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Fioravanzo, Lara (2009) Study of role of A-Beta peptides on angiogenesis related to Alzheimer's disease. [Tesi di dottorato]

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Abstract (inglese)

One of the main difficulties for the success of tissue and organ transplantation is the deficiency of techniques that ensure regular and stable vascularisation. The knowledge of mechanisms involved in the formation of new vessels could allow application of clinic strategies to facilitate tissue and organ transplantation on the one hand and the reduction of excessive vascular growth in many pathologies on the other.
Angiogenesis is a complex process, where several cell types and mediators interact to establish a specific microenvironment suitable for the formation of new vessels (Bouïs et al, 2006). It occurs in both physiological (embryo development, menstrual cycle, pregnancy, wound healing) and pathological conditions (inflammation, psoriasis, metastasis growth).
In the last years an increasing amount of evidence suggests that Alzheimer's disease (AD) is not only a neurodegenerative disease characterized by neuronal degeneration and loss of synaptic connections but it is also a vascular pathology. In fact cerebral microvessel vasoconstriction, degeneration of vessel smooth muscle cells and alteration of the basal membrane have been observed in cerebral capillary AD brains. In particular the possible connection between angiogenesis and AD starts from the observations of increased vascular density close to senile plaques in AD patients in comparison to older non-demented people (Vagnucci and Li, 2003). Senile plaques (extracellular and vascular deposits of proteinaceous material where the major component is β-amyloid peptide, Aβ) together with neuro fibrillary tangles (NFT) (composed of iperphophorylated form of τ protein) represent the main neuropathological alterations founded in AD brains (Robertson et al., 2003).
A large amount of in vivo, ex-vivo and in vitro experiments show contradictory data on Aβ peptide angiogenic capability (Paris et al., 2004; Cantara et al., 2004). The aim of the present research is the attempt to understand some of the mechanisms that favour angiogenesis.
The research deonstrates that Aβ peptides are pro-angiogenic in vivo, ex-vivo and in vitro on human cerebral endothelial cells while they are anti-angiogenic on rat cerebral endothelial cells, probably as consequence of their cytotoxic effect on these cells. Moreover Aβ1-42 peptide does not affect the release of VEGF-A and the expression of mRNA of VEGF-A on rat cerebral endothelial cells and rat hippocampal astrocytes cultures. Astrocytes release factors that induce angiogenesis and release of IL-6 on rat cerebral endothelial cells independently of the presence of Aβ1-42 peptide. Finally
the presence of hVEGF165 inhibits toxic and anti-angiogenic effects of the peptide probably taking away Aβ1-42 through the binding A?-hVEGF165.
Other experiments are necessary to understand mechanisms connected to AD although it seems that the result could depend on numerous different cell types in response of Aβ effects. Nevertheless capillary alteration and the hypoxia seem to contribute to the process.

Abstract (italiano)

Uno dei principali impedimenti alla ricostruzione di tessuti ed organi nell'ingegneria tessutale è la mancanza di strategie che assicurino costante e stabile vascolarizzazione. L'acquisizione di nuove conoscenze sui meccanismi che intervengono nel processo angiogenico potrebbe permettere una efficace applicazione clinica nella ricostruzione di tessuti e organi da una parte e nella riduzione della eccessiva crescita vascolare in patologie angiogenesi-dipendente dall'altra.
L'angiogenesi è un processo complesso in cui numerosi cellule e mediatori cellulari interagiscono per stabilire un microambiente specifico per la crescita di nuovi vasi sanguigni (Bouïs et al, 2006). Questo processo si verifica sia in condizioni fisiologiche (sviluppo embrionale, ciclo mestruale, gravidanza, guarigione di ferite) che patologiche (processi infiammatori, psoriasi, crescita di metastasi).
Negli ultimi tempi si sta affermando l'ipotesi che la demenza di Alzheimer (AD) non sia solamente una malattia neurodegenerativa caratterizzata da degenerazione neuronale e perdita delle connessioni sinaptiche ma anche una patologia vascolare. Numerose infatti sono le alterazioni riscontrabili a livello dei capillari cerebrali nei soggetti AD: vasocostrizione dei microvasi cerebrali, degenerazione delle cellule muscolari lisce vasali e alterazione della membrana basale. In particolare desta vasto interesse il possibile legame tra angiogenesi e AD derivante dall'osservazione di una aumentata densità  vascolare in prossimità delle placche senili nei pazienti AD rispetto a soggetti anziani non dementi (Vagnucci and Li, 2003). Le placche senili, depositi extracellulari e vascolari di materiale proteico costituite in prevalenza dal peptide β-amiloide (Aβ), assieme ai grovigli neurofibrillari (NFT), costituiti dalla forma iperfosforilata della proteina τ, rappresentano le principali alterazioni neuro-patologiche riscontrabili nel cervello di soggetti AD (Robertson et al., 2003).
Numerosi studi in vivo, ex-vivo e in vitro riportano risultati contraddittori sulla capacità  del peptide Aβ di stimolare il processo angiogenico (Paris et al., 2004; Cantara et al.,
2004). Scopo di questo lavoro di ricerca è il tentativo di comprendere alcuni meccanismi che favoriscono questo processo.
Da questo lavoro di ricerca è emerso che il peptide Aβ è pro-angiogenico in vivo, ex-vivo e in vitro su cellule endoteliali cerebrali umane e anti-angiogenico su cellule endoteliali cerebrali di ratto, probabilmente come conseguenza dell'effetto citotossico effettuato dal peptide su queste ultime. Inoltre il Aβ1-42 non interferisce sulla via di espressione e rilascio di VEGF da parte di cellule endoteliali cerebrali e di astrociti di ippocampo di ratto in coltura. Gli astrociti secernono sostanze che inducono angiogenesi e rilascio di IL-6 da parte delle cellule endoteliali cerebrali di ratto in coltura indipendentemente dal trattamento con Aβ1-42. Infine la presenza di hVEGF165 inibisce l'effetto tossico e anti-angiogenico del peptide probabilmente in seguito al sequestramento del Aβ1-42 ad opera del legame Aβ1-42-hVEGF165.
Altri studi sono necessari al fine di comprendere i meccanismi di angiogenesi correlata con la AD sebbene sembri evidente che essa possa derivare dall'interazione di diversi tipi cellulari in risposta all'effetto del peptide Aβ su di esse. Sembra tuttavia che l'alterazione dell'endotelio capillare e la conseguente ipossia contribuiscano all'attivazione di tale processo.

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Tipo di EPrint:Tesi di dottorato
Relatore:Folin, Marcella
Dottorato (corsi e scuole):Ciclo 21 > Scuole per il 21simo ciclo > BIOLOGIA E MEDICINA DELLA RIGENERAZIONE > INGEGNERIA DEI TESSUTI E DEI TRAPIANTI
Data di deposito della tesi:28 Gennaio 2009
Anno di Pubblicazione:Gennaio 2009
Parole chiave (italiano / inglese):angiogenesis, Alzheimer, beta-amyloid, astrocytes, endothelial cells
Settori scientifico-disciplinari MIUR:Area 05 - Scienze biologiche > BIO/13 Biologia applicata
Struttura di riferimento:Dipartimenti > Dipartimento di Scienze Farmaceutiche
Codice ID:1528
Depositato il:28 Gen 2009
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