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Translational pancreatic cancer research:a comparative study on patient-derived xenograft models

World Journal of Gastroenterology 2018年第7期24卷 794-809页

作者： Mercedes Rubio-Manzanares Dorado Luis Miguel Marín Gómez Daniel Aparicio Sánchez Sheila Pereira Arenas Juan Manuel Praena-Fernández Juan Jose Borrero Martín Francisco Farfán López Miguel ángel Gómez Bravo jordi muntané relat Javier Padillo Ruiz Department of Hepatobiliary and Pancreatic Surgery Virgen del Rocio University Hospital Oncology Surgery Cell Therapy and Organ Transplantation Group Institute of Biomedicine of Seville (IBiS) Virgen del Rocio University Hospital University of Seville Statistics Methodology and Evaluation of Research Unit Virgen del Rocio University Hospital Pathology Department Virgen del Rocio University Hospital

AIM To assess the viability of orthotopic and heterotopic patient-derived pancreatic cancer xenografts implanted into nude mice.METHODS This study presents a prospective experimental analytical follow-up of the development of tumours in mice upon implantation of human pancreatic adenocarcinoma samples. Specimens were obtained surgically from patients with a pathological diagnosis of pancreatic adenocarcinoma. Tumour samples from pancreatic cancer patients were transplanted into nude mice in three different locations(intraperitoneal, subcutaneous and pancreatic). Histological analysis(haematoxylin-eosin and Masson's trichrome staining) and immunohistochemical assessment of apoptosis(TUNEL), proliferation(Ki-67), angiogenesis(CD31) and fibrogenesis(α-SMA) were performed. When a tumour xenograft reached the target size, it was reimplanted in a new nude mouse. Three sequential tumour xenograft generations were generated(F1, F2 and F3).RESULTS The overall tumour engraftment rate was 61.1%. The subcutaneous model was most effective in terms of tissue growth(69.9%), followed by intraperitoneal(57.6%) and pancreatic(55%) models. Tumour development was faster in the subcutaneous model(17.7 ± 2.6 wk) compared with the pancreatic(23.1 ± 2.3 wk) and intraperitoneal(25.0 ± 2.7 wk) models(P = 0.064). There was a progressive increase in the tumour engraftment rate over successive generations for all three models(F1 28.1% vs F2 71.4% vs F3 80.9%, P < 0.001). There were no significant differences in tumour xenograft differentiation and cell proliferation between human samples and the three experimental models among the sequential generations of tumour xenografts. However, a progressive decrease in fibrosis, fibrogenesis, tumour vascularisation and apoptosis was observed in the three experimental models compared with the human samples. All three pancreatic patient-derived xenograft models presented similar histological and immunohistochemical characteristics.CONCLUSION In our experience, the faster development andgreatest number o

关键词： Immunohistological analysis Pancreatic cancer Patient-derived xenograft Animal model Nude mice

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Nitric oxide and cancer

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World Journal of Hepatology 2010年第9期2卷 337-344页

作者： jordi muntané Manuel De la Mata Liver Research Unit Maimonides Institute for Biomedical Research of Cordoba (IMIBIC)"Reina Sofia" University Hospital Biomedical Research Network Center for Liver and Digestive Diseases (CIBERehd) Digestive Clinical Unit Maimonides Institute for Biomedical Research of Cordoba (IMIBIC)"Reina Sof ia" University Hospital

Nitric oxide (NO) is a lipophilic,highly diffusible and short-lived physiological messenger which regulates a variety of important physiological responses including va sodilation,respiration,cell migration,immune resp onse and apoptosis.NO is synthesized by three differ entially gene-encoded NO synthase (NOS) in mammals: neuronal NOS (nNOS or NOS-1),inducible NOS (iNOS or NOS-2) and endothelial NOS (eNOS or NOS-3).All isof or ms of NOS catalyze the reaction of L-arginine,NA DPH and oxygen to NO,L-citrulline and NADP.NO may exert its cellular action by cGMP-dependent as well as by cGMP-independent pathways including postranslational modifications in cysteine (S-nitrosylation or S-nit rosation) and tyrosine (nitration) residues,mixed disulf ide formation (S-nitrosoglutathione or GSNO) or prom ot ing further oxidation protein stages which have been related to altered protein function and gene transcription,genotoxic lesions,alteration of cell-cycle check points,apoptosis and DNA repair.NO sensitizes tumor cells to chemotherapeutic compounds.The expression of NOS-2 and NOS-3 has been found to be increased ina variety of human cancers.The multiple actions of NO in the tumor environment is related to heterogeneous cell responses with particular attention in the regulation of the stress response mediated by the hypoxia inducible factor-1 and p53 generally leading to growth arrest,apoptosis or adaptation.

关键词： Nitric oxide Cancer Carcinogenesis

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Mitochondrial protection by low doses of insulin-like growth factor-Ⅰin experimental cirrhosis

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World Journal of Gastroenterology 2008年第17期14卷 2731-2739页

作者： Raquel Pérez María García-Fernández Matías Díaz-Sánchez Juan E Puche Gloria Delgado Marian Conchillo jordi muntané Inma Castilla-Cortázar Departments of Human Physiology and Internal Medicine University of Navarra Department of Medical Physiology School of Medicine University of Málaga Department of Internal Medicine Liver Unit University of Córdoba

AIM: To characterize the mitochondrial dysfunction in experimental cirrhosis and to study whether insulin-like growth factor-Ⅰ(IGF-Ⅰ) therapy (4 wk) is able to induce beneficial effects on damaged mitochondria leading to cellular protection. METHODS: Wistar rats were divided into three groups: Control group, untreated cirrhotic rats and cirrhotic rats treated with IGF-Ⅰtreatment (2 μg/100 g bw/d). Mitochondrial function was analyzed by flow cytometry in isolated hepatic mitochondria, caspase 3 activation was assessed by Western blot and apoptosis by TUNEL in the three experimental groups. RESULTS: Untreated cirrhotic rats showed a mitochon- drial dysfunction characterized by a significant reduction of mitochondrial membrane potential (in status 4 and 3); an increase of intramitochondrial reactive oxigen species (ROS) generation and a significant reduction of ATPase activity. IGF-Ⅰtherapy normalized mitochondrial func-tion by increasing the membrane potential and ATPase activity and reducing the intramitochondrial free radical production. Activity of the electron transport complexes Ⅰand Ⅲ was increased in both cirrhotic groups. In addition, untreated cirrhotic rats showed an increase of caspase 3 activation and apoptosis. IGF-Ⅰtherapy reduced the expression of the active peptide of caspase 3 and resulted in reduced apoptosis.

关键词：胰岛素增长因素细胞凋亡症状实验研究

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Conversion from calcineurin inhibitors to mTOR inhibitors stabilizes diabetic and hypertensive nephropathy after liver transplant

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World Journal of Transplantation 2015年第1期5卷 19-25页

作者： José M álamo Claudia Olivares Lydia Barrera Luis M Marín Gonzalo Suarez Carmen Bernal Juan Serrano jordi muntané Francisco J Padillo Miguel A Gómez Unit of Liver Transplantation Department of Surgery University of Seville41013 Seville Spain Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBEREH o Ciberehd) Instituto de Salud Carlos Ⅲ 08036 Barcelona Spain Department of General Surgery Hospital Universitario Virgen del Rocío/CSIC/Universidad de Sevilla Seville 41013 Spain

AIM: To investigate if conversion to the mammalian target of rapamycin inhibitors(mTORi) improves renal function in diabetic and/or hypertensive liver transplant patients immunosuppressed with tacrolimus or cyclosporine.METHODS: The study included 86 liver graft recipients immunosuppressed with mTORi treatment after orthotopic liver transplantation(OLT), including all liver recipients with worsening renal function before conversion to mTORi(n = 55 patients) and recipients with normal renal function who converted to m TORi for other reasons(n = 31 patients). We identified patients with diabetes mellitus(n = 28), arterial hypertension(n = 27), proteinuria(n = 27) and all three factors(n = 8)(some patients have hypertension and diabetes and no proteinuria). The primary endpoint was evolution in renal function defined as the development in plasma creatinine as a function of diabetes mellitus(DM), hypertension(HT) or proteinuria. We required elevated serum creatinine for at least two weeks to define renal dysfunction.RESULTS: Only patients that converted because of renal failure with plasma creatinine levels > 1.5 mg/dL showed an improvement of renal function(2.14 to 1.77 mg/dL)(P = 0.02). Patients with DM showed no improvement of serum creatinine levels(1.31 mg/dL to 1.37 mg/dL) compared with non DM patients(1.31 mg/dL to 1.15 mg/dL)(P = 0.01), HT patients(1.48 mg/dL to 1.5 mg/dL) with non HT patients(1.21mg/d L to 1.08 mg/dL) and patients with proteinuria(1.44 mg/dL to 1.41 mg/dL) and no proteinuria(1.31 mg/dL to 1.11 mg/dL). CONCLUSION: In OLT recipients with diabetes or hypertensive nephropathy, conversion to m TORi does not improve renal function but stabilizes plasma levels of creatinine. Proteinuria is not a contraindication to conversion to m TORi; it also stabilizes renal function. Conversion to m TORi should only be avoided in patients with diabetes, hypertension and proteinuria.

关键词： Mammalian target of rapamycin inhibitors Liver transplant Renal dysfunction Hypertension Diabetes

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Improving outcomes in pancreatic cancer: Key points in perioperative management

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World Journal of Gastroenterology 2014年第39期20卷 14237-14245页

作者： José M álamo Luis M Marín Gonzalo Suarez Carmen Bernal Juan Serrano Lydia Barrera Miguel A Gómez jordi muntané Francisco J Padillo Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBEREH o Ciberehd) Instituto de Salud Carlos Ⅲ 28029 Madrid Spain Francisco J Padillo Department of General Surgery Hospital Universitario Virgen del Rocío/CSIC/Universidad de Sevilla 41013 Seville Spain

This review focused in the perioperative management of patients with pancreatic cancer in order to improve the outcome of the disease. We consider that the most controversial points in pancreatic cancer management are jaundice management, vascular resection and neoadjuvant therapy. Preoperative biliary drainage is recommended only in patients with severe jaundice, as it can lead to infectious cholangitis, pancreatitis and delay in resection, which can lead to tumor progression. The development of a phase Ⅲ clinical trial is mandatory to clarify the role of neo-adjuvant radiochemotherapy in pancreatic adenocarcinoma. Venous resection does not adversely affect postoperative mortality and morbidity, therefore, the need for venous resection should not be a contraindication to surgical resection in selected patients. The data on arterial resection alone, or combined with vascular resection at the time of pancreatectomy are more heterogeneous, thus, patient age and comorbidity should be evaluated before a decision on operability is made. In patients undergoing R0 resection, arterial resection can also be performed.

关键词： Pancreatic cancer Obstructive jaundice Preoperativ

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