Assistant Professor – BIO/14 Pharmacology
Reasearch activity: Pharmacology and Toxicology
Tel. +39 0382 986445
Fax +39 0382 986328
Assistant Professor (RTDB) – BIO/05 Zoology
Fax: + 39 382 986079
Contact for students: appointment by E-mail
Assistant Professor (RTDB) – BIO/04 Plant Physiology
Fax: + 39 382 987240
Contact for students:
Tel. ++39 0382 98 6309; 6310
Fax +39 0382 98 6406
Assistant Professor – BIO/05 Zoology
Tel. ++39 0382 986023
Contact for students: appointment by E-mail
Assistant Professor – BIO/04 Plant Physiology
Tel. +39 0382 985583 (Office)
Fax +39 0382 528496
Contact for students: appointment by E-mail
The call for application to the PhD Course for the academic year 2017/2018 (XXXIII Cycle) is now open The deadline is July 14, 2017 – 17:00 local time.
For informations about the PhD Programme in Genetics Molecular and Cellular Biology see the web site page
To download the documentation and for all the informations and details to file an application see also the University web page
Erasmus + / International mobility
Incoming Erasmus students at our department are invited to contact Prof. Rosanna Nano, Chief Erasmus + and International Mobility delegate for Biology and Biotechnology in order to settle their learning agreement.
For more information useful to incoming Erasmus students see the following University of page web page: Erasmus Incoming Students
Erasmus contacts at the University of Pavia
ERASMUS INCOMING STUDENTS
Chantal Riccardi, firstname.lastname@example.org
+39 0382 984601
OUTGOING ERASMUS STUDENTS
Irene De Stefani, email@example.com
+39 0382 984004
Erasmus Traineeship Students Incoming at DBB
The DBB Erasmus Traineeship coordinator is Prof. Sergio Comincini.
Students that are going to apply for available positions within DBB are invited to directly interact with the group leaders of the research groups listed here: http://dbb.unipv.it/eng/research-topics/
Additional information and contact details for the group leaders can be found in their personal pages, that you can reach using the search box in the homepage or by looking into the DBB people list.
Erasmus Traineeship contacts at the University of Pavia
The UNIPV Erasmus Traineeship coordinator is Dr. Francesca Montagna Napoleone, firstname.lastname@example.org
Assistant Professor – BIO/11 Molecular Biology (05/E2)
Elena Giulotto: Molecular and Cellular Biology
Elena Raimondi: Molecular Cytogenetics
Solomon Nergadze: Epigenomics and Bioinformatcs
Eleonora Cappelletti (PhD student), Marco Corbo (PhD student), Riccardo Gamba (PhD student), Francesco Gozzo (PhD student), Lela Khoriauli (Post-doctoral fellow), Francesca Piras (Post-doctoral fellow), Annalisa Roberti (PhD student), Marco Santagostino (Post-doctoral fellow)
The complementary competences of the research groups involved allow them to approach the scientific topics described below taking advantage of different experimental procedures encompassing a wide range of cellular, biomolecular (including next generation sequencing), cytogenetic (including the analysis of single chromatin molecules) and bioinformatic methodologies.
The research projects are focused on the molecular mechanisms involved in the maintenance of the integrity of the mammalian genome which play a central role in carcinogenesis and in evolution. The main structures needed for the maintenance of functional and structural chromosome integrity are centromeres and telomeres.
1) The centromere and its epigenetic nature
Centromeres are essential for the separation of chromosomes during cell division. They are enigmatic structures since, contrary to other genetic loci, the function of centromeres is not determined by the underlying DNA sequence, but depends on epigenetic factors. In particular, centromeric chromatin is peculiar for the presence of a modified H3 histone, CENP-A, and of a number of proteins forming a complex named CCAN (Constitutive Centromere Associated Network). Moreover, centrochromatin is characterised by histone modifications whose role in centromere function is still debated.
Mammalian centromeres are typically associated to highly repetitive DNA (satellite DNA); this pattern of organisation hampered a detailed molecular analysis of centromeric chromatin.
The equid model system
Our research group demonstrated, for the first time, the existence in nature of satellite-free centromeres which are stable and fully functional (Wade et al Science 2009). These centromeres are present in a number of chromosomes in the genus Equus (horses, donkeys, zebras), they represent a unique model system for the study of the epigenetic control of centromere function and of its role in karyotype evolution (Piras et al PLoS Genetics 2010; Purgato et al Chromosoma 2015).
It is well known that alterations in chromosome segregation are responsible for the occurrence of numerical chromosomal abnormalities observed in the large majority of tumours.
The equid model system offers the opportunity to investigate the mechanisms involved in the numerical chromosome instability associated to the development and progression of many different types of cancer.
The absence of repetitive DNA at the centromere of a number of equid centromeres makes it possible to study the architectural organisation of centrochromatin and, in particular, the role of DNA methylation, transcription and histone modifications.
Collaborations: Kevin Sullivan (University of Galway, Ireland), Aurora Ruiz-Herrera (Universitat Autonoma de Barcelona, Spain), Giulio Pavesi (Università di Milano), Mariano Rocchi (Università di Bari), Doug Antczak (Cornell University, Ithaca New York), The Horse Genome Project, The Horse FAANG Project.
2) The telomere and its transcription
Telomeres are nucleoproteic structures located at the termini of linear chromosomes. In mammals, telomeric DNA, consisting of the tandem repetition of the hexasanucleotide TTAGGG, is bound to a multiproteic complex named shelterin. Telomeres protect chromosome ends from degradation and from fusion with other telomeres. Telomere malfunctioning causes chromosome rearrangements that can lead to tumorigenesis. Too short telomeres cannot be distinguished from accidental DNA breaks and cause genomic instability. Our group contributed to the demonstration that the telomeres are transcribed in non-coding RNA molecules named TElomeric Repeat-containing RNA (TERRA) (Azzalin et al. Science 2007). We also demonstrated that telomere transcription is strictly regulated (Nergadze et al. RNA 2009) and is probably involved in the maintenance of telomere integrity.
Telomeric transcripts in cancer
It has been demonstrated that telomeric RNA transcription deregulation is deleterious for telomere metabolism and causes genome instability. Therefore, defects in the regulation of telomeric RNA expression may contribute to carcinogenesis and tumour progression. We intend to study the transcriptional state of the telomeres in normal and cancer cells. The analysis of telomeric RNA expression in tissue samples obtained from patients suffering from different forms of cancer will allow us to study the link among the deregulation of telomeric RNA transcription, carcinogenesis and cancer progression.
Interstitial telomeres consist of repetitions of telomeric sequences located in internal chromosome sites. Our previous work demonstrated that, in mammals, interstitial telomeres have been inserted, in the course of evolution, during telomerase mediated repair of double strand breaks or by the capture of double stranded telomeric DNA molecules (Nergadze et al Genome Res. 2004; Nergadze et al Genome Biol. 2007). We have recently demonstrated that some interstitial telomeres play a role in the regulation of gene expression. Our next goal is to analyse the evolution and the function of interstitial telomeres combining molecular, cytogenetic and bioinformantic approaches.
Ionizing radiation induced telomere instability
Ionizing radiations cause severe damages to the DNA double helix, due to the formation of free radicals and to oxidative stress. Telomeres are particularly susceptible to the damages induced by ionising radiations causing an acceleration of their shortening.
We intend to identify the factors involved in telomere metabolism focusing our attention on modifications of the regulation of transcription following the exposure to ionizing radiations.
Collaborations: Claus Azzalin (ETH Zurich, Switzerland), Camillo Porta (IRCCS San Matteo, Pavia), Giovanni Barosi e Vittorio Rosti (IRCCS San Matteo, Pavia), Aurora Ruiz-Herrera (Universitat Autonoma de Barcelona, Spain), Andrea Ottolenghi (Università di Pavia)
3) Production of recombinant proteins in human cells
The production of human recombinant proteins is particularly relevant in various applied fields (drug production, diagnosis) as well as in basic research (biochemical and structural studies). Recombinant proteins can be produced in prokaryotic or eukaryotic expression systems. Prokaryotic expression systems are commonly used for large scale production of some proteic drugs such as insulin. Other molecules, for example erythropoietin, need post-translational modifications that can correctly occur only in mammalian cells.
We developed a vector-host cell system which allows a high production efficiency compared to the mammalian cell systems traditionally employed. With this system we can get hundreds of copies of exogenous genes in human cell lines resulting in overexpression of the encoded protein. The system allows to increase the production efficiency compared to conventional systems thereby decreasing the production costs. The adoption of a production system in human cells opens new prospects for improving the therapeutic efficacy of the protein drugs for human use also reducing side effects. Projects for the construction of human cell lines productive of specific proteins of pharmacological and diagnostic interest are ongoing.
Collaborations: Mark Pepys (University College London), Federico Forneris (Università di Pavia), Vittorio Bellotti e Sofia Giorgetti (Università di Pavia)
Groups leaders: Cinzia Calvio, Davide Sassera, Claudio Seppi
Collaborators: Giulia Barbieri (Postdoctoral fellow), Paolo Gabrieli (Postdoctoral fellow), Elisabetta Andreoli (technician), Giuliano Gasperi (adjunct professor), Luca Longanesi (assegnista), Francesco Comandatore (Postdoctoral fellow), Stefano Gaiarsa (PhD student), Leone De Marco (PhD student), Emanuela Clementi (technician), Luciano Sacchi (adjunct professor)
Bacillus subtilis STRAIN IMPROVEMENT FOR γ-PGA PRODUCTION
C. Calvio, C. Seppi; in collaboration with P. Mustarelli, Chemistry Dept. and P. Magni, Electrical Computer and Biomedical Engineering (University of Pavia), and G. Mazzini (IGM-CNR, Pavia)
The need of safer raw material, derived from renewable sources is the motor driving the growing interest towards natural biopolymers. γ-PGA is an anionic polymer produced by Bacilli, formed by thousands glutamic acid units. Due to its non-toxicity, the water solubility and biodegradability it finds application in several biotechnological fields as: flocculant for heavy metal removal, cryoprotectant, humectant, thickening additive in cosmetics and food industries, as biological glue, as drug or vaccine carrier or scaffold for biomedical engineering. However, for its full industrial exploitation it is mandatory to reduce production costs, both increasing bacterial productivity and reducing fermentation costs. Our Lab has obtained a producer strain derived from the B. subtilis lab strain, 168, fully characterized. The availability of a well defined strain, which is genetically amendable, offers the opportunity to apply genetic engineering to improve productivity and rationalize metabolic pathways for lowering fermentation costs. By introducing specific mutations we already obtained strains that show high product yield. Now, the aim is to obtain a producer able to ferment organic components contained in some agro-industrial by-products. We plan to take advantage of both rice straw, an abundant biomass currently under-exploited, and raw glycerol, a co-product in the biodiesel industry as bacterial feedstock. The meeting of the above objectives will not only lead to cheaper γ-PGA, but will also contribute to the valorization of the rice and biodiesel production chains and reinforce the development of new bio-economy sectors. This research line is currently funded by two CARIPLO FOUNDATION grants.
γ-PGA-HYDROLASES AS ANTIBACTERIAL TOOLS
We identified and characterized four new genes of B. subtilis that code for γ-PGA hydrolases. We found that those gene are phage-derived and spread across bacteria through horizontal gene transfer. We also identified γ-PGA coding capacity in several microbial species, among which several pathogens. We are now tryng to analyze the role of γ-PGA in the virulence behavior in some of those species and the possible use of the γ-PGA hydrolase and antibacterial agents.
THE ROLE OF SwrA, A RECENTLY DICOVERED Bacillus subtilis REGULATORY FACTOR
In Bacillus subtilis the two-component system DegS-DegU controls the expression of one hundred of genes involved in the exponential-to stationary phase of growth transition, coordinates single cells differentiation in multicellular communities and in pathogenic species, as Listeria monocytogenes or Bacillus anthracis, is involved in virulence. It has been shown that DegU regulates B. subtilis motility in a complex way. Also SwrA a protein which has no similarity to previously characterized proteins, is involved in such complex regulation. We have shown that there is a functional and molecular interaction between the two proteins, DegU and SwrA, in motility and now would like to extend our analyses to other genetic pathways DegU-regulated.
GENOMIC EPIDEMIOLOGY AND COMPARATIVE GENOMICS OF NOSOCOMIAL PATHOGENS
D. Sassera; in partnership with P. Marone (IRCCS Policlinico S. Matteo in Pavia), S. Brisse (Institut Pasteur, Paris), E. Feil (University of Bath)
This research line is focused on whole-genome sequencing of large numbers of strains of nosocomial pathogens. The genomes thus generated are analyzed using bioinformatics tools, some developed ad-hoc, for the comparative genomic and genomic epidemiology analysis. The genomics of nosocomial pathogens allows to evaluate the genomic variability, the presence and mobility of virulence factors and antibiotic resistance, describe recombination events and overall architecture of genomic variations. This approach leads to applicative outcomes, such as the ability to reconstruct epidemic events and to identify transmission patterns, to characterize strains of interest based on the resistance and virulence profiles, and to approach the objective of using genomics in diagnostic microbiology.
ROLE OF SYMBIOSIS IN PARASITES
D. Sassera; in collaboration with C. Bandi (Statale University of Milan), B. Makepeace (University of Liverpool), O. Plantard (INRA, Nantes), G. Favia (University of Camerino)
Symbiotic microorganisms are widespread in many organisms, where they assume different roles, from parasitism to obligate mutualism. In the context of this research line, we use an interdisciplinary approach to identify new symbionts and to understand their role in the biology of the hosts. Integrating methods of light and electron microscopy, molecular biology, genomics and transcriptomics we are currently working on different microorganisms:
Midichloria mitochondrii: bacterium order Rickettsiales, symbiont of the tick Ixodes ricinus, has the unique ability to localize within the mitochondria of host cells. We are currently working to understand the role of bacteria in the physiology of the tick, through an integrated approach of electron microscopy, transcriptomics and proteomics. In parallel we are looking for bacteria phylogenetically related to M. mitochondrii other host with genomic techniques.
Symbiotic yeasts: yeast strains may have a role of symbionts in different species of arthropods, including vectors of important diseases. With a multidisciplinary project coupling electron microscopy and comparative genomics, we are investigating the presence and role of the yeast species Wickeramomyces anomalus and Meyerozyma guilliermondii in mosquitoes and sandflies.
TRANSCRIPTOMICS OF ARTHROPOD VECTORS
D. Sassera; in collaboration with S. Epis (University of Milan), S. Urbanelli and D. Porretta (University of Rome La Sapienza) , Favia G. and I. Ricci (University of Camerino)
The use of the RNA-seq methodology allows the evaluation of changes in gene expression at the whole transcriptome level. In this line of research we investigate these changes in arthropods that are vectors of diseases of great medical interest, mosquitoes and ticks, during the course of the life cycle or in response to specific stimuli, such as the presence of insecticide molecules. The goal is to understand fundamental aspects of the biology of these vectors, as well as specific responses to treatments, in order to design innovative control strategies.
Group leaders: Prof. Mariangela Bonizzoni, Prof. Ludvik Gomulski, Prof. Lino Ometto, Giuliamo Gasperi (Contract Professor), Anna Rodolfa Malacrida (Contract Professor)
Collaborators: Francesca Scolari (Assistant Professor), Paolo Gabrieli (Postdoc), Patrizia Chiari (Technician), Liliana Marcorio (Technician), Alessandro Di Cosimo (PhD student), Grazia Savini (PhD student)
Climate and demographic change, the increase of trade, urbanization and human travel have facilitated the spread of invasive insect species of economic and sanitary importance. These species include the Asian tiger mosquito, a vector of many viruses pathogenic to humans, and species of fruit flies that are important agricultural pests. The risks due to the invasion of these insects are compounded by the lack of vaccines and treatments for the pathogens that they transmit (i.e. Dengue, Chikungunya and Zika viruses) and the emergence of resistance to insecticides, which are currently the most used tool for their control.
The research group deals with genomics, transcriptomics, proteomics and metabolomics of insects of agricultural interest and public health relevance with the aim of developing innovative control strategies that are environmentally sustainable. Additionally, we study their population genetics and evolution in order to understand and prevent their further spread and we use transgenic approaches to study the mechanisms underlying their reproduction in order to improve current genetic control methods.
The insect species currently studied are:
Vectors of disease pathogens:
Fruit flies (Diptera, Tephritidae) of African or Asian origin, that are highly invasive and that have recently been introduced into Southern Europe and the Americas where they have produced significant economic damage:
Main Lines of Research
Genome sequence of the tsetse fly (Glossina morsitans): vector of African trypanosomiasis. Science (2014) 344: 380-6.
Genome sequence of the Asian Tiger mosquito, Aedes albopictus, reveals insights into its biology, genetics, and evolution. Proc Natl Acad Sci USA (2015) 112: E5907-15.
A draft genome sequence of an invasive mosquito: an Italian Aedes albopictus. Pathog Glob Health (2015) 109:207-20.
Presence of extensive Wolbachia symbiont insertions discovered in the genome of its host Glossina morsitans morsitans. PLoS Negl Trop Dis. 2014 Apr 24;8(4):e2728.
We characterized high resolution molecular markers, such as microsatellites and SNPs, in Ae. albopictus, C. capitata and other fruit flies, and we have applied them to study the genetic links between ancestral and derived populations and to characterize the species invasion processes. We also applied mitochondrial markers, in combination with microsatellites, to study the origin of Ae. albopictus in California and, in collaboration with prof. Torroni, to study the variability of ancestral and derived populations.
Relevant genetic differentiation among Brazilian populations of Anastrepha fraterculus (Diptera, Tephritidae). Zookeys. 2015 Nov 26;(540):157-73.
Molecular markers for analyses of intraspecific genetic diversity in the Asian Tiger mosquito, Aedes albopictus. Parasit Vectors. 2015 Mar 28;8:188.
The oriental fruitfly Bactrocera dorsalis s.s. in East Asia: disentangling the different forces promoting the invasion and shaping the genetic make-up of populations. Genetica. 2014 Jun;142(3):201-13.
A new threat looming over the Mediterranean basin: emergence of viral diseases transmitted by Aedes albopictus mosquitoes. PLoS Negl Trop Dis. 2012;6(9):e1836.
The utility of microsatellite DNA markers for the evaluation of area-wide integrated pest management using SIT for the fruit fly, Bactrocera dorsalis (Hendel), control programs in Thailand. Genetica. 2011 Jan;139(1):129-40.
Uncovering the tracks of a recent and rapid invasion: the case of the fruit fly pest Bactrocera invadens (Diptera: Tephritidae) in Africa. Mol Ecol. 2009 Dec;18(23):4798-810.
Isolation and characterization of microsatellite markers in the newly discovered invasive fruit fly pest in Africa, Bactrocera invadens (Diptera: Tephritidae). Mol Ecol Resour. 2008 Nov;8(6):1509-11.
Globalization and fruitfly invasion and expansion: the medfly paradigm. Genetica. 2007 Sep;131(1):1-9.
The Spermatophore in Glossina morsitans morsitans: Insights into Male Contributions to Reproduction. Sci Rep. 2016 Feb 5;6:20334.
How functional genomics will impact fruit fly pest control: the example of the Mediterranean fruit fly, Ceratitis capitata. BMC Genet. 2014;15 Suppl 2:S11.
Transcriptome profiling of sexual maturation and mating in the Mediterranean fruit fly, Ceratitis capitata. PLoS One. 2012;7(1):e30857.
Gene discovery in an invasive tephritid model pest species, the Mediterranean fruit fly, Ceratitis capitata. BMC Genomics. 2008 May 23;9:243.
Polyandry in the medfly – shifts in paternity mediated by sperm stratification and mixing. BMC Genet. 2014;15 Suppl 2:S10.
Towards mosquito sterile insect technique programmes: exploring genetic, molecular, mechanical and behavioural methods of sex separation in mosquitoes. Acta Trop. 2014 Apr;132 Suppl:S178-87.
Polyandry is a common event in wild populations of the Tsetse fly Glossina fuscipes fuscipes and may impact population reduction measures. PLoS Negl Trop Dis. 2011 Jun;5(6):e1190.
Safe and fit genetically modified insects for pest control: from lab to field applications. Genetica. 2011 Jan;139(1):41-52.
Sperm storage and use in polyandrous females of the globally invasive fruitfly, Ceratitis capitata. J Insect Physiol. 2010 Nov;56(11):1542-51.
Site-specific recombination for the modification of transgenic strains of the Mediterranean fruit fly Ceratitis capitata. Proc Natl Acad Sci U S A (2009) 106:18171.
Fluorescent sperm marking to improve the fight against the pest insect Ceratitis capitata (Wiedemann; Diptera: Tephritidae). N Biotechnol. 2008 Jun;25(1):76-84.
Sniffing out chemosensory genes from the Mediterranean fruit fly, Ceratitis capitata. PLoS One (2014) 9:e85523.
Identification of pheromone components and their binding affinity to the odorant binding protein CcapOBP83a-2 of the Mediterranean fruit fly, Ceratitis capitata. Insect Biochem Mol Biol (2014) 48:51-62.
RNA-seq analyses of changes in the Anopheles gambiae transcriptome associated with resistance to pyrethroids in Kenya: identification of candidate-resistance genes and candidate-resistance SNPs. Parasit Vectors (2015) 8:474.
In particular, by merging evolutionary genomics and transcriptomics with ecological, behavioural and morphological data, my research aims at understanding the mechanisms underlying the evolution of invading pest insects and insects of insects of agricultural and medical importance.
Active Research Grants
Prof. Daniela Curti, Prof. Maurizia Dossena, Prof. Ornella Pastoris, Prof. Roberto Federico Villa
Collaborators: Sara Arnica, Alice Buzzella, Barbara Balestra, Daniela Buonocore, Federica Ferrari, Silvia Molino, Manuela Verri.
Main research topics
– Neuropharmacology (see “Neurobiology”)
– Pharmacobiochemistry and Toxicology
1) Quest for novel antitumor compounds.
Rational design and evaluation of in vitro and in vivo efficacy of newly synthesized monomeric and dimeric ligands binding sigma-1 and sigma-2 receptors. The research is aimed to identify and optimize new candidate drugs with antitumor activity. In collaboration with: S. Collina and M. Paolillo (Dept. Drug Sciences, UNIPV) (contact: D.Curti).
2) Prevention of oxiplatin-induced peripheral neuropathy
Randomized phase II study on the efficacy of Superoxide Dismutase + Vitamin E supplementation in patients with colorectal cancer. Oxaliplatin, an antineoplastic platinum-derived compound, is the standard drug used for the treatment of colorectal cancer. However, chronic treatment leads to oxaliplatin-induced peripheral neuropathy. The project aims to investigate the efficacy of patients supplementation with Superoxide Dismutase and Vitamin E to prevent/reduce the oxaliplatin-induced neurological complications. The oxidative status of the patients over time and the metabolomic profile will be evaluated to identify the molecules involved in the induction and development of the peripheral neuropathy. In collaboration with: S. Brugnatelli (S.C. Oncologia, Fondazione I.R.C.C.S. Policlinico San Matteo di Pavia); J. Á. Rufián Henares (Dept. Nutrition and Bromatology, University of Granada, Spain (contact: M. Dossena)
3) Assessment of the toxicity, antioxidant, antiradical, antinflammatory, immunomodulatory properties and bioavailability of bioactive compounds extracted from vegetables and microalgae.
The study aims to characterize: 1) naringenin, naringin, as well other polyphenols’ content in grapefruit (C. paradisi) zest, rind, pulp, and juice extract; 2) bioactive compounds extracted from fruit/vegetable waste and microalgae such as Scenedesmus and Aphanizomenon flos-aquae (AFA). AFA, also called Klamath Algae, is a cyanobacterium characterized by a unique nutritional profile, containing specific immunomodulatory, anti-inflammatory and antioxidants molecules. In collaboration with: J. Á. Rufián Henares (Dept. Nutrition and Bromatology, University of Granada, Spain); AOP Unolombardia sacpa and Regione Lombardia (contact: M. Dossena).
4) Metabolism and physical exercise
The energy demands of the human body depend on energy expenditure and basal metabolic rate. The basal metabolic rate is the minimum energy needed to maintain vital functions. During physical exercise, particularly in sports, our metabolism increases and remains at high level for several hours after the end of the training (up to 12 hours). Exercise increases muscle metabolism indirectly through the secretion of anabolic hormones and the accumulation of muscle mass. However, athletes, in comparison with sedentary people, are more exposed to psychophysical stress that leads to release of cortisol and hormonal imbalance; to the increase of free radicals and to muscle, tendon and ligament damage. Our study will evaluate: 1) energy expenditure associated with various types of physical activities; 2) salivary and blood concentrations of hormones and other molecules related to physical activity or sports (e.g. cortisol, testosterone, growth hormone, inflammatory cytokines, ROS); 3) effects of sport food and supplements on exercise performance and post-exercise muscle recovery; 4) change in body composition by the modulation of diet programs (relationship between macronutrients and glycemic load). In collaboration with: G. D’Antona, M. Negro (Centro di Medicina dello Sport,Voghera, UNIPV); I. Fabbri (FIT e MOTOGPTM) (contact: M. Dossena).
5) Gastrointestinal neurobiopathology
Functional gastrointestinal neurobiopathology is studied by evaluating the neuronal control, and its modulation by neurotransmitters, of intestinal motility. The molecular aspects are investigated by analysing miRNA expression, pharmacogenetics and the influence of the intestinal microbiota in major gastrointestinal disorders such as Celiac disease, Irritable Bowel Syndrome (IBS). Our research interest is also focused on topics related to the pathophysiology of skeletal muscle, especially in the Neuroleptic Malignant Syndrome (NMS). In particular, studies are designed to evaluate the composition of intestinal microbiota in celiac patients using next generation sequencing and to perform genetic and epigenetic analyses in patients with NMS. In collaboration with: Unità di Medicina Interna e Gastroenterologia, Dip. Medicina Interna e Terapia medica UNIPV, Fondazione IRCCS Policlinico San Matteo Unità di Medicina Interna Dip. di Scienze Mediche e Chirurgiche, Università di Bologna, Policlinico S. Orsola-Malpighi; Laboratorio di Tossicologia Clinica e Sperimentale, Centro Antiveleni – IRCCS Fondazione Salvatore Maugeri, Pavia (contact: O.Pastoris).
Physiology is a discipline that covers an extremely wide extension of research fields relevant to the basis and mechanisms of normal cellular and systemic functions of animal organisms, with approaches typically characterized by the application of the highest possible levels of quantitative exactness.
The physiologists of the Dept. of Biological Sciences and Biotechnology “L. Spallanzani” are mainly active in the fields of Neurophysiology, Cellular and molecular Physiology, Cardiovascular Physiololgy, and some other application-directed ones.
Group leaders: Prof. Mauro Toselli, Prof. Gerardo Biella, Prof.ssa Paola Rossi, Prof. Franco Tanzi, Prof.ssa Laura Botta, Prof. Francesco Moccia.
Coworkers: Elisabetta Cesana (postdoc), Claudia Maniezzi (Ph.D. student), Luigi Salvioli (Ph.D. student), Francesca Talpo (postdoc).
Main research topics:
– Neurophysiology (see in the descriction of the research topic “Neurobiology”)
– Cellular and molecular Physiology
(i) Roles of ion channels in tumor cells
It has long been known than ion channels, particularly Ca2+-permeable channels, are involved in the cancer hallmarks described by Hanahan and Weinberg, such as uncontrolled proliferation, apoptosis resistance, and metastasis. The present research project intends to study the role played by intracellular Ca2+ signals in the processes of proliferation, apoptosis resistance and migration in primary cell lines established from patients suffering from two different solid cancers, i.e. renal cellular carcinoma and colorectal carcinoma. Our attention is particularly focused on Orai1-3 and TRPC1-77 channel families.
Research Supervisor: F. Moccia
Collaborations: U. Laforenza, Dipartimento di Medicina Molecolare, UNIPV; D. Montagna, P. Pedrazzoli, C. Porta, IRCCS Policlinico S. Matteo, Pavia
– Cardiovascular Physiology
(i) Role of Ca2+ signaling in physiological and pathological vasculogenesis
Recent studies showed that vasculogenesis, i.e. the formation of blood vessels de novo by endothelial progenitor cells (EPCs), not only takes place during embryonic development, but also during the angiogenic switch in solid tumors. Our group showed for the first time that Ca2+ signals play a key role in the activation of healthy EPCs and EPCs deriving from several types of tumor (renal cellular carcinoma, infantile hemangioma and primary myelofibrosis). The present research project intends to: 1) keep on investigating on the Ca2+ toolkit in healthy EPCs to shed further light on their basic physiological mechanisms; 2) assess whether the Ca2+ toolkit i also dis-regulated in breast cancer-derived patients; 3) investigate whether and how the remodeling of the Ca2+ toolkit underpins tumor EPC resistance to chemotherapeutics and anti-angiogenic drugs; and 4) investigate the molecular mechanisms by which tumor microenvironment alters EPC’s Ca2+ toolkit. The present project takes advantage from the collaboration with IRCCS Policlinico San Matteo in Pavia, which ensures the continuous availability of EPCs deriving from either healthy donors and tumor-affected patients.
Research Supervisor: F. Moccia
Collaborations A. Balduini, U. Laforenza, Dipartimento di Medicina Molecolare, UNIPV; D. Lim, Università del Piemonte Orientale, Novara; G. Guerra, Università del Molise, Campobasso; M. Della Porta, Humanitas Research University, Rozzano; M. Massa, P. Pedrazzoli, C. Porta, V. Rosti, IRCCS Policlinico S. Matteo, Pavia; L. Munaron, Università di Torino
– Other application-directed projects
(i) Antibiofilm profile of Pistacia lentiscus L. Oil. and Lentinula edodes
This research project studies the antioxidant and antimicrobial ability of extracts of Pistacia lentiscus and Lentinula edodes on human (oral) microbiota and, in particular on lingual biofilm. With the antibiotic resistance phenomenon in expansion the search for new antibacterial strategies rises great interest in the clinical context.
The activity will include the isolation of lingual biofilm and the assessment of the extent of oxydative stress with spectrophotometric measurements.
Research Supervisor: P. Rossi
The research activities are focused on the the morphology, physiology, genetics, immunology, pathology, pharmacology of the central nervous system at molecular, cellular and integrated system levels and on the development of innovative strategies to counteract neurodegenerative diseases.
Main research fields:
Group leaders: Prof. Daniela Curti, Prof. Maurizia Dossena, Prof. Ornella Pastoris, Prof. Roberto F. Villa
Coworkers: Sara Arnica (research fellow), A. Buzzella (research fellow), Barbara Balestra (graduated technician), Daniela Buonocore (research fellow), Federica Ferrari (voluntary laboratory attendant)), Silvia Molino (Ph.D. student), Manuela Verri (graduated technician)
(i) Role of Sigma-1 receptor (S1R) isoforms and oligomerisation in motoneuron (MN) survival and death.
S1Rs are ubiquitary proteins, particularly enriched in MNs where they sit on mitochondrial associated endoplasmic reticulum (ER)membranes (MAMs) and in cisternae localized under the post-synaptic densities of cholinergic C-terminals. The physiological role of the enigmatic S1R and its oligomerisation state and the potential as therapeutic target in MN diseases will be studied in cellular and animal models of amyotrophic lateral sclerosis (ALS), in lymphocytes and autoptic spinal cord samples of patients affected by ALS with the aim of developing novel strategies to counteract the disease. In collaboration with M. Peviani (Harvard Medical School, Boston); C. Bendotti (IRCCS M. Negri, Milano); S. Collina (Dip. Scienze del Farmaco, UNIPV); M. Zorzetto, (IRCCS Policlinico San Matteo, Pavia); M. Ceroni (IRCCS C. Mondino e UNIPV) .
Research supervisor: D. Curti
Collaborations: M. Peviani (Harvard Medical School, Boston); C. Bendotti (IRCCS M. Negri, Milano); S. Collina (Dip. Scienze del Farmaco, UNIPV); M. Zorzetto, (IRCCS Policlinico San Matteo, Pavia); M. Ceroni IRCCS C. Mondino e UNIPV.
(ii) Evaluation of metabolic alterations in patients with dementias.
role of insulin-resistance The study aims to evaluate potential alterations of cerebral, liver and skeletal muscle energy metabolism in subjects with dementia, considering blood and cerebrospinal fluid as biological matrices. Potential alterations in the aminoacidic and metabolomic blood profile, in the concentration of plasma acylcarnitines, together with insulin-resistance condition are considered. Pharmacological treatments in combination with nutritional and/or neurological rehabilitation are evaluated. In collaboration with: A. Costa (IRCCS “C. Mondino”, Pavia); A. Navarra (IRCCS, S. Maugeri Pavia, Servizio di Medicina di Laboratorio); M. Daglia (Dip. Scienze del Farmaco, UNIPV); Prof. Luisa Mannina (Dipartimento di Chimica e Tecnologie del Farmaco, Università Sapienza di Roma); Prof. Bruno Solerte (Dipartimento di Medicina Interna e Terapia Medica – IDR S. Margherita, Pavia).
Research supervisor: M. Dossena
Collaborations: A. Costa (IRCCS “C. Mondino”, Pavia); A. Navarra (IRCCS, S. Maugeri Pavia, Servizio di Medicina di Laboratorio); M. Daglia (Dip. Scienze del Farmaco, UNIPV); L. Mannina (Dip. Chimica e Tecnologie del Farmaco, Università Sapienza di Roma); B. Solerte (Dip. Medicina Interna e Terapia Medica – IDR S. Margherita, Pavia).
(iii) Correlation between oxidative stress and cytotoxicity in the progression of Anorexia Nervosa (AN): new biochemical and morphological approaches
Determination of the cytotoxicity of altered lipoproteins and modified amount/activity of hormones in blood of patients with eating disorder and in selected neural cell lines. Investigation of the molecular mechanisms mediating neural damage [In collaboration with MG. Bottone (Dept. Biology and Biotechnology, UNIPV); U. Balottin (IRCCS C. Mondino, UNIPV)].
Research supervisors: M. Dossena, M.G. Bottone
Collaborations: U. Balottin (IRCCS C. Mondino, Pavia).
(iv) Gastrointestinal neurobiopathology
Functional gastrointestinal neurobiopathology is studied by evaluating the neuronal control, and its modulation by neurotransmitters, of intestinal motility. The molecular aspects are investigated by analysing miRNA expression, pharmacogenetics and the influence of the intestinal microbiota. In particular, studies are designed to evaluate: 1) intestinal permeability, glial activation and intestinal microbiota in patients with Parkinson’s disease; 2) the dysfunction of gastrointestinal motility in an animal model of Parkinson’s disease. In collaboration with: F. Blandini (Laboratorio di Neurochimica Funzionale, IRCCS Istituto Neurologico “C. Mondino”, Pavia ).
Research supervisor: O. Pastoris
Collaborations: F. Blandini (Laboratorio di Neurochimica Funzionale, IRCCS Istituto Neurologico “C. Mondino”, Pavia ).
(v) Functional Proteomics and Metabolomics of enzyme bioenergetics
The experimental project regards molecular studies investigating in vivo the mechanisms underlying the biochemical changes in Functional Proteomics of the enzymatic systems linked to energy transduction of cerebral tissue. The analysis of the biochemical connections (Metabolomics) existing between the catalytic properties of some energy linked enzymes during acute or chronic and neurodegenerative Diseases of neurological and psychiatric interest, are examined in the perspective of a neuropharmacological approach for evaluating potential therapies to improve the Central Nervous System functional efficiency.
Research supervisor: R.F. Villa.
National and international collaborations and the outcome of the scientific research carried out so far can be found in R. F. Villa’s curriculum and Publication list, respectively: please refer to these two documents.
– Cellular and integrated Neurophysiology
Group leaders: Prof. Jacopo Magistretti, Prof. Mauro Toselli, Prof. Gerardo Biella, Prof. Paola Rossi, Prof. Franco Tanzi, Prof. Laura Botta, Prof. Francesco Moccia
Coworkers: Elisabetta Cesana (postdoc), Claudia Maniezzi (Ph.D. student), Luigi Salvioli (Ph.D. student), Francesca Talpo (postdoc)
(i) Actions of oxytocin in the central nervous system
Oxytocin, the hormone synthesized in the hypothalamus, acts both peripherally and centrally, thus regulating several behavioral functions. Recently, a correlation between the oxytonergic system and autism has been suggested to exist. Oxytocin receptors are widely distributed throughout the hippocampus, and various electrophysiological studies have been carried out in rodents to identify the oxytonergic circuits and to understand the neurodulatory mechanisms exerted by the neuropeptide in this structure. Oxytocin has turned out to enhance, in the hippocampal CA1 field, inhibitory transmission, by increasing the discharge frequency of a particular class of GABAergic neurons synaptically contacting pyramidal neurons. However, the mechanism through which oxytocin increaseses these interneurons’ discharge frequency has not been clarified yet, neither is it known how this increase influences pyramidal neurons’ excitability. Moreover, the mechanisms through which oxytocin is able to improve the symptoms of the autistic disease are also unknown. Our study focuses on the characterization of oxytocin-operated neuromodulation in the mouse hippocampus, in order to fill the knowledge gaps still existing in this field.
Research supervisors: M. Toselli, G. Biella
Collaborations: B. Chini, Istituto di Neuroscienze – CNR, Milano); M. Parenti, Dip. Medicina Sperimentale Università di Milano Bicocca, Milano
(ii) Analysis of synaptic activity in animal models of Huntington Disease (HD)
HD is a neurodegenerative disease caused by a dominant, autosomal mutation in the IT-15 gene, which codes for a protein named huntingtin (Htt). The normal, functional role(s) of Htt are still unknown. The protein is ubiquitous, it is essential for normal embryogenesis, normal neuronal development and survival, and is also involved in synaptic activity. The expansion of the nucleotidic triplet CAG in exon 1 of the IT-65 gene (with > 36 repetitions) generates a mutated form of Htt (mHtt) which somehow becomes toxic to neurons and causes a wide loss of cerebral neurons, especially in the cortex and the striatum. mHtt brings about, besides neuronal death in the late stages of the disease, also progressive alterations in the morphology, excitability, and synaptic connectivity and function of cortical pyramidal neurons and striatal medium spiny neurons (MSN). Hence, the earliest behavioral and cognitive symptoms of HD anticipate, rather than being a consequence of, neuronal death. Derangements in synaptic connectivity and plasticity at the CPN/MSN synapses, as well as excitotoxicity, mainly mediated by alterations of NMDA receptors, appear to be crucial in HD pathogenesis and progression. By resorting to multidisciplinary experimental approaches (e.g., electrophysiological, behavioral, etc.), we plan to analyze the basis of the progressive de-coupling of excitatory cortico-striatal synapses in knock-in mouse models of HD, both in the pre-symptomatic and symptomatic stages of the disease.
Research supervisors: M. Toselli, G. Biella
(iii) Functional evaluation of striatal medium spiny neurons differentiated from embryonal stem cells and re-programmed from fibroblasts of patients with Huntington disease (HD)
This research project focuses on the functional characterization of a specific class of triatal neurons, namely medium spiny neurons (MSN). By means of a specific protocol of differentiation form human embryonal stem cells (hES, H9 strain), 2D or 3D MSN cultures will be obtained. Some cell lines will also exposed to induced over-expression of factors like GSX2 and EBF1 to enhance cell yield and quality. Moreover, given the devastating MSN degeneration typical of HD, our interest is directed towards the electrophysiological characterization of striatal neurons differentiated from induced, pluripotent stem cells (hiPS) obtained from fibroblasts of both normal and HD human subjects. In this way, it becomes possible to model HD in an in vitro system, thus providing a major contribution to understanding of the multiple and still unclear or unknown molecular mechanisms underlying the neurodegeneration occurring in HD. Moreover, the electrophysiological investigations will contribute to validate and consolidate the cell differentiation protocol, in sight of the ultimate aim of generating genuine in vitro MSN, usable for cell therapy, which currently appears to be an excellent candidate in neurodegenerative disease management.
Research supervisors: M. Toselli, G. Biella
Collaborations: E. Cattaneo, Dip. Bioscienze, UNIMI, Milano; M. Onorati , Dept. Neurobiology, Yale School of Medicine, New Haven, Connecticut, USA
(iv) Mechanisms and neuromodulation of membrane excitability in the neurons of parahippocampal cortices and the perirhinal cortex
Parahippocampal cortices (PHCs) establish bidirectional synaptic interactions with the hippocampus that are of fundamental importance for the memory functions and spatial orientation functions typical of the medial-temporal lobe memory system. This project focuses on the study of the mechanisms governing the intrinsic excitable properties of PHC neurons, of their functional implications as far as communications between the parahippocampal region and the hippocampus are concerned, and of the neuromodulatory systems controlling them. In particular, the following aspects are studied: 1) the intrinsic membrane mechanisms which determine the specific discharge properties of medial entorhinal cortex (mEC) neurons; 2) membrane resonance phenomena, their underlying mechanisms, and functional implications in mEC and perirhinal-cortex (PRC) neurons; 3) the control mechanisms by which PRC is able to operate its characteristic function of selecting input signals of cortical origin and directed towards the hippocampus via the mEC, due to the intervention of a complex intrinsic synaptic network in which both projection pyramidal neurons and several different classes of GABAergic interneurons that generate a local inhibitiory action on pyramidal neurons are involved; 4) whether and how various physiological neuromodulatory systems, such as those relying on neurotransmitters like acetylcholine, dopamine, norepinephrine, etc., influence the above mentioned properties of single neurons and local circuits.
Research supervisors: J. Magistretti, M. Toselli, G. Biella
(v) Role of endothelial Ca2+ signals in neurovascular coupling
Neurovascular coupling is the mechanism through which brain blood flux increases or decreases in response to modifications of neural activity of the same sign. The most widely accepted view attributes the ability to release vasoactive mediators, in response to synaptic activity, to neurons and astrocytes. This research project is currently examining, for the first time, the ability of brain vessel endothelium to directly sense the synaptic release of neurotransmitters. Our attention is focusing on the sensitivity of bEND.5 cells, a widely validated model for the study of brain endothelium, to the neurotransmitters, glutamate and acetylcholine, and the neuropeptide, catestatin. Specifically, the present project intends to: 1) study the mechanisms responsible for the generation of the intracellular Ca2+ oscillations which emerge after exposure to each one of the three above-mentioned agonists; 2) to assess whether and how such oscillations trigger the synthesis nitrogen monoxide (NO), the main vasodilating agent of the brain; and 3) evaluating whether and how the NO released by brain endothelial cells can act not only on vascular tone, but also, retroactively, on synaptic activity.
Research supervisor: F. Moccia
Collaborations: T. Angelone, Università della Calabria, Cosenza; E. D’Angelo, Dipartimento di Scienze del Cervello e del Sistema Nervoso, UNIPV; S. Dragoni, University College London, London; G. Guerra, Università del Molise, Campobasso; D. Lim, Università del Piemonte Orientale, Novara
(vi) Study on the effects of oral integration with the medicinal mushroom, Hericium erinaceus (Lion’s mane) on neurogenesis and cognitive functions in wild-type mice
Hericium (Bull.) Pers. Is a medicinal mushroom that is able to modulate the immune system and enhance cognitive functions in humans. In the last decade several in vitro and in vivo studies have been carried out to evaluate the effects Hericium after oral administration. In humans, in particular, the data available in the literature report a neuroprotective effect against brain stroke and effects of partial cognitive recovery in neurodegenerative diseases, such as dementia and Alzheimer disease (AD). In a mouse model of pharmacologically-induced AD development, food supplementation with H. erinaceus prevents impairment of short-term as well as visual recognition memory. No data are available as far as wild-type animals are concerned.
This research line studies the effects of oral administration of this mycotherapic agent in wild-type mice. In particular, two aspects are dealt with: the ability of supplementation to improve some cognitive performances related to memory and the effects on hippocampal neurogenesis.
The patch-clamp technique is applied to hippocampal slice, and in vivo behavioral studies are carried out.
Research supervisor: P. Rossi
Collaborations: F. Brandalise, Brain Research Institute, University of Zurich; A. Gregori, MycoMedica d.o.o., Kranjska Gora, Slovenia; G. Orrù, Dipartimento di Scienze Chirurgiche, Università di Cagliari; M.L. Guglielminetti, M.Rodolfi ed E. Savino, Dipartimento della Terra e dell’Ambiente, Università di Pavia.
(vii) Study on plastic phenotypic modifications and of the neural olfactory network in eggs and tadpole embryos exposed to predation signals
Phenotypic plasticity is the ability of organisms to modify their phenotype in response to environmental changes. In particular, the prey-predator interaction is studied by exposing eggs and tadpole larvae to various chemical signals produced by different kinds of predators (specific, aspecific, indigenous, non-indigenous, etc.), and evaluating the ensuing phenotypic modifications and response changes in the olfactory system’s mitral cells. The physiological mechanisms underlying such changes are studied in the attempt to understand how the information contained in predator-produced signals (kairomones) are coded at the various levels of the prey’s sensory system.
The study exploits the application of the patch-clamp technique to the olfactory system (mitral cells) of in vivo tadpoles as well as the documentation of behavioral changes in tadpoles.
Research supervisor: P. Rossi
Collaborations: P. Galeotti e A. Gazzola, Dipartimento della Terra e dell’Ambiente, Università di Pavia; F. Brandalise, Brain Research Institute, University of Zurich; D. Rubolini, Dipartimento di di Bioscienze, Università di Milano
(viii) Expression of inward-rectifier potassium currents in the vermis and the cerebellar hemispheres of the mouse
Inward-rectifier potassium currents coupled to G proteins (GIRK) play a crucial role during migration and maturation of cerebellar granule cells in the cerebellar vermis. Consistently with such a role, the weaver mouse model, which is characterized by a spontaneous point mutation of a GIRK2-type channel, shows deep alterations in motor function, with a pattern of severe cerebellar ataxia. In cerebellar hemispheres, however, the ontogenesis of GIRK currents during cerebellar granule migration and differentiation shows a completely different pattern. This research line analyzes the regional differences in ontogenetic development of voltage-dependent currents.
The study is based on the application of the patch-clamp technique in mouse cerebellar slices and immunohistochemical measurements with specific antibodies.
Research supervisor: P. Rossi
Collaborations: U. Gerber e F. Brandalise, Brain Research Institute, University of Zurich; R. Lujan, Instituto de Investigaciónen Discapacidades Neurológicas (IDINE), Dept. Ciencias Médicas, Facultad de Medicina, Universidad Castilla-La Mancha, Albacete, Spain
Group leaders: Prof.ssa Maria Grazia Bottone, Prof. Sergio Comincini, Prof.ssa Rosanna Nano
Collaborators: Violetta Insolia (Ph.D. student), Irene Masiello (Ph.D. student), Marco Biggiogera (full professor)
(i) Study of the development of the central nervous system of mice deficient in prolidase: histochemical alterations in cytoarchitecture areas of NCS
The gene prolidase (PEPD) encodes for the unique Mn (II)-dependent imminodipeptidase responsible for the hydrolysis of dipeptides containing proline or hydroxyproline at the C-terminal. Mutations in this gene cause prolidase deficiency (PD), a rare, autosomal recessive disease. PD patients show reduced or absent enzyme ctivity and a wide spectrum of phenotypes: dermatologic manifestations as incurable leg ulcers; susceptibility to recurrent respiratory infections and varying degrees of mental retardation. In our laboratory, on a mouse model of PD, the dark-like mutant (dal), we assess the correlation between PD and the brain neuroarchitecture and histochemical alterations. In particular, we analyse postnatal alterations in the cerebellum, hippocampus and cerebral cortex. In three different mouse genotypes (wt, dal/+ and dal/dal) we shall evaluate the disorganization of the basement membrane and meninges; the alterations in the migration processes and neuronal differentiation; the imbalance between cell proliferation and cell death; impairment in the major neurotransmitter systems; neurodegeneration and angiogenesis. Morphological, histochemical and microscopic techniques will be utilized together with Western Blotting for quantitative analyses.
Research supervisor: M.G. Bottone
Collaborations: M. Biggiogera, (Dip. Biologia e Biotecnologie, UNIPV); A. Forlino (Dip. Medicina Molecolare, UNIPV)
(i) Astrocytic tumors
ia) Identification of biomarkers to aid in the histopathologic diagnosis of astrocytic tumors, important in the stratification of patients (molecular subtyping) and, in general, for personalized medicine. Development of preclinical protocols for the modulation of autophagic programmed cell death. Translational medicine studies for the development of preclinical therapeutic protocols adjuvant to chemotherapy or radiation treatments.
Research supervisor: S. Comincini
Collaborations: T. Florio (Dept. Internal Medicine and Medical Specialties, University of Genoa) and G. Fossati (ITALFARMACO spa, Milan).
-ib) Development of preclinical anticancer therapeutic strategies based on the use of exosomes and the modification of their molecular content.
Nano-vesicular systems, represented by exosomes, are important systems of communication between the cells and thus may also constitute, given their ability to reach cell districts even very distant, an efficient system for vehicular bioactive molecules to specific target cells, such as cancer cells. Consequently, the isolation and molecular manipulation of exosomial carriers may represent a novel tumor-interference system, useful to improve the effectiveness of current conventional treatment strategies.
Research supervisor: S. Comincini
Collaborations: M. Biggiogera, F. Moccia, G.F. Guidetti (Dept. Biology and Biotechnology, UNIPV).
(iii) Behavioral neurogenetics
In this context, we are focusing on DNA polymorphisms (SNPs) to identify new genetic determinants associated with particular social, psychological, behavioral (behavior genetics). Currently a study is in progress on the identification of polymorphisms of the oxytocin receptor gene associated with the neurological disorder in humans of prosopagnosia (difficulty in recognizing faces).
Research supervisor: S. Comincini
Collaborations: Z. Cattaneo (Dept. Psychiatry, University of Milan Bicocca).
(iv) Study of novel radiopharmaceuticals for the diagnosis of gliomas and of glial cells in physiological and pathological conditions.
The differential diagnosis between tumor recurrence (gliomas and brain metastases) and radiation necrosis phenomena (RN), plays an important role both as a prognostic and therapeutic point of view. Despite significant technological advances of current imaging modalities (PET-CT, MRI, ..) is not yet possible an accurate diagnosis to distinguish areas of active neoplastic proliferation from necrotic or neuro-inflammation areas. Our current research involves the study of new radiopharmaceuticals (18F-FCH, 18F- FET) specific to distinguish low and high grade gliomas. As experimental model various lines of human glioblastoma (T98G, U251, U87) are used to evaluate the in vitro uptake of radiopharmaceuticals mimicking the in vivo situation.
Research supervisor: R. Nano
Collaborations: L. Lodola, M. Persico, F. Buroni (S.C. Medicina Nucleare, IRCCS Policlinico San Matteo, Pavia); F. Corbella, F. Pasi (S.C. Radioterapia Oncologica, IRCCS Policlinico San Matteo, Pavia); A. Facoetti, C. Aprile (Fondazione CNAO, Pavia); E. Giombelli (Divisione Neurochirurgia, Ospedale di Parma); A. Messina (Div. Neurochirurgia, Ospedale di Sondalo); E. Benericetti (Div. Neurochirurgia, Maria Cecilia Hospital GVM, Cotignola, BO).
Group leaders: Prof. Antonio Torroni, Prof. Ornella Semino, Prof. Luca Ferretti, Prof. Anna Olivieri, Prof. Alessandro Achilli
Collaborators: Vincenza Battaglia, Marco Rosario Capodiferro (PhD student), Alessandro Raveane (Postdoc)
One of the main objectives of the research group is the reconstruction, based on genetic and genomic data, of the evolutionary history and demographic events that have involved present and past human populations (at both micro and macro-geographic levels) and some animals (especially domestic animals and those living in close contact with our species). Genetic findings can be easily employed also in multidisciplinary studies involving apparently far away scientific and cultural contexts, from forensics to history, archaeology, linguistics, anthropology, education and public health.
The genetic systems under study are the autosomes as well as the uniparentally transmitted mitochondrial DNA (mtDNA) and male-specific portion of the Y chromosome (MSY). The latter two are not reshuffled by recombination and thus constitute a molecular archive of the history and migration of females and males, respectively, who transmitted them to subsequent generations. Our studies has also implications in biomedicine. For instance, the mitochondrial DNA contains 37 genes that are coding for components of respiratory chain enzyme complexes and are essential for cellular energy production. Thus, one of the aims of the research of this group is also to define the role of the “normal” sequence variation of mtDNA in some diseases (Parkinson’s, Alzheimer’s, diabetes, reduced fertility), in aging and athletic performance.
1) Origin of populations from Europe and the Mediterranean area
The demographic and genetic history of Europe and surrounding regions is extremely complex. The purpose of this research is to investigate, through the analysis of modern and ancient DNA genomes, what aspects of the variability of European populations can be traced back to the first colonization of the continent by modern humans, to post – glacial expansions, the Neolithic diffusion and (or) more recent events of gene flow. Over the years our studies on this general topic have been very productive and have used as the main tool of investigation the two non-recombinant genetic systems (mtDNA and MSY), which we are now integrating with genome-wide analyses.
Collaborations: Prof. Fulvio Cruciani (Sapienza Univ. di Roma); Dr. Martin Bodner, Prof. Walther Parson (Innsbruck Medical Univ., Austria); Dr. Francesca Gandini, Prof. Maria Pala, Prof. Martin B. Richards (Univ. of Huddersfield, UK); Prof. Vincent Macaulay (Univ. of Glasgow, UK); Prof. Andrea Novelletto (Univ. Tor Vergata, Roma); Prof. Antonio Salas (Univ. of Santiago de Compostela, Spain); Prof. Richard Villems (Estonian Biocentre, Tartu, Estonia).
2) The genetic origin of Italian populations
Despite its small geographic area, the Italian peninsula, for geographical, climatic and historical reasons, has occupied for several millennia a central position in the flow of genetic exchange and commercial trades among ancient populations. The objective of this research line is to reconstruct the prehistorical and historical genetics of the early Italy at a very high level of genetic resolution in a very large number of subjects (covering all Italian regions). To accomplish this task, a genome-wide scan on rare and common SNPs as well as analyses of both uniparental genetic systems (MSY and mtDNA) are going to be performed.
Collaborations: Prof. Andrea Angius (Istituto di Ricerca Genetica e Biomedica – CNR, Cagliari); Prof. Cristian Capelli (Univ. of Oxford, UK); Prof. David Caramelli (Univ. di Firenze); Prof. Cristina Cattaneo (Univ. di Milano); Dr. Hovirag Lancioni, Dr. Irene Cardinali (Univ. di Perugia); Prof. Carlo Previderè (Dip. di Medicina Legale, Univ. di Pavia); Prof. Mait Metspalu (Estonian Biocentre, Tartu, Estonia).
3) The peopling of Sardinia
Sardinian people are a “genetic outlier” in the European contest. Archaeological data point to a first evidence of modern human presence on the island dating back to about 13,000 years ago (Upper Palaeolithic). Distinctive genetic traits of Sardinians have been retrieved with all genetic systems, due to a combination of founder effects, geographic isolation, and selection. Recent paleo-genomic data based on nuclear genome markers have revealed that, among present-day populations, Sardinians retain the highest levels of genetic similarity with Early Neolithic farmers across Europe. However, this scenario appears in contrast with data provided by uniparentally-transmitted genetic systems. We are currently investigating on this topic, also from an ancient DNA perspective.
Collaborations: Prof. Andrea Angius, Prof. Francesco Cucca, Dr. Carlo Sidore (Istituto di Ricerca Genetica e Biomedica – CNR, Cagliari); Prof. Johannes Krause (Max Planck Institute for the Science of Human History, Germania).
4) The peopling of the Americas: a genetic perspective
The first peopling of the New World is a hot topic in genetics and anthropology. Archaeology, linguistics and genetics agree on the Asian origin of Native Americans. However, the arrival times of the first settlers, the number of migrations / expansions involved in the process and in the subsequent colonization of Central and South America, as well as the genetic consequences of the flourishing of the great Mesoamerican and Andean civilizations are still extensively debated. To clarify these issues, sequence analyses of the two non-recombinant genetic systems (MSY and mtDNA), both at the micro- and macro-geographic levels, are carried out in several modern populations and some ancient samples. In some contexts, these studies are integrated by genome-wide analyses at the nuclear level.
Collaborations: Dr. Martin Bodner, Prof. Walther Parson (Innsbruck Medical Univ., Austria); Prof. Richard Cooke (Smithsonian Tropical Research Institute, Panama); Prof. Ripan Malhi (Univ. of Illinois at Urbana-Champaign, US); Dr. Jorge A. Motta (Secretario Nacional de Ciencia y Tecnología e Innovación, Panama); Dr. Juan Miguel Pascale (Instituto Conmemorativo Gorgas de Estudios de la Salud, Panama); Dr. Ugo A. Perego (Genetic Genealogy Consultant, US).
5) A comparative analysis of historical-cultural and genetic data: conquest, commerce, crisis, culture and the Panamanian junction (1513-1671) (ERC CoG 648,535 – ArtEmpire)
The narrow Isthmus of Panama was a strategic node of the Spanish Empire and a crucial site for early modern globalization. This project combines analyses of historical documents with archaeological and archaeogenetic approaches to highlight the cultural and biological consequences of encounters between populations and goods from Europe, America, Africa and Asia in the Isthmus of Panama area during the sixteenth and seventeenth centuries.
Collaborations: Dr. Bethany Arhan (Universidad Pablo de Olavide, Spain); Dr. Corina Knipper (Curt-Engelhorn-Zentrum Archäometrie, Mannheim, Germany); Dr. Juan Martin, Dr. Javier Rivera (Universidad del Norte, Colombia).
6 ) Identification of new pathological mtDNA mutations and the roles of mitochondrial background (haplogroups) in the expression of diseases / phenotypes
The mitochondrial ATP production by the oxidative phosphorylation is essential for the maintenance of normal functions of organs and tissues. MtDNA mutations, by interfering with ATP synthesis, can cause serious maternally-transmitted diseases. The search for new disease-causing mtDNA mutations is carried out mainly on Leber’s Optic Neuropathy (LHON). In recent years, an important role of the “neutral” sequence variation of mtDNA has also been postulated for many complex diseases and other phenotypes (aging, athletic performance). To evaluate also these aspects, we are sequencing entire mitogenomes from numerous human populations and building up a database that includes representatives of each mtDNA haplogroup and sub-haplogroup present in our species.
Collaborations: Prof. Valerio Carelli (Univ. di Bologna); Dr. Cristina Cereda (IRCCS Istituto Neurologico Nazionale C. Mondino, Pavia); Prof. Alessandra Montecucco (Istituto di Genetica Molecolare – CNR, Pavia).
7) The genetics of taste
The ability to perceive flavors is genetically determined and the perception of flavors is the main factor driving the choice of foods, causing their acceptance or rejection. On the other hand, the choice of foods is an important factor that can modulate the risk of common diseases, such as obesity, diabetes, metabolic syndrome, but also cardiovascular diseases and cancer. The aim of our work is to correlate the variations of genes involved in taste perception, and thus in food choices, with diet and health state, in particular with tumors of the digestive system.
Collaborations: Prof. G. Nadia Ranzani (Dip. di Biologia e Biotecnologie, Univ. di Pavia).
8) Origin and spread of the Asian tiger mosquito (Aedes albopictus)
Over the last 40 years, the tiger mosquito Aedes albopictus, indigenous to East Asia, has colonized every continent except Antarctica. Its spread has become a growing public health concern, being a competent vector for many arboviruses including those causing West Nile fever, yellow fever, Dengue fever, Saint Louis encephalitis and the recently emerged Zika fever.
The present study involves the definition and analysis of mitogenome sequence variation in samples from numerous African, European and American populations, in order to identify the ancestral Asian source(s) of Ae. albopictus adventive populations and their possible selective advantages as well as migration routes and expansion times.
Collaborations: Dr. Paolo Gabrieli, Prof. Giuliano Gasperi, Prof. Ludvik Gomulski, Prof. Anna Malacrida (Department of Biology and Biotechnology, Univ. of Pavia).
9) Modes and times of domestication of some mammal species (Bos taurus, Bubalus bubalis, Ovis aeries, Capra hircus, Equus caballus)
Our first phylogeographic studies on the entire mitogenome of cattle, horse, goat and sheep provided new information on the domestication process(es). Most of the domestic species have been involved in a single domestication event that occurred in the Neolithic period (about 10,000 years ago) in the Fertile Crescent. On the contrary, the high number of haplogroups found in modern equine breeds suggests that the domestication of wild horses took place in several centres around Eurasia. The analysis is currently extended to the nuclear genome (in particular to the Y chromosome), focusing on the origin of local breeds (especially in Italy). The overall aim is to highlight parallel genetic histories of humans and domestic animals useful to further extend the current knowledge of human populations.
Collaborations: Dr. Hovirag Lancioni, Dr. Irene Cardinali, Dr. Emiliano Lasagna, Prof. Francesca Maria Sarti, Dr. Katia Cappelli, Prof. Maurizio Silvestrelli and Prof. Emidio Albertini (Univ. di Perugia); Dr. Licia Colli and Prof. Paolo Ajmone Marsan (Univ. Cattolica del S. Cuore, Piacenza).
10) Genetic origins of Italian wild mammals (Sus scrofa)
The same phylogeographic approach employed to analyze the domestic species is now employed for studying the variability of some wild species that are related or depending on human activities. In particular, virtually all European wild boars belong to only one mtDNA haplogroup, named D1, with the notable exception of the Italian-specific haplogroup D4. A phylogeographic study based on complete mitogenomes is in progress to clarify the origin and spread of this “Italian” lineage.
Collaborations: Dr. Hovirag Lancioni and Dr. Francesca Vercillo (Univ. of Perugia); Dr. Massimo Scandura (Univ. of Sassari).
1) Identification of the cellular target of new antitubercular compounds
Prof. Giovanna Riccardi, Prof. Maria Rosalia Pasca, Dr. Laurent Chiarelli, Dr. José Camilla Sammartino
Mycobacterium tuberculosis, the etiological agent of tuberculosis, (TB) is responsible for an estimated 1.5 million deaths worldwide annually. The emergence and spread of M. tuberculosis multidrug-resistant (MDR) and extensively drug-resistant (XDR) strains and, more recently, totally drug-resistant (TDR) strains is an extremely dangerous threat to public health, in particular to immunocompromised patients. In the new pathway for the discovery of drugs to combat the increasing menace of drug-resistant microorganisms, the crucial first step is to generate a set of target genes, from whole-genome data, that offer the potential for effective therapeutic intervention. Our objectives in the project “More Medicines for Tuberculosis – MM4TB” (VII Framework programme) are to find the molecular targets of prioritized molecules, already available, as well as natural products and synthetic compounds (hits) effective in models of active or latent TB. Among these molecules, we are studying the mechanism of activation of the TP053 prodrug (Albesa-Jove D et al., 2014, ACS Chem. Biol.) and we are searching for its possible cellular target. We have identified the mechanism of inactivation of the Ty38c quinoxaline derivative (Neres J et al., 2014, ACS Chem. Biol.), targeting the DprE1 enzyme identified within the previous “New Medicines for Tuberculosis – NM4TB” (VI Framework programme) as cellular target of Benzothiazinones (BTZ). Recently, a BTZ derivative entered human clinical trials. Lastly, the PyrG CTP synthase enzyme has been identified as the common cellular target of two different prodrugs (Mori G et al., 2015, Chem. Biol.) and it is considered a new promising vulnerable target in M. tuberculosis.
Collaborators: Cole ST (EPFL, Lausanne, Switzerland), Makarov V (Bakh Institute of Biochemistry, Russian Academy of Science, Moscow, Russia), Baltas M (CNRS, Toulouse, France), Mikusova K (Comenius University in Bratislava, Department of Biochemistry, Bratislava, Slovakia), Guerin M (Biophysics Unit, University of the Basque Country, Bilbao, Basque Country, Spain), Marengo E (Università del Piemonte Orientale, Alessandria, Italy).
2) New drugs against Burkholderia cenocepacia (FFC 2015)
Prof. Giovanna Riccardi, Dr. Silvia Buroni, Dr. Laurent Chiarelli, Dr. Viola Camilla Scoffone, Dr. Gabriele Trespidi
Burkholderia cenocepacia is a Gram-negative bacterium that infects the airways of patients with cystic fibrosis (CF). Eradication of this infection is complicated by the intrinsic resistance of this microorganism to different antibiotics. In this way, the development of novel antibiotics remains a major issue for the treatment of infectious lung disease, such as that in CF. We are currently pursuing two different ways:
Collaborations: Makarov V (Bakh Institute of Biochemistry, Russian Academy of Science, Moscow, Russia), Cardona S (University of Manitoba, Winnipeg, Canada), Coenye T (University of Gent, Belgium), Ungaro F (Università di Napoli Federico II), Bragonzi A (Ospedale San Raffaele, Milano), Drevinek P (Motol Hospital, Prague, Czech Republic).
3) Bacterial endophytes from leguminosae seeds for searching new antibacterial molecules, promoting seed germination and phytoremediation of soils
Prof. Edda De Rossi
Endophytic bacteria have been found in virtually every plant studied. Most endophytes appear to originate from the rhizosphere or phyllosphere; however, some may be transmitted through the seed. Endophytic bacteria can promote plant growth and yield and can act as biocontrol agents. Endophytes can also be beneficial to their host by producing a range of natural products that could be harnessed for potential use in medicine, agriculture or industry. In addition, it has been shown that they have the potential to remove soil contaminants by enhancing phytoremediation. So a major reason for the increasing interest in endophytes is the potential for biotechnological applications.
All classes of antibiotics have seen the emergence of resistance compromising their use. The emergence and dissemination of multi-drug-resistant bacteria and the lack of new antibiotics with new modes of action are today one of the major challenges to treat infectious diseases. Endophytes are capable of synthesizing bioactive compounds that are used by plants for defense against pathogens; thus, these microorganisms may represent an underexplored reservoir of novel species of potential interest in the discovery of novel lead compounds. Natural product drug discovery has regained interest due to low production costs, structural diversity, and multiple uses of active compounds to treat various diseases.
Seed germination is a vital phase to plants’ survival and reproduction in either optimal or stressful environmental conditions. Thus, developing methods by which seedling emergence can be enhanced and predicted under the limitations of heat or drought is valuable. The use of endophytic symbionts is a promising method by which seed germination can be enhanced and plants protected from environmental stressors.
The exploitation of plant–endophyte partnerships for the remediation of contaminated soils is a promising area. For example, in the case of phytoremediation of toxic metals, endophytes able to produce natural metal chelators can reduce metal toxicity for their host plant.
The aim of this recently activated project is to isolate the cultivable, endophytic bacteria from the seeds of commercial genotypes of leguminosae belonging to genus Medicago and Trifolium to characterize bacterial strains for the production of new antibacterial agents, germination of seeds and phytoremediation of soils.
Collaborations: Prof.ssa Alma Balestrazzi (Dip. di Biologia e Biotecnologie, Pavia)
4) Study of the membrane protein MmpL3 as cellular target for new antitubercular agents
Prof. Edda De Rossi
The number of multidrug resistant (MDR) strains of Mycobacterium tuberculosis (Mtb) is increasing and without effective treatments; the fear is that the number of infections caused by MDR strains will increase out of control. Therefore, new drugs of different classes against diverse targets are urgently needed. Pyrrole derivatives show a potent activity against Mtb and MDR clinical isolates of Mtb as well as on intracellular bacilli. On the basis of these results, BM212 was considered the lead compound of this new class of derivatives. The Pavia team has identified the membrane protein MmpL3 as target for this compound. MmpL3 is required for the export of mycolic acids in the form of trehalose monomycolates (TMM) to the periplasmic space or outer membrane of Mtb. Recently other researchers have identified seven series of inhibitors that abolish MmpL3-mediated TMM export confirming this essential protein as a novel target for new antitubercular agents. The aim of this project is to characterize the MmpL3 protein for both molecular modeling studies and synthesis of new and more efficient inhibitors.
5) New drugs against Mycobacterium abscessus and other nontuberculous mycobacteria
Prof.ssa Maria Rosalia Pasca
Collaborators: Dr. Laurent Chiarelli, Dr. José Camilla Sammartino
Nontuberculous mycobacteria (NTM) are emerging as important pathogens in cystic fybrosis (CF) lung disease worldwide with an estimated prevalence of about 9%; the Mycobacterium abscessus strains are the most spread in Europe. The M. abscessus treatments are further complicated by the diffusion of strains with inducible macrolide resistance. In these cases, the surgical resection of infected lung tissue could be beneficial in selected patients. Moreover, unsuccessful M. abscessus eradication is considered a contraindication for lung transplantation, being associated with treatment failure and increased mortality.
Consequently, new more active drugs are urgently needed, in particular against M. abscessus. In this project, taking advantage of our experience in tuberculosis research, we would like to move all our knowledge in the new challenging NTM field, by pursuing the following goals:
Collaboration: Makarov V (Bakh Institute of Biochemistry, Russian Academy of Science, Moscow, Russia).