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// Eugenio Gaudio 1,2,3,* , Francesco Paduano 3,* , Apollinaire Ngankeu 1 , Francesco Ortuso 4 , Francesca Lovat 1 , Sandra Pinton 2 , Sabrina D’Agostino 3 , Nicola Zanesi 1 , Rami I. Aqeilan 1,5 , Pietro Campiglia 6 , Ettore Novellino 7 , Stefano Alcaro 4 , Carlo M. Croce 1 and Francesco Trapasso 3 1 Department of Molecular Immunology, Virology and Medical Genetics, The Ohio State University, Columbus, Ohio, USA 2 Lymphoma & Genomics Research Program, IOR Institute of Oncology Research, Bellinzona, Switzerland 3 Dipartimento di Medicina Sperimentale e Clinica, University Magna Graecia, Campus “S. Venuta”, Catanzaro, Italy 4 Dipartimento di Scienze della Salute, University Magna Graecia, Campus “S. Venuta”, Catanzaro, Italy 5 The Lautenberg Center for Immunology and Cancer Research, Institute for Medical Research, The Hebrew University, Jerusalem, Israel 6 Dipartimento di Farmacia, Universita di Salerno, Fisciano, Italy 7 Dipartimento di Farmacia, Universita degli Studi di Napoli “Federico II”, Napoli, Italy * These authors have contributed equally to this work Correspondence to: Carlo M. Croce, email: // Francesco Trapasso, email: // Keywords : fragile histidine triad, FHIT, annexin A4, ANXA4, mimetic peptides Received : January 07, 2016 Accepted : April 11, 2016 Published : May 04, 2016 Abstract We recently reported that Fhit is in a molecular complex with annexin A4 (ANXA4); following to their binding, Fhit delocalizes ANXA4 from plasma membrane to cytosol in paclitaxel-resistant lung cancer cells, thus restoring their chemosensitivity to the drug. Here, we demonstrate that Fhit physically interacts with A4 through its N-terminus; molecular dynamics simulations were performed on a 3D Fhit model to rationalize its mechanism of action. This approach allowed for the identification of the QHLIKPS heptapeptide (position 7 to 13 of the wild-type Fhit protein) as the smallest Fhit sequence still able to preserve its ability to bind ANXA4. Interestingly, Fhit peptide also recapitulates the property of the native protein in inhibiting Annexin A4 translocation from cytosol to plasma membrane in A549 and Calu-2 lung cancer cells treated with paclitaxel. Finally, the combination of Tat-Fhit peptide and paclitaxel synergistically increases the apoptotic rate of cultured lung cancer cells and blocks in vivo tumor formation. Our findings address to the identification of chemically simplified Fhit derivatives that mimic Fhit tumor suppressor functions; intriguingly, this approach might lead to the generation of novel anticancer drugs to be used in combination with conventional therapies in Fhit-negative tumors to prevent or delay chemoresistance.

PTPRJ, a receptor protein tyrosine phosphatase strongly downregulated in human cancer, displays tumor suppressor activity by negatively modulating several proteins involved in proliferating signals. Here, through a proteomic-based approach, we identified a list of potential PTPRJ-interacting proteins and among them we focused on CD98hc, a type II glycosylated integral membrane protein encoded by SLC3A2, corresponding to the heavy chain of a heterodimeric transmembrane amino-acid transporter, including LAT1. CD98hc is widely overexpressed in several types of cancers and contributes to the process of tumorigenesis by interfering with cell proliferation, adhesion, and migration. We first validated PTPRJ-CD98hc interaction, then demonstrated that PTPRJ overexpression dramatically reduces CD98hc protein levels in A549 lung cancer cells. In addition, following to the treatment of PTPRJ-transduced cells with MG132, a proteasome inhibitor, CD98hc levels did not decrease compared to controls, indicating that PTPRJ is involved in the regulation of CD98hc proteasomal degradation. Moreover, PTPRJ overexpression combined with CD98hc silencing consistently reduced cell proliferation and triggered apoptosis of lung cancer cells. Interestingly, by interrogating the can Evolve database, we observed an inverse correlation between PTPRJ and SLC3A2 gene expression. Indeed, the non-small cell lung cancers (NSCLCs) of patients showing a short survival rate express the lowest and the highest levels of PTPRJ and SLC3A2, respectively. Therefore, the results reported here contribute to shed lights on PTPRJ signaling in cancer cells: moreover, our findings also support the development of a novel anticancer therapeutic approach by targeting the pathway of PTPRJ that is usually downregulated in highly malignant human neoplasias.

Sleep disorders are very common in elderly institutionalized people with dementia and acupressure recently has been associated with conventional medicine in their treatment.Exploring the effectiveness of acupressure for the treatment of insomnia and other sleep disturbances and we want to show that the acupressure treatment is feasible also in elderly resident patients.We enrolled institutionalized patients suffering from Alzheimer's disease with mild cognitive impairment and insomnia. A daily acupressure on HT7 point (H7 Insomnia Control(®)) was performed for a 8-week period. We administered the following scales: the mini mental state examination, the global deterioration scale, the neuropsychiatric inventory, the state-trait-anxiety inventory, the activity daily living and the instrumental activity daily living, the global health quality of life, and the Pittsburgh sleep quality index.After receiving the acupressure treatment, patients saw a significant decrease of sleep disorders. The number of hours of effective sleep was perceived as increased. Furthermore, the time necessary to fall asleep decreased significantly and also the quality of sleep increased. Additionally, also the quality of life was bettered. Sedative drugs have been reduced in all patients involved in the study.Acupressure can be recommended as a complementary, effective, and non-intrusive method to reduce sleep disturbances in old resident patients affected by cognitive disorders. A limitation of the study is the small sample size. More studies are needed to further validate the results of our study.

Among the proteins putatively interacting with PTPRJ, a tyrosine phosphatase with tumor suppressor activity, we focused on the oncoprotein CD98hc as a very interesting candidate for the development of innovative targeted drugs. In fact, CD98hc, representing the heavy chain of a transmembrane aminoacid transporter, is overexpressed in several human cancers. Furthermore, CD98hc higher expression is associated with poor prognosis in lung cancer patients. CD98hc is linked to light chains (LATs, xCT) by disulfide bridge, polar and hydrophobic interactions. The light chain confers substrate specificity, and ERK, AKT, FAK and mTOR pathways are involved in CD98hc-LATs/xCT downstream signals. Moreover, CD98hc is a coreceptor of b-integrins and it is involved in cell proliferation, migration and invasion. We identified two peptides and 15 small molecules (putatively targeting the disulfide bridge) as candidate CD98hc inhibitors by phage display and in silico screenings, respectively, and validated in vitro the binding between peptides and transmembrane fraction of CD98hc by cytofluorimetric assay. Then, we tested the capability of both types of compounds to affect cell viability and proliferation through MTT and CFSE assays, respectively. We observed that the targeting of CD98hc through both peptides and small molecules reduced cell proliferation in A549 human lung cancer cells, strongly encouraging a deeper characterization of these candidate anticancer molecules. Indeed, our next goals will be the assessment of biochemical activity of CD98hc following to its inhibition, as well as the evaluation of compounds toxicity, both in vitro and in vivo. The final aim is to identify lead compounds inhibiting CD98hc, in order to develop novel molecules to be translated in the clinical setting and to be used as monotherapy and/or in a combinatorial approaches for the treatment of cancer patients. Citation Format: Delia Lanzillotta, Enrico Iaccino, Anna Artese, Selena Mimmi, Sabrina D'Agostino, Isabella Romeo, Patrizia Cantafio, Vincenzo Dattilo, Giosuè Costa, Carolina Brescia, Eugenio Gaudio, Stefano Alcaro, Francesco Trapasso. The CD98hc oncoprotein as a target of new anticancer therapy [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr B122. doi:10.1158/1535-7163.TARG-19-B122

// Anna Bilotta 1 , Vincenzo Dattilo 2 , Sabrina D'Agostino 1 , Stefania Belviso 1 , Stefania Scalise 1 , Mariaconcetta Bilotta 1 , Eugenio Gaudio 1, 3 , Francesco Paduano 1, 4 , Nicola Perrotti 2 , Tullio Florio 5 , Alfredo Fusco 6 , Rodolfo Iuliano 1 , Francesco Trapasso 1 1 Department of Medicina Sperimentale e Clinica, University Magna Graecia of Catanzaro, Catanzaro, Italy 2 Department of Scienze della Salute, University Magna Graecia of Catanzaro, Catanzaro, Italy 3 Lymphoma and Genomics Research Program, Institute of Oncology Research (IOR), Bellinzona, Switzerland 4 Tecnologica Research Institute, Biomedical Section, Crotone, Italy 5 Laboratory of Pharmacology, Dept. of Internal Medicine, and Center of Excellence for Biomedical Research (CEBR), University of Genova, Genova, Italy 6 Istituto di Endocrinologia e Oncologia Sperimentale - CNR c/o Dipartimento di Medicina Molecolare e Biotecnologie Mediche, University Federico II of Napoli, Napoli, Italy Correspondence to: Rodolfo Iuliano, email: iuliano@unicz.it Francesco Trapasso, email: trapasso@unicz.it Keywords: protein tyrosine phosphatase, soluble isoform, angiogenesis, glioblastoma, angiogenic factor Received: September 20, 2016 Accepted: December 13, 2016 Published: December 29, 2016 ABSTRACT PTPRJ is a receptor protein tyrosine phosphatase with tumor suppressor activity. Very little is known about the role of PTPRJ ectodomain, although recently both physiological and synthetic PTPRJ ligands have been identified. A putative shorter spliced variant, coding for a 539 aa protein corresponding to the extracellular N-terminus of PTPRJ, is reported in several databases but, currently, no further information is available. Here, we confirmed that the PTPRJ short isoform (named sPTPRJ) is a soluble protein secreted into the supernatant of both endothelial and tumor cells. Like PTPRJ, also sPTPRJ undergoes post-translational modifications such as glycosylation, as assessed by sPTPRJ immunoprecipitation. To characterize its functional activity, we performed an endothelial cell tube formation assay and a wound healing assay on HUVEC cells overexpressing sPTPRJ and we found that sPTPRJ has a proangiogenic activity. We also showed that sPTPRJ expression down-regulates endothelial adhesion molecules, that is a hallmark of proangiogenic activity. Moreover, sPTPRJ mRNA levels in human high-grade glioma, one of the most angiogenic tumors, are higher in tumor samples compared to controls. Further studies will be helpful not only to clarify the way sPTPRJ works but also to supply clues to circumvent its activity in cancer therapy.

Hepatitis C virus (HCV) infection represents a great public healthcare challenge as it affects nearly 170 million individuals worldwide. Therefore, the deep investigation of the mechanisms involved in the pathogenesis of chronic hepatitis induced by HCV is a crucial step in the design of novel targeted therapies for the treatment of this condition. However, techniques of molecular biology to characterize HCV proteins can suffer of intrinsic limitations due to high mutation rates of the virus genome. In this study, we propose a novel strategy to synthesize a viral cDNA sequence corresponding to the p7 gene in HCV genome-free conditions. Our approach consists of a three-steps polymerase chain reactions (PCRs) by using a set of four large overlapping synthetic oligonucleotides aimed to separately amplify both 5’ and 3’ ends of the p7 gene; 5’ and 3’ products, overlapping themselves, were then used as a template in a third PCR amplification in order to get a full-length p7 cDNA. Our methodology represents an interesting proof-of-principle as it allows for the safe manipulation of short viral genes. Moreover, this new technique overcomes the elevated genetic variability of HCV genomes without affecting the antigenic characteristics of the putative viral protein.