Introduction: The translocator protein 18 kDa (TSPO), a biochemical marker of neuroinflammation, is highly expressed in the brain activated microglia and it is also expressed by peripheral inflammatory cells and normal peripheral tissues. Thus, development of radioligands for the TSPO may contribute to further understanding the in vivo TSPO function in central and peripheral inflammatory processes and other pathologies. Here, we report the biodistribution, the specific binding and the radiometabolites of [(18)F]DPA-714, a promising fluorinated PET radiotracer, in normal mice using a microPET/CT scanner.

Methods: The in vivo biodistribution and kinetics of [(18)F]DPA-714 were measured in mice brain and peripheral tissues. Specific binding to TSPO sites was assessed using pharmacological competitive studies by means of saturation experiments performed by i.v. injection of 1mg/kg of unlabeled DPA-714 or 3mg/kg of unlabeled PK11195. A region of interest analysis was performed to generate time-activity curves in the brain, heart, lung, kidney, spleen and liver. Metabolites assay was performed in the plasma and peripheral organs by radio-HPLC.

Results: [(18)F]DPA-714 reached high concentration in lung, heart, kidney and spleen, tissues well known to be rich in TSPO sites. [(18)F]DPA-714 kinetics were faster in the lung and slower in the kidney. Pre-injection of unlabeled DPA-714 or PK11195 inhibited about 80% of [(18)F]DPA-714 uptake in the lung and heart (p<0.0005). The percentage of inhibition in the kidney was lower and achieved at later times only with DPA-714 (p<0.05) but not with PK11195. Sixty minutes after radiotracer injection only unmetabolized radioligand was found in the brain, lung, heart and spleen.

Conclusion: These results suggest that [(18)F]DPA-714 is a suitable PET ligand for imaging in mice brain and peripheral tissues since it binds with high specificity TSPO binding sites and it is almost unchanged at 60 minutes after radiotracer injection in the brain and TSPO-rich regions.
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http://dx.doi.org/10.1016/j.nucmedbio.2014.11.009DOI Listing
March 2015

Evaluation of growth patterns and body composition in C57Bl/6J mice using dual energy X-ray absorptiometry.

Biomed Res Int 2014 10;2014:253067. Epub 2014 Jul 10.

CEINGE-Biotecnologie Avanzate Scarl, Via G. Salvatore 486, 80145 Naples, Italy ; Department of Advanced Biomedical Sciences, Federico II University, Via Pansini 5, 80131 Naples, Italy.

The normal growth pattern of female C57BL/6J mice, from 5 to 30 weeks of age, has been investigated in a longitudinal study. Weight, body surface area (BS), and body mass index (BMI) were evaluated in forty mice. Lean mass and fat mass, bone mineral content (BMC), and bone mineral density (BMD) were monitored by dual energy X-ray absorptiometry (DEXA). Weight and BS increased linearly (16.15 ± 0.64-27.64 ± 1.42 g; 51.13 ± 0.74-79.57 ± 2.15 cm(2), P < 0.01), more markedly from 5 to 9 weeks of age (P < 0.001). BMD showed a peak at 17 weeks (0.0548 ± 0.0011 g/cm(2) ∗ m, P < 0.01). Lean mass showed an evident gain at 9 (15.8 ± 0.8 g, P < 0.001) and 25 weeks (20.5 ± 0.3 g, P < 0.01), like fat mass from 13 to 17 weeks (2.0 ± 0.4-3.6 ± 0.7 g, P < 0.01). BMI and lean mass index (LMI) reached the highest value at 21 weeks (3.57 ± 0.02-0.284 ± 0.010 g/cm(2), resp.), like fat mass index (FMI) at 17 weeks (0.057 ± 0.009 g/cm(2)) (P < 0.01). BMI, weight, and BS showed a moderate positive correlation (0.45-0.85) with lean mass from 5 to 21 weeks. Mixed linear models provided a good prediction for lean mass, fat mass, and BMD. This study may represent a baseline reference for a future comparison of wild-type C57BL/6J mice with models of altered growth.
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http://dx.doi.org/10.1155/2014/253067DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4119710PMC
March 2015

Genetic deletion in uncoupling protein 3 augments 18F-fluorodeoxyglucose cardiac uptake in the ischemic heart.

BMC Cardiovasc Disord 2014 Aug 8;14:98. Epub 2014 Aug 8.

Department of Advanced Biomedical Sciences, University Federico II, Via Pansini 5, 80131 Naples, Italy.

Background: We investigated the effects of uncoupling protein 3 (UCP3) genetic deletion on 18F-fluorodeoxyglucose (FDG) cardiac uptake by positron emission tomography (PET)/computed tomography (CT) dedicated animal system after permanent coronary artery ligation.

Methods: Cardiac 18F-FDG PET/CT was performed in UCP3 knockout (UCP3-/-) and wild-type (WT) mice one week after induction of myocardial infarction or sham procedure.

Results: In sham-operated mice no difference in left ventricular (LV) volume was detectable between WT and UCP3-/-. After myocardial infarction, LV volume was higher in both WT and UCP3-/- compared to sham animals, with a significant interaction (p < 0.05) between genotype and myocardial infarction. In sham-operated animals no difference in FDG standardized uptake value (SUV) was detectable between WT (1.8 ± 0.6) and UCP3-/- (1.8 ± 0.6). After myocardial infarction SUV was significantly higher in remote areas than in infarcted territories in both UCP3-/- and WT mice (both p < 0.01). Moreover, in remote areas, SUV was significantly higher (p < 0.001) in UCP3-/- as compared to WT, while in the infarcted territory SUV was comparable (p = 0.29). A significant relationship (r = 0.68, p < 0.001) between LV volume and SUV was found.

Conclusions: In a mice model of permanent coronary occlusion, UCP3 deficiency results in a metabolic shift that favored glycolytic metabolism and increased FDG uptake in remote areas.
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http://dx.doi.org/10.1186/1471-2261-14-98DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4127083PMC
August 2014

Monitoring reversal of MET-mediated resistance to EGFR tyrosine kinase inhibitors in non-small cell lung cancer using 3'-deoxy-3'-[18F]-fluorothymidine positron emission tomography.

Clin Cancer Res 2014 Sep 22;20(18):4806-15. Epub 2014 Jul 22.

Institute of Biostructures and Bioimages, National Research Council, Naples, Italy. CEINGE-Advanced Biotechnologies, Naples Italy. Department of Advanced Biomedical Sciences, University "Federico II," Naples, Italy.

Purpose: MET amplification is one of the mechanisms underlying acquired resistance to EGFR tyrosine kinase inhibitors (TKI) in non-small cell lung cancer (NSCLC). Here, we tested whether 3'-deoxy-3'-[(18)F]-fluorothymidine ([(18)F]FLT) positron emission tomography/computerized tomography (PET/CT) can detect MET-mediated resistance to EGFR TKIs and monitor the effects of MET inhibitors in NSCLC.

Experimental Design: H1993 and H820 NSCLC cells with high and low levels of MET amplification, respectively, and HCC827-expressing MET, but without gene amplification, were tested for the effects of MET inhibitors on the EGFR pathway and proliferation both in vitro and in vivo. Nude mice bearing NSCLCs with and without MET amplification were subjected to [(18)F]FLT PET/CT before and after treatment with crizotinib or erlotinib (50 mg/kg and 100 mg/kg p.o. for 3 days).

Results: H1993 cells showed high responsiveness to MET inhibitors and were resistant to erlotinib. Conversely, HCC827 cells showed high sensitivity to erlotinib and were resistant to MET inhibitors. Accordingly, H1993 tumors bearing MET amplification showed a mean reduction in [(18)F]FLT uptake of 28% and 41% after low- and high-dose treatment with crizotinib for 3 days, whereas no posttherapy changes of [(18)F]FLT uptake were observed in HCC827 tumors lacking MET amplification. Furthermore, a persistently high [(18)F]FLT uptake was observed in H1993 tumors after treatment with erlotinib, whereas HCC827 tumors showed up to 39% reduction of [(18)F]FLT uptake following erlotinib treatment. Imaging findings were confirmed by Ki67 immunostaining of tumor sections.

Conclusions: [(18)F]FLT PET/CT can detect MET-mediated resistance to EGFR TKIs and its reversal by MET inhibitors in NSCLC.
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http://dx.doi.org/10.1158/1078-0432.CCR-14-0264DOI Listing
September 2014

FKBP51 increases the tumour-promoter potential of TGF-beta.

Clin Transl Med 2014 Jan 27;3(1). Epub 2014 Jan 27.

Department of Molecular Medicine and Medical Biotechnologies, Federico II University, via Pansini, Naples 5, 80131, Italy.

Background: FKBP51 (FKBP5 Official Symbol) is a large molecular weight component of the family of FK506 binding proteins (FKBP). In recent years, research studies from our laboratory highlighted functions for FKBP51 in the control of apoptosis and melanoma progression. FKBP51 expression correlated with the invasiveness and aggressiveness of melanoma. Since a role for TGF-β in the enhanced tumorigenic potential of melanoma cells is widely described, we hypothesized a cooperative effect between FKBP51 and TGF-β in melanoma progression.

Methods: SAN and A375 melanoma cell lines were utilized for this study. Balb/c IL2γ NOD SCID served to assess the ability to colonize organs and metastasize of different cell lines, which was evaluated by in vivo imaging. Realtime PCR and western blot served for measurement of mRNA and protein expression, respectively.

Results: By comparing the metastatic potential of two melanoma cell lines, namely A375 and SAN, we confirmed that an increased capability to colonize murine organs was associated with increased levels of FKBP51. A375 melanoma cell line expressed FKBP51 mRNA levels 30-fold higher in comparison to the SAN mRNA level and appeared more aggressive than SAN melanoma cell line in an experimental metastasis model. In addition, A375 expressed, more abundantly than SAN, the TGF-β and the pro angiogenic TGF-β receptor type III (TβRIII) factors. FKBP51 silencing produced a reduction of TGF-β and TβRIII gene expression in A375 cell line, in accordance with previous studies. We found that the inducing effect of TGF-β on Sparc and Vimentin expression was impaired in condition of FKBP51 depletion, suggesting that FKBP51 is an important cofactor in the TGF-β signal. Such a hypothesis was supported by co immunoprecipitation assays, showing that FKBP51 interacted with either Smad2,3 and p300. In normal melanocytes, FKBP51 potentiated the effect of TGF-β on N-cadherin expression and conferred a mesenchymal-like morphology to such round-shaped cells.

Conclusions: Overall, our findings show that FKBP51 enhances some pro oncogenic functions of TGF-β, suggesting that FKBP51-overexpression may help melanoma to take advantage of the tumor promoting activities of the cytokine.
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http://dx.doi.org/10.1186/2001-1326-3-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3906759PMC
January 2014

Effects of some anesthetic agents on skin microcirculation evaluated by laser Doppler perfusion imaging in mice.

BMC Vet Res 2013 Dec 17;9:255. Epub 2013 Dec 17.

Institute of Biostructures and Bioimages of the National Council of Research, Via T, De Amicis 95, Naples 80145, Italy.

Background: Anesthetic agents alter microcirculation, influencing tissue oxygenation and delivery of vital substrates. Laser Doppler perfusion imaging is a widespread technique in the field of microvascular research that can evaluate noninvasively and in real time the effects of environmental conditions, physical manipulations, diseases and treatments on peripheral perfusion. This study aims to evaluate laser Doppler perfusion imaging as a means to detect changes in skin microcirculation induced by some popular anesthetic agents in a murine model. Twenty-four age- and gender-matched healthy CD1 mice were examined by laser Doppler perfusion imaging. The skin microcirculatory response was measured at the level of plantar surfaces during isoflurane anesthesia with or without subsequent dexmedetomidine or acepromazine. At the end of the procedure, dexmedetomidine was reversed by atipamezole administration.

Results: In all mice, skin blood flow under isoflurane anesthesia did not show significant differences over time (P = 0.1). The serial perfusion pattern and values following acepromazine or dexmedetomidine administration differed significantly (P < 0.05).

Conclusions: We standardized a reliable laser Doppler perfusion imaging protocol to non-invasively assess changes in skin microcirculation induced by anesthesia in mice, considering the advantages and drawbacks of this technique and its translational value.
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http://dx.doi.org/10.1186/1746-6148-9-255DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3878498PMC
December 2013