Neoplasie della vescica urinaria negli animali: modello di studio ?

Prof. Pippo Borzacchiello Dipartimento Medicina veterinaria e Produzioni animali

Università degli Studi di Napoli Federico II borzacch@unina.it

Neoplasie della vescica urinaria dei bovini

Virus: Papillomavirus bovino Alimentazione: Felce

Oncogeni E5 – E7 Ptaquiloside: sostanza oncogena

CANCRO

Neoplasie uroteliali e mesenchimali (emangiomi ed emangiosarcomi)

Neoplasie della vescica urinaria dei bovini: il ruolo dell’oncogene E5

Oncogene, 2006

ORIGINAL ARTICLE

Bovine papillomavirus E5 oncoprotein binds to the activated form of theplatelet-derived growth factor b receptor in naturally occurring bovineurinary bladder tumours

G Borzacchiello1, V Russo1, F Gentile2, F Roperto1, A Venuti3, L Nitsch2, M S Campo4

and S Roperto1

1Department of Pathology and Animal health, Faculty of Veterinary M edicine, Naples University ‘Federico I I ’, Naples, I taly;2Department of Cellular and M olecular Biology and Pathology ‘L .Califano’ c/o Istituto di Endocrinologia e Oncologia SperimentaleIEOS CNR, Naples, I taly; 3Regina Elena Cancer Institute, Laboratory of Virology, Rome, I taly and 4Institute of ComparativeM edicine, Division of Pathological Sciences, University of Glasgow, Glasgow, UK

Studies regarding the functions of the bovine papilloma-virus (BPV) E5 oncoprotein in vivo are lacking and no E5-mediated mechanism underlying epithelial carcinogenesisis known. We have shown that BPV-2 DNA is present inthe majority of naturally occurring urinary bladdertumours of cattle and that E5 is expressed in the cancercells. Here we show that the interaction between theplatelet-der ived growth factor (PDGF) b receptor andBPV E5, described in vitro in cultured cells, takes place invivo in bovine urinary bladder cancers. In these cancers,E5 and PDGF b receptor colocalize, as shown by confocalmicroscopy, and physically interact, as shown by coim-munoprecipitation. Furthermore, the PDGF b receptorassociated with E5 is highly phosphorylated, suggestingthe functional activation of the receptor upon E5interaction. Our results demonstrate, for the first time,that E5–PDGF b receptor interaction occurs during thenatural history of bovine urinary bladder tumours,suggesting an important role for E5 in carcinogenesis.Finally, the system provides a suitable animal model ofpapillomavirus-associated cancer to test therapeuticvaccination against E5. Successful bladder tumourregression would provide a valuable model for therapeuticvaccination against papillomavirus-associated tumours.Oncogene advance online publication, 3 October 2005;doi:10.1038/sj.onc.1209152

Keywords: bovine papillomavirus type 2; E5; PDGF breceptor; urinary bladder tumours

Introduction

Urinary bladder tumours are known to occur rarely inanimals. However, cattle grazing on bracken fern-

infested lands and suffering from chronic enzootichaematuria (CEH) frequently develop bladder cancerinvolving both the epithelium and mesenchyme(Pamukcu et al., 1976).

CEH is a severe syndrome due to prolonged ingestionof bracken fern (Pteridium spp), theonly plant proven tocause cancer in animals (Smith, 1997). This syndromeoccurs in several areas worldwide and is very commonin Southern I taly, where bracken is widespread(Borzacchiello et al., 2003b).

I t was first suggested in the 1960s that a papilloma-virus is associated with bladder cancer in cattle (Olsonet al., 1965), and since then Campo and co-workersestablished a strong relationship between bovine papil-lomavirus (BPV)-2 and bracken fern in both experi-mental and naturally occurring bovine bladder cancer(Campo et al., 1992). Although the synergism betweenthe virus and bracken is still poorly understood, it islikely that BPV-2 infects the bladder mucosa, producingan abortive latent infection, as no structural proteins orvirionsare found in theurinary bladder mucosa (Campoet al., 1992; Borzacchiello et al., 2003c). Immunosup-pressants and chemical carcinogens from bracken actsynergistically with the viral proteins, inducing neoplas-tic disease, and it has been suggested that latent BPV isactivated by bracken-induced immunosuppression, thusinitiating progression to malignancy (Campo, 1997).BPV-2 DNA has also been detected in lymphocytesfrom field cases of cattle, both healthy and sufferingfrom CEH and urinary bladder cancer (S Roperto,unpublished observation; Campo et al., 1994; Stoccodos Santos et al., 1998), suggesting that the peripheralblood cells can function as a reservoir for latent virus. I thas been proposed that papillomavirus is involved alsoin human urinary bladder cancers (Shibutani et al.,1992; Agliano et al., 1994; De Gaetani et al., 1999), butthis suggestion is often not corroborated (Oliver et al.,1998).

The papillomavirus E5 proteins are short hydropho-bic polypeptides (from 83 amino-acid residues in humanpapillomavirus type 16 (HPV-16) to 42 residues inBPV-4), many of which have transforming activity. E5

Received 15 M arch 2005; revised 26 August 2005; accepted 30 August2005

Correspondence: Dr G Borzacchiello, Department of Pathology andAnimal Health, Faculty of Veterinary M edicine, University of Naples‘Federico I I ’, Via F Delpino, 1, Naples 80137, I taly.E-mail: borzacch@unina.it

Oncogene (2005), 1–10

& 2005 Nature Publishing Group All rights reserved 0950-9232/05 $30.00

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Neoplasie della vescica urinaria dei bovini: tutta colpa di E5?

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Analogie con la controparte umana: 1.Istopatologiche (ca. uroteliale invasivo) 2.Molecolari 3.Comportamento biologico – metastasi 4.Risposta alla terapia 5.Prognosi

Neoplasie della vescica urinaria dei cani

Linee cellulari derivate da carcinoma vescicale di cane: 1. Caratterizzazione (CK, E-cadherin +) 2. Tumorigeniche nei topi nudi 3. Crescita ancoraggio indipendente 4. Caratteristiche molecolari: p53; cox-2 pRb

MODELLO CANINO IN VITRO

Neoplasie della vescica urinaria dei cani

Linee cellulari stabilizzate

Caratteristiche citogenetiche, morfologiche e biochimiche molto diverse dalle neoplasie uroteliali naive

Modelli murini

Fino a 12 mesi per lo sviluppo delle neoplasie

Modelli murini ortotopici Modelli murini eterotopici

Topi KO per delezioni specifiche uroteliali p53 and pRb

Caratteristiche di un modello ideale di studio

1. Facilmente disponibile e poco costoso 1. Deve avere caratteristiche biologiche comparabili con l’uomo 1. Profilo cellulare e molecolare comparabile con l’uomo 1. Responsivo alle terapie 1. Gli effetti avversi delle terapie e l’effetto antitumorale DEVE essere predittivo

dell’esito nell’uomo

E’ tempo di dare delle linee guida

1. Ideale per ca. transizione invasivo

2. Ideale per testare nuove terapie antineoplastiche

1. Individuazione fattori di rischio indoor

1. Ideale per lo studio della cancerogenesi da papillomavirus

2. Ideale per testare nuove terapie farmacologiche e vaccini

3. Ideale per lo studio della morfologia

Neoplasie della vescica urinaria degli animali domestici

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“Un modello è una bugia che aiuta a vedere la verità.”

Howard Skipper

M.PATLAK “Targeting leukemia: from bench to bedside”

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