Miguel Hernández

Hypermethylation of apoptotic genes as independent prognostic factor in neuroblastoma disease

Neuroblastoma (NB) is an embryonal tumour of neuroectodermal cells, and its prognosis is based on patient age at diagnosis, tumour stage and MYCN amplification, but it can also be classified according to their degree of methylation. Considering that epigenetic aberrations could influence patient survival, we studied the methylation status of a series of 17 genes functionally involved in different cellular pathways in patients with NB and their impact on survival. We studied 82 primary NB tumours and we used methylation‐specific‐PCR to perform the epigenetic analysis. We evaluated the putative association among the evidence of hypermethylation with the most important NB prognostic factors, as well as to determine the relationship among methylation, clinical classification and survival. CASP8 hypermethylation showed association with relapse susceptibility and, TMS1 and APAF1 hypermethylation are associated with bad prognosis and showed high influence on NB overall survival. Hypermethylation of apoptotic genes has been identified as a good candidate of prognostic factor. We propose the simultaneous analysis of hypermethylation of APAF1, TMS1 and CASP8 apoptotic genes on primary NB tumour as a good prognostic factor of disease progression. Mol. Carcinog.

Value of Deoxyribonucleic Acid Ploidy and Nuclear Morphometry for Prediction of Disease Progression in Renal Cell Carcinoma

PURPOSE A retrospective study was performed on 108 patients with localized renal cell carcinoma (pT1 to 3a N0M0) to determine whether ploidy and nuclear morphometry are independent predictive factors in addition to stage and grade. MATERIALS AND METHODS Deoxyribonucleic acid (DNA) content was analyzed by flow cytometry and nuclear morphometry characterized by 5 nuclear descriptors. A Cox proportional hazards regression model was used to identify significant prognostic factors for disease progression. RESULTS A model combining tumor stage and grade, DNA ploidy and nuclear minor axis was chosen as optimal with risk of disease progression increased with increasing tumor stage and grade, DNA aneuploidy and increasing nuclear minor axis. CONCLUSIONS This improved ability to predict disease progression in localized renal cell carcinoma may have important clinical use.

Prognostic value of DNA ploidy and nuclear morphometry in prostate cancer treated with androgen deprivation.

OBJECTIVES To assess the prognostic value of flow cytometry and nuclear morphometry in prostate cancer after androgen deprivation treatment. METHODS A total of 127 patients with a prostate cancer diagnosis who had undergone androgen suppression were retrospectively studied. The DNA content by flow cytometry and nuclear morphometry was studied from biopsy specimens. In the patients with Stage M0, two multivariate analyses by the Cox proportional regression model were performed to determine whether the experimental variables (DNA content and nuclear area) added independent information to the classic prognostic factors (Gleason score and stage). Using the statistical analysis results, risk groups were created. RESULTS T and M categories, Gleason score, DNA ploidy, and mean nuclear area proved to have prognostic value in the univariate analysis. For the group of patients free of metastasis (M0), it was possible to create low, intermediate, and high-risk groups using stage and Gleason score with statistically significant differences in survival. Multivariate analysis, combining the classic and experimental variables, selected Gleason score and DNA content as prognostic independent factors. Also, risk groups with statistically significant differences in survival were created. However, the net result of combining both kinds of factors was at least as valuable as the combination of stage and Gleason score in predicting survival. CONCLUSIONS The determination of DNA ploidy and mean nuclear area do not add enough independent information to improve the predictive value to justify their use in this group of patients treated with hormonal therapy.

miR-200c and phospho-AKT as prognostic factors and mediators of osteosarcoma progression and lung metastasis

Lung metastasis is the major cause of death in osteosarcoma patients. However, molecular mechanisms underlying this metastasis remain poorly understood. To identify key molecules related with pulmonary metastasis of pediatric osteosarcomas, we analyzed high‐throughput miRNA expression in a cohort of 11 primary tumors and 15 lung metastases. Results were further validated with an independent cohort of 10 primary tumors and 6 metastases. In parallel, we performed immunohistochemical analysis of activated signaling pathways in 36 primary osteosarcomas. Only phospho‐AKT associated with lower overall survival in primary tumors, supporting its role in osteosarcoma progression. CTNNB1 expression also associated with lower overall survival but was not strong enough to be considered an independent variable. Interestingly, miR‐200c was overexpressed in lung metastases, implicating an inhibitory feed‐back loop to PI3K‐AKT. Moreover, transfection of miR200c‐mimic in U2‐OS cells reduced phospho‐AKT levels but increased cellular migration and proliferation. Notably, miR‐200c expression strongly correlated with miR‐141 and with the osteogenic inhibitor miR‐375, all implicated in epithelial to mesenchymal transition. These findings contrast epithelial tumors where reduced miR‐200c expression promotes metastasis. Indeed, we noted that osteosarcoma cells in the lung also expressed the epithelial marker CDH1, revealing a change in their mesenchymal phenotype. We propose that miR‐200c upregulation occurs late in osteosarcoma progression to provide cells with an epithelial phenotype that facilitates their integration in the metastatic lung niche. Thus, our findings identify phospho‐AKT in the primary tumor and miR‐200c later during tumor progression as prognostic molecules and potential therapeutic targets to prevent progression and metastasis of pediatric osteosarcomas.

Subtelomeric analysis of pediatric astrocytoma: subchromosomal instability is a distinctive feature of pleomorphic xanthoastrocytoma

Astrocytic neoplasms are genetically heterogeneous; however a low frequency of genomic changes has been found in juvenile pilocytic astrocytoma (PA) in molecular studies. Concerning pleomorphic xanthoastrocytomas (PXA), recent studies have given heterogeneous results for chromosomal alterations. We studied the subtelomeric regions of 19 primary astrocytoma tumors. Results were near normality for the PA group with relative scarcity of chromosomal imbalances, except for the duplication of 3pter in 4/15 and deletion of 21qter in 5/15 of them. In contrast, a specific profile was observed in the 4 PXA tumoral samples. This involved 3pter, 14qter and 19pter duplication and 4qter, 6qter, 9qter, 13cen, 17pter, 18qter and 21qter deletion. Our results indicate that the chromosomal and genetic aberrations in PXAs differed from those typically associated with the diffusely infiltrating astrocytic and oligodendroglial gliomas. These genetic differences would likely contribute to the more favorable behavior of PXAs and may be helpful for molecular differential diagnosis of pediatric cerebral tumors.

Rare chromosomal complement of trisomy 21 in a boy conceived by IVF and cryopreservation

This report describes a case of mosaic Down syndrome due to an unusual karyotype in a patient conceived by assisted reproductive techniques and cryopreservation. The chromosomal complement consists of two different cell lines, one predominantly trisomic with a derivative chromosome due to a Robertsonian translocation (21;21) and another carrying a ring chromosome 21. The present work analyses the different mechanisms that could have led to mosaicism.