Oncohematología y Genética

Biología molecular del cáncer

Amancio Carnero Moya
IBiS
Campus Hospital Universitario Virgen del Rocío
Avda. Manuel Siurot, s/n.
41013 · Sevilla

Laboratorio: 214

Amancio Carnero Moya

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  1. Miembros del grupo
  2. Áreas de trabajo
  3. Publicaciones

Grupo: Biología molecular del cáncer

Directorio
Miembros del grupo Biología molecular del cáncer
  • Carnero Moya, Amancio.Investigador Cientifico CSIC
  • Chaves Conde, Manuel.Licenciado en Medicina. Facultativo clínico
  • Estévez García, Purificación.Oncólogo Clínico
  • Felipe Abrio, Blanca.Estudiante de doctorado
  • Fuentes Pradera, José.FEA HUV Valme
  • García Heredia, José Manuel.Profesor ayudante doctor
  • Jiménez García, Manuel Pedro .Investigador en Formación
  • Lucena Cacace, Antonio.Estudiante Doctorado
  • Marín López, Juan José.Doctor en Biología. Profesor Asociado.
  • Muñoz Galván, Sandra.Investigadora Postdoctoral.
  • Navas Maldonado, Lola Eufemia.Investigadora en Formación
  • Otero Albiol, Daniel.Estudiante Predoctoral. Ldo. en Biología
  • Peinado Serrano, Javier.Ldo. en Medicina
  • Perez, Marco.BIR.
  • Suárez Martínez, Elisa.Lda. en Biomedicina. Predoctoral.
  • Verdugo Sivianes, Eva Mª.Investigadora en Formación

Áreas de trabajo

 

 LOGO FEDER

 

 

TÍTULO DEL PROYECTO: Identificación y caracterización de factores genéticos y fisiológicos involucrados en senescencia celular. Relevancia clínica en cáncer.

 

RESUMEN: (Objetivo, diseño, ámbito del estudio, sujetos de estudio, instrumentación, resultados, conclusiones)

La senescencia celular es un mecanismo supresor de tumores que debe ser anulado durante el proceso de tumorigénesis. Todos los tumores llevan alteraciones que contribuyen a evitar la senescencia. Entender el proceso de senescencia es importante no sólo para descubrir la fisiología de las células somáticas, y su contraparte normal inmortal, las células madre, sino más relevante para ser capaz de identificar nuevas alteraciones genéticas y epigenéticas que proporcionan nuevos genes supresores de tumores u oncogenes que pueden actuar como biomarcadores de diferenciación entre tumores benignos y malignos, nuevos biomarcadores con valor pronóstico o predictivo. Para generar nuevos métodos para identificar tumores en etapas tempranas, para identificar nuevas dianas moleculares y fisiológicas adecuadas para la intervención terapéutica y el control de la enfermedad. Por lo tanto en nuestro proyecto planeamos un estudio en profundidad de los nuevos procesos fisiológicos implicados en la senescencia celular a través de los nuevos genes descubiertos en nuestro laboratorio. Estos nuevos genes pertenecen a familias funcionales implicadas en transportadores y canales iónicos de membrana, sistemas de degradación de proteínas y autofagia, oxidación e hipoxia, metabolismo y long non coding RNAs. Tenemos la intención de estudiar estas nuevas alteraciones genéticas, su papel en la inmortalización y la tumorigénesis, su papel como biomarcadores pronósticos o predictivos y su viabilidad para ser utilizado como nueva diana para el tratamiento contra el cáncer. También planeamos estudiar la pertinencia de la senescencia celular en la carcinogénesis medioambiental ycómo podemos revertir el fenotipo utilizando la senescencia como punto final para la terapia antitumoral.

 

ÁREAS DE TRABAJO GENERALES 

 

Nuestro laboratorio ha centrado sus esfuerzos en 2 vías paralelas. Por un lado la medicina de precisión, con especial hincapié en biomarcadores predictivos de eficacia y biomarcadores pronóstico en distintos modelos tumorales. Por otro lado hemos seguido desarrollando nuestra búsqueda y caracterización de nuevas alteraciones genéticas y epigenéticas que tengan relevancia en cáncer. En estos sentidos hemos avanzado en las siguientes areas.

  • NAMPT Is a Potent Oncogene in Colon Cancer Progression that Modulates Cancer Stem Cell Properties and Resistance to Therapy through Sirt1 and PARP.

Purpose: Colorectal cancer is the second most common cancer in women and the third most common in men worldwide. However, despite current progress, many patients with advanced and metastatic tumors still die from the malignancy. Refractory disease often relies on nicotinamide adenine dinucleotide (NAD)-dependent mechanisms. NAD metabolism and a stable NAD regeneration circuit are required to maintain tissue homeostasis and metabolism. However, high levels of NAD confer therapy resistance to tumors.Experimental Design: Ectopic overexpression of nicotinamide phosphoribosil transferase (NAMPT) and shRNAs in colorectal cancer cell lines, tumorigenic and stemness properties and transcription measurement in culture and in vivo Transcriptional analysis in public databases. Therapeutic approaches.Results: NAMPT, the rate-limiting enzyme responsible for the highest source of physiologic NAD biosynthesis, increases tumorigenic properties and induces cancer stem cell-like properties through pathways that control stem cell signaling, thus enriching the cancer-initiating cell (CIC) population. Furthermore, NAMPT expression correlated with high levels of CIC-like cells in colon tumors directly extracted from patients, and transcription meta-analysis revealed that NAMPT is also a key factor that induces cancer stem pathways in colorectal cancer tumors. This effect is mediated by PARP and SIRT1. In addition, we report a novel NAMPT-driven signature that stratifies prognosis from high to low expression groups. The NAMPT signature contained SIRT1 and PARP1 levels as well as other cancer stem cell-related genes. Finally, NAMPT inhibition increased the sensitivity to apoptosis in both NAMPT-expressing cells and tumorspheres.Conclusions: NAMPT represents a novel therapeutic target in colon cancer progression and relapse, particularly the CIC subset of human colon cancers.

  • NAMPT overexpression induces cancer stemness and defines a novel tumor signature for glioma prognosis.

Gliomas are the most prevalent primary malignant brain tumors associated with poor prognosis. NAMPT, a rate-limiting enzyme that boosts the nicotinamide adenine dinucleotide (NAD) regeneration in the salvage pathway, is commonly expressed in these tumors. NAD metabolism is required to maintain tissue homeostasis. To maintain metabolism, cancer cells require a stable NAD regeneration circuit. However, high levels of NAD confer resistance to therapy to these tumors, usually treated with Temozolomide (TMZ). We report that NAMPT overexpression in glioma cell lines increases tumorigenic properties controlling stem cell pathways and enriching the cancer-initiating cell (CIC) population. Furthermore, NAMPT expression correlated with high levels of Nanog, CD133 and CIC-like cells in glioblastoma directly extracted from patients. Meta-analysis reveals that NAMPT is also a key factor inducing cancer stem pathways in glioma cells. Furthermore, we report a novel NAMPT-driven signature which stratify prognosis within tumor staging. NAMPT signature also correlates directly with EGFR positive and IDH negative tumors. Finally, NAMPT inhibition increases sensitivity to apoptosis in both NAMPT-expressing cells and tumorspheres. Therefore, NAMPT represents a novel therapeutic target in Glioma progression and relapse.

  • The Cargo Protein MAP17 (PDZK1IP1) Regulates the Cancer Stem Cell Pool Activating the Notch Pathway by Abducting NUMB.

Purpose: Cancer stem cells (CSC) are self-renewing tumor cells, with the ability to generate diverse differentiated tumor cell subpopulations. They differ from normal stem cells in the deregulation of the mechanisms that normally control stem cell physiology. CSCs are the origin of metastasis and highly resistant to therapy. Therefore, the understanding of the CSC origin and deregulated pathways is important for tumor control.Experimental Design: We have included experiments in vitro, in cell lines and tumors of different origins. We have used patient-derived xenografts (PDX) and public transcriptomic databases of human tumors.Results: MAP17 (PDZKIP1), a small cargo protein overexpressed in tumors, interacts with NUMB through the PDZ-binding domain activating the Notch pathway, leading to an increase in stem cell factors and cancer-initiating-like cells. Identical behavior was mimicked by inhibiting NUMB. Conversely, MAP17 downregulation in a tumor cell line constitutively expressing this gene led to Notch pathway inactivation and a marked reduction of stemness. In PDX models, MAP17 levels directly correlated with tumorsphere formation capability. Finally, in human colon, breast, or lung there is a strong correlation of MAP17 expression with a signature of Notch and stem cell genes.Conclusions: MAP17 overexpression activates Notch pathway by sequestering NUMB. High levels of MAP17 correlated with tumorsphere formation and Notch and Stem gene transcription. Its direct modification causes direct alteration of tumorsphere number and Notch and Stem pathway transcription. This defines a new mechanism of Notch pathway activation and Stem cell pool increase that may be active in a large percentage of tumors.

 

 

  • The cargo protein MAP17 (PDZK1IP1) regulates the immune microenvironment.

Inflammation is a complex defensive response activated after various harmful stimuli allowing the clearance of damaged cells and initiating healing and regenerative processes. Chronic, or pathological, inflammation is also one of the causes of neoplastic transformation and cancer development. MAP17 is a cargo protein that transports membrane proteins from the endoplasmic reticulum. Therefore, its overexpression may be linked to an excess of membrane proteins that may be recognized as an unwanted signal, triggering local inflammation. Therefore, we analyzed whether its overexpression is related to an inflammatory phenotype. In this work, we found a correlation between MAP17 expression and inflammatory phenotype in tumors and in other inflammatory diseases such as Crohn's disease, Barrett's esophagus, COPD or psoriasis. MAP17 expression correlated also with the markers of inflammation HLAs, BBS10, HERC2, ADNP and PYCARD. Furthermore, we found that MAP17 expression directly regulates NFAT2 and IL-6 activation, inducing the differentiation of monocytes to dendritic cells and suggesting a causal role of MAP17 in inflammation. Immunohistochemistry confirms local inflammation, mainly CD45+ cells, at the site of expression of MAP17, at least in tumors, Crohn's and psoriasis. Therefore, our data indicates that the overexpression of the protein MAP17 plays important role in diseases involving chronic inflammation.

  • Inflammation and stem markers association to PIM1/PIM2 kinase-induced tumors in breast and uterus.

The PIM family of Ser/Thr kinase proteins has been implicated in tumorigenesis at different levels. PIM proteins are overexpressed in several tumor types and have been associated with chemoresistance. However, their role in hormone-dependent female tissues has not been explored, especially in the uterus, breast and ovary. We generated conditional transgenic mice with confined expression of human PIM1 or PIM2 genes in these tissues. We characterized the tumoral response to these genetic alterations corroborating their role as oncogenes since they induce hyperproliferation in all tissues and tumors in mammary gland and uterus. Furthermore, we observed a high degree of inflammatory infiltration in these tissues of transgenic mice accompanied by NFAT and mTOR activation and IL6 expression. Moreover, PIM1/2 were overexpressed in human breast, uterine and ovarian tumors, correlating with inflammatory features and stem cell markers. Our data suggest that PIM1/2 kinase overexpression provoke tissue alterations and a large IL6-dependent inflammatory response that may act synergistically during the process of tumorigenesis. The possible end-point is an increased percentage of cancer stem cells, which may be partly responsible for the therapy resistance found in tumors overexpressing PIM kinases.

  • Genome-Wide miRNA Screening for Genes Bypassing Oncogene-Induced Senescence.

MicroRNAs are small noncoding RNAs that regulate gene expression by binding to sequences within the 3'-UTR of mRNAs. Genome-wide screens have proven powerful in associating genes with certain phenotypes or signal transduction pathways and thus are valuable tools to define gene function. Here we describe a genome-wide miRNA screening strategy to identify miRNAs that are required to bypass oncogene-induced senescence.

  • Coordinated downregulation of Spinophilin and the catalytic subunits of PP1, PPP1CA/B/C, contributes to a worse prognosis in lung cancer.

The scaffold protein Spinophilin (Spinophilin, PPP1R9B) is one of the regulatory subunits of phosphatase-1 (PP1), directing it to distinct subcellular locations and targets. The loss of Spinophilin reduces PP1 targeting to pRb, thereby maintaining higher levels of phosphorylated pRb. Spinophilin is absent or reduced in approximately 40% of human lung tumors, correlating with the malignant grade. However, little is known about the relevance of the coordinated activity or presence of Spinophilin and its reported catalytic partners in the prognosis of lung cancer. In the present work, we show that the downregulation of Spinophilin, either by protein or mRNA, is related to a worse prognosis in lung tumors. This effect is more relevant in squamous cell carcinoma, SCC, than in adenocarcinoma. Downregulation of Spinophilin is related to a decrease in the levels of its partners PPP1CA/B/C, the catalytic subunits of PP1. A decrease in these subunits is also related to prognosis in SCC and, in combination with a decrease in Spinophilin, are markers of a poor prognosis in these tumors. The analysis of the genes that correlate to Spinophilin in lung tumors showed clear enrichment in ATP biosynthesis and protein degradation GO pathways. The analysis of the response to several common and pathway-related drugs indicates a direct correlation between the Spinophilin/PPP1Cs ratio and the response to oxaliplatin and bortezomib. This finding indicates that this ratio may be a good predictive biomarker for the activity of the drugs in these tumors with a poor prognosis.

  • Histology-dependent prognostic role of pERK and p53 protein levels in early-stage non-small cell lung cancer.

Lung tumors represent a major health problem. In early stage NSCLC tumors, surgical resection is the preferred treatment, but 30-55% of patients will relapse within 5 years after surgery. Thus, the identification of prognostic biomarkers in early stage NSCLC patients, especially those which are therapeutically addressable, is crucial to enhance survival of these patients. We determined the immunohistochemistry expression of key proteins involved in tumorigenesis and oncogenic signaling, p53, EGFR, pAKT and pERK, and correlated their expression level to clinicopathological characteristics and patient outcome. We found EGFR expression is higher in the squamous cell carcinomas than in adenocarcinomas (p=0.043), and that nuclear p53 staining correlated with lower differentiated squamous tumors (p=0.034). Regarding the prognostic potential of the expression of these proteins, high pERK levels proved to be an independent prognostic factor for overall (p<0.001) and progression-free survival (p<0.001) in adenocarcinoma patients, but not in those from the squamous histology, and high p53 nuclear levels were identified as independent prognostic factor for progression-free survival (p=0.031) only in squamous cell carcinoma patients. We propose a role as early prognostic biomarkers for pERK protein levels in adenocarcinoma, and for nuclear p53 levels in squamous cell lung carcinoma. The determination of these potential biomarkers in the adequate histologic context may predict the outcome of early stage NSCLC patients, and may offer a therapeutic opportunity to enhance survival of these patients.

Revistas Internacionales
Garcia-Carbonero R, Carnero A, Paz-Ares L.
Inhibition of HSP90 molecular chaperones: moving into the clinic.
Lancet Oncol. 2013 Aug;14(9):e358-69. doi: 10.1016/S1470-2045(13)70169-4. Review.
Estevez-Garcia P, Lopez-Calderero I, Molina-Pinelo S, Muñoz-Galvan S, Salinas A, Gomez-Izquierdo L, Lucena-Cacace A, Felipe-Abrio B, Paz-Ares L, Garcia-Carbonero R, Carnero A.
Spinophilin loss correlates with poor patient prognosis in advanced stages of colon carcinoma.
Clin Cancer Res. 2013 Jul 15;19(14):3925-35. doi: 10.1158/1078-0432.CCR-13-0057. Epub 2013 May 31.
Parro T, Medrano MA, Cubo L, Muñoz-Galván S, Carnero A, Navarro-Ranninger C, Quiroga AG.
The second generation of iodido complexes: trans-[PtI2(amine)(amine')] bearing different aliphatic amines.
. J Inorg Biochem. 2013 Oct;127:182-7. doi:10.1016/j.jinorgbio.2013.04.010. Epub 2013 Apr 28
Rodríguez-Rodero S, Fernández AF, Fernández-Morera JL, Castro-Santos P, Bayon GF, Ferrero C, Urdinguio RG, Gonzalez-Marquez R, Suarez C, Fernández-Vega I, Fresno Forcelledo MF, Martínez-Camblor P, Mancikova V, Castelblanco E, Perez M, Marrón PI, Mendiola M, Hardisson D, Santisteban P, Riesco-Eizaguirre G,Matías-Guiu X, Carnero A, Robledo M, Delgado-Álvarez E, Menéndez-Torre E, Fraga MF.
DNA methylation signatures identify biologically distinct thyroid cancer subtypes.
J Clin Endocrinol Metab. 2013 Jul;98(7):2811-21. doi: 10.1210/jc.2012-3566. Epub 2013 May 10.
Pastor MD, Nogal A, Molina-Pinelo S, Meléndez R, Salinas A, González De la Peña M, Martín-Juan J, Corral J, García-Carbonero R, Carnero A, Paz-Ares L.
Identification of proteomic signatures associated with lung cancer and COPD.
. J Proteomics. 2013 Aug 26;89:227-37. doi: 10.1016/j.jprot.2013.04.037.
Pastor MD, Nogal A, Molina-Pinelo S, Carnero A, Paz-Ares L.
Proteomic biomarkers in lung cancer.
Clin Transl Oncol. 2013 Sep;15(9):671-82. doi: 10.1007/s12094-013-1034-0. Epub 2013 Apr 20.
Medrano MÁ, Álvarez-Valdés A, Perles J, Lloret-Fillol J, Muñoz-Galván S, Carnero A, Navarro-Ranninger C, Quiroga AG.
Oxidation of anticancer Pt(II) complexes with monodentate phosphane ligands: towards stable but active Pt(IV) prodrugs.
Chem Commun (Camb). 2013 May 25;49(42):4806-8. doi: 10.1039/c3cc38416k.
Narlik-Grassow M, Blanco-Aparicio C, Carnero A.
The PIM Family of Serine/Threonine Kinases in Cancer.
Med Res Rev. 2014 Jan;34(1):136-59. doi: 10.1002/med.21284. Epub 2013 Apr 10.
Narlik-Grassow M, Blanco-Aparicio C, Cecilia Y, Perez M, Muñoz-Galvan S, Cañamero M, Renner O, Carnero A.
Conditional transgenic expression of PIM1 kinase in prostate induces inflammation-dependent neoplasia.
PLoS One. 2013;8(4):e60277. doi:0.1371/journal. 2013 Apr 2.
Perez M, Praena-Fernandez JM, Felipe-Abrio B, Lopez-Garcia MA, Lucena-Cacace A, Garcia A, Lleonart M, Roncador G, Marin JJ, Carnero A.
MAP17 and SGLT1 protein expression levels as prognostic markers for cervical tumor patient survival
PLoS One. 2013;8(2):e56169. doi: 10.1371/journal.pone.0056169. Epub 2013 Feb 13.
Pastor MD, Nogal A, Molina-Pinelo S, Meléndez R, Romero-Romero B, Mediano MD, López-Campos JL, García-Carbonero R, Sanchez-Gastaldo A, Carnero A, Paz-Ares L.
Identification of oxidative stress related proteins as biomarkers for lung cancer and chronic obstructive pulmonary disease in bronchoalveolar lavage.
Int J Mol Sci. 2013 Feb 6;14(2):3440-55. doi: 10.3390/ijms14023440
Carnero A.
Markers of cellular senescence.
Methods Mol Biol. 2013;965:63-81. doi: 10.1007/978-1-62703-239-1_4. Review.