Almost 2% of Spanish breast cancer families are associated to germline pathogenic mutations in the ATM gene.

PURPOSE: There is still a considerable percentage of hereditary breast and ovarian cancer (HBOC) cases not explained by BRCA1 and BRCA2 genes. In this report, next-generation sequencing (NGS) techniques were applied to identify novel variants and/or genes involved in HBOC susceptibility. METHODS: Using whole exome sequencing, we identified a novel germline mutation in the moderate-risk gene ATM (c.5441delT; p.Leu1814Trpfs*14) in a family negative for mutations in BRCA1/2 (BRCAX). A case-control association study was performed to establish its prevalence in Spanish population, in a series of 1477 BRCAX families and 589 controls further screened, and NGS panels were used for ATM mutational screening in a cohort of 392 HBOC Spanish BRCAX families and 350 patients affected with diseases not related to breast cancer. RESULTS: Although the interrogated mutation was not prevalent in case-control association study, a comprehensive mutational analysis of the ATM gene revealed 1.78% prevalence of mutations in the ATM gene in HBOC and 1.94% in breast cancer-only BRCAX families in Spanish population, where data about ATM mutations were very limited. CONCLUSION: ATM mutation prevalence in Spanish population highlights the importance of considering ATM pathogenic variants linked to breast cancer susceptibility.

Sobreexpresión proteica y amplificación génica del oncogén HER2/neu en cáncer de mama: estudio de 125 casos / No disponible

Objetivo: estudiar una serie de 125 casos de cáncer demama procedentes de diferentes hospitales de la ComunidadValenciana, analizados de forma homogénea y protocolizadaen un mismo centro, comprobando el porcentaje real de casosamplificados para HER2/neu en los distintos grupos de expresiónproteica: sin sobreexpresión (0, 1+) y con sobreexpresión(2+, 3+). Determinar asimismo, el porcentaje de casos aneuploidespara el cromosoma 17 y la relación existente entre laaneuploidía y la amplificación HER2/neu. Comparar nuestrosresultados de inmunohistoquímica con los resultados en loscentros de origen.Pacientes y métodos: los métodos para la valoración delestado de HER2/neu utilizados fueron la determinación mediantetécnica inmunohistoquímica (kit HerceptestTM automatizado)para el análisis de la expresión proteica e hibridación insitu fluorescente (HER2 FISH pharmDxTM kit) para la amplificacióngénica.Resultados: de los casos con expresión proteica 3+(45/125), un 87% eran amplificados y un 13% no amplificados.De los casos 2+ (43/125) un 28% aparecían amplificadosy un 72% no amplificados. En los casos 1+/0 (37/125)un 3% estaban amplificados (1/7) y un 97% no amplificados.Adicionalmente se detectaron 36 casos aneuploides para elcromosoma 17 (29% del total), de los cuales un 31% estabanasimismo amplificados y un 69% no amplificados. La concordanciaen la determinación del Herceptest inter-centro presentaun coeficiente de correlación r = 0,527 (p < 0,01).Conclusiones: la técnica de hibridación in situ fluorescentese revela como técnica de referencia o gold standard para ladeterminación del estado del oncogen Her2/neu permitiendodiferenciar una sobreexpresión proteica producida por una“verdadera” amplificación génica, de la producida por unaaneuploidía...(AU)

Objectives: one hundred and twenty five breast tumors fromdifferent centers of the Valencian community were assessed bya consensus HER2/neu protocol in a single center. To determinateboth amplification and overexpression HER2/neu status,we investigate the real percentage of amplificated cases in thenon-overexpression (0, 1+) and overexpression (2+, 3+)groups. Furthermore, to determine the percentage of the chromosome17 aneusomy and its relationships with HER2/neuamplification. Finally, to compare our immunohistochemistry(IHC) results from those results obtained in the origin center.Patients and methods: all cases were assessed by IHC (HerceptestTMautomatized protocol) and fluorescence in situ hybridization(FISH) (HER2 FISH pharmDxTM kit) to detect HER2/neuexpression and HER2/neu amplification, respectively.Results: HER2/neu 3+ overexpression was observed in45/125 cases, showing amplification in 87% and 13% nonamplificated.The 2+ cases (43/125) showing amplification in28% and 72% non-amplificated. Finally, the 1+/0 cases wereobserved in 37/125 cases, showing amplification in 3% andin 97% non-amplificated. In addition, 36 (29%) cases showeda c17 aneusomy, 31% of those were amplificated and 69%non-amplificated. The inter-center IHC concordance displaysa correlation coefficient of r = 0.527 (p < 0.01).Conclusions: the FISH assay is the gold standard to determinatethe Her2/neu gene status, distinguishing between aHER2/neu protein overexpression due to a gene amplificationand those produced by an aneusomy. It is important to unify acommon criterion for the IHC classification to avoid differencesinter-observer as a consequence of different evaluation approaches(AU)

Renal donor implication in the origin of BK infection: analysis of genomic viral subtypes.

UNLABELLED: BK virus (BKV) reactivation in immunocompromised kidney transplant patients can produce a tubulointerstitial nephropathy (BKVN). Molecular tools that test for DNA-BKV provide early detection and assist in management, but some aspects of the pathogenesis of this infection, such as donor causality, remain unclear. MATERIALS AND METHODS: Between November 2004 and January 2006, 55 Spanish kidney donors were studied for BK infection. A quantitative PCR assay was performed on urine and serum to detect BKV. To determine the origin of the viral infection, a transcription control region of the BK polymorphism sequence was designed to identify the viral subtype. RESULTS: Fifteen of 55 (27%) donors were BK-PCR positive: 13 in urine and 2 in serum and urine. Moreover, monitoring of recipient pairs detected BK-PCR positivity in 14 of 73 recipients. We studied eight BK-PCR positive recipients (corresponding to four pairs) and their respective donors. The same viral genome was observed in the four pairs, namely, the A250-1-a, WW-like, AS, and JL genotypes. Interestingly, one of the four pairs showed the donor and the two recipients to display exactly the same JL genotype. CONCLUSION: On the basis of our preliminary results analyzing the molecular fingerprints of donor and recipient pairs, we have presented new data implicating the donor, in at least some cases, as the source of BK infection.