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Pinna, Claudia Maria Assunta (2017) Epithelial-stromal interaction in the pathogenesis of colorectal cancer. [Ph.D. thesis]

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Abstract (english)

Colorectal cancer (CRC) is not a homogenous disease. Recent molecular classification of established tumours based on gene expression and (epi)genetic mutation burden, has revealed considerable disease heterogeneity. The relative importance of the epithelial and stromal tissue compartments varies between different tumour subtypes and this contributes to the observed clinical and molecular heterogeneity of CRC.
The AIM of this study was to explore the role of the stroma in different precancerous pathologies (polyps) and in CRC.
Stromal gene expression varies considerably between the different polyp subtypes (SSA and TVA) with a comparatively greater number of differentially expressed genes in serrated lesion stroma, suggesting the hypothesis that SSA lesions, usually initiated by BRAF mutations and methylation, require the recruitment of pro-tumorigenic stroma to enable lesion progression. In contrast, TVA are initiated by epithelial mutations that disrupt Wnt signaling (such as APC) and this is sufficient to drive tumourigenesis, irrespective of stromal influences.
Moreover, transwell tissue culture 3D techniques and animal models revealed that fibroblasts support the cross species growth of mouse epithelial organoids and abrogate the normal media requirement for Noggin and EGF. Interestingly, mouse epithelium grown in this co culture system develop as spheroids rather than the branching organoids seen with media morphogen supplementation, indicating a phenotype modulating effect of the fibroblasts.
Preliminary results revealed that fibroblasts have effects on cell proliferation and cell cycle regulation by upregulation of genes involved with cell cycle progression, DNA synthesis/repair, protein translation, vesicles mediated transport and lipid metabolism.
Primary stromal cell cultures isolated from adenoma and colon cancer (CMS2 and CMS4) might in part represent the corresponding cancer microenvironment, thus providing a useful complement to the current cellular biochemistry and therapeutic research in CRC.

Abstract (italian)

Il cancro del colon-retto (CRC) non è una malattia omogenea. Una recente classificazione molecolare sul CRC basata sull’ espressione genica e mutazioni (epi)genetiche, ha rivelato una notevole eterogeneità nella malattia. L' importanza dei compartimenti tissutali, epitelialiali e stromali, varia tra i diversi sottotipi di CRC e questo contribuisce all’ eterogeneità clinica e molecolare osservata nel CRC.
Lo scopo di questo studio è stato quello di esplorare il ruolo dello stroma in diverse forme pretumorali (polipi) e nel CRC.
Dalle analisi eseguite, e’ emerso che il profilo genetico dello stroma nelle due condizioni pre cancerogene di CRC oggetto di studio (TVA e SSA), differisce in maniera significativa, suggerendo l’ipotesi che i polipi di tipo SSA, in genere promossi da mutazioni nel gene BRAF, richiedono l'assunzione di stroma pro-cancerogeno per consentire la progressione della lesione. Al contrario, in lesioni di tipo TVA, le mutazioni epiteliali che interrompono la via di segnalazione Wnt (es. APC), siano sufficienti per promuovere la tumorigenesi a prescindere dall’ influenza dello stroma.
Inoltre, con l’ausilio di tecniche di coltura 3D, inserti e modelli animali, e’ emerso che i fibroblasti sostengono la crescita degli organoidi murine senza l’impiego dei fattori di crescita Noggin e EGF. In particolare, i fibroblasti causano lo sviluppo di sferoidi piuttosto che di organoidi, indicando un effetto modulante sul fenotipo.
I risultati preliminary, hanno rivelato che i fibroblasti hanno effetti sulla proliferazione cellulare e la regolazione del ciclo cellulare mediante la regolazione in maniera positiva dei geni coinvolti nella progressione del ciclo cellulare, nella sintesi e/o riparazione del DNA, nella traduzione di proteine, nel trasporto mediato da vescicole e nel metabolismo dei lipidi.
I fibroblasti isolati da adenoma e CRC (CMS2 e CMS4) potrebbero in parte rappresentare il microambiente tumorale, fornendo così un utile complemento biochimico e cellulare sulla ricerca terapeutica nel tumore del colon.

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EPrint type:Ph.D. thesis
Tutor:Sturniolo, Giacomo Carlo
Supervisor:Russo, Francesco Paolo
Ph.D. course:Ciclo 29 > Corsi 29 > BIOMEDICINA
Data di deposito della tesi:27 January 2017
Anno di Pubblicazione:27 January 2017
Key Words:CAFs in CRC pathogenesis
Settori scientifico-disciplinari MIUR:Area 06 - Scienze mediche > MED/12 Gastroenterologia
Struttura di riferimento:Dipartimenti > Dipartimento di Scienze Chirurgiche Oncologiche e Gastroenterologiche
Codice ID:9975
Depositato il:24 Nov 2017 09:49
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