65 results on '"Iaccarino, E"'
Search Results
2. Cloverleaf skull anomaly and de novo trisomy 4p
- Author
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DE BRASI, D, PERONE, L, DI MICCO, P, ANDRIA, G, SEBASTIO, G, IACCARINO, E, PINTO, L, and ALIBERTI, F
- Published
- 1999
3. Targeting VEGF receptors with mini-factors
- Author
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CALVANESE, LUISA, CAPORALE, ANDREA, FALCIGNO, LUCIA, D'AURIA, GABRIELLA, Iaccarino, E., Focà, G., Sandomenico, A., Doti, N., Ruvo, M., Giancarlo Morelli - Paolo Grieco - Michele Saviano - Menotti Ruvo Eds, Calvanese, Luisa, Caporale, Andrea, Iaccarino, E., Focà, G., Sandomenico, A., Doti, N., Falcigno, Lucia, D'Auria, Gabriella, and Ruvo, M.
- Published
- 2016
4. Novel highly constrained peptide ligands for modulating the activity of Cripto-1 and Activin-like receptors
- Author
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Iaccarino, E., Sandomenico, A., Ruvo, M., CALVANESE, LUISA, CAPORALE, ANDREA, D'AURIA, GABRIELLA, FALCIGNO, LUCIA, Giancarlo Morelli - Paolo Grieco - Michele Saviano - Menotti Ruvo Eds, Iaccarino, E., Sandomenico, A., Calvanese, Luisa, Caporale, Andrea, D'Auria, Gabriella, Falcigno, Lucia, and Ruvo, M.
- Published
- 2016
5. Computational design of short linear D-tripeptides as binding moieties for protein pockets
- Author
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Ruvo, M., Incisivo, M., Cross, S., Doti, N., Focà, A., Focà, G., Raimondo, D., Sandomenico, A., Chambery, A., Rega, C., Cruciani, G., Iaccarino, E., Mascanzoni, F., CAPORALE, ANDREA, FALCIGNO, LUCIA, CALVANESE, LUISA, D'AURIA, GABRIELLA, Giancarlo Morelli - Paolo Grieco - Michele Saviano - Menotti Ruvo Eds, Ruvo, M., Caporale, Andrea, Incisivo, M., Cross, S., Doti, N., Focà, A., Focà, G., Raimondo, D., Falcigno, Lucia, Calvanese, Luisa, D'Auria, Gabriella, Sandomenico, A., Chambery, A., Rega, C., Cruciani, G., Iaccarino, E., and Mascanzoni, F.
- Published
- 2016
6. Molecular interactions of the transforming growth factor Nodal and its receptors
- Author
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CALVANESE, LUISA, D'AURIA, GABRIELLA, FALCIGNO, LUCIA, Sandomenico, A., Iaccarino, E., Focà, A., Focà, G., Caporale, A., Ruvo, M., Società Chimica Italiana - DCSB, Calvanese, Luisa, D'Auria, Gabriella, Sandomenico, A., Iaccarino, E., Focà, A., Focà, G., Caporale, A., Ruvo, M., and Falcigno, Lucia
- Published
- 2016
7. Novel bi-cyclic CFC-like peptides for modulating the activity of Cripto-1 and ALK4 receptors
- Author
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Iaccarino, E., Sandomenico, A., Caporale, A., Ruvo, M., CALVANESE, LUISA, D'AURIA, GABRIELLA, FALCIGNO, LUCIA, Società Chimica Italiana - DCSB, Iaccarino, E., Sandomenico, A., Calvanese, Luisa, Caporale, A., D'Auria, Gabriella, Falcigno, Lucia, and Ruvo, M.
- Published
- 2016
8. Immunological disorder and Hirschsprung disease in round femoral inferior epiphysis dysplasia
- Author
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LECORA, M., primary, PARENTI, G., additional, IACCARINO, E., additional, SCARANO, G., additional, CUCCHIARA, S., additional, and ANDRIA, G., additional
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- 1995
- Full Text
- View/download PDF
9. Reduced bone mineral content and normal serum osteocalcin in non-steroid-treated patients with juvenile rheumatoid arthritis.
- Author
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Polito, C, primary, Strano, C G, additional, Rea, L, additional, Alessio, M, additional, Iammarrone, C S, additional, Todisco, N, additional, Marotta, A, additional, Iaccarino, E, additional, and Pirozzi, M, additional
- Published
- 1995
- Full Text
- View/download PDF
10. Cow\'s milk protein intolerance: evaluation of clinical, histological, endoscopic and laboratory criteria for post-challenge diagnosis
- Author
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CUCCHIARA S, DE RITIS G, FOLLO D, IACCARINO E, ROMANIELLO G, LATTE F, AURICCHIO S., STAIANO, ANNAMARIA, Cucchiara, S, DE RITIS, G, Follo, D, Iaccarino, E, Romaniello, G, Latte, F, Staiano, Annamaria, and Auricchio, S.
- Abstract
I
- Published
- 1983
11. Genotyping of Toxoplasma gondii strain directly from human CSF samples of congenital toxoplasmosis clinical case
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Pagliuca, C., Pastore, G., Scaglione, E., Migliucci, A., Maruotti, G. M., Cicatiello, A. G., Elena SALVATORE, Picardi, M., Sammartino, J. C., Buonocore, M. C., Martinelli, P., Iaccarino, E., Colicchio, R., Salvatore, P., Pagliuca, Chiara, Pastore, G, Scaglione, Elena, Migliucci, A, Maruotti, GIUSEPPE MARIA, Cicatiello, ANNUNZIATA GAETANA, Salvatore, Elena, Picardi, Marco, Camilla Sammartino, J, Consiglio Buonocore, M, Martinelli, Pasquale, Iaccarino, E, Colicchio, Roberta, and Salvatore, Paola
- Subjects
Genotyping ,Congenital toxoplasmosi ,Nested-PCR-RFLP ,Toxoplasma gondii - Abstract
This report describes a case of congenital toxoplasmosis in a newborn in Southern Italy. A pregnant mother had been admitted at the 20th week of her pregnancy on account of pharyngodynia and laterocervical lymphadenopathy. Although serological testing of the mother's serum documented a seroconversion with positive IgG and IgM anti-Toxoplasma antibodies during II trimester, the woman refused to perform prenatal diagnosis for congenital toxoplasmosis. Fetal ultrasound scan already showed mild asymmetrical triventricular hydrocephaly and cerebral calcifications. After birth, real-time PCR on cerebrospinal fluid and blood samples of the newborn showed a positive result for 529bp-repeat element DNA of T. gondii, In addition brain magnetic resonance imaging and computed tomography showed a characteristic diffuse brain tissue loss associated with hydrocephalus. For the first time molecular characterization of T. gondii isolate was performed directly from the newborn's CSF samples by using nested-PCR-RFLP of sag-2 and pk1 genes. The PCR-RLFP analysis revealed that the isolate belongs to the clonal type II, the predominant lineage causing human toxoplasmosis, as confirmed by DNA sequencing.
12. Cloverleaf skull anomaly and de novo trisomy 4p [3]
- Author
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Daniele De Brasi, Perone, L., Di Micco, P., Andria, G., Sebastio, G., Iaccarino, E., Pinto, L., and Aliberti, F.
13. Cloverleaf skull anomaly and de novo trisomy 4p
- Author
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Brasi, D. de, Perone, L., Micco, P. di, Andria, G., Sebastio, G., Iaccarino, E., Pinto, L., and Aliberti, F.
- Published
- 1999
14. Structure-based design of EGF-like bicyclic peptide inhibitors of protein Nodal activity
- Author
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E. Iaccarino, L. Falcigno, D. delle Cave, E. Lonardo, M. Ruvo, A. Sandomenico, G. Morelli, P. Grieco, M. Saviano, M. Ruvo, Iaccarino, E., Falcigno, L., delle Cave, D., Lonardo, E., Ruvo, M., and Sandomenico, A.
- Subjects
bicyclic peptides, Nodal, Cripto-1, EGF-like, TBMB scaffold - Published
- 2022
15. Development of a New Highly Selective Monoclonal Antibody against Preferentially Expressed Antigen in Melanoma (PRAME) and Identification of the Target Epitope by Bio-Layer Interferometry
- Author
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Jwala Priyadarsini Sivaccumar, Domenica Capasso, Antonio Leonardi, Giusy Corvino, Annamaria Sandomenico, Rosita Russo, Nunzianna Doti, Luca Sanguigno, Mariangela Valletta, Angela Chambery, Debora Latino, Menotti Ruvo, Emanuela Iaccarino, Sivaccumar, J. P., Leonardi, A., Iaccarino, E., Corvino, G., Sanguigno, L., Chambery, A., Russo, R., Valletta, M., Latino, D., Capasso, D., Doti, N., Ruvo, M., and Sandomenico, A.
- Subjects
0301 basic medicine ,Bio-layer interferometry ,Epitope ,lcsh:Chemistry ,Epitopes ,Mice ,Antineoplastic Agents, Immunological ,0302 clinical medicine ,Antibody Specificity ,Molecular Targeted Therapy ,lcsh:QH301-705.5 ,Melanoma ,Spectroscopy ,biology ,Chemistry ,Antibodies, Monoclonal ,General Medicine ,Recombinant Protein ,epitope identification ,Flow Cytometry ,Recombinant Proteins ,Computer Science Applications ,Blot ,030220 oncology & carcinogenesis ,Antibody ,Human ,Protein Binding ,medicine.drug_class ,Enzyme-Linked Immunosorbent Assay ,Monoclonal antibody ,Article ,Catalysis ,Inorganic Chemistry ,03 medical and health sciences ,Drug Development ,Antigen ,Antigens, Neoplasm ,PRAME ,medicine ,Animals ,Humans ,bio-layer interferometry ,Physical and Theoretical Chemistry ,Molecular Biology ,Kinetic ,mAb ,Dose-Response Relationship, Drug ,Animal ,Organic Chemistry ,Molecular biology ,Kinetics ,Interferometry ,030104 developmental biology ,lcsh:Biology (General) ,lcsh:QD1-999 ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,biology.protein ,Cancer biomarkers - Abstract
Background: Monoclonal antibodies (mAbs) against cancer biomarkers are key reagents in diagnosis and therapy. One such relevant biomarker is a preferentially expressed antigen in melanoma (PRAME) that is selectively expressed in many tumors. Knowing mAb’s epitope is of utmost importance for understanding the potential activity and therapeutic prospective of the reagents. Methods: We generated a mAb against PRAME immunizing mice with PRAME fragment 161–415, the affinity of the antibody for the protein was evaluated by ELISA and SPR, and its ability to detect the protein in cells was probed by cytofluorimetry and Western blotting experiments. The antibody epitope was identified immobilizing the mAb on bio-layer interferometry (BLI) sensor chip, capturing protein fragments obtained following trypsin digestion and performing mass spectrometry analyses. Results: A mAb against PRAME with an affinity of 35 pM was obtained and characterized. Its epitope on PRAME was localized on residues 202–212, taking advantage of the low volumes and lack of fluidics underlying the BLI settings. Conclusions: The new anti-PRAME mAb recognizes the folded protein on the surface of cell membranes suggesting that the antibody’s epitope is well exposed. BLI sensor chips can be used to identify antibody epitopes.
- Published
- 2021
- Full Text
- View/download PDF
16. Insights into the Interaction Mechanism of DTP3 with MKK7 by Using STD-NMR and Computational Approaches
- Author
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Lorenzo Di Rienzo, Emanuela Iaccarino, Rosita Russo, Marco D'Abramo, Lucia Falcigno, Luisa Calvanese, Edoardo Milanetti, Menotti Ruvo, Domenico Raimondo, Annamaria Sandomenico, Guido Franzoso, Angela Chambery, Gabriella D'Auria, Laura Tornatore, Sandomenico, A., Di Rienzo, L., Calvanese, L., Iaccarino, E., D'Auria, G., Falcigno, L., Chambery, A., Russo, R., Franzoso, G., Tornatore, L., D'Abramo, M., Ruvo, M., Milanetti, E., Raimondo, D., Sandomenico, Annamaria, Di Rienzo, Lorenzo, Calvanese, Luisa, Iaccarino, Emanuela, D'Auria, Gabriella, Falcigno, Lucia, Chambery, Angela, Russo, Rosita, Franzoso, Guido, Tornatore, Laura, D'Abramo, Marco, Ruvo, Menotti, Milanetti, Edoardo, and Raimondo, Domenico
- Subjects
0301 basic medicine ,GADD45 beta ,Biochemistry & Molecular Biology ,MKK7 ,Medicine (miscellaneous) ,Computational biology ,Drug resistance ,Research & Experimental Medicine ,01 natural sciences ,General Biochemistry, Genetics and Molecular Biology ,Article ,03 medical and health sciences ,In vivo ,GADD45β ,STD-NMR ,multiple myeloma ,protein-ligand interaction ,Pharmacology & Pharmacy ,Binding site ,Mode of action ,lcsh:QH301-705.5 ,Science & Technology ,010405 organic chemistry ,Mechanism (biology) ,Chemistry ,In vitro ,0104 chemical sciences ,Bioavailability ,030104 developmental biology ,lcsh:Biology (General) ,Medicine, Research & Experimental ,Apoptosis ,Life Sciences & Biomedicine - Abstract
GADD45&beta, /MKK7 complex is a non-redundant, cancer cell-restricted survival module downstream of the NF-kB survival pathway, and it has a pathogenically critical role in multiple myeloma, an incurable malignancy of plasma cells. The first-in-class GADD45&beta, /MKK7 inhibitor DTP3 effectively kills MM cells expressing its molecular target, both in vitro and in vivo, by inducing MKK7/JNK-dependent apoptosis with no apparent toxicity to normal cells. DTP3 combines favorable drug-like properties, with on-target-specific pharmacology, resulting in a safe and cancer-selective therapeutic effect, however, its mode of action is only partially understood. In this work, we have investigated the molecular determinants underlying the MKK7 interaction with DTP3 by combining computational, NMR, and spectroscopic methods. Data gathered by fluorescence quenching and computational approaches consistently indicate that the N-terminal region of MKK7 is the optimal binding site explored by DTP3. These findings further the understanding of the selective mode of action of GADD45&beta, /MKK7 inhibitors and inform potential mechanisms of drug resistance. Notably, upon validation of the safety and efficacy of DTP3 in human trials, our results could also facilitate the development of novel DTP3-like therapeutics with improved bioavailability or the capacity to bypass drug resistance.
- Published
- 2020
- Full Text
- View/download PDF
17. Lab on fiber nano-cavity integrated with charge responsive microgels for biosensing
- Author
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A. Aliberti, Andrea Cusano, Alberto Micco, E. Iaccarino, M. Giaquinto, F. Gambino, A.M. Cusano, P. Cicatiello, A. Iele, Menotti Ruvo, Armando Ricciardi, Gambino, F., Cicatiello, P., Giaquinto, M., Cusano, A. M., Aliberti, A., Micco, A., Iele, A., Iaccarino, E., Ruvo, M., Ricciardi, A., and Cusano, A.
- Subjects
Materials science ,Optical fiber ,Nanotechnology ,Fabry-Perot cavity ,Smart material ,law.invention ,Planar ,law ,Materials Chemistry ,Miniaturization ,Electrical and Electronic Engineering ,Instrumentation ,Lab-on-fiber technology ,Microgels ,Biosensing ,business.industry ,Smart materials ,Metals and Alloys ,Polyelectrolytes ,Condensed Matter Physics ,Surfaces, Coatings and Films ,Electronic, Optical and Magnetic Materials ,Optical cavity ,Optode ,Photonics ,business ,Biosensor - Abstract
The development of photonic-based point of care (POC) devices requires the use of compact integrated biosensing platforms enabling reliable detection performances in small volumes. The sensing system should be designed to overcome limitations of benchtop technologies in terms of portability and the easy-of-use. The Lab-on-Fiber (LOF) technology is continuously trying to develop miniaturized, lightweight, and light-coupled optrodes with unprecedented detection performances associated with portable, plug-and-play and user-friendly features. The sensing platforms, yet successfully developed in planar configurations, are being reimagined and suitably scaled down to be directly integrated onto optical fibers. In the wake of this ‘chip-to-fiber translation’ vision, we have realized a micro-sized optical cavity made of smart materials based on synthetic microgels and integrated on the top of an optical fiber. As a specific case study, the optrode has been here tested within a ‘general purpose’ detection assay for biochemical applications. Specifically, we detected nanomolar concentrations of homopolyarginines and positively charged linear polymers that are frequently exploited as surplus in indirect binding assays. The high degree of miniaturization of our probe enables a significant improvement in the minimum detectable concentration (at least 1 nM) compared to those achieved with similar devices having planar configurations. The probe sensitivity and overall performance are strongly influenced by the molecular weight and number of positive charges of the polymer. Overall, our findings represent a step forward in establishing LOF optrodes as sensing units for optical POC, further enlarging their applicability for the detection of a broad range of relevant biomarkers.
- Published
- 2022
18. Exploring the interaction between the swi/snf chromatin remodeling complex and the zinc finger factor ctcf
- Author
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Ilaria Baglivo, Sara Iachettini, Angela Chambery, Claudia Angelini, Annamaria Sandomenico, Mariangela Valletta, Italia De Feis, Rosita Russo, Paolo V. Pedone, Emanuela Iaccarino, Sara Ragucci, Annamaria Biroccio, Menotti Ruvo, Veronica Russo, Valletta, M., Russo, R., Baglivo, I., Russo, V., Ragucci, S., Sandomenico, A., Iaccarino, E., Ruvo, M., De Feis, I., Angelini, C., Iachettini, S., Biroccio, A., Pedone, P. V., and Chambery, A.
- Subjects
CCCTC-Binding Factor ,Chromosomal Proteins, Non-Histone ,Cell Cycle Proteins ,Biology ,Article ,Catalysis ,Chromatin remodeling ,Chromodomain ,Inorganic Chemistry ,BRG1 ,Tandem Mass Spectrometry ,Humans ,Protein Interaction Maps ,Physical and Theoretical Chemistry ,Molecular Biology ,Transcription factor ,Spectroscopy ,Adenosine Triphosphatases ,Transcriptionally active chromatin ,Zinc finger ,Binding Sites ,Mass spectrometry ,Genome, Human ,Organic Chemistry ,DNA Helicases ,Nuclear Proteins ,Zinc Fingers ,General Medicine ,Chromatin Assembly and Disassembly ,CTCF ,SWI/SNF ,Chromatin ,Computer Science Applications ,Cell biology ,Protein–protein interaction ,Gene Expression Regulation ,Multiprotein Complexes ,BRK ,biological phenomena, cell phenomena, and immunity ,mass spectrometry ,protein-protein interaction ,transcription factor ,chromatin ,Transcription Factors - Abstract
The transcription factor CCCTC-binding factor (CTCF) modulates pleiotropic functions mostly related to gene expression regulation. The role of CTCF in large scale genome organization is also well established. A unifying model to explain relationships among many CTCF-mediated activities involves direct or indirect interactions with numerous protein cofactors recruited to specific binding sites. The co-association of CTCF with other architectural proteins such as cohesin, chromodomain helicases, and BRG1, further supports the interplay between master regulators of mammalian genome folding. Here, we report a comprehensive LC-MS/MS mapping of the components of the switch/sucrose nonfermentable (SWI/SNF) chromatin remodeling complex co-associated with CTCF including subunits belonging to the core, signature, and ATPase modules. We further show that the localization patterns of representative SWI/SNF members significantly overlap with CTCF sites on transcriptionally active chromatin regions. Moreover, we provide evidence of a direct binding of the BRK-BRG1 domain to the zinc finger motifs 4&ndash, 8 of CTCF, thus, suggesting that these domains mediate the interaction of CTCF with the SWI/SNF complex. These findings provide an updated view of the cooperative nature between CTCF and the SWI/SNF ATP-dependent chromatin remodeling complexes, an important step for understanding how these architectural proteins collaborate to shape the genome.
- Published
- 2020
19. Generation and testing of engineered multimeric Fabs of trastuzumab
- Author
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Annamaria Sandomenico, Silvia Scaramuzza, Fabio Selis, Emanuela Iaccarino, Luisa Calvanese, Andrea Esperti, Emanuela Truppo, Paolo Dell'Omo, Maria Cantile, Sandro De Falco, Lucia Falcigno, Menotti Ruvo, Andrea Caporale, Riccardo Sanna, Giancarlo Tonon, Annalia Focà, Selis, F., Sandomenico, A., Cantile, M., Sanna, R., Calvanese, L., Falcigno, L., Dell'Omo, P., Esperti, A., De Falco, S., Foca, A., Caporale, A., Iaccarino, E., Truppo, E., Scaramuzza, S., Tonon, G., and Ruvo, M.
- Subjects
Models, Molecular ,Immunoconjugates ,Protein Conformation ,Receptor, ErbB-2 ,Peptide ,02 engineering and technology ,Protein Engineering ,Biochemistry ,law.invention ,Structural Biology ,law ,Cytotoxic T cell ,Internalization ,media_common ,chemistry.chemical_classification ,Transglutamination ,0303 health sciences ,biology ,Chemistry ,General Medicine ,Antibody fragment ,Bioconjugation ,Trastuzumab ,Recombinant Fab ,021001 nanoscience & nanotechnology ,Recombinant Proteins ,Recombinant DNA ,Cystine ,Female ,Antibody ,0210 nano-technology ,DNA, Complementary ,media_common.quotation_subject ,Breast Neoplasms ,03 medical and health sciences ,Immunoglobulin Fab Fragments ,Cell Line, Tumor ,Escherichia coli ,Humans ,Amino Acid Sequence ,Molecular Biology ,030304 developmental biology ,Fluorescent Dyes ,Transglutaminases ,Carcinoma ,Periplasmic space ,Surface Plasmon Resonance ,Peptide Fragments ,Drug Design ,biology.protein ,Drug Screening Assays, Antitumor ,Protein Multimerization ,Conjugate - Abstract
Recombinant antibodies fragments in several new formats are routinely investigated and used in diagnostic and therapeutic applications as anti-cancers molecules. New antibody formats are generated to compensate the need for multispecificity and site-specific introduction of fluorescent dyes, cytotoxic payloads or for generating semisynthetic multimeric molecules. Fabs of trastuzumab bearing transglutaminase (MTG) reactive sites were generated by periplasmic expression in E. coli and purified. Multimeric Fabs were generated by either disulfide bridge formation or by using MTG-sensitive peptide linkers. Binding to receptor was assessed by ELISA and SPR methods. Internalization and growth inhibition assays were performed on BT-474 and SKBR3 Her2+ cells. Fabs were successfully produced and dimerized or trimerized using MTG and suitably designed peptide linkers. Site-specific derivatizations with fluorophores were similarly achieved. The monomeric, dimeric and trimeric variants bind the receptor with affinities similar or superior to the full antibody. Fab and Fab2 are rapidly internalized in Her2+ cells and exhibit growth inhibition abilities similar to the full antibody. Altogether, the data show that the recombinant Fabs can be produced in E. coli and converted into multimeric variants by MTG-based bioconjugation. Similar approaches are extendable to the introduction of cytotoxic payloads for the generation of novel Antibody Drug Conjugates.
- Published
- 2020
20. Investigating the oxidative refolding mechanism of Cripto-1 CFC domain
- Author
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Giuseppina Focà, Angela Chambery, Andrea Caporale, Giusy Corvino, Josephine Alba, Domenico Raimondo, Menotti Ruvo, Annamaria Sandomenico, Rosita Russo, Emanuela Iaccarino, Marco D'Abramo, Valeria Severino, Iaccarino, E., Sandomenico, A., Corvino, G., Foca, G., Severino, V., Russo, R., Caporale, A., Raimondo, D., D'Abramo, M., Alba, J., Chambery, A., and Ruvo, M.
- Subjects
CFC domain ,Cripto ,disulfide bridges ,oxidative folding ,In silico ,02 engineering and technology ,Molecular Dynamics Simulation ,GPI-Linked Proteins ,Biochemistry ,Protein Refolding ,Serine ,03 medical and health sciences ,Protein Domains ,Structural Biology ,Molecule ,Humans ,Amino Acid Sequence ,Disulfides ,Disulfide bridge ,Molecular Biology ,030304 developmental biology ,0303 health sciences ,Chemistry ,Oxidative folding ,Wild type ,General Medicine ,021001 nanoscience & nanotechnology ,Peptide Fragments ,Neoplasm Proteins ,Folding (chemistry) ,Kinetics ,Intramolecular force ,Biophysics ,Intercellular Signaling Peptides and Proteins ,0210 nano-technology ,Oxidation-Reduction - Abstract
Using a combined approach based on MS, enzyme digestion and advanced MD studies we have determined the sequential order of formation of the three disulfide bridges of the Cripto-1 CFC domain. The domain has a rare pattern of bridges and is involved in the recognition of several receptors. The bridge formation order is C1-C4, C3-C5, C2-C6, however formation of C1-C4 plays no roles for the formation of the others. Folding is driven by formation of the C3-C5 bridge and is supported by residues lying within the segment delimited by these cysteines. We indeed observe that variants CFC-W123A and CFC-ΔC1/C4, where C1 and C4 are replaced by serines, are able to refold in the same time window as the wild type, while CFC-K132A and CFC-W134A are not. A variant where cysteines of the second and third bridge are mutated to serine, convert slowly to the monocyclic molecule. Data altogether support a mechanism whereby the Cripto-1 CFC domain refolds by virtue of long-range intramolecular interactions that involve residues close to cysteines of the second and third bridge. These findings are supported by the in silico study that shows how distant parts of the molecules come into contact on a long time scale. Using a combined approach based on MS, enzyme digestion and advanced MD studies we have determined the sequential order of formation of the three disulfide bridges of the Cripto-1 CFC domain. The domain has a rare pattern of bridges and is involved in the recognition of several receptors. The bridge formation order is C1-C4, C3-05, C2-C6, however formation of C1-C4 plays no roles for the formation of the others. Folding is driven by formation of the C3-05 bridge and is supported by residues lying within the segment delimited by these cysteines. We indeed observe that variants CFC-W123A and CFC-Delta C1/C4, where C1 and C4 are replaced by serines, are able to refold in the same time window as the wild type, while CFC-K132A and CFC-W134A are not. A variant where cysteines of the second and third bridge are mutated to serine, convert slowly to the monocyclic molecule. Data altogether support a mechanism whereby the Cripto-1 CFC domain refolds by virtue of long-range intramolecular interactions that involve residues close to cysteines of the second and third bridge. These findings are supported by the in silica study that shows how distant parts of the molecules come into contact on a long time scale. (C) 2019 Elsevier B.V. All rights reserved.
- Published
- 2019
21. Intrinsic structural versatility of the highly conserved 412-423 epitope of the Hepatitis C Virus E2 protein
- Author
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Alfonso De Simone, Daniela Barone, Menotti Ruvo, Luigi Vitagliano, Emanuela Iaccarino, Annamaria Sandomenico, Nicole Balasco, Balasco, N., Barone, D., Iaccarino, E., Sandomenico, A., De Simone, A., Ruvo, M., and Vitagliano, L.
- Subjects
0301 basic medicine ,Viral Hepatitis Vaccines ,Sequence (biology) ,Computational biology ,Hepacivirus ,Molecular Dynamics Simulation ,Biochemistry ,Virus ,Epitope ,03 medical and health sciences ,Molecular dynamics ,Epitopes ,Viral Proteins ,Structural Biology ,Protein flexibility ,Molecular Biology ,Protein secondary structure ,Flexibility (engineering) ,030102 biochemistry & molecular biology ,Chemistry ,Replica exchange molecular dynamics ,General Medicine ,Amyloid-like aggregate ,030104 developmental biology ,Interaction with host ,Proteome ,Pathogen-host recognition - Abstract
HCV infection is a major threaten for human health as it affects hundreds of million people worldwide. Here we investigated the conformational properties of the 412–423 fragment of the envelope E2 protein, one of the most immunogenic regions of the virus proteome whose characterization may provide interesting insights for anti-HCV vaccine development. The spectroscopic characterization of the polypeptide unravels its unexpected tendency to form amyloid-like aggregates. When kept in monomeric state, it shows a limited tendency to adopt regular secondary structure. Enhanced molecular dynamics simulations, starting from four distinct conformational states, highlight its structural versatility. Interestingly, all multiform conformational states of the polypeptide detected in crystallographic complexes with antibodies are present in the structural ensemble of all simulations. This observation corroborates the idea that known antibodies recognize this region through a conformational selection mechanism. Accordingly, the design of effective anti-HCV vaccines should consider the intrinsic flexibility of this region. The structural versatility of the 412–423 region is particularly puzzling if its remarkable sequence conservation is considered. It is likely that flexibility and sequence conservation are important features that endow this epitope with the ability to accomplish distinct functions such as immunity escape and interaction with host receptors.
- Published
- 2018
22. A study of mAb 3D1-Nodal peptides interaction by STD NMR spectroscopy
- Author
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L. Calvanese, A. Focà, A. Sandomenico, G. Focà, A. Caporale, E. Iaccarino, A. Leonardi, N. Doti, G. D’Auria, M. Ruvo, L. Falcigno, Giancarlo Morelli, Paolo Grieco, Menotti Ruvo, Calvanese, L., Focà, A., Sandomenico, A., Focà, G., Caporale, A., Iaccarino, E., Leonardi, A., Doti, N., D’Auria, G., Ruvo, M., and Falcigno, L.
- Published
- 2018
23. Therapeutic potential of novel Cripto-1 CFC small peptide mimetics in melanoma
- Author
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E. Iaccarino, A. Sandomenico, L. Calvanese, A. Caporale, G. D’Auria, L. Falcigno, L. Strizzi, M. Ruvo, Giancarlo Morelli, Paolo Grieco, Menotti Ruvo, Iaccarino, E., Sandomenico, A., Calvanese, L., Caporale, A., D’Auria, G., Falcigno, L., Strizzi, L., and Ruvo, M.
- Published
- 2018
24. Mini-factors revealing VEGF receptors in imaging applications
- Author
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L. Calvanese, A. Caporale, G. Focà, E. Iaccarino, A. Sandomenico, N. Doti, I. Apicella, G. M. Incisivo, S. De Falco, L. Falcigno, G. D’Auria, M. Ruvo, Giancarlo Morelli, Paolo Grieco, Menotti Ruvo, Calvanese, L., Caporale, A., Focà, G., Iaccarino, E., Sandomenico, A., Doti, N., Apicella, I., Incisivo, G. M., De Falco, S., Falcigno, L., D’Auria, G., and Ruvo, M.
- Published
- 2018
25. Segno, simbolo e marchio
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R. Saccoia, L. Iaccarino e M. Tregua, and Saccoia, R.
- Published
- 2017
26. Unbalanced translocation (3;5)(q26.1;p14): A clinical report
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Lucia Perone, Vito Guzzetta, Emilia Iaccarino, Generoso Andria, Maria Vittoria Andreucci, Maria Grazia Marzano, Daniele De Brasi, Pasqua Di Micco, Massimiliano Rossi, Giovanna Roberta Vega, Rossi, M, Di Micco, P, Perone, L, De Brasi, D, Guzzetta, V, Andreucci, Mv, Vega, Gr, Marzano, Mg, Iaccarino, E, and Andria, Generoso
- Subjects
Male ,congenital, hereditary, and neonatal diseases and abnormalities ,Monosomy ,medicine.medical_specialty ,Pathology ,Chromosomal translocation ,Biology ,Translocation, Genetic ,Diagnosis, Differential ,Fatal Outcome ,Intellectual Disability ,Internal medicine ,Gene duplication ,medicine ,Humans ,Abnormalities, Multiple ,translocation (3,5)(q26.1,p14) ,Genetics (clinical) ,Infant ,Karyotype ,medicine.disease ,Smith-Lemli-Opitz Syndrome ,Endocrinology ,Smith–Lemli–Opitz syndrome ,Child, Preschool ,Karyotyping ,dup ,Chromosomes, Human, Pair 5 ,Chromosomes, Human, Pair 3 ,Differential diagnosis ,Trisomy - Abstract
A patient with a multiple congenital anomalies/mental retardation (MCA/MR) syndrome had an unbalanced translocation (3;5)(q26.1;p14), causing partial 5p monosomy and partial 3q trisomy. The phenotype observed in this patient results from the combination of those described in the isolated dup(3q) and del(5p) syndromes. Some clinical features of this patient are shared by the Smith-Lemli-Opitz syndrome (SLOS), a well-known MCA/MR syndrome due to the deficiency of 7-dehydrocholesterol reductase (DHCR7). We review the previously reported cases of chromosomal anomalies with clinical features suggesting SLOS.
- Published
- 2002
27. Metabolic Alkalosis resulting from a Congenital Duodenal Diaphragm
- Author
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A, Passariello, Y, Maddalena, M, Malamisura, S, Rojo, N, Aragione, E, Iaccarino, G, Franco, P, Giliberti, Passariello, A, Maddalena, Y, Malamisura, M, Rojo, S, Aragione, N, Iaccarino, E, Franco, G, and Giliberti, P
- Subjects
Urinary electrolytes concentration ,Neonatal surgery ,Metabolic alkalosi ,Pediatrics, Perinatology and Child Health ,lcsh:RJ1-570 ,lcsh:Surgery ,Case Report ,lcsh:Pediatrics ,Surgery ,lcsh:RD1-811 ,musculoskeletal system ,Metabolic alkalosis - Abstract
Duodenal diaphragm is an unusual cause of upper intestinal obstruction. We present here a neonate with duodenal diaphragm who presented with features of metabolic alkalosis. Further, an algorithm of management of metabolic alkalosis in a newborn is suggested.
- Published
- 2014
28. Cloverleaf skull anomaly and de novo trisomy 4p
- Author
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D, de Brasi, L, Perone, P, di Micco, G, Andria, G, Sebastio, E, Iaccarino, L, Pinto, F, Aliberti, de Brasi, D, Perone, L, di Micco, P, Andria, Generoso, Sebastio, G, Iaccarino, E, Pinto, L, and Aliberti, F.
- Subjects
Male ,Craniosynostoses ,Cloverleaf ,Skull ,Infant, Newborn ,trisomy 4p ,Humans ,Abnormalities, Multiple ,Trisomy ,Chromosomes, Human, Pair 4 ,Letters to the Editor ,In Situ Hybridization, Fluorescence - Published
- 1999
29. General catalogue of Italian earthquakes
- Author
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Iaccarino, E
- Published
- 1973
30. Italian seismicity in the past centuries
- Author
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Iaccarino, E
- Published
- 1973
31. Immunological disorder and Hirschsprung disease in round femoral inferior epiphysis dysplasia
- Author
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E Iaccarino, M. Lecora, G Scarano, Giancarlo Parenti, Salvatore Cucchiara, Generoso Andria, Lecora, M, Parenti, Giancarlo, Iaccarino, E, Scarano, G, Cucchiara, S, and Andria, Generoso
- Subjects
Male ,congenital, hereditary, and neonatal diseases and abnormalities ,Pathology ,medicine.medical_specialty ,Disease ,Osteochondrodysplasias ,Pathology and Forensic Medicine ,medicine ,Cartilage–hair hypoplasia ,Humans ,Femur ,epiphysis dysplasia ,Hirschsprung Disease ,Immunological disorder ,Genetics (clinical) ,business.industry ,Infant, Newborn ,General Medicine ,medicine.disease ,Infant newborn ,medicine.anatomical_structure ,Immune System Diseases ,Epiphysis ,Dysplasia ,Pediatrics, Perinatology and Child Health ,Infantile onset ,Anatomy ,business ,Epiphyses - Abstract
We describe a case of semi-lethal chondrodysplasia with skeletal manifestations, detectable at birth, typical of the round femoral inferior epiphysis dysplasia (RFIED) or 'Glasgow' variant. Immunological abnormalities and Hirschsprung disease, so far described only in cartilage hair hypoplasia (CHH), were documented during the first months of life in our patient. These findings confirm the hypothesis that RFIED is a form of CHH with early infantile onset.
- Published
- 1995
32. CD38 restrains the activity of extracellular cGAMP in a model of multiple myeloma.
- Author
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Cuollo L, Di Cristofano S, Sandomenico A, Iaccarino E, Oliver A, Zingoni A, Cippitelli M, Fionda C, Petillo S, Kosta A, Tassinari V, Petrucci MT, Fazio F, Ruvo M, Santoni A, Raimondo D, and Soriani A
- Abstract
2'3'-cyclic guanosine monophosphate-adenosine monophosphate (cGAMP) is the endogenous agonist of STING; as such, cGAMP has powerful immunostimulatory activity, due to its capacity to stimulate type I interferon-mediated immunity. Recent evidence indicates that cancer cells, under certain conditions, can release cGAMP extracellularly, a phenomenon currently considered important for therapeutic responses and tumor rejection. Nonetheless, the mechanisms that regulate cGAMP activity in the extracellular environment are still largely unexplored. In this work, we collected evidence demonstrating that CD38 glycohydrolase can inhibit extracellular cGAMP activity through its direct binding. We firstly used different cell lines and clinical samples to demonstrate a link between CD38 and extracellular cGAMP activity; we then performed extensive in silico molecular modeling and cell-free biochemical assays to show a direct interaction between the catalytic pocket of CD38 and cGAMP. Altogether, our findings expand the current knowledge about the regulation of cGAMP activity., Competing Interests: The authors declare no competing interests., (© 2024 The Author(s).)
- Published
- 2024
- Full Text
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33. Recombinant humanized Fab fragments targeting the CFC domain of human Cripto-1.
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Sandomenico A, Selis F, Sivaccumar JP, Olimpieri P, Iaccarino E, Cicatiello V, Cantile M, Sanna R, Leonardi A, De Falco S, and Ruvo M
- Subjects
- Animals, Humans, Mice, Antibodies, GPI-Linked Proteins metabolism, Intercellular Signaling Peptides and Proteins, Neoplasm Proteins metabolism, Immunoglobulin Fab Fragments chemistry, Neoplasms
- Abstract
In the era of immunotherapy, the targeting of disease-specific biomarkers goes hand in hand with the development of highly selective antibody-based reagents having optimal pharmacological/toxicological profiles. One interesting and debated biomaker for several types of cancers is the onco-fetal protein Cripto-1 that is selectively expressed in many solid tumours and has been actively investigated as potential theranostic target. Starting from previously described anti-CFC/Cripto-1 murine monoclonal antibodies, we have moved forward to prepare the humanized recombinant Fabs which have been engineered so as to bear an MTGase site useful for a one-step site-specific labelling. The purified and bioconjugated molecules have been extensively characterized and tested on Cripto-1-positive cancer cells through in vitro binding assays. These recombinant Fab fragments recognize the target antigen in its native form on intact cells suggesting that they can be further developed as reagents for detecting Cripto-1 in theranostic settings., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: MENOTTI RUVO reports financial support was provided by Italy Ministry of University. MENOTTI RUVO reports financial support was provided by Campania Region. Maria Cantile, Riccardo Sanna reports financial support was provided by Italy Ministry of University. Maria Cantile and Riccardo Sanna are employees of BIOVIIIx srl., (Copyright © 2023 Elsevier Inc. All rights reserved.)
- Published
- 2024
- Full Text
- View/download PDF
34. Production in Bacteria and Characterization of Engineered Humanized Fab Fragment against the Nodal Protein.
- Author
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Sivaccumar JP, Iaccarino E, Oliver A, Cantile M, Olimpieri P, Leonardi A, Ruvo M, and Sandomenico A
- Abstract
Drug development in recent years is increasingly focused on developing personalized treatments based on blocking molecules selective for therapeutic targets specifically present in individual patients. In this perspective, the specificity of therapeutic targets and blocking agents plays a crucial role. Monoclonal antibodies (mAbs) and their surrogates are increasingly used in this context thanks to their ability to bind therapeutic targets and to inhibit their activity or to transport bioactive molecules into the compartments in which the targets are expressed. Small antibody-like molecules, such as Fabs, are often used in certain clinical settings where small size and better tissue penetration are required. In the wake of this research trend, we developed a murine mAb (3D1) neutralizing the activity of Nodal, an oncofetal protein that is attracting an ever-increasing interest as a selective therapeutic target for several cancer types. Here, we report the preparation of a recombinant Fab of 3D1 that has been humanized through a computational approach starting from the sequence of the murine antibody. The Fab has been expressed in bacterial cells (1 mg/L bacterial culture), biochemically characterized in terms of stability and binding properties by circular dichroism and bio-layer interferometry techniques and tested in vitro on Nodal-positive cancer cells.
- Published
- 2023
- Full Text
- View/download PDF
35. Unveiling CD59-Antibody Interactions to Design Paratope-Mimicking Peptides for Complement Modulation.
- Author
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Sandomenico A, Ruggiero A, Iaccarino E, Oliver A, Squeglia F, Moreira M, Esposito L, Ruvo M, and Berisio R
- Subjects
- Humans, Binding Sites, Antibody, CD59 Antigens metabolism, Complement Membrane Attack Complex metabolism, Complement Inactivating Agents, Complement System Proteins metabolism, HIV-1 physiology
- Abstract
CD59 is an abundant immuno-regulatory human protein that protects cells from damage by inhibiting the complement system. CD59 inhibits the assembly of the Membrane Attack Complex (MAC), the bactericidal pore-forming toxin of the innate immune system. In addition, several pathogenic viruses, including HIV-1, escape complement-mediated virolysis by incorporating this complement inhibitor in their own viral envelope. This makes human pathogenic viruses, such as HIV-1, not neutralised by the complement in human fluids. CD59 is also overexpressed in several cancer cells to resist the complement attack. Consistent with its importance as a therapeutical target, CD59-targeting antibodies have been proven to be successful in hindering HIV-1 growth and counteracting the effect of complement inhibition by specific cancer cells. In this work, we make use of bioinformatics and computational tools to identify CD59 interactions with blocking antibodies and to describe molecular details of the paratope-epitope interface. Based on this information, we design and produce paratope-mimicking bicyclic peptides able to target CD59. Our results set the basis for the development of antibody-mimicking small molecules targeting CD59 with potential therapeutic interest as complement activators.
- Published
- 2023
- Full Text
- View/download PDF
36. Oxidized Substrates of APEH as a Tool to Study the Endoprotease Activity of the Enzyme.
- Author
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Sandomenico A, Gogliettino M, Iaccarino E, Fusco C, Caporale A, Ruvo M, Palmieri G, and Cocca E
- Subjects
- Humans, Models, Molecular, Oxidation-Reduction, Substrate Specificity, Peptide Hydrolases metabolism, Peptides metabolism
- Abstract
APEH is a ubiquitous and cytosolic serine protease belonging to the prolyl oligopeptidase (POP) family, playing a critical role in the processes of degradation of proteins through both exo- and endopeptidase events. Endopeptidase activity has been associated with protein oxidation; however, the actual mechanisms have yet to be elucidated. We show that a synthetic fragment of GDF11 spanning the region 48-64 acquires sensitivity to the endopeptidase activity of APEH only when the methionines are transformed into the corresponding sulphoxide derivatives. The data suggest that the presence of sulphoxide-modified methionines is an important prerequisite for the substrates to be processed by APEH and that the residue is crucial for switching the enzyme activity from exo- to endoprotease. The cleavage occurs on residues placed on the C-terminal side of Met(O), with an efficiency depending on the methionine adjacent residues, which thereby may play a crucial role in driving and modulating APEH endoprotease activity.
- Published
- 2021
- Full Text
- View/download PDF
37. Members of the GADD45 Protein Family Show Distinct Propensities to form Toxic Amyloid-Like Aggregates in Physiological Conditions.
- Author
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Smaldone G, Caruso D, Sandomenico A, Iaccarino E, Focà A, Ruggiero A, Ruvo M, and Vitagliano L
- Subjects
- Amyloid chemistry, Cell Survival, Cells, Cultured, Humans, Intracellular Signaling Peptides and Proteins genetics, MAP Kinase Kinase 7 metabolism, Protein Aggregation, Pathological metabolism, Protein Binding, Protein Conformation, beta-Strand, Protein Stability, Recombinant Proteins, Thermodynamics, GADD45 Proteins, Amyloid metabolism, Intracellular Signaling Peptides and Proteins chemistry, Intracellular Signaling Peptides and Proteins metabolism, Protein Aggregates
- Abstract
The three members (GADD45α, GADD45β, and GADD45γ) of the growth arrest and DNA damage-inducible 45 (GADD45) protein family are involved in a myriad of diversified cellular functions. With the aim of unravelling analogies and differences, we performed comparative biochemical and biophysical analyses on the three proteins. The characterization and quantification of their binding to the MKK7 kinase, a validated functional partner of GADD45β, indicate that GADD45α and GADD45γ are strong interactors of the kinase. Despite their remarkable sequence similarity, the three proteins present rather distinct biophysical properties. Indeed, while GADD45β and GADD45γ are marginally stable at physiological temperatures, GADD45α presents the Tm value expected for a protein isolated from a mesophilic organism. Surprisingly, GADD45α and GADD45β, when heated, form high-molecular weight species that exhibit features (ThT binding and intrinsic label-free UV/visible fluorescence) proper of amyloid-like aggregates. Cell viability studies demonstrate that they are endowed with a remarkable toxicity against SHSY-5Y and HepG2 cells. The very uncommon property of GADD45β to form cytotoxic species in near-physiological conditions represents a puzzling finding with potential functional implications. Finally, the low stability and/or the propensity to form toxic species of GADD45 proteins constitute important features that should be considered in interpreting their many functions.
- Published
- 2021
- Full Text
- View/download PDF
38. The role of Nodal and Cripto-1 in human oral squamous cell carcinoma.
- Author
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Daraghma H, Untiveros G, Raskind A, Iaccarino E, Sandomenico A, Ruvo M, Arnouk H, Ciancio MJ, Cuevas-Nunez M, and Strizzi L
- Subjects
- Adult, Cell Line, Tumor, Humans, Squamous Cell Carcinoma of Head and Neck, Carcinoma, Squamous Cell, Head and Neck Neoplasms, Mouth Neoplasms
- Abstract
Oral squamous cell carcinoma (OSCC) is a common epithelial malignancy of the oral cavity. Nodal and Cripto-1 (CR-1) are important developmental morphogens expressed in several adult cancers and are associated with disease progression. Whether Nodal and CR-1 are simultaneously expressed in the same tumor and how this affects cancer biology are unclear. We investigate the expression and potential role of both Nodal and CR-1 in human OSCC. Immunohistochemistry results show that Nodal and CR-1 are both expressed in the same human OSCC sample and that intensity of Nodal staining is correlated with advanced-stage disease. However, this was not observed with CR-1 staining. Western blot analysis of lysates from two human OSCC line experiments shows expression of CR-1 and Nodal, and their respective signaling molecules, Src and ERK1/2. Treatment of SCC25 and SCC15 cells with both Nodal and CR-1 inhibitors simultaneously resulted in reduced cell viability and reduced levels of P-Src and P-ERK1/2. Further investigation showed that the combination treatment with both Nodal and CR-1 inhibitors was capable of reducing invasiveness of SCC25 cells. Our results show a possible role for Nodal/CR-1 function during progression of human OSCC and that targeting both proteins simultaneously may have therapeutic potential., (© 2020 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd. All rights reserved.)
- Published
- 2021
- Full Text
- View/download PDF
39. A structure-based approach for the development of a bicyclic peptide acting as a miniaturized anti-CD55 antibody.
- Author
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Moreira M, Ruggiero A, Iaccarino E, Barra G, Sandomenico A, Ruvo M, and Berisio R
- Subjects
- Amino Acid Sequence, Binding Sites, Antibody immunology, CD55 Antigens chemistry, Cyclization, Humans, Kinetics, Models, Molecular, Molecular Docking Simulation, Antibodies pharmacology, CD55 Antigens immunology, Miniaturization, Peptides, Cyclic chemistry
- Abstract
CD55 is a major regulator of the complement system, a complex network of proteins that cooperate to clear tissue and blood pathogens from the organism. Indeed, overexpression of CD55 is associated with many diseases and is connected to the resistance mechanisms exhibited by several cancers towards immunotherapy approaches. High level of CD55 expression on tumour cells renders it a good target for both imaging and immunotherapy. Indeed, a conceivable approach to tackle disease is to interfere with CD55-mediated complement regulation with the use of CD55-targeting antibodies. However, the large size and poor tissue penetration together with to the high costs of antibodies often limits their widespread therapeutic use. Here, we employed bioinformatic and chemical approaches to design and synthesize molecules of small dimensions able to mimic a CD55 blocking antibody. As a result, a bicyclic peptide, named as miniAB55, proved to bind CD55 with nanomolar affinity. This molecule represents an attracting chemical scaffold for CD55-directed diagnostic tools in diseases associated with CD55 overproduction. To further support the applicative potential of miniAB55, we prove that the miniAB55 binds CD55 on the same region involved in inactivation of the complement C3 and C5 convertases, thus opening promising scenarios for the development of complement-modulating tools., (Copyright © 2021. Published by Elsevier B.V.)
- Published
- 2021
- Full Text
- View/download PDF
40. Identification and validation of viral antigens sharing sequence and structural homology with tumor-associated antigens (TAAs).
- Author
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Ragone C, Manolio C, Cavalluzzo B, Mauriello A, Tornesello ML, Buonaguro FM, Castiglione F, Vitagliano L, Iaccarino E, Ruvo M, Tagliamonte M, and Buonaguro L
- Subjects
- Amino Acid Sequence, Antigens, Neoplasm chemistry, Antigens, Viral chemistry, Cells, Cultured, Cross Reactions, Databases, Protein, Enzyme-Linked Immunospot Assay, Epitope Mapping, Host-Pathogen Interactions, Humans, Interferon-gamma metabolism, Interferon-gamma Release Tests, Memory T Cells metabolism, Memory T Cells virology, Models, Immunological, Protein Conformation, Sequence Homology, Amino Acid, Antigens, Neoplasm immunology, Antigens, Viral immunology, Epitopes, Immunologic Memory, Memory T Cells immunology
- Abstract
Background: The host's immune system develops in equilibrium with both cellular self-antigens and non-self-antigens derived from microorganisms which enter the body during lifetime. In addition, during the years, a tumor may arise presenting to the immune system an additional pool of non-self-antigens, namely tumor antigens (tumor-associated antigens, TAAs; tumor-specific antigens, TSAs)., Methods: In the present study, we looked for homology between published TAAs and non-self-viral-derived epitopes. Bioinformatics analyses and ex vivo immunological validations have been performed., Results: Surprisingly, several of such homologies have been found. Moreover, structural similarities between paired TAAs and viral peptides as well as comparable patterns of contact with HLA and T cell receptor (TCR) α and β chains have been observed. Therefore, the two classes of non-self-antigens (viral antigens and tumor antigens) may converge, eliciting cross-reacting CD8
+ T cell responses which possibly drive the fate of cancer development and progression., Conclusions: An established antiviral T cell memory may turn out to be an anticancer T cell memory, able to control the growth of a cancer developed during the lifetime if the expressed TAA is similar to the viral epitope. This may ultimately represent a relevant selective advantage for patients with cancer and may lead to a novel preventive anticancer vaccine strategy., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2021. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)- Published
- 2021
- Full Text
- View/download PDF
41. Development of a New Highly Selective Monoclonal Antibody against Preferentially Expressed Antigen in Melanoma (PRAME) and Identification of the Target Epitope by Bio-Layer Interferometry.
- Author
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Sivaccumar JP, Leonardi A, Iaccarino E, Corvino G, Sanguigno L, Chambery A, Russo R, Valletta M, Latino D, Capasso D, Doti N, Ruvo M, and Sandomenico A
- Subjects
- Animals, Antibodies, Monoclonal chemistry, Antibodies, Monoclonal immunology, Antibody Specificity, Antineoplastic Agents, Immunological chemistry, Antineoplastic Agents, Immunological immunology, Dose-Response Relationship, Drug, Enzyme-Linked Immunosorbent Assay, Epitopes chemistry, Flow Cytometry, Humans, Kinetics, Melanoma, Mice, Molecular Targeted Therapy, Protein Binding immunology, Recombinant Proteins, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Antibodies, Monoclonal pharmacology, Antigens, Neoplasm immunology, Antineoplastic Agents, Immunological pharmacology, Drug Development, Epitopes immunology, Interferometry
- Abstract
Background: Monoclonal antibodies (mAbs) against cancer biomarkers are key reagents in diagnosis and therapy. One such relevant biomarker is a preferentially expressed antigen in melanoma (PRAME) that is selectively expressed in many tumors. Knowing mAb's epitope is of utmost importance for understanding the potential activity and therapeutic prospective of the reagents., Methods: We generated a mAb against PRAME immunizing mice with PRAME fragment 161-415; the affinity of the antibody for the protein was evaluated by ELISA and SPR, and its ability to detect the protein in cells was probed by cytofluorimetry and Western blotting experiments. The antibody epitope was identified immobilizing the mAb on bio-layer interferometry (BLI) sensor chip, capturing protein fragments obtained following trypsin digestion and performing mass spectrometry analyses., Results: A mAb against PRAME with an affinity of 35 pM was obtained and characterized. Its epitope on PRAME was localized on residues 202-212, taking advantage of the low volumes and lack of fluidics underlying the BLI settings., Conclusions: The new anti-PRAME mAb recognizes the folded protein on the surface of cell membranes suggesting that the antibody's epitope is well exposed. BLI sensor chips can be used to identify antibody epitopes.
- Published
- 2021
- Full Text
- View/download PDF
42. Selective inhibition of acylpeptide hydrolase in SAOS-2 osteosarcoma cells: is this enzyme a viable anticancer target?
- Author
-
Gogliettino M, Cocca E, Sandomenico A, Gratino L, Iaccarino E, Calvanese L, Rossi M, and Palmieri G
- Subjects
- Apoptosis genetics, Cell Line, Tumor, Cell Proliferation genetics, Cell Survival genetics, Enzyme Inhibitors therapeutic use, Humans, Osteosarcoma drug therapy, Osteosarcoma pathology, Peptide Hydrolases chemistry, Peptide Hydrolases drug effects, Proteasome Endopeptidase Complex genetics, Substrate Specificity, Enzyme Inhibitors chemistry, Molecular Targeted Therapy, Osteosarcoma genetics, Peptide Hydrolases genetics
- Abstract
Serine hydrolases play crucial roles in many physiological and pathophysiological processes and a panel of these enzymes are targets of approved drugs. Despite this, most of the human serine hydrolases remain poorly characterized with respect to their biological functions and substrates and only a limited number of in vivo active inhibitors have been so far identified. Acylpeptide hydrolase (APEH) is a member of the prolyl-oligopeptidase class, with a unique substrate specificity, that has been suggested to have a potential oncogenic role. In this study, a set of peptides was rationally designed from the lead compound SsCEI 4 and in vitro screened for APEH inhibition. Out of these molecules, a dodecapeptide named Ala 3 showed the best inhibitory effects and it was chosen as a candidate for investigating the anti-cancer effects induced by inhibition of APEH in SAOS-2 cell lines. The results clearly demonstrated that Ala 3 markedly reduced cell viability via deregulation of the APEH-proteasome system. Furthermore, flow cytometric analysis revealed that Ala 3 anti-proliferative effects were closely related to the activation of a caspase-dependent apoptotic pathway. Our findings provide further evidence that APEH can play a crucial role in the pathogenesis of cancer, shedding new light on the great potential of this enzyme as an attractive target for the diagnosis and the quest for selective cancer therapies.
- Published
- 2021
- Full Text
- View/download PDF
43. Insights into the Interaction Mechanism of DTP3 with MKK7 by Using STD-NMR and Computational Approaches.
- Author
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Sandomenico A, Di Rienzo L, Calvanese L, Iaccarino E, D'Auria G, Falcigno L, Chambery A, Russo R, Franzoso G, Tornatore L, D'Abramo M, Ruvo M, Milanetti E, and Raimondo D
- Abstract
GADD45β/MKK7 complex is a non-redundant, cancer cell-restricted survival module downstream of the NF-kB survival pathway, and it has a pathogenically critical role in multiple myeloma, an incurable malignancy of plasma cells. The first-in-class GADD45β/MKK7 inhibitor DTP3 effectively kills MM cells expressing its molecular target, both in vitro and in vivo, by inducing MKK7/JNK-dependent apoptosis with no apparent toxicity to normal cells. DTP3 combines favorable drug-like properties, with on-target-specific pharmacology, resulting in a safe and cancer-selective therapeutic effect; however, its mode of action is only partially understood. In this work, we have investigated the molecular determinants underlying the MKK7 interaction with DTP3 by combining computational, NMR, and spectroscopic methods. Data gathered by fluorescence quenching and computational approaches consistently indicate that the N-terminal region of MKK7 is the optimal binding site explored by DTP3. These findings further the understanding of the selective mode of action of GADD45β/MKK7 inhibitors and inform potential mechanisms of drug resistance. Notably, upon validation of the safety and efficacy of DTP3 in human trials, our results could also facilitate the development of novel DTP3-like therapeutics with improved bioavailability or the capacity to bypass drug resistance.
- Published
- 2020
- Full Text
- View/download PDF
44. Generation and testing of engineered multimeric Fabs of trastuzumab.
- Author
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Selis F, Sandomenico A, Cantile M, Sanna R, Calvanese L, Falcigno L, Dell'Omo P, Esperti A, De Falco S, Focà A, Caporale A, Iaccarino E, Truppo E, Scaramuzza S, Tonon G, and Ruvo M
- Subjects
- Amino Acid Sequence, Breast Neoplasms pathology, Carcinoma pathology, Cell Line, Tumor, Cystine chemistry, DNA, Complementary genetics, Drug Design, Drug Screening Assays, Antitumor, Escherichia coli, Female, Fluorescent Dyes, Humans, Immunoconjugates immunology, Immunoglobulin Fab Fragments genetics, Immunoglobulin Fab Fragments immunology, Models, Molecular, Peptide Fragments chemical synthesis, Protein Conformation, Protein Engineering, Protein Multimerization, Receptor, ErbB-2 immunology, Recombinant Proteins chemistry, Recombinant Proteins genetics, Recombinant Proteins immunology, Surface Plasmon Resonance, Trastuzumab immunology, Immunoconjugates chemistry, Immunoglobulin Fab Fragments chemistry, Transglutaminases immunology, Trastuzumab chemistry
- Abstract
Recombinant antibodies fragments in several new formats are routinely investigated and used in diagnostic and therapeutic applications as anti-cancers molecules. New antibody formats are generated to compensate the need for multispecificity and site-specific introduction of fluorescent dyes, cytotoxic payloads or for generating semisynthetic multimeric molecules. Fabs of trastuzumab bearing transglutaminase (MTG) reactive sites were generated by periplasmic expression in E. coli and purified. Multimeric Fabs were generated by either disulfide bridge formation or by using MTG-sensitive peptide linkers. Binding to receptor was assessed by ELISA and SPR methods. Internalization and growth inhibition assays were performed on BT-474 and SKBR3 Her2+ cells. Fabs were successfully produced and dimerized or trimerized using MTG and suitably designed peptide linkers. Site-specific derivatizations with fluorophores were similarly achieved. The monomeric, dimeric and trimeric variants bind the receptor with affinities similar or superior to the full antibody. Fab and Fab2 are rapidly internalized in Her2+ cells and exhibit growth inhibition abilities similar to the full antibody. Altogether, the data show that the recombinant Fabs can be produced in E. coli and converted into multimeric variants by MTG-based bioconjugation. Similar approaches are extendable to the introduction of cytotoxic payloads for the generation of novel Antibody Drug Conjugates., Competing Interests: Declaration of competing interest The authors declare no conflicts of interests., (Copyright © 2020 Elsevier B.V. All rights reserved.)
- Published
- 2020
- Full Text
- View/download PDF
45. Exploring the Interaction between the SWI/SNF Chromatin Remodeling Complex and the Zinc Finger Factor CTCF.
- Author
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Valletta M, Russo R, Baglivo I, Russo V, Ragucci S, Sandomenico A, Iaccarino E, Ruvo M, De Feis I, Angelini C, Iachettini S, Biroccio A, Pedone PV, and Chambery A
- Subjects
- Adenosine Triphosphatases genetics, Binding Sites genetics, Cell Cycle Proteins genetics, Gene Expression Regulation genetics, Genome, Human genetics, Humans, Multiprotein Complexes genetics, Protein Interaction Maps genetics, Tandem Mass Spectrometry, Cohesins, CCCTC-Binding Factor genetics, Chromatin Assembly and Disassembly genetics, Chromosomal Proteins, Non-Histone genetics, DNA Helicases genetics, Nuclear Proteins genetics, Transcription Factors genetics, Zinc Fingers genetics
- Abstract
The transcription factor CCCTC-binding factor (CTCF) modulates pleiotropic functions mostly related to gene expression regulation. The role of CTCF in large scale genome organization is also well established. A unifying model to explain relationships among many CTCF-mediated activities involves direct or indirect interactions with numerous protein cofactors recruited to specific binding sites. The co-association of CTCF with other architectural proteins such as cohesin, chromodomain helicases, and BRG1, further supports the interplay between master regulators of mammalian genome folding. Here, we report a comprehensive LC-MS/MS mapping of the components of the switch/sucrose nonfermentable (SWI/SNF) chromatin remodeling complex co-associated with CTCF including subunits belonging to the core, signature, and ATPase modules. We further show that the localization patterns of representative SWI/SNF members significantly overlap with CTCF sites on transcriptionally active chromatin regions. Moreover, we provide evidence of a direct binding of the BRK-BRG1 domain to the zinc finger motifs 4-8 of CTCF, thus, suggesting that these domains mediate the interaction of CTCF with the SWI/SNF complex. These findings provide an updated view of the cooperative nature between CTCF and the SWI/SNF ATP-dependent chromatin remodeling complexes, an important step for understanding how these architectural proteins collaborate to shape the genome.
- Published
- 2020
- Full Text
- View/download PDF
46. Structure-based design of small bicyclic peptide inhibitors of Cripto-1 activity.
- Author
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Iaccarino E, Calvanese L, Untiveros G, Falcigno L, D'Auria G, Latino D, Sivaccumar JP, Strizzi L, Ruvo M, and Sandomenico A
- Subjects
- Activin Receptors, Type I genetics, Activin Receptors, Type I metabolism, Amino Acid Motifs, Cell Line, Tumor, Cell Proliferation drug effects, Cell Survival drug effects, Drug Design, GPI-Linked Proteins chemistry, GPI-Linked Proteins genetics, GPI-Linked Proteins metabolism, Humans, Intercellular Signaling Peptides and Proteins chemistry, Intercellular Signaling Peptides and Proteins genetics, Intercellular Signaling Peptides and Proteins metabolism, Magnetic Resonance Spectroscopy, Neoplasm Proteins chemistry, Neoplasm Proteins genetics, Neoplasm Proteins metabolism, Peptides pharmacology, GPI-Linked Proteins antagonists & inhibitors, Neoplasm Proteins antagonists & inhibitors, Peptides chemistry
- Abstract
Bicyclic peptides assembled around small organic scaffolds are gaining an increasing interest as new potent, stable and highly selective therapeutics because of their uncommon ability to specifically recognize protein targets, of their small size that favor tissue penetration and of the versatility and easiness of the synthesis. We have here rationally designed bicyclic peptides assembled around a common tri-bromo-methylbenzene moiety in order to mimic the structure of the CFC domain of the oncogene Cripto-1 and, more specifically, to orient in the most fruitful way the hot spot residues H120 and W123. Through the CFC domain, Cripto-1 binds the ALK4 receptor and other protein partners supporting uncontrolled cell growth and proliferation. Soluble variants of CFC have the potential to inhibit these interactions suppressing the protein activity. A CFC analog named B3 binds ALK4 in vitro with an affinity in the nanomolar range. Structural analyses in solution via NMR and CD show that B3 has rather flexible conformations, like the parent CFC domain. The functional effects of B3 on the Cripto-1-positive NTERA cancer cell line have been evaluated showing that both CFC and B3 are cytotoxic for the cells and block the Cripto-1 intracellular signaling. Altogether, the data suggest that the administration of the soluble CFC and of the structurally related analog has the potential to inhibit tumor growth., (© 2020 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.)
- Published
- 2020
- Full Text
- View/download PDF
47. Oral Delivery of a Tetrameric Tripeptide Inhibitor of VEGFR1 Suppresses Pathological Choroid Neovascularization.
- Author
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Tarallo V, Iaccarino E, Cicatiello V, Sanna R, Ruvo M, and De Falco S
- Subjects
- Administration, Oral, Animals, Intravitreal Injections, Lasers, Mice, Inbred C57BL, Oligopeptides chemistry, Spectroscopy, Fourier Transform Infrared, Trifluoroacetic Acid chemistry, Vascular Endothelial Growth Factor Receptor-1 metabolism, Choroidal Neovascularization drug therapy, Choroidal Neovascularization pathology, Oligopeptides administration & dosage, Oligopeptides therapeutic use, Vascular Endothelial Growth Factor Receptor-1 antagonists & inhibitors
- Abstract
Age-related macular degeneration (AMD) is the primary cause of blindness in advanced countries. Repeated intravitreal delivery of anti-vascular endothelial growth factor (VEGF) agents has represented an important advancement for the therapy of wet AMD with significative results in terms of blindness prevention and partial vision restore. Nonetheless, some patients are not responsive or do not attain significant visual improvement, intravitreal injection may cause serious complications and important side effects have been reported for the prolonged block of VEGF-A. In order to evaluate new anti-angiogenic strategies, we focused our attention on VEGF receptor 1 (VEGFR1) developing a specific VEGFR-1 antagonist, a tetrameric tripeptide named inhibitor of VEGFR 1 (iVR1). We have evaluated its anti-angiogenic activity in the preclinical model of AMD, the laser-induced choroid neovascularization (CNV). iVR1 is able to potently inhibit CNV when delivered by intravitreal injection. Surprisingly, it is able to significantly reduce CNV also when delivered by gavage. Our data show that the specific block of VEGFR1 in vivo represents a valid alternative to the block of VEGF-A and that the inhibition of the pathological neovascularization at ocular level is also possible by systemic delivery of compounds not targeting VEGF-A.
- Published
- 2020
- Full Text
- View/download PDF
48. Investigating the oxidative refolding mechanism of Cripto-1 CFC domain.
- Author
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Iaccarino E, Sandomenico A, Corvino G, Focà G, Severino V, Russo R, Caporale A, Raimondo D, D'Abramo M, Alba J, Chambery A, and Ruvo M
- Subjects
- Amino Acid Sequence, Disulfides chemistry, GPI-Linked Proteins metabolism, Humans, Intercellular Signaling Peptides and Proteins metabolism, Kinetics, Molecular Dynamics Simulation, Neoplasm Proteins metabolism, Oxidation-Reduction, Peptide Fragments chemistry, Protein Domains, GPI-Linked Proteins chemistry, Intercellular Signaling Peptides and Proteins chemistry, Neoplasm Proteins chemistry, Protein Refolding
- Abstract
Using a combined approach based on MS, enzyme digestion and advanced MD studies we have determined the sequential order of formation of the three disulfide bridges of the Cripto-1 CFC domain. The domain has a rare pattern of bridges and is involved in the recognition of several receptors. The bridge formation order is C1-C4, C3-C5, C2-C6, however formation of C1-C4 plays no roles for the formation of the others. Folding is driven by formation of the C3-C5 bridge and is supported by residues lying within the segment delimited by these cysteines. We indeed observe that variants CFC-W123A and CFC-ΔC1/C4, where C1 and C4 are replaced by serines, are able to refold in the same time window as the wild type, while CFC-K132A and CFC-W134A are not. A variant where cysteines of the second and third bridge are mutated to serine, convert slowly to the monocyclic molecule. Data altogether support a mechanism whereby the Cripto-1 CFC domain refolds by virtue of long-range intramolecular interactions that involve residues close to cysteines of the second and third bridge. These findings are supported by the in silico study that shows how distant parts of the molecules come into contact on a long time scale., (Copyright © 2019 Elsevier B.V. All rights reserved.)
- Published
- 2019
- Full Text
- View/download PDF
49. Development of conformational antibodies targeting Cripto-1 with neutralizing effects in vitro.
- Author
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Focà G, Iaccarino E, Focà A, Sanguigno L, Untiveros G, Cuevas-Nunez M, Strizzi L, Leonardi A, Ruvo M, and Sandomenico A
- Subjects
- Activin Receptors, Type I immunology, Activin Receptors, Type I metabolism, Animals, Antibodies, Monoclonal, Murine-Derived immunology, Antibodies, Neoplasm immunology, Antibodies, Neutralizing immunology, Cell Line, Tumor, Endoplasmic Reticulum Chaperone BiP, Heat-Shock Proteins immunology, Heat-Shock Proteins metabolism, Humans, Mice, Mice, Inbred BALB C, Protein Domains, Antibodies, Monoclonal, Murine-Derived chemistry, Antibodies, Neoplasm chemistry, Antibodies, Neutralizing chemistry, Flow Cytometry, GPI-Linked Proteins immunology, GPI-Linked Proteins metabolism, Intercellular Signaling Peptides and Proteins immunology, Intercellular Signaling Peptides and Proteins metabolism, Neoplasm Proteins immunology, Neoplasm Proteins metabolism
- Abstract
Human Cripto-1 (Cripto-1), the founding member of the EGF-CFC superfamily, is a key regulator of many processes during embryonic development and oncogenesis. Cripto-1 is barely present or even absent in normal adult tissues while it is aberrantly re-expressed in various tumors. Blockade of the CFC domain-mediated Cripto-1 functions is acknowledged as a promising therapeutic intervention point to inhibit the tumorigenic activity of the protein. In this work, we report the generation and characterization of murine monoclonal antibodies raised against the synthetic folded CFC [112-150] domain of the human protein. Through subtractive ELISA assays clones were screened for the ability to specifically recognize "hot spot" residues on the CFC domain, which are crucial for the interaction with Activin Type I receptor (ALK4) and GRP78. On selected antibodies, SPR and epitope mapping studies have confirmed their specificity and have revealed that recognition occurs only on a conformational epitope. Furthermore, FACS analyses have confirmed the ability of 1B4 antibody to recognize the membrane-anchored and soluble native Cripto-1 protein in a panel of human cancer cells. Finally, we have evaluated its functional effects through in vitro cellular signaling assays and cell cycle analysis. These findings suggest that the selected anti-CFC mAbs have the potential to neutralize the protein oncogenic activity and may be used as theranostic molecules suitable as tumor homing agents for Cripto-1-overexpressing cancer cells and tissues and to overcome drug-resistance in routine cancer therapies., (Copyright © 2019 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.)
- Published
- 2019
- Full Text
- View/download PDF
50. Intrinsic structural versatility of the highly conserved 412-423 epitope of the Hepatitis C Virus E2 protein.
- Author
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Balasco N, Barone D, Iaccarino E, Sandomenico A, De Simone A, Ruvo M, and Vitagliano L
- Subjects
- Epitopes chemistry, Hepacivirus chemistry, Molecular Dynamics Simulation, Viral Hepatitis Vaccines chemistry, Viral Proteins chemistry
- Abstract
HCV infection is a major threaten for human health as it affects hundreds of million people worldwide. Here we investigated the conformational properties of the 412-423 fragment of the envelope E2 protein, one of the most immunogenic regions of the virus proteome whose characterization may provide interesting insights for anti-HCV vaccine development. The spectroscopic characterization of the polypeptide unravels its unexpected tendency to form amyloid-like aggregates. When kept in monomeric state, it shows a limited tendency to adopt regular secondary structure. Enhanced molecular dynamics simulations, starting from four distinct conformational states, highlight its structural versatility. Interestingly, all multiform conformational states of the polypeptide detected in crystallographic complexes with antibodies are present in the structural ensemble of all simulations. This observation corroborates the idea that known antibodies recognize this region through a conformational selection mechanism. Accordingly, the design of effective anti-HCV vaccines should consider the intrinsic flexibility of this region. The structural versatility of the 412-423 region is particularly puzzling if its remarkable sequence conservation is considered. It is likely that flexibility and sequence conservation are important features that endow this epitope with the ability to accomplish distinct functions such as immunity escape and interaction with host receptors., (Copyright © 2018 Elsevier B.V. All rights reserved.)
- Published
- 2018
- Full Text
- View/download PDF
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