Open Access
Impact of FLT3-ITD mutation status and its ratio in a cohort of 2901 patients undergoing upfront intensive chemotherapy: a PETHEMA registry study
(MDPI, 2022-11-24) Ayala, Rosa; Carreño-Tarragona, Gonzalo; Barragán, Eva; Boluda, Blanca; Larráyoz, María José; Chillón, María Carmen; Carrillo-Cruz, Estrella; Bilbao, Cristina; Sánchez-García, Joaquín; Bernal, Teresa; Martínez-Cuadrón, David; Gil, Cristina; Serrano, Josefina; Rodríguez-Medina, Carlos; Bergua, Juan; Pérez-Simón, José A.; Calbacho, María; Alonso-Domínguez, Juan M.; Labrador, Jorge; Tormo, Mar; Amigo, María Luz; Herrera-Puente, Pilar; Rapado, Inmaculada; Sargas, Claudia; Vázquez Urio, Iria; Calasanz, María José; Gómez-Casares, Teresa; García-Sanz, Ramón; Sanz, Miguel A.; Martínez-López, Joaquín; Montesinos, Pau; Ciencias; Zientziak
FLT3-ITD results in a poor prognosis in terms of overall survival (OS) and relapse-free survival (RFS) in acute myeloid leukemia (AML). However, the prognostic usefulness of the allelic ratio (AR) to select post-remission therapy remains controversial. Our study focuses on the prognostic impact of FLT3-ITD and its ratio in a series of 2901 adult patients treated intensively in the pre-FLT3 inhibitor era and reported in the PETHEMA registry. A total of 579 of these patients (20%) harbored FLT3-ITD mutations. In multivariate analyses, patients with an FLT3-ITD allele ratio (AR) of >0.5 showed a lower complete remission (CR rate) and OS (HR 1.47, p = 0.009), while AR > 0.8 was associated with poorer RFS (HR 2.1; p < 0.001). Among NPM1/FLT3-ITD-mutated patients, median OS gradually decreased according to FLT3-ITD status and ratio (34.3 months FLT3-ITD-negative, 25.3 months up to 0.25, 14.5 months up to 0.5, and 10 months ≥ 0.5, p < 0.001). Post-remission allogeneic transplant (allo-HSCT) resulted in better OS and RFS as compared to auto-HSCT in NPM1/FLT3-ITD-mutated AML regardless of pre-established AR cutoff (≤0.5 vs. >0.5). Using the maximally selected log-rank statistics, we established an optimal cutoff of FLT3-ITD AR of 0.44 for OS, and 0.8 for RFS. We analyzed the OS and RFS according to FLT3-ITD status in all patients, and we found that the group of FLT3-ITD-positive patients with AR < 0.44 had similar 5-year OS after allo-HSCT or auto-HSCT (52% and 41%, respectively, p = 0.86), but worse RFS after auto-HSCT (p = 0.01). Among patients with FLT3-ITD AR > 0.44, allo-HSCT was superior to auto-HSCT in terms of OS and RFS. This study provides more evidence for a better characterization of patients with AML harboring FLT3-ITD mutations.
Open Access
Loss of PPP2R2A inhibits homologous recombination DNA repair and predicts tumor sensitivity to PARP inhibition
(American Association for Cancer Research, 2012-10-18) Kalev, Peter; Simicek, Michal; Vázquez Urio, Iria; Munck, Sebastian; Chen, Liping; Soin, Thomas; Danda, Natasha; Chen, Wen; Sablina, Anna; Ciencias; Zientziak
Reversible phosphorylation plays a critical role in DNA repair. Here, we report the results of a loss-of-function screen that identifies the PP2A heterotrimeric serine/threonine phosphatases PPP2R2A, PPP2R2D, PPP2R5A, and PPP2R3C in double-strand break (DSB) repair. In particular, we found that PPP2R2A-containing complexes directly dephosphorylated ATM at S367, S1893, and S1981 to regulate its retention at DSB sites. Increased ATM phosphorylation triggered by PPP2R2A attenuation dramatically upregulated the activity of the downstream effector kinase CHK2, resulting in G1 to S-phase cell-cycle arrest and downregulation of BRCA1 and RAD51. In tumor cells, blocking PPP2R2A thereby impaired the high-fidelity homologous recombination repair pathway and sensitized cells to small-molecule inhibitors of PARP. We found that PPP2R2A was commonly downregulated in non-small cell lung carcinomas, suggesting that PPP2R2A status may serve as a marker to predict therapeutic efficacy to PARP inhibition. In summary, our results deepen understanding of the role of PP2A family phosphatases in DNA repair and suggest PPP2R2A as a marker for PARP inhibitor responses in clinic.
Open Access
NGS-based molecular karyotyping of multiple myeloma: results from the GEM12 clinical trial
(MDPI, 2022-10-21) Rosa-Rosa, Juan Manuel; Cuenca, Isabel; Medina, Alejandro; Vázquez Urio, Iria; Sánchez-delaCruz, Andrea; Buenache, Natalia; Sánchez Pérez, Ricardo; Jiménez, Cristina; Rosiñol, Laura; Gutiérrez, Norma C.; Ruiz-Heredia, Yanira; Barrio Santiago; Oriol, Albert; Martín-Ramos, María-Luisa; Blanchard, María-Jesús; Ayala, Rosa; Ríos-Tamayo, Rafael; Sureda, Anna; Hernández, Miguel-Teodoro; Rubia, Javier de la; Alkorta Aranburu, Gorka; Agirre, Xabier; Bladé, Joan; Mateos, María-Victoria; Lahuerta, Juan-José; San-Miguel, Jesús F.; Calasanz, María José; García-Sanz, Ramón; Martínez-López, Joaquín; Ciencias; Zientziak
Next-generation sequencing (NGS) has greatly improved our ability to detect the genomic aberrations occurring in multiple myeloma (MM); however, its transfer to routine clinical labs and its validation in clinical trials remains to be established. We designed a capture-based NGS targeted panel to identify, in a single assay, known genetic alterations for the prognostic stratification of MM. The NGS panel was designed for the simultaneous study of single nucleotide and copy number variations, insertions and deletions, chromosomal translocations and V(D)J rearrangements. The panel was validated using a cohort of 149 MM patients enrolled in the GEM2012MENOS65 clinical trial. The results showed great global accuracy, with positive and negative predictive values close to 90% when compared with available data from fluorescence in situ hybridization and whole-exome sequencing. While the treatments used in the clinical trial showed high efficacy, patients defined as high-risk by the panel had shorter progression-free survival (p = 0.0015). As expected, the mutational status of TP53 was significant in predicting patient outcomes (p = 0.021). The NGS panel also efficiently detected clonal IGH rearrangements in 81% of patients. In conclusion, molecular karyotyping using a targeted NGS panel can identify relevant prognostic chromosomal abnormalities and translocations for the clinical management of MM patients.
Open Access
EVI1 controls proliferation in acute myeloid leukaemia through modulation of miR-1-2
(Nature Publishing Group, 2010-09-14) Gómez-Benito, María; Conchillo, Ana; García, M. A.; Vázquez Urio, Iria; Maicas, Miren; Vicente, Carmen; Cristobal, I.; Marcotegui, Nerea; García-Ortí, L.; Bandrés Elizalde, Eva; Calasanz, María José ; Alonso, M. M.; Odero, María D.; Ciencias; Zientziak; Ciencias de la Salud; Osasun Zientziak; Gobierno de Navarra / Nafarroako Gobernua
Bakground: the EVI1(ecotropic virus integration site 1) gene codes for a zinc-finger transcription factor, whose transcriptional activation leads to a particularly aggressive form of acute myeloid leukaemia (AML). Although, EVI1 interactions with key proteins in hematopoiesis have been previously described, the precise role of this transcription factor in promoting leukaemic transformation is not completely understood. Recent works have identified specific microRNA (miRNA) signatures in different AML subgroups. However, there is no analysis of miRNAs profiles associated with EVI1 overexpression in humans. Methods: we performed QT-RT-PCR to assess the expression of 250 miRNAs in cell lines with or without EVI1 overexpression and in patient samples. We used ChIP assays to evaluated the possible binding of EVI1 binding to the putative miRNA promoter. Proliferation of the different cell lines transfected with the anti- or pre-miRs was quantified by MTT. Results: our data showed that EVI1 expression was significantly correlated with the expression of miR-1-2 and miR-133-a-1 in established cell lines and in patient samples. ChIP assays confirmed that EVI1 binds directly to the promoter of these two miRNAs. However, only miR-1-2 was involved in abnormal proliferation in EVI1 expressing cell lines. Conclusions: our data showed that EVI1 controls proliferation in AML through modulation of miR-1-2. This study contributes to further understand the transcriptional networks involving transcription factors and miRNAs in AML.
Open Access
The tyrosine phosphatase PTPRO sensitizes colon cancer cells to anti-EGFR therapy through activation of SRC-mediated EGFR signaling
(Impact Journals, 2014-10-11) Asbagh, Layka Abbasi; Vázquez Urio, Iria; Vecchione, Loredana; Budinska, Eva; De Vriendt, Veerle; Baietti, Maria Francesca; Steklov, Mikhail; Jacobs, Bart ; Hoe, Nicholas ; Singh, Sharat; Imjeti, Naga-Sailaja; Zimmermann, Pascale; Sablina, Anna; Tejpar, Sabine; Ciencias; Zientziak
Inappropriate activation of epidermal growth factor receptor (EGFR) plays a causal role in many cancers including colon cancer. The activation of EGFR by phosphorylation is balanced by receptor kinase and protein tyrosine phosphatase activities. However, the mechanisms of negative EGFR regulation by tyrosine phosphatases remain largely unexplored. Our previous results indicate that protein tyrosine phosphatase receptor type O (PTPRO) is down-regulated in a subset of colorectal cancer (CRC) patients with a poor prognosis. Here we identified PTPRO as a phosphatase that negatively regulates SRC by directly dephosphorylating Y416 phosphorylation site. SRC activation triggered by PTPRO down-regulation induces phosphorylation of both EGFR at Y845 and the c-CBL ubiquitin ligase at Y731. Increased EGFR phosphorylation at Y845 promotes its receptor activity, whereas enhanced phosphorylation of c-CBL triggers its degradation promoting EGFR stability. Importantly, hyperactivation of SRC/EGFR signaling triggered by loss of PTPRO leads to high resistance of colon cancer to EGFR inhibitors. Our results not only highlight the PTPRO contribution in negative regulation of SRC/EGFR signaling but also suggest that tumors with low PTPRO expression may be therapeutically targetable by anti-SRC therapies.
Open Access
The genomic profiling of high-risk smoldering myeloma patients treated with an intensive strategy unveils potential markers of resistance and progression
(Springer Nature, 2024-04-29) Medina-Herrera, Alejandro; Vázquez Urio, Iria; Cuenca, Isabel ; Rosa-Rosa, Juan Manuel; Ariceta, Beñat; Jiménez, Cristina; Fernández-Mercado, Marta; Larráyoz, María José; Gutiérrez, Norma C.; Fernández-Guijarro, M.; González-Calle, Verónica; Rodríguez-Otero, Paula; Oriol, Albert; Rosiñol, Laura; Alegre, Adrián; Escalante, Fernando; Rubia, Javier de la; Teruel, A. I.; Arriba, Felipe de; Hernández, Miguel-Teodoro; López-Jiménez, Javier; Ocio, Enrique María; Puig, Noemí; Paiva, Bruno David Lourenço; Lahuerta, Juan-José; Bladé, Joan; San-Miguel, Jesús F.; Mateos, María-Victoria; Martínez-López, Joaquín; Calasanz, María José; García-Sanz, Ramón; GEM/PETHEMA (Grupo Español de Mieloma/Programa para el Estudio de la Terapéutica en Hemopatías Malignas) cooperative study group; Ciencias; Zientziak
Smoldering multiple myeloma (SMM) precedes multiple myeloma (MM). The risk of progression of SMM patients is not uniform, thus different progression-risk models have been developed, although they are mainly based on clinical parameters. Recently, genomic predictors of progression have been defined for untreated SMM. However, the usefulness of such markers in the context of clinical trials evaluating upfront treatment in high-risk SMM (HR SMM) has not been explored yet, precluding the identification of baseline genomic alterations leading to drug resistance. For this reason, we carried out next-generation sequencing and fluorescent in-situ hybridization studies on 57 HR and ultra-high risk (UHR) SMM patients treated in the phase II GEM-CESAR clinical trial (NCT02415413). DIS3, FAM46C, and FGFR3 mutations, as well as t(4;14) and 1q alterations, were enriched in HR SMM. TRAF3 mutations were specifically associated with UHR SMM but identified cases with improved outcomes. Importantly, novel potential predictors of treatment resistance were identified: NRAS mutations and the co-occurrence of t(4;14) plus FGFR3 mutations were associated with an increased risk of biological progression. In conclusion, we have carried out for the first time a molecular characterization of HR SMM patients treated with an intensive regimen, identifying genomic predictors of poor outcomes in this setting. © The Author(s) 2024.
Open Access
Integrated flow cytometry and sequencing to reconstruct evolutionary patterns from dysplasia to acute myeloid leukemia
(American Society of Hematology, 2023-01-10) Simoes, Catia; Chillón, María Carmen; Martínez-Cuadrón, David; Calasanz, María José; Vridiales, María-Belén; Vázquez Urio, Iria; Hernández-Ruano, Montserrat; Ariceta, Beñat; Aguirre-Ruiz, Paula; Burgos, Leire; Alignani, Diego; Sarvide, Sarai; Villar, Sara; Alfonso Pierola, Ana; Prósper, Felipe; Ayala, Rosa; Martínez-López, Joaquín; Bergua Burgués, Juan Miguel; Vives, Susana; Pérez-Simón, José A.; García-Fortes, María; Bernal del Castillo, Teresa; Colorado, Mercedes; Olave, Mayte; Rodríguez-Gutiérrez, Juan I.; Labrador, Jorge; Gonzalez Díaz, Marcos; San-Miguel, Jesús F.; Sanz, Miguel A.; Montesinos, Pau; Paiva, Bruno David Lourenço; Ciencias; Zientziak
Clonal evolution in acute myeloid leukemia (AML) originates long before diagnosis and is a dynamic process that may affect survival. However, it remains uninvestigated during routine diagnostic workups. We hypothesized that the mutational status of bone marrow dysplastic cells and leukemic blasts, analyzed at the onset of AML using integrated multidimensional flow cytometry (MFC) immunophenotyping and fluorescence-activated cell sorting (FACS) with next-generation sequencing (NGS), could reconstruct leukemogenesis. Dysplastic cells were detected by MFC in 285 of 348 (82%) newly diagnosed patients with AML. Presence of dysplasia according to MFC and World Health Organization criteria had no prognostic value in older adults. NGS of dysplastic cells and blasts isolated at diagnosis identified 3 evolutionary patterns: stable (n = 12 of 21), branching (n = 4 of 21), and clonal evolution (n = 5 of 21). In patients achieving complete response (CR), integrated MFC and FACS with NGS showed persistent measurable residual disease (MRD) in phenotypically normal cell types, as well as the acquisition of genetic traits associated with treatment resistance. Furthermore, whole-exome sequencing of dysplastic and leukemic cells at diagnosis and of MRD uncovered different clonal involvement in dysplastic myelo-erythropoiesis, leukemic transformation, and chemoresistance. Altogether, we showed that it is possible to reconstruct leukemogenesis in ~80% of patients with newly diagnosed AML, using techniques other than single-cell multiomics.
Open Access
Down-regulation of EVI1 is associated with epigenetic alterations and good prognosis in patients with acute myeloid leukemia
(Ferrata Storti Foundation, 2012-10-18) Vázquez Urio, Iria; Maicas, Miren; Cervera, José; Agirre, Xabier; Marin-Béjar, Oskar; Marcotegui, Nerea; Vicente, Carmen; Lahortiga, Idoya; Gómez-Benito, María; Carranza, Claudia; Valencia, Ana; Brunet, Salut; Lumbreras, Eva; Prósper, Felipe; Gómez-Casares, Teresa; Hernández-Rivas, Jesús M.; Calasanz, María José; Sanz, Miguel A.; Sierra, Jorge; Odero, María D.; Ciencias; Zientziak; Gobierno de Navarra / Nafarroako Gobernua
Background The EVI1 gene (3q26) codes for a zinc finger transcription factor with important roles in both mammalian development and leukemogenesis. Over-expression of EVI1 through either 3q26 rearrangements, MLL fusions, or other unknown mechanisms confers a poor prognosis in acute myeloid leukemia. Design and Methods We analyzed the prevalence and prognostic impact of EVI1 over-expression in a series of 476 patients with acute myeloid leukemia, and investigated the epigenetic modifications of the EVI1 locus which could be involved in the transcriptional regulation of this gene. Results Our data provide further evidence that EVI1 over-expression is a poor prognostic marker in acute myeloid leukemia patients less than 65 years old. Moreover, we found that patients with no basal expression of EVI1 had a better prognosis than patients with expression/over-expression (P=0.036). We also showed that cell lines with over-expression of EVI1 had no DNA methylation in the promoter region of the EVI1 locus, and had marks of active histone modifications: H3 and H4 acetylation, and trimethylation of histone H3 lysine 4. Conversely, cell lines with no expression of EVI1 have DNA hypermethylation and are marked by repressive trimethylation of histone H3 lysine 27 at the EVI1 promoter. Conclusions Our results identify EVI1 over-expression as a poor prognostic marker in a large, independent cohort of acute myeloid leukemia patients less than 65 years old, and show that the total absence of EVI1 expression has a prognostic impact on the outcome of such patients. Furthermore, we demonstrated for the first time that an aberrant epigenetic pattern involving DNA methylation, H3 and H4 acetylation, and trimethylation of histone H3 lysine 4 and histone H3 lysine 27 might play a role in the transcriptional regulation of EVI1 in acute myeloid leukemia. This study opens new avenues for a better understanding of the regulation of EVI1 expression at a transcriptional level.
Open Access
Assessment of minimal residual disease by next generation sequencing in peripheral blood as a complementary tool for personalized transplant monitoring in myeloid neoplasms
(MDPI, 2020) Aguirre-Ruiz, Paula; Ariceta, Beñat; Viguria Alegría, María Cruz; Zudaire, María Teresa; Blasco-Iturri, Zuriñe; Arnedo, Patricia; Aguilera-Diaz, Almudena; Jauregui, Axier; Mañú, Amagoia; Prósper, Felipe; Mateos, María Carmen; Fernández-Mercado, Marta; Larráyoz, María José; Redondo, Margarita; Calasanz, María José; Bandrés Elizalde, Eva; Vázquez Urio, Iria; Ciencias de la Salud; Osasun Zientziak; Gobierno de Navarra / Nafarroako Gobernua
Patients with myeloid neoplasms who relapsed after allogenic hematopoietic stem cell transplant (HSCT) have poor prognosis. Monitoring of chimerism and specific molecular markers as a surrogate measure of relapse is not always helpful; therefore, improved systems to detect early relapse are needed. We hypothesized that the use of next generation sequencing (NGS) could be a suitable approach for personalized follow-up post-HSCT. To validate our hypothesis, we analyzed by NGS, a retrospective set of peripheral blood (PB) DNA samples previously evaluated by high-sensitive quantitative PCR analysis using insertion/deletion polymorphisms (indel-qPCR) chimerism engraftment. Post-HCST allelic burdens assessed by NGS and chimerism status showed a similar time-course pattern. At time of clinical relapse in 8/12 patients, we detected positive NGS-based minimal residual disease (NGS-MRD). Importantly, in 6/8 patients, we were able to detect NGS-MRD at time points collected prior to clinical relapse. We also confirmed the disappearance of post-HCST allelic burden in non-relapsed patients, indicating true clinical specificity. This study highlights the clinical utility of NGS-based post-HCST monitoring in myeloid neoplasia as a complementary specific analysis to high-sensitive engraftment testing. Overall, NGS-MRD testing in PB is widely applicable for the evaluation of patients following HSCT and highly valuable to personalized early treatment intervention when mixed chimerism is detected.
Open Access
Overexpression of GATA2 predicts an adverse prognosis for patients with acute myeloid leukemia and it is associated with distinct molecular abnormalities
(Springer Nature, 2012) Vicente, Carmen; Vázquez Urio, Iria; Conchillo, Ana; García-Sánchez, M. A.; Marcotegui, Nerea; Fuster, Óscar; González, Marcos; Calasanz, María José; Lahortiga, Idoya; Odero, María D.; Ciencias; Zientziak; © 2011 Springer Nature Limited.
Acute myeloid leukemia (AML) represents a heterogeneous group of hematological neoplasms associated with the accumulation of acquired genetic and epigenetic aberrations in hematopoietic progenitor cells. Myeloid leukemogenesis is directly linked to the disruption of expression of transcription factors that regulate the proliferation, survival and differentiation of myeloid progenitors. Moreover, overexpression of particular genes has been associated with prognosis in AML, enabling patients with different survival rates to be identified within cytogenetic risk groups. The GATA2 gene encodes a transcription factor that mediates essential functions in hematopoietic stem cell and progenitor cell (HSC/HPC) compartments. The essential role of GATA2 in hematopoiesis is evident in mice lacking GATA2, which are anemic, have fewer HSC/HPC cells and that die by day 10–11 of gestation.