P.M. Biava, M. Basevi, L. Biggiero, A. Borgonovo, E. Borgonovo and F. Burigana
The recent tumor research has lead scientists to recognize the central role played by cancer stem cells in sus- taining malignancy and chemo-resistance. A model of cancer presented by one of us describes the mechanisms that give rise to the different kinds of cancer stem-like cells and the role of these cells in cancer diseases. The model implies a shift in the conceptualization of the disease from reductionism to complexity theory. By exploiting the link between the agent- based simulation technique and the theory of complexity, the medical view is here translated into a corresponding compu- tational model. Two main categories of agents characterize the model, 1) cancer stem-like cells and 2) stem cell differen- tiation stage factors. Cancer cells agents are then distinguished based on the differentiation stage associated with the ma- lignancy. Differentiation factors interact with cancer cells and then, with varying degrees of fitness, induce differentiation or cause apoptosis. The model inputs are then fitted to experimental data and numerical simulations carried out. By per- forming virtual experiments on the model’s choice variables a decision-maker (physician) can obtains insights on the pro- gression of the disease and on the effects of a choice of administration frequency and or dose. The model also paves the way to future research, whose perspectives are discussed.
1Maria Cecilia Hospital, Gruppo Villa Maria (GVM) Care & Research and Ettore Sansavini Health Science Foundation, Cotignola and Lugo, Ravenna, Italy; and 2Department of
Experimental, Diagnostic and Specialty Medicine, University of Bologna, Italy
Margherita Maioli, Salvatore Rinaldi, Sara Santaniello, Alessandro Castagna,
Gianfranco Pigliaru, Sara Gualini, Claudia Cavallini, Vania Fontani, and Carlo Ventura
Department of Biomedical Sciences, University of Sassari, Sassari, Italy
Laboratory of Molecular Biology and Stem Cell Engineering, National Institute of Biostructures and Biosystems, Bologna, Italy
Rinaldi Fontani Institute, Florence, Italy
§Cardiovascular Department, S. Orsola-Malpighi Hospital, University of Bologna, Bologna, Italy
Somatic cells can be directly reprogrammed to alternative differentiated fates without first becoming stem/
progenitor cells. Nevertheless, the initial need for viral-mediated gene delivery renders this strategy unsafe in
humans. Here, we provide evidence that exposure of human skin fibroblasts to a Radio Electric Asymmetric
Conveyer (REAC), an innovative device delivering radio electric conveyed fields at a radiofrequency of 2.4 GHz,
afforded remarkable commitment toward cardiac, neuronal, and skeletal muscle lineages. REAC induced the
transcription of tissue-restricted genes, including Mef2c, Tbx5, GATA4, Nkx2.5, and prodynorphin for cardiac
reprogramming, as well as myoD, and neurogenin 1 for skeletal myogenesis and neurogenesis, respectively.
Conversely, REAC treatment elicited a biphasic effect on a number of stemness-related genes, leading to early
transcriptional increase of Oct4, Sox2, cMyc, Nanog, and Klf4 within 6–20 h, followed by a downregulation at
later times. The REAC action bypassed a persistent reprogramming toward an induced pluripotent stem celllike
state and involved the transcriptional induction of the NADPH oxidase subunit Nox4. Our results show for
the first time the feasibility of using a physical stimulus to afford the expression of pluripotentiality in human
adult somatic cells up to the attainment of three major target lineages for regenerative medicine.
Received for publication, November 20, 2009, and in revised form, January 15, 2010 Published, JBC Papers in Press, January 22, 2010, DOI 10.1074/jbc.M109.087254
Vincenzo Lionetti, Silvia Cantoni, Claudia Cavallini, Francesca Bianchi, Sabrina Valente, Irene Frascari,
Elena Olivi, Giovanni D. Aquaro, Francesca Bonavita, Ignazio Scarlata, Margherita Maioli, Valentina Vaccari,
Riccardo Tassinari, Antonietta Bartoli, Fabio A. Recchia, Gianandrea Pasquinelli, and Carlo Ventura
From the Sector of Medicine, Scuola Superiore S. Anna, I-56124 Pisa, Italy, the Laboratory of Molecular Biology and Stem Cell
Engineering, Cardiovascular Department-National Institute of Biostructures and Biosystems, S. Orsola-Malpighi Hospital,
University of Bologna, I-40138 Bologna, Italy, the Bioscience Institute, RSM-47891 Falciano, Republic of San Marino, the §Institute
of Clinical Physiology, Consiglio Nazionale delle Ricerche Fondazione G. Monasterio, I-56124 Pisa, Italy, the ‡‡Department of
Biomedical Sciences, University of Sassari, I-07100 Sassari, Italy, the §§Department of Physics, University of Pisa, I-56124 Pisa, Italy,
the Department of Physiology, New York Medical College, Valhalla, New York 10595, and the **Department of Hematology,
Oncology, and Clinical Pathology, University of Bologna, I-40138 Bologna, Italy
A NOVEL DIFFERENTIATING GLYCOCONJUGATE AFFORDING A HIGH THROUGHPUT OF CARDIOGENESIS IN EMBRYONIC STEM CELLS
Received for publication, February 20, 2004, and in revised form, March 16, 2004
Published, JBC Papers in Press, March 24, 2004, DOI 10.1074/jbc.M401869200
Carlo Ventura, Margherita Maioli, Yolande Asara, Daniela Santoni, Ignazio Scarlata,
Silvia Cantoni, and Alberto Perbellini
From the Laboratory of Molecular Biology and Stem Cell Engineering, National Institute of Biostructures and
Biosystems, University of Bologna, I-40138 Bologna, Italy, the Department of Biomedical Sciences, University of Sassari,
I-07100 Sassari, Italy, and the Department of Biochemistry, Biophysics, and Chemistry of Macromolecules, University of
Trieste, I-34127 Trieste, Italy
Received for publication, October 3, 2006, and in revised form, March 9, 2007 Published, JBC Papers in Press, March 15, 2007, DOI 10.1074/jbc.M609350200
Carlo Ventura, Silvia Cantoni, Francesca Bianchi, Vincenzo Lionetti, Claudia Cavallini, Ignazio Scarlata,
Laura Foroni, Margherita Maioli, Laura Bonsi, Francesco Alviano, Valentina Fossati, Gian Paolo Bagnara,
Gianandrea Pasquinelli, Fabio A. Recchia, and Alberto Perbellini
From the Laboratory of Molecular Biology and Stem Cell Engineering, Institute of Cardiology, National Institute of Biostructures
and Biosystems, University of Bologna, I-40138 Bologna, Italy, Department of Experimental Pathology, University of Bologna,
I-40138 Bologna, Italy, Department of Histology, Embryology, and Applied Biology, University of Bologna, I-40138 Bologna, Italy,
Sector of Medicine, Scuola Superiore S. Anna, CNR Institute of Clinical Physiology, I-56124 Pisa, Italy and Department of
Physiology, New York Medical College, Valhalla, New York 10595, and Department of Biomedical Sciences,
University of Sassari, I-07100 Sassari, Italy
Carlo Ventura, Elisabetta Zinellu, Emiliana Maninchedda, Margherita Maioli
Abstract—The cardiac differentiation of embryonic stem (ES) cells was found to involve prodynorphin gene and dynorphin
B expression and was associated with the interaction of secreted dynorphin B with cell surface opioid receptors coupled
with protein kinase C (PKC) signaling and complex subcellular redistribution patterning of selected PKC isozymes.
Here, confocal microscopy revealed the presence of immunoreactive dynorphin B–like material in GTR1 ES cells,
suggesting that dynorphin peptides may also act intracellularly. Opioid binding sites were identified in ES cell nuclei,
with a single dissociation constant in the low nanomolar range. A significant increase in Bmax for a opioid receptor
ligand was observed in nuclei isolated from ES-derived cardiomyocytes compared with nuclei from undifferentiated
cells. Direct exposure of nuclei isolated from undifferentiated ES cells to dynorphin B or U-50,488H, a synthetic
opioid receptor agonist, time- and dose-dependently activated the transcription of GATA-4 and Nkx-2.5, 2 cardiac
lineage–promoting genes. Nuclear exposure to dynorphin B also enhanced the rate of prodynorphin gene transcription.
These responses were abolished in a stereospecific fashion by the incubation of isolated nuclei with selective opioid
receptor antagonists. Nuclei isolated from undifferentiated cells were able to phosphorylate the acrylodan-labeled
MARCKS peptide, a high-affinity fluorescent PKC substrate. Exposure of isolated nuclei to dynorphin B induced a
remarkable increase in nuclear PKC activity, which was suppressed by opioid receptor antagonists. Nuclear treatment
with PKC inhibitors abolished the capability of dynorphin B to prime the transcription of cardiogenic genes. (Circ Res.
Carlo Ventura, Elisabetta Zinellu, Emiliana Maninchedda, Marina Fadda, Margherita Maioli
Abstract—The prodynorphin gene and its product, dynorphin B, have been found to promote cardiogenesis in embryonic
cells by inducing the expression of GATA-4 and Nkx-2.5, two transcription factor– encoding genes essential for
cardiogenesis. The molecular mechanism(s) underlying endorphin-induced cardiogenesis remain unknown. In the
present study, we found that GTR1 embryonic stem (ES) cells expressed cell surface opioid receptors, as well as
protein kinase C (PKC)-, -1, -2, -, -, and -. Cardiac differentiation was associated with a marked increase in the
Bmax value for a selective opioid receptor ligand and complex subcellular redistribution of selected PKC isozymes.
PKC-, -1, -2, -, and - all increased in the nucleus of ES-derived cardiac myocytes, compared with nuclei from
undifferentiated cells. In both groups of cells, PKC- and - were mainly expressed at the nuclear level. The nuclear
increase of PKC-, -1, and -2 was due to a translocation from the cytosolic compartment. In contrast, the increase of
both PKC- and PKC- in the nucleus of ES-derived cardiomyocytes occurred independently of enzyme translocation,
suggesting changes in isozyme turnover and/or gene expression during cardiogenesis. No change in PKC- expression
was observed during cardiac differentiation. Opioid receptor antagonists prevented the nuclear increase of PKC-,
PKC-1, and PKC-2 and reduced cardiomyocyte yield but failed to affect the nuclear increase in PKC- and -. PKC
inhibitors prevented the expression of cardiogenic genes and dynorphin B in ES cells and abolished their development
into beating cardiomyocytes. (Circ Res. 2003;92:617-622.)
Key Words: protein kinase C cardiac differentiation embryonic stem cells gene expression endorphins
LO STRESS E L’ANSIA NON SONO SOLO UN EFFETTO COLLATERALE EMOTIVO DELLA DIAGNOSI, MA COSTITUISCONO ANCHE UN FATTORE IN GRADO DI RIDURRE SIGNIFICATIVAMENTE L’EFFICACIA DEI FARMACI CONTRO IL CANCRO ALLA PROSTATA E DI ACCELERARE LO SVILUPPO DELLA MALATTIA.
Una nuova ricerca condotta da ricercatori della Wake Forest Baptist Medical Center dimostra che lo stress e l’ansianon sono solo un effetto collaterale emotivo della diagnosi, ma costituiscono anche un fattore in grado di ridurre significativamente l’efficacia dei farmaci contro il cancro alla prostata e di accelerare lo sviluppo della malattia.
In un primo studio sono stati utilizzati topi cui sono state impiantate cellule umane “malate” e trattate con un farmaco attualmente in sperimentazione clinica per il trattamento del cancro della prostata. Quando gli animali erano tenuti in una condizione di calma e assenza di stress, il farmaco è risultato efficace nell’inibizione della crescita tumorale. Viceversa, nella conduzione in cui i topi venivano sottoposti a elevati livelli di stress, il farmaco non è stato in grado di bloccare lo sviluppo tumorale e di debellare le cellule malate.
In un secondo studio, alcuni topi geneticamente modificati per sviluppare il cancro alla prostata sono stati ripetutamente sottoposti a stress: questi animali hanno sviluppato tumori di dimensioni maggiori, e nel momento della terapia il farmaco antitumorale bicalutamide (attualmente in uso) non ha prodotto alcun effetto rispetto al gruppo di topi non stressati.
Sembrerebbero in gioco processi biologici attraverso cui l’epifrenina innesca una reazione a catena che inibisce la morte cellulare delle cellule malate. Quindi in presenza di stress e ansia, ovviamente conseguente a tali diagnosi, si può innescare un circolo vizioso tra emotività negativa e progressione della patologia.
Tuttavia, nel momento in cui si somministrano ai topi farmaci beta-bloccanti (che inibiscono la reazione a cascata elicitata dall’epifrenina che impedisce la morte cellulare delle cellule tumorali) anche livelli elevati di stress non hanno favorito l’avanzamento del tumore. I ricercatori hanno intenzione di verificare la generalizzabilità di tali risultati su un campione umano.
Pertanto identificare i pazienti particolarmente colpiti da stress e ansia può essere utile per evitare che tali variabili psicofisiologiche costituiscano un ostacolo all’efficacia terapeutica, intervenendo sia a livello farmacologico (introduzione di betabloccanti) che psicoterapico per la gestione dell’ansia e dello stress.