Littérature scientifique sur le sujet « Adenocarcinoma duttale del pancreas »

Descripción del caso: perro de nueve años de edad, macho, mestizo (Terranova/chow chow) con historia de diarrea crónica, vómito y sin respuesta a diferentes tratamientos; fue recibido en el Hospital Veterinario de Especialidades de la Universidad Nacional Autónoma de México para continuar con el proceso diagnóstico de una probable neoplasia intestinal.Hallazgos clínicos: condición corporal 2/5, 6 % de deshidratación, depresión, vómito, diarrea, dolor abdominal y anorexia. El diagnóstico fue pancreatitis aguda y un proceso infiltrativo (neoplásico o inflamatorio). En los estudios sanguíneos se observó anemia ligera a moderada relacionada con inflamación crónica y posible pérdida entérica; hipoproteinemia por pérdidas entéricas (diarrea) y anorexia prolongada, y en el leucograma hubo inflamación activa de forma persistente.Pruebas de laboratorio: ultrasonido que mostró cambios en el páncreas y el duodeno. Se realizaron varios estudios sanguíneos y una gasometría arterial, las alteraciones fueron asociadas a dolor.Tratamiento y evolución: el paciente fue hospitalizado para realizar biopsia quirúrgica y colocarle una sonda de yeyunostomía; tres días después mostró dolor, melena, empeoramiento del estado mental y efusión abdominal. El perro fue diagnosticado con peritonitis séptica. Debido a la condición y riesgo del paciente, se le practicó la eutanasia.Relevancia clínica: las neoplasias intestinales en perros son raras, el adenocarcinoma es el segundo más común. Como factores de pobre pronóstico se mencionan, la localización, el tamaño y la pobre diferenciación histológica. Este reporte es relevante por la limitada información e incidencia de este tipo de neoplasia, principalmente por la localización y manejo médico y quirúrgico. Duodenal adenocarcinoma in a dog: diagnostic approach and complications Abstract Case description: nine-year-old male dog, mix breed (Terranova/chow chow) with a history of chronic diarrhea and vomiting without response to different treatments. It continues in diagnosis process due to probable intestinal neoplasia.Clinical findings: body condition 2/5, 6 % dehydration, depression, vomiting, diarrhea, abdominal pain and anorexia. The diagnosis was acute pancreatitis and infiltrative intestinal process (neoplastic or inflammatory). The blood counts observed mild to moderate anemia related to chronic inflammation and possible enteric losses, hypoproteinemia due to enteric losses (diarrhea) and prolonged anorexia, and in the leukogram it had active inflammation persistently.Lab test: ultrasound showed changes in the pancreas and duodenum. Several blood counts were performed, and an arterial blood gas, the alterations were associated with pain.Treatment and evolution: the patient was hospitalized for surgical biopsy and placement of a jejunostomy tube; three days later there was pain, mane, worsening mental state and abdominal effusion. The dog was diagnosed with septic peritonitis was. Due to the patient´s risk, euthanasia was practiced.Clinical relevance: intestinal neoplasm in dogs are rare, the adenocarcinoma is the second most common. As factors of poor prognosis, the location, size and poor histological differentiation are mentioned. It is important to know the available tools to make a proper diagnosis, establish the prognosis and take therapeutic decisions. The relevance of this case report is the limited information and incidence of this type of neoplasm, mainly the location, which implies a medical-surgical management different from others tumours.Key words: duodenal adenocarcinoma, effusion citology, lactate, glucose, intestinal neoplasia.

Introduzione. L’adenocarcinoma duttale (PDAC) rappresenta circa l’85% delle neoplasie maligne pancreatiche. È un tipo di neoplasia molto aggressiva e ha prognosi infausta. Il PDAC è una malattia con una elevata mortalità, spesso diagnosticata in uno stadio avanzato per il quale esistono poche e inefficaci terapie. L’alta incidenza di ricadute locali unita alla precoce metastatizzazione sono le caratteristiche cliniche più tipiche di questo tumore. Inoltre la nota resistenza del tumore alla chemio e alla radioterapia limita l’efficacia di questi approcci terapeutici. Scopo. Identificare e validare nuovi marcatori biologici associati a caratteristiche di aggressività dell’adenocarcinoma pancreatico al fine di utilizzarli per comprendere proprietà biologiche del tumore stesso o in clinica per una corretta valutazione prognostica. Metodi e risultati. Abbiamo studiato n=17 linee cellulari umane immortalizzate di PDAC per alcune caratteristiche di aggressività cellulare: per la clonogenicità e la chemioresistenza alla gemcitabina in vitro e, in vivo, per la capacità di crescita in topi immunocompromessi (CD1-nude). Tutte le 17 linee cellulari sono state caratterizzate per l’espressione di classi di marcatori molecolari: recettori delle chemochine (CCR1-CCR10; CXCR1-CXCR6; CX3CR1; XCR1) e putativi marcatori staminali tumorali (ESA+CD24+CD44+; CD133+, CXCR4+) mediante citometria a flusso, secrezione di fattori solubili (n=48) tramite la tecnologia luminex e l’espressione di geni (n=11) coinvolti nello sviluppo pancreatico con Real Time PCR. Usando l’analisi statistica inter-linea (regressione lineare o di cox) abbiamo cercato una correlazione tra i fenotipi biologici e le caratteristiche di malignità cellulare individuando nuovi marcatori. Questi marcatori sono stati validati su tessuti tumorali primari in casistiche di pazienti affetti da PDAC: l’espressione del marcatore identificato è stata correlata con l’esisto clinico della neoplasia. In una prima analisi inter-linea n=35 fattori sono risultati statisticamente associati ad una o più caratteristiche di aggressività. È seguita una classificazione per priorità che, avvalendosi della sola correlazione con la tumorigenicità in vivo, ha ridotto a n=20 i fattori di rischio da validare. Abbiamo quindi approfondito lo studio su 4 marcatori molecolari di sviluppo pancreatico (ISL1, PDX1, PAX6, KRT19), sui fenotipi staminali e su 2 recettori delle chemochine (CCR5, CXCR3). L’espressione genica dell’mRNA di ISL1, PDX1, PAX6 e KRT19 è stata valutata in sezioni criostatiche di n=42 resezioni chirurgiche di pazienti affetti da PDAC. Non sono emerse correlazioni significative tra l’espressione di questi fattori e la sopravvivenza globale. Tuttavia alti livelli dell’mRNA di KRT19 predicono una progressione più precoce e di tipo metastatico. Più elevati livelli di PDX1 e PAX6 sono associati con una più alta probabilità di recidiva locale. Inoltre combinando i marcatori è stato individuato un fenotipo più aggressivo correlato con una minor sopravvivenza: si tratta dei pazienti che esprimono ad elevati livelli sia PDX1 che KRT19. La nostra strategia di screening ha mostrato essere fattori di rischio per lo sviluppo tumorale nel topo, non i classici fenotipi staminali descritti in letteratura (ESA+/CD24+/CD44+, CD133+, CD133+/CXCR4+) ma la combinazione ESA+/CD24-/CD44+ e la sola espressione di CXCR4 ed ESA: tuttavia la validazione clinica, condotta su una coorte di 39 pazienti affetti da adenocarcinoma duttale, non ha confermato che questi marcatori, né i classici già descritti, correlino in maniera statisticamente significativa con la sopravvivenza o con la progressione nel tempo e nemmeno con il sito di recidiva. Il fenotipo ESA+/CD24+/CD44- è invece risultato un fattore di rischio prognostico indipendente sia per la sopravvivenza (HR=4,166 p=0,001) che per la progressione (HR=2,208 p=0,019). CCR5 e CXCR3 sono risultati espressi su tessuti tumorali processati a fresco (n=6) ed analizzati in citometria a flusso a fronte di una negatività del pancreas di donatori d’organo (n=10). Essi sono espressi rispettivamente nell’11,9% e nel 17,6% delle cellule CA19.9+ del tumore, mentre solo nello 0,4% e il 0,34% nel tessuto sano. L’aumentata espressione di CCR5 e CXCR3 sembra essere una caratteristica tipica del PDAC. Conclusioni. La nostra strategia ha identificato marcatori biologici capaci di distinguere differenti comportamenti clinici del tumore in termini di progressione e sede di recidiva. La differente espressione di questi predittori potrebbe essere la causa di differenze biologiche che hanno un effetto sui meccanismi di progressione e diffusione tumorale. Al fine di confermare i nostri risultati stiamo realizzando un tissue microarray di resezioni chirurgiche di pazienti affetti da adenocarcinoma duttale. Inoltre in futuro il modello statistico sviluppato potrà essere applicato per testare ogni nuovo potenziale marcatore; caratterizzata la sua espressione sulle linee cellulari, si procederà con l’analisi inter-linea per verificare se sia un potenziale indicatore di aggressività in vitro o in vivo nel modello murino; quindi si potrà confermare il suo reale ruolo diagnostico, prognostico o predittivo in ambito clinico.

Abstract Epithelial-to-Mesenchymal Transition (EMT) is a stepwise and complex biological mechanism which plays an important role in tumor progression. During EMT, epithelial cells undergo a phenotypical switch to mesenchymal cells, acquiring motility and invasion capability. We aimed at clarifying the role of this complex mechanism in the Pancreatic Ductal Adenocarcinoma (PDAC) focusing on the expression of EMT markers in different cell culture conditions, such as monolayer and spheroids. For this study we used immunofluorescence and confocal microscopy, zymography, differential proteomics, real-time pcr and western blot to analyse and characterize three commercial PDAC cell lines ( HPAF-II, HPAC and PL45) cultured as monolayer or 3D- spheroids. Our analyses revealed that PDAC cells cultured in monolayer show different epithelial characteristics, such as retention of the E-cadherin/β-catenin complex – and, consequently, keep zonula adherens junctions – as well as some mesenchymal and EMT markers, such as high levels of matrix metalloproteinases (MMPs). The analysis of 3D-spheroids revealed that the E-cadherin/ β-catenin complex was similarly expressed at the cell boundaries on the plasma membrane in 2D-monolayers as well as in the 3D-spheroids, but cleavage fragments were detectable in lysates obtained from monolayers. By contrast, N-cadherin expression was observed in few HPAC cells grown in monolayers but increased in 3D-spheroids, and some cells expressing collagen type I were observed in 3D-spheroids. Podoplanin and α-smooth muscle actin were similarly expressed in both experimental conditions, as well as Snail, Slug and Zeb1. Our results confirm that EMT plays a role in PDAC. In particular, the concomitant retention of an epithelial phenotype based on the retention of zonula adherens junctions together with the expression of mesenchymal markers and proteins involved in actin cytoskeleton reorganization, strongly suggest a collective migration mechanism for PDAC cells. Moreover, the overall information provided by this study support the use of 3D cultures in biomedical research to bridge the gap between traditional cell cultures and in vivo settings. 3D cultures seem provide powerful information on PDAC cells phenotype possibly better reflecting the in vivo organization than monolayer cell culture systems, and could therefore represent a pre-clinical model to identify and validate tumor markers and to study new molecular tools to inhibit signalling pathways, or to target EMT transcription factors.

Serum levels of several molecules associated to pancreatic adenocarcinoma (PDAC) pathophysiology are evaluated in this work, in order to determine their diagnostic value, distinguishing between PDAC patients and healthy controls (HC), different gastrointestinal tumours (GIT) and chronic pancreatitis (CP). Plasma mRNA levels in plasma of sialyltransferases (ST3Gal III and ST3Gal IV) could differentiate between HC and PDAC. Moreover, lower levels of ST3Gal III in early stages of PDAC compared to PDAC advanced stages were reported. The Glasgow Prognostic Score (GPS), inflammatory response quantification, differentiated between all the study groups. IGF-1 levels were lower in neoplasic groups of patient vs HC and CP. We assessed the diagnostic capacity of different markers alone or in combination and compared with that of CA 19.9. The best diagnostic capacity was found combining CEA, CA 19.9 and IGF-1 compared with CA 19.9 and it could be useful to distinguish PDAC from CP.
En este trabajo se analizan distintos parámetros séricos relacionados con la fisiopatología del adenocarcinoma pancreático (PDAC), para evaluar su uso como marcadores que permitan distinguir entre pacientes con PDAC y controles sanos (C), otras neoplasias del tracto gastrointestinal (ONEOS) y pancreatitis crónica (PC). La expresión en plasma de sialiltransferasas, ST3Gal III y ST3Gal IV diferencia entre controles y PDAC. Así mismo, los niveles de ST3Gal III permiten diferenciar en PDAC entre estadíos iniciales y metastáticos. El Glasgow Prognostic Score (GPS), medida de la respuesta inflamatoria, diferencia entre todos los grupos del estudio. Los valores de IGF-1 disminuyen en procesos neoplásicos vs C y PC.Se analiza la capacidad diagnóstica de los distintos parámetros individualmente y combinados entre sí respecto al CA19.9. La combinación de CA 19.9, CEA, IGF-1 aumenta la precisión diagnóstica frente al CA19.9 y podría ser útil para distinguir PDAC de PC.

OBJECTIVES: CD4+CD25+FOXP3+ T cells are an heterogeneous population of cells playing a key role in maintenance of the immune system’s tolerance of both foreign and self-antigens. They can inibit immune responses mediated by T effector cells. Patients with a variety of cancers have an enlarged pool of Treg in their peripheral blood, in tumor-draining lymph nodes and in the tumor microenvironment. Pancreatic cancer remains a major unsolved health problem, with conventional cancer treatement having little impact on disease course. Poorly prognosis in pancreatic cancer (PC) patients has been correlated with increase of Treg in peripheral blood, lymph nodes et tumor tissue. Despite the advanced understanding of the mechanism of cancer evasion, the exact mechanism and clinical significance of Treg elevation in PC patients remains unclear. Further work is required to investigate of wich of Treg specific subpopulation is involved in pancreatic cancer developement to provide greater insights into potential mechanisms of cancer evasion and to develope an array of treatements that will inhibit spacific pathways that mediate evolution and progression of ACP. To evaluate the distibution of Tregs in peripheral blood of PC patients, in the present prospectic study the percentage of circulating rTregs and aTreg has been compared to that of healthy volunteers, colon cancer patients and inflammatory diseases patients. Furthemore, we analyse the balance of the different subsets of Tregs in peripheral blood in PC patients and the correlation between the prevalence of circulating Tregs in PC patients and their clinico-pathological findings and outcomes. METHODS: Among a total of 29 patients with ductal adenocarcinoma of the pancreas, 24 underwent pancreatectomy and 5 biliopancreatic diversion. Their peripheral blood mononuclear cells (PBMCs) were analysed to determine the proportion of CD4+CD25+Foxp3+ T cells (Tregs), as a percentage of the total CD4+ cells, by FACScan analysis after labeling with anti-CD4, anti-CD25, anti-CD45 and anti-Foxp3 antibodies. Clinical stages were classified according to the TNM classification. RESULTS: The percentage of CD4+CD25+++CD45RA- Foxp3hi (aTreg) cells among the PBMCs was significatively increased in PC patients compared with healty donors (HD) (P<0,001), colon cancer patients and patients with inflammatory diseases (P<0,001). The analys of rTreg/aTreg balance showed an evolutive inflammatory profil in PC patients compared to that of healthy donors and confirm the Treg cell differentiation dynamics and interactions. A positive correlation was found between the percentage of CD4+CD25++CD45RA+FoxP3low (nTreg), CD4+CD25+++CD45-RAFoxp3hi (aTreg) and the patients age. No correlation was found between the percentage of circulating Treg and clinical stages and survival. CONCLUSIONS: The increase in CD4+CD25+++CD45RA-FoxP3hi Treg cells may be related to immunosuppression and tumor progression in patients with ductal pancreatic cancer. Their removal can boost the antitumor immunity with the potential to achieve therapeutic impacts. Further studies are required: - To determine the mechanisms exact of tumor evasion and of Treg cells elevation in peripheral blood, draining lymph nodes and tumor tissue in cancer patients. - to evaluated dynamic interactions between different subsets of Treg cells and their clinical significance. The immunological monitoring of Treg may be useful to predict the prognosis for patients with pancreatic cancer. Additionally, an increase in our understanding of exact mechanism of Treg-mediated suppression will offer insights into potential alternatives for inhibiting, in a selective way, specific Treg activity and enhance immunotherapeutic potentials.