Estudi citogenètic de líquid amniòtic i vellositats corials : valor de la implementació del cribatge combinat de primer trimestre de gestació en el diagnòstic prenatal d'anomalies cromosòmiques

La implementació del cribatge combinat de primer trimestre de gestació ha repercutit en els estudis citogenètics, disminuïnt el nombre de proves invasives i augmentant la taxa de detecció d'anomalies cromosòmiques. Es presenten els resultats citogenètics de mostres de líquid amniòtic i de vellositats corials realitzats del 1999 al 2010 al servei laboratori d'Hematologia de l'Hospital Germans Trias i Pujol per valorar la implementació d'aquest cribatge

Associació de l'estat mutacional de JAK2 i les anomalies citogenètiques en les neoplàsies mieloproliferatives cròniques Filadèlfia negatives : Correlació amb les dades clíniques i analítiques

L'estudi correlaciona l'estat mutacional de JAK2V617F amb les alteracions citogenètiques, algunes dades clíniques i paràmetres de laboratori i valorar l'impacte sobre la progressió i pronòstic de la malaltia de 526 casos de NMPc Ph-. Es conclou que la freqüència d'alteracions citogenètiques, l'esplenomegàlia, els símptomes constitucionals i la progressió són independents de la mutació JAK2V617F, però la majoria de pacients en progressió i cariotip alterat eren JAK2 mutat. Els casos JAK2+ es correlacionen amb les complicacions en el curs de la malaltia i l'augment de la xifra de leucòcits i d'hemoglobina en la TE i de plaquetes en la PV

Targeted deep sequencing improves outcome stratification in chronic myelomonocytic leukemia with low risk cytogenetic features

Clonal cytogenetic abnormalities are found in 20-30% of patients with chronic myelomonocytic leukemia (CMML), while gene mutations are present in >90% of cases. Patients with low risk cytogenetic features account for 80% of CMML cases and often fall into the low risk categories of CMML prognostic scoring systems, but the outcome differs considerably among them. We performed targeted deep sequencing of 83 myeloid-related genes in 56 CMML patients with low risk cytogenetic features or uninformative conventional cytogenetics (CC) at diagnosis, with the aim to identify the genetic characteristics of patients with a more aggressive disease. Targeted sequencing was also performed in a subset of these patients at time of acute myeloid leukemia (AML) transformation. Overall, 98% of patients harbored at least one mutation. Mutations in cell signaling genes were acquired at time of AML progression. Mutations in ASXL1, EZH2 and NRAS correlated with higher risk features and shorter overall survival (OS) and progression free survival (PFS). Patients with SRSF2 mutations associated with poorer OS, while absence of TET2 mutations (TET2 wt) was predictive of shorter PFS. A decrease in OS and PFS was observed as the number of adverse risk gene mutations (ASXL1, EZH2, NRAS and SRSF2) increased. On multivariate analyses, CMML-specific scoring system (CPSS) and presence of adverse risk gene mutations remained significant for OS, while CPSS and TET2wt were predictive of PFS. These results confirm that mutation analysis can add prognostic value to patients with CMML and low risk cytogenetic features or uninformative CC

A pediatric regimen for adolescents and young adults with Philadelphia chromosome-negative acute lymphoblastic leukemia : Results of the ALLRE08 PETHEMA trial

Altres ajuts: Supported in part by grants from Fundació La Caixa and CIBERONC (JMHR and AO).Pediatric-based or -inspired trials have improved the prognosis of adolescents and young adults (AYA) with Philadelphia chromosome-negative (Ph-neg) acute lymphoblastic leukemia (ALL). This study reports the results of treatment of the ALLRE08 trial, a full pediatric trial for AYA aged 15-30 years with standard-risk (SR) ALL. From 2008 to 2018, 89 patients (38 adolescents [15-18 years] and 51 young adults [YA, 19-30 years], median age: 20 [15-29] years) were enrolled in the ALLRE08 trial. The complete response (CR) was 95%. Twenty-two patients were transferred to a high-risk (HR) protocol because of poor marrow response on day 14 (n = 20) or high-level of end-induction minimal residual response (MRD ≥ 0.25%, n = 2). Cumulative incidence of relapse (CIR) at 5 years was 35% (95%CI: 23%-47%), with significant differences between adolescents and YA: 13% (4%-28%) vs 52% (34%-67%), P = .012. No treatment-related mortality was observed in 66/66 patients following the ALLRE08 trial vs 3/23 patients moved to a HR trial. The estimated 5-year overall survival (OS) was 74% (95%CI: 63%-85%), with significantly higher rates for adolescents vs YA: 87% (95%CI: 74%-100%) vs 63% (46%-80%), P = .021. Although CIR or OS were lower in patients who were transferred to a HR trial, the differences were not statistically significant (CIR: 34% [21%-47%] vs 37% [14%-61%]; OS: 78% [66%-90%] vs 61% [31%;91%]). A full pediatric trial is feasible and effective for AYA with Ph-neg, SR-ALL, with better results for adolescents than for YA. Outcome of patients with poor early response rescued with a HR trial was not significantly inferior. A full pediatric trial is feasible and effective for adolescent and young adults with acute lymphoblastic leukemia, with better results for adolescents than for young adults. The outcome of patients showing poor early response was not significantly inferior than that observed for good responders after being transferred to a high-risk trial

Frequency and clinical impact of CDKN2A/ARF/CDKN2B gene deletions as assessed by in-depth genetic analyses in adult T cell acute lymphoblastic leukemia

Altres ajuts: This project was supported by the Asociación Española Contra el Cáncer, AECC (project ref.: GC16173697BIGA), Obra Social "La Caixa" and by Celgene Spain. A. Gonzalez-Perez is supported by a Ramon y Cajal fellowship (RYC-2013-14554) of the Educational Ministry (Madrid, Spain). This work was also partially supported by FEDER funds from CIBERONC (CB16/12/00284 and CB16/12/00400), Madrid, Spain).Recurrent deletions of the CDKN2A/ARF/CDKN2B genes encoded at chromosome 9p21 have been described in both pediatric and adult acute lymphoblastic leukemia (ALL), but their prognostic value remains controversial, with limited data on adult T-ALL. Here, we investigated the presence of homozygous and heterozygous deletions of the CDKN2A/ARF and CDKN2B genes in 64 adult T-ALL patients enrolled in two consecutive trials from the Spanish PETHEMA group. Alterations in CDKN2A/ARF/CDKN2B were detected in 35/64 patients (55%). Most of them consisted of 9p21 losses involving homozygous deletions of the CDKNA/ARF gene (26/64), as confirmed by single nucleotide polymorphism (SNP) arrays and interphase fluorescence in situ hybridization (iFISH). Deletions involving the CDKN2A/ARF/CDKN2B locus correlated with a higher frequency of cortical T cell phenotype and a better clearance of minimal residual disease (MRD) after induction therapy. Moreover, the combination of an altered copy-number-value (CNV) involving the CDKN2A/ARF/CDKN2B gene locus and undetectable MRD (≤ 0.01%) values allowed the identification of a subset of T-ALL with better overall survival in the absence of hematopoietic stem cell transplantation

Perfil molecular en cáncer escamoso de pulmón

Dentro de los cánceres de pulmón de célula no pequeña se ha concentrado una gran cantidad de esfuerzos en los adenocarcinomas, tanto en la práctica clínica diaria como en la investigación básica. Por ello, este tipo histológico es uno de los tumores mejor caracterizados desde el punto de vista molecular y se han desarrollado diversas terapias dirigidas contra alteraciones genéticas frecuentes en el mismo, terapias que han conseguido mejoras objetivas en supervivencia y calidad de vida en subgrupos seleccionados de pacientes. En comparación, el carcinoma escamoso ha recibido una atención relativa y no existen fármacos específicos contra el mismo en uso clínico, ni se han identificado claros marcadores moleculares con valor predictivo o pronóstico. En este trabajo, se incluyeron 95 pacientes diagnosticados de carcinoma escamoso de pulmón del Hospital Universitario Quirón Dexeus y del Hospital Universitari General Germans Trias i Pujol. De todos ellos se recogieron los datos clínicos de edad, sexo, performance status (estado general), habito tabáquico, estadio clínico y TNM al diagnóstico. Los patólogos de ambos centros hicieron una selección de muestras tumorales a estudiar y tras valoración y macro o microdisección de las mismas, se analizaron 19 genes mediante cuatro técnicas de biología molecular buscando: mutaciones, amplificaciones y niveles de expresión génica, con el objetivo de realizar una caracterización molecular en este tipo de tumor analizando genes clave, así como identificar marcadores moleculares pronósticos o predictivos. Los resultados de la presente tesis apoyan la hipótesis que el carcinoma escamoso constituye una entidad diferenciada del adenocarcinoma desde el punto de vista molecular. Muchas alteraciones genéticas se hallan en porcentajes distintos en ambos tipos histológicos (como las mutaciones de EGFR o PIK3CA) y en algunos casos los subgrupos de alteraciones genéticas también difieren (como es el caso de las mutaciones de K-RAS, siendo la G12C mucho más frecuente en adenocarcinomas de fumadores). Finalmente, ciertas alteraciones parecen ser casi exclusivas del carcinoma escamoso (como la amplificación de FGFR1, PIK3CA o SOX2). En este trabajo se han identificado diversos marcadores con posible valor pronóstico en carcinoma escamoso que sería interesante validar en una cohorte más amplia de pacientes lo que permitiría, además, correlacionarlos con distintos tratamientos. En estadios I-IIIA se ha correlacionado la alta expresión de los genes SOX2 y EZH2 con una mejor Overall Survival (OS) y Progression Free Survival (PFS) de forma estadísticamente significativa. Por otro lado, en estadios avanzados, no mutaciones de K-RAS, co-amplificación de SOX2 + PIK3CA y elevada expresión de los genes SIAH2, BRCA1 y EZH2 correlacionaban, de forma estadísticamente significativa o quasi-significativa, con OS más larga. Estos tres últimos genes se asociaban también a mejor PFS. Todos estos marcadores se podrían integrar en firmas genéticas específicas con valor pronóstico para carcinoma escamoso. Por lo general, se considera al carcinoma escamoso como un subtipo histológico sin marcadores predictivos definidos y con un arsenal de terapias dirigidas limitado en comparación con el adenocarcinoma. Esta tesis demuestra que esta visión no se corresponde con la realidad. En efecto, un porcentaje muy elevado de pacientes con esta patología presenta alteraciones moleculares susceptibles de tratamiento con fármacos dirigidos en uso clínico (como los inhibidores de la tirosina quinasa) o en fases avanzadas de ensayos clínicos, entre los que podemos citar amplificaciones de FGFR1, AXL y MET o las mutaciones de PIK3CA. Sería interesante que se iniciaran más ensayos clínicos para determinar el beneficio terapéutico de tratamientos dirigidos e identificar biomarcadores de respuesta adicionales en carcinoma escamoso.Within non-small-cell lung cancer, a great deal of research efforts have focused on adenocarcinomas, both in daily clinical practice and basic research. This histological type is therefore one of the best characterized tumors from the molecular point of view, and targeted therapies against common genetic alterations have been developed which have achieved improvements in survival and quality of life in selected subgroups of patients. In comparison, less attention has been paid to squamous cell carcinoma, and no targeted therapies exist against this subtype for clinical use; neither have any molecular markers with clear predictive or prognostic value been identified. In the present study, we included 95 patients diagnosed with squamous cell carcinoma of the lung from the Hospital Universitario Quirón Dexeus and Hospital Universitario Germans Trias i Pujol. Clinical data was collected for all patients: age, sex, performance status, smoking habits, and TNM stage at diagnosis. Pathologists from both hospitals selected the tumor samples to be studied and, after evaluation and macro- or micro-dissection, 19 genes were analyzed using four different molecular biology techniques to look for mutations, amplifications and gene expression levels, with the aim of molecularly characterizing this tumor type, analyzing key genes and identifying prognostic or predictive molecular markers. The results of this thesis support the hypothesis that squamous cell carcinoma of the lung is a distinct entity from adenocarcinoma from the molecular point of view. Many genetic alterations (such as EGFR or PIK3CA mutations) have different rates of detection between both histological types, and in some cases the subgroups of genetic alterations also differ (as in the case of K-RAS mutations, G12C being much more frequent in adenocarcinoma in smokers). Finally, some alterations appear to be almost exclusive to squamous cell carcinoma (such as FGFR1, PIK3CA and SOX2 amplification). In this paper we have identified several markers with potential prognostic value in squamous cell carcinoma which it would be interesting to validate in a larger cohort of patients, something which would also permit correlation of these markers with different treatments. In stages I-IIIA, high expression of EZH2 and SOX2 genes has been correlated with statistical significance with better overall survival (OS) and progression free survival (PFS). Similarly, in advanced stages, K-RAS wild type, co-amplification of PIK3CA + SOX2 and high expression of SIAH2, BRCA1 and EZH2 correlated, also statistically significantly or quasi-significantly, with longer OS. These last three genes were also associated with better PFS. All these markers could be integrated into specific genetic signatures with prognostic value in squamous cell carcinoma. In general, squamous cell carcinoma is considered to be a histological subtype without defined predictive markers and with a limited arsenal of targeted therapies when compared to adenocarcinoma. This thesis demonstrates that this view does not correspond to the reality. In fact, a high percentage of patients with this disease have molecular alterations which are susceptible to treatment with targeted drugs currently in clinical use (such as tyrosine kinase inhibitors) or under investigation in advanced phase clinical trials, such as FGFR1, AXL and MET amplifications or PIK3CA mutations, among others. It would be interesting to initiate more clinical trials to determine the therapeutic benefit of targeted therapies and identify additional biomarkers of response in squamous cell carcinoma of the lung

Associació de l'estat mutacional de JAK2 i les anomalies citogenètiques en les neoplàsies mieloproliferatives cròniques Filadèlfia negatives : Correlació amb les dades clíniques i analítiques

L'estudi correlaciona l'estat mutacional de JAK2V617F amb les alteracions citogenètiques, algunes dades clíniques i paràmetres de laboratori i valorar l'impacte sobre la progressió i pronòstic de la malaltia de 526 casos de NMPc Ph-. Es conclou que la freqüència d'alteracions citogenètiques, l'esplenomegàlia, els símptomes constitucionals i la progressió són independents de la mutació JAK2V617F, però la majoria de pacients en progressió i cariotip alterat eren JAK2 mutat. Els casos JAK2+ es correlacionen amb les complicacions en el curs de la malaltia i l'augment de la xifra de leucòcits i d'hemoglobina en la TE i de plaquetes en la PV

Frequency and clinical impact of CDKN2A/ARF/CDKN2B gene deletions as assessed by in-depth genetic analyses in adult T cell acute lymphoblastic leukemia

Abstract Recurrent deletions of the CDKN2A/ARF/CDKN2B genes encoded at chromosome 9p21 have been described in both pediatric and adult acute lymphoblastic leukemia (ALL), but their prognostic value remains controversial, with limited data on adult T-ALL. Here, we investigated the presence of homozygous and heterozygous deletions of the CDKN2A/ARF and CDKN2B genes in 64 adult T-ALL patients enrolled in two consecutive trials from the Spanish PETHEMA group. Alterations in CDKN2A/ARF/CDKN2B were detected in 35/64 patients (55%). Most of them consisted of 9p21 losses involving homozygous deletions of the CDKNA/ARF gene (26/64), as confirmed by single nucleotide polymorphism (SNP) arrays and interphase fluorescence in situ hybridization (iFISH). Deletions involving the CDKN2A/ARF/CDKN2B locus correlated with a higher frequency of cortical T cell phenotype and a better clearance of minimal residual disease (MRD) after induction therapy. Moreover, the combination of an altered copy-number-value (CNV) involving the CDKN2A/ARF/CDKN2B gene locus and undetectable MRD (≤ 0.01%) values allowed the identification of a subset of T-ALL with better overall survival in the absence of hematopoietic stem cell transplantation