Oncohematology and Genetics

Molecular Pathology of Sarcomas and other Tumors

Enrique  de Álava Casado
Enrique de Álava Casado
IBiS
Campus Hospital Universitario Virgen del Rocío
Avda. Manuel Siurot, s/n.
41013 · Sevilla

Laboratory: 203

Enrique  de Álava Casado

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  1. Group members
  2. Research areas
  3. Publications

Group: Molecular Pathology of Sarcomas and other Tumors

Directory
Group members  Molecular Pathology of Sarcomas and other Tumors
  • Aguado Domínguez, Elena.Investigadora Postdoctoral
  • Biscuola, Michele.Ldo. En Biología
  • Carmona Berraquero, Fernando.Licenciando en Ciencias Ambientales
  • Castilla Ramírez, Carolina.Doctora en Biología
  • de Álava Casado, Enrique .Investigador Responsable
  • Díaz Martín, Juan.Doctor en Biología
  • García Domínguez, Daniel José.Investigador Posdoctoral
  • Hernández Barrera, Mª José .Técnico Sup. Lab. Anatomía Patológica (FP2)
  • Hontecillas Prieto, Lourdes .Investigadora Posdoctoral
  • Jordán Pérez, Mª del Carmen.Técnico Anatomía Patológica
  • Monteiro Amaral, Ana Teresa.Investigadora Posdoctoral
  • Puerto Camacho, Pilar.Investigadora Predoctoral
  • Robles Frías, Mª José .Doctora en Medicina.
  • Rodríguez Núñez , Pablo.Investigador Predoctoral
  • Salguero Aranda, Carmen.Dra. Biotecnología.

Research areas

Ewing Sarcoma (ES) is a malignant bone and soft-tissue tumor mainly affecting children and young adults. Advances in terms of  multimodal therapies in patients presenting localized disease, lead to a surprising overall survival of 70% at 5-year diagnostic. Despite this, patients presenting multifocal/metastatic or refractory disease present disappointing overall survival rates of around 20% .

From a molecular point of view, ES is characterized by the presence of reciprocal chromosomal translocations, given rise to fusion genes which in turn are translated into chimerical proteins (mostly involving EWSR1-ETS members). These functional chimerical proteins represent the major oncogenic event in ES. They typically involve the EWSR1 gene and a member of the ETS family, mostly FLI1, in about 85% of the cases. This aberrant transcription factor deregulates the transcription of target genes such as IGF1/IGF1R related genes, among many others.

 Despite the evident role of the ES fusion protein, recent evidence has demonstrated that other secondary alterations also play an enormous part in terms of tumor development, progression and even clinical prognostic.

The main research line of our group is based on the definition and characterization of these secondary alterations and how they may affect ES biology. Our goal is to establish new therapeutic targets as well as biomarkers of response to treatment based on preclinical studies focused on alternative/novel therapies.

 Detailed Objectives

 Discovery and validation of molecular mechanisms of tumor initiation and progression in sarcomas, namely in Ewing Sarcoma, our model of study.

 Translate these molecular mechanisms to the clinic in the form of biomarkers either of response to treatment or therapeutic monitoring.

 Establish stable collaborations with academic/ pharmaceutical groups interested in our research

 Research Lines:

 1.     Discovery and validation of molecular mechanisms of tumor initiation and progression in Ewing Sarcoma.

 The mechanisms of tumor initiation of Ewing Sarcoma remain unclear until this day. Despite this, the mesenchymal stem cell has been postulated as the most probable cell of origin of this tumor entity. Our group, developed an Ewing sarcoma cell line where the EWSR1-FLI1 stable interference led to a decrease of the intrinsic tumor characteristics followed by an increase of mesenchymal stem cells characteristics, namely differentiation capacity and phenotype. This study revealed a new target of the chimerical protein, TOPK1. Also, the in vivo implantation of the interfered tumor cells resulted in the production of smaller tumors in comparison to the parental cell line. (Herrero-Martín D et al, Br J Cancer. 2009 Jul 7;101(1):80-90.)

More recently, in collaboration with other European groups and within the Europoan Project EuroBONET, we have studied the phenotype of Mesenchymal stem cells derived from Ewing Sarcoma patients in comparison to Ewing Sarcoma cells and Ewing Sarcoma samples. We observed that Ewing Sarcoma patients are similar to other mesenchymal stem cells and show no differential phenotype. However, Ewing sarcoma cells present some phenotypic features of mesenchymal stem cells.

Also, the detailed study of CD99 expression in a large set of mesenchymal stem cells revealed that the inhibition of this receptor showed no impairment on this cell type survival. (Amaral AT et al, PLoS One, 2014, )

2. Discovery, functional validation and further clinical translation of new therapeutic targets from genomic, integrative proteomic studies in Ewing Sarcoma.

 Pre-clinical in vitro and in vivo studies described the protein HSP90 (Heat Shock Protein 90) as a biomarker of response to treatment with anti-IGF1R (Insulin-like growth factor receptor 1) inhibitors (namely ADW). Integrative proteomic analysis revealed that ES cell lines resistant to treatment with ADW, expressed higher levels of HSP90. This fact was later validated by RNA interference and in vivo studies in xenograft models. Also, Histopathological analysis of Ewing sarcoma samples demonstrated that in fact, in patients, the differential expression of HSP90 was a marker of worst response to treatment. (Martins AS et al, Cancer Res. 2008 Aug 1;68(15):6260-70.)

[T1] 

More recently, in a multicentre retrospective study, we demonstrated through CGH arrays the prognostic value of a particular secondary alteration. The gain of chromosome long arm (1qG) was present in approximately 30% of a total of 67 tumor samples. Furthermore, we performed a transcriptomic study with 38 tumor samples and finally established the 1qG fingerprint. This fingerprint was characterized by the differential expression of 74 genes located on 1q. Cases presenting this 1qG fingerprint were correlated with cell cycle deregulation, relapse and worst survival. Some genes with described over expression resulting from this 1qG in these cases were CDT2 and PARP1. (Mackintosh C et al, Oncogene. 2012 Mar 8;31(10):1287-98)

 Design and development of diagnostic tools which will allow the selection of patients for specific treatments, with particular emphasis to FISH and molecular signatures related with the over expression of 1q located genes.

 Retrospective Studies showed the clinical relevance of secondary alterations in Ewing Sarcoma, namely, alterations of copy number (CNAs), 1q and 8q Gains, 16q losses, among others. The next objective is to establish in prospective studies the prognostic value of CNAs in Ewing Sarcoma patients, through the development of FISH probes (for the evaluation of 1qG and 16q losses) and genes such as CDT2 and PARP1.

 Preclinical evaluation of experimental/commercial drugs on Ewing Sarcoma cell lines (previously validated in the European Project EuroBonet) and in xenograft models.

 a)     Taking together that CDT2 belongs to a subtype of complexes with  ubiquitin ligase activity (known as Cullin-RING ligases, CRL) and that these complexes con be inhibited with a potent specific new drug (MLN4924) we decided to study the effectiveness of MLN4924 in Ewing Sarcoma cell lines. Our results showed a high sensitivity of Ewing Sarcoma cell lines, with a dose dependent mechanism of response. At concentrations lower than the IC50 of proliferation (concentration needed to achieve the death of 50% of the cell population), we observed moderate apoptosis, with a delay in the G2-phase mostly due to the accumulation of WEE1. On the other hand, at concentrations higher than the IC50 of proliferation we observed a delay in the S-phase and high apoptotic index. More importantly, an efficient antitumoral effect was observed in our in vivo model. (Mackintosh C et al, Oncogene. 2013 Mar 14; 32(11):1441-51.)

 b)    PARP proteins are crucial for cell survival and are involved in several mechanisms such as DNA damage repair, cell cycle, apoptosis induction, angiogenesis and tumor formation and tumor progression. This fact evidently renders PARP proteins as interesting therapeutic targets. Although studies in other solid tumors suggested that PARP inhibitors were mostly active in tumors with deficiencies/mutations in DNA repair mechanisms, and that in Ewing Sarcoma these have not been described, the use of PARP inhibitors in combination with other chemotherapeutic agents (Temozolamide and radiotherapy) has shown great results in recent in vitro / in vivo studies. In our lab, we are focused in studying possible drug combinations of PARP inhibitors and other DNA-related agents in in vitro and in vivo models in Ewing Sarcoma.

 c)     Another key mechanism related to malignancy is the over expression of the IGF1-IGF1R axis in Ewing Sarcoma. Initially, we studied the effects of small molecules (ADW), which by competition of IGF1 exert an antagonistic action, blocking IGF1R activation. This tyrosine kynase inhibitor is an important regulator of cell proliferation in Ewing Sarcoma. ADW demonstrated an efficient anti-tumoral activity in comparison to traditional chemotherapeutic agents such as Vincristine or Doxorubicin. Interestingly, resistance to anti-IGF1R therapies has been related to the IGF2/IR dependency as well as with HSP90 over expression. So, more recently, and within the European project EuroSARC, we have studied the effects of a dual inhibitor of IGF1R/IR in Ewing Sarcoma cell lines, Linsitinb. In fact, we combined Linsitinib with DNA damage inducing agent, Trabectedin, and observed high levels of synergism.

 Development of epigenetic studies based in the activity of the chimerical protein EWSR1/FLI1 in models of inducible ectopic expression. These studies include chromatin remodeling, non-coding RNAs and pre-clinical in vitro and in vivo studies with epigenetic-based drugs.

 Secondary genetic alterations and epigenetic alterations might help to explain the most aggressive forms of Ewing Sarcoma and reveal new promising therapeutic targets. In this study we propose the stratification of Ewing sarcoma patients according to their epigenetic profile. We are currently developing and validating diagnostic tools which will allow us to molecularly characterize Ewing Sarcoma patients through techniques which can be easily translated to the clinical reality. Finally, we plan to further study the targets found through in vivo and in vitro studies and according to their effectiveness applied them to future clinical trials in Ewing Sarcoma patients.

 

 

 

 

 


 [T1]En la versión en castellano, falta referencia a este trabajo.

International Journals
Martin-Broto J, Stacchiotti S, Lopez-Pousa A, Redondo A, Bernabeu D, de Alava E, Casali PG, Italiano A, Gutierrez A, Moura DS, Peña-Chilet M, Diaz-Martin J, Biscuola M, Taron M, Collini P, Ranchere-Vince D, Garcia Del Muro X, Grignani G, Dumont S, Martinez-Trufero J, Palmerini E, Hindi N, Sebio A, Dopazo J, Dei Tos AP, LeCesne A, Blay JY, Cruz J.
Pazopanib for treatment of advanced malignant and dedifferentiated solitary fibrous tumour: a multicentre, single-arm, phase 2 trial.
Lancet Oncol
María del Valle Enguix-Riego, Jon Cacicedo Fernández de Bobadilla, Blas David Delgado León, Jose María Nieto-Guerrero Gómez, Daniel Herrero Rivera, Marco Perez, Juan Manuel Praena-Fernández, Gerardo Sanchez Carmona, Eleonor Rivin del Campo, María José Ortiz Gordillo, Jose Luis Lopez Guerra.
The single nucleotide variant rs2868371 associates with the risk of mortality in non-small cell lung cancer patients: a multicenter prospective validation.
Radiotherapy & Oncology
Sandra Muñoz-Galván, Antonio Lucena-Cacace, Marco Perez, Daniel Otero-Albiol, Julian Gomez-Cambronero and Amancio Carnero.
Tumor cell-secreted PLD increases tumor stemness by senescence-mediated communication with microenvironment.
Oncogene
Irene Ferrer; Alvaro Quintanal-Villalonga, Sonia Molina-Pinelo, Jose Manuel Garcia- Heredia, Marco Perez, Rocío Suárez, Santiago Ponce-Aix, Luis Paz-Ares, Amancio Carnero.
Lung cancer treatments efficacy in tumors with high MAP17 (PDZK1IP1) expression levels.
J Exp Clin Cancer Res
Díaz-Martín J, Biscuola M, Benoit J, Marcilla D, Civantos G, de Álava E.
What's in a name? Molecular subclassification of sarcomas creates fresh challenges.
J Pathol
Sáez-Freire MDM, Blanco-Gómez A, Castillo-Lluva S, Gómez-Vecino A, Galvis-Jiménez JM, Martín-Seisdedos C, Isidoro-García M, Hontecillas-Prieto L, García-Cenador MB, García-Criado FJ, Patino-Alonso MC, Galindo-Villardón P, Mao JH, Prieto C, Castellanos-Martín A, Kaderali L, Pérez-Losada J.
Supplementary data for the biological age linked to oxidative stress modifies breast cancer aggressiveness.
Data Brief
Puerto-Camacho P, Amaral AT, Lamhamedi-Cherradi SE, Menegaz BA, Castillo-Ecija H, Ordóñez JL, Domínguez S, Jordan-Perez C, Diaz-Martin J, Romero-Pérez L, Lopez-Alvarez M, Civantos-Jubera G, Robles-Frías MJ, Biscuola M, Ferrer C, Mora J, Cuglievan B, Schadler K, Seifert O, Kontermann R, Pfizenmaier K, Simón L, Fabre M, Carcaboso ÁM, Ludwig JA, de Álava E.
Preclinical Efficacy of Endoglin-Targeting Antibody-Drug Conjugates for the Treatment of Ewing Sarcoma.
Clin Cancer Res
García-Domínguez DJ, Hontecillas-Prieto L, Rodríguez-Núñez P, Pascual-Pasto G, Vila-Ubach M, García-Mejías R, Robles MJ, Tirado OM, Mora J, Carcaboso AM, de Álava E.
The combination of epigenetic drugs SAHA and HCI-2509 synergistically inhibits EWS-FLI1 and tumor growth in Ewing sarcoma
Oncotarget
Grünewald TGP, Cidre-Aranaz F, Surdez D, Tomazou EM, de Álava E, Kovar H, Sorensen PH, Delattre O, Dirksen U.
Ewing sarcoma.
Nat Rev Dis Primers
Sáez-Freire MDM, Blanco-Gómez A, Castillo-Lluva S, Gómez-Vecino A, Galvis-Jiménez JM, Martín-Seisdedos C, Isidoro-García M, Hontecillas-Prieto L, García-Cenador MB, García-Criado FJ, Patino-Alonso MC, Galindo-Villardón P, Mao JH, Prieto C, Castellanos-Martín A, Kaderali L, Pérez-Losada J.
The biological age linked to oxidative stress modifies breast cancer aggressiveness
Free Radic Biol Med
Hajji N, García-Domínguez DJ, Hontecillas-Prieto L, O'Neill K, de Álava E, Syed N.
The bitter side of epigenetics: variability and resistance to chemotherapy.
Epigenomics.
Baldauf MC, Gerke JS, Orth MF, Dallmayer M, Baumhoer D, de Alava E, Hartmann W, Kirchner T, Grünewald TGP.
Are EWSR1-NFATc2-positive sarcomas really Ewing sarcomas?
Mod Pathol
Garcia-Monclús S, López-Alemany R, Almacellas-Rabaiget O, Herrero-Martín D, Huertas-Martinez J, Lagares-Tena L, Alba-Pavón P, Hontecillas-Prieto L, Mora J, de Álava E, Rello-Varona S, Giangrande PH, Tirado OM.
EphA2 receptor is a key player in the metastatic onset of Ewing sarcoma
Int J Cancer
Hontecillas-Prieto L, García-Domínguez DJ, García-Mejías R, Ramírez-Villar GL, Sáez C, de Álava E.
HMGA2 overexpression predicts relapse susceptibility of blastemal Wilms tumor patients.
Oncotarget
de Alava E.
Ewing Sarcoma, an Update on Molecular Pathology with Therapeutic Implications
Surg Pathol Clin
Jose Garcia-Heredia, Antonio Lucena-Cacace, Eva Verdugo-Sivianes, Marco Perez, and Amancio Carnero.
The cargo protein MAP17 (PDZK1IP1) regulates the cancer stem cell pool activating the Notch pathway by abducting NUMB.
Clinical Cancer Research
Ordóñez JL, Amaral AT, Carcaboso AM, Herrero-Martín D, García-Macías MDC, Sevillano V, Alonso D, Pascual-Pasto G, San-Segundo L, Vila-Ubach M, Rodrigues T, Fraile S, Teodosio C, Mayo-Iscar A, Aracil M, Galmarini CM, Tirado OM, Mora J, de Álava E.
Correction: The PARP inhibitor olaparib enhances the sensitivity of Ewing sarcoma to trabectedin.
Oncotarget
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DNA methylation heterogeneity defines a disease spectrum in Ewing sarcoma.
Nat Med
Hontecillas-Prieto L, Garcia-Dominguez DJ, Vaca DP, Garcia-Mejias R, Marcilla D, Ramirez-Villar GL, Saez C, de Álava E.
Multidrug resistance transporter profile reveals MDR3 as a marker for stratification of blastemal Wilms tumour patients.
Oncotarget
Huertas-Martínez J, Court F, Rello-Varona S, Herrero-Martín D, Almacellas-Rabaiget O, Sáinz-Jaspeado M, Garcia-Monclús S, Lagares-Tena L, Buj R, Hontecillas-Prieto L, Sastre A, Azorin D, Sanjuan X, López-Alemany R, Moran S, Roma J, Gallego S, Mora J, García Del Muro X, Giangrande PH, Peinado MA, Alonso J, de Alava E, Monk D, Esteller M, Tirado OM.
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Cancer Lett
Hernandez-Muñoz I, Figuerola E, Sanchez-Molina S, Rodriguez E, Fernández-Mariño AI, Pardo-Pastor C, Bahamonde MI, Fernández-Fernández JM, García-Domínguez DJ, Hontecillas-Prieto L, Lavarino C, Carcaboso AM, de Torres C, Tirado OM, de Alava E, Mora J.
RING1B contributes to Ewing sarcoma development by repressing the NaV1.6 sodium channel and the NF-κB pathway, independently of the fusion oncoprotein.
Oncotarget
Blanco-Gómez A, Castillo-Lluva S, Del Mar Sáez-Freire M, Hontecillas-Prieto L, Mao JH, Castellanos-Martín A, Pérez-Losada J.
Missing heritability of complex diseases: Enlightenment by genetic variants from intermediate phenotypes.
Bioessays
Marco Perez Javier Peinado-Serrano, Jose Manuel Garcia-Heredia, Irene Felipe Abrio, Cristina Tous, Irene Ferrer, Javier Martin-Broto, Carmen Saez, Amancio Carnero.
Efficacy of bortezomib in sarcomas with high levels of MAP17 (PDZK1IP1).
Oncotarget
de Alava E, Marcilla D.
Birth and evolution of the desmoplastic small round-cell tumor.
Semin Diagn Pathol
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Loss of the tumor suppressor spinophilin (PPP1R9B) increases the cancer stem cell population in breast tumors.
Oncogene
Marco Perez, Antonio Lucena-Cacace, Luis Miguel MarínGómez, Javier Padillo-Ruiz, Maria Jose Robles, Carmen Saez , Rocio Garcia Carbonero and Amancio Carnero.
Dasatinib, a Src inhibitor, sensitizes liver metastatic colorectal carcinoma to oxaliplatin in tumors with high phospho-Src.
Oncotarget.
Ordóñez JL, Amaral AT, Carcaboso AM, Herrero-Martín D, del Carmen García-Macías M, Sevillano V, Alonso D, Pascual-Pasto G, San-Segundo L, Vila-Ubach M, Rodrigues T, Fraile S, Teodosio C, Mayo-Iscar A, Aracil M, Galmarini CM, Tirado OM, Mora J, de Álava E.
The PARP inhibitor olaparib enhances the sensitivity of Ewing sarcoma to trabectedin
Oncotarget.
Romero-Pérez L, Garcia-Sanz P, Mota A, Leskelä S, Hergueta-Redondo M, Díaz-Martín J, López-García MA, Castilla MA, Martínez-Ramírez A, Soslow RA, Matias-Guiu X, Moreno-Bueno G, Palacios J.
A role for the transducer of the Hippo pathway, TAZ, in the development of aggressive types of endometrial cancer.
Mod Pathol.
Perez M, Muñoz-Galván S, Jiménez-García MP, Marín JJ, Carnero A.
Efficacy of CDK4 inhibition against sarcomas depends on their levels of CDK4 mRNA and p16Ink4a.
Oncotarget
Alfranca A, Martinez-Cruzado L, Tornin J, Abarrategi A, Amaral T, de Alava E, Menendez P, Garcia-Castro J, Rodriguez R.
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Cell Mol Life Sci
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Endocr Relat Cancer
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Unraveling heterogeneous susceptibility and the evolution of breast cancer using a systems biology approach.
Genome Biol
Castillo-Lluva S, Hontecillas-Prieto L, Blanco-Gómez A, Del Mar Sáez-Freire M, García-Cenador B, García-Criado J, Pérez-Andrés M, Orfao A, Cañamero M, Mao JH, Gridley T, Castellanos-Martín A, Pérez-Losada J.
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Oncogene.
Castillo-Lluva S, Hontecillas-Prieto L, Blanco-Gómez A, Del Mar Sáez-Freire M, García-Cenador B, García-Criado J, Pérez-Andrés M, Orfao A, Cañamero M, Mao JH, Gridley T, Castellanos-Martín A, Pérez-Losada J.
A new role of SNAI2 in postlactational involution of the mammary gland links it to luminal breast cancer development.
Oncogene
Muñoz Galvan S, Gutierrez G, Perez M, Carnero A.
MAP17 (PDZKIP1) .Expression Determines Sensitivity to the Proteasomal Inhibitor Bortezomib by Preventing Cytoprotective Autophagy and NFκB Activation in Breast Cancer.
Mol Cancer Ther
Amaral AT, Garofalo C, Frapolli R, Manara MC, Mancarella C, Uboldi S, Di Giandomenico S, Ordóñez JL, Sevillano V, Malaguarnera R, Picci P, Hassan AB, De Alava E, D'Incalci M, Scotlandi K.
Trabectedin efficacy in Ewing sarcoma is greatly increased by combination with anti-IGF signaling agents.
Clin Cancer Res
Díaz-López A, Díaz-Martín J, Moreno-Bueno G, Cuevas EP, Santos V, Olmeda D, Portillo F, Palacios J, Cano A.
Zeb1 and Snail1 engage miR-200f transcriptional and epigenetic regulation during EMT.
Int J Cancer