Intratumoral heterogeneity and clonal evolution in blood malignancies and solid tumors

This meeting held at the University of Barcelona in March 2017, brought together scientists and clinicians worldwide to discuss current and future clinico-biological implications of intratumoral heterogeneity (ITH) and subclonal evolution in cancer diagnosis, patient stratification, and treatment resistance in diagnosis, treatment and follow-up. There was consensus that both longitudinal and tumor multi-region studies in matched samples are needed to better understand the dynamics of ITH. The contribution of the epigenome and microenvironment to ITH and subclone evolution remains understudied. It was recommended to combine computational, pathology and imaging tools to study the role of the microenvironment in subclone selection/ evolution

Tratamiento de la pseudoartrosis de fracturas intertrocantéreas: a propósito de dos casos

Las fracturas intertrocantéreas se tratan en la mayoría de los casos mediante fijación interna con una tasa de éxitos cercana al 100%. En el caso de fracaso se puede optar por la re-osteosíntesis o por la reconversión a artroplastia de cadera. Se presenta un caso de cada solución. Caso 1: paciente de 69 años que presenta fractura pertrocantérea intervenida mediante sistema cefalomedular con defectos técnicos; pseudoartrosis a los 10 meses, rescate con el mismo sistema de fijación interna a foco cerrado y percutáneo con buen resultado y sin necrosis aséptica tras cuatro años de seguimiento. Caso 2: paciente de 43 años con fractura pertrocantérea intervenida mediante sistema cefalomedular que presenta "cutout" a los 16 meses, es reintervenido mediante la colocación de una artroplastia total de cadera híbrida. Se realiza revisión de la bibliografía existente sobre el tema.Most intertrochanteric fractures can be treated successfully with internal fixation. Nevertheless, in a small percentage of patients, the fracture fails to heal. The two mains treatment options are internal fixation of the ununited fracture or salvage treatment with a hip arthroplasty. We pre- sent one case of each treatment. Case 1: female 69 years old with intertrochanteric fracture treated by cephalomedullary internal fixation system, ten months later the fracture didn ́t heal. She was re-operated with same internal fixation system without fracture exposure and fracture healed. Case 2: male 43 years old with intertrochanteric fracture treated with cepa- lomedullary system, 16 months later it was ununited. It was salvaged with hybrid total hip arthroplasty. A is done biblio- graphy rewiew

Clavo Gamma 3 (Stryker) para el tratamiento de fracturas trocantéreas: nuestra experiencia durante el año 2005

Objetivos: estudiar los resultados del sistema de enclavado Gamma 3 en el tratamiento de las fracturas trocantéreas durante el año 2005. Material y Métodos: 73 pacientes con fractura trocantérea mayores de 60 años tratados mediante enclavado céfalo-medular Gamma 3 durante el año 2005 con un seguimiento de 8 a18 meses. Edad media 80 años, 40% de los pacientes deambulantes de forma independiente previo a la fractura, 50% ASA III y IV. Resultados: tiempo quirúrgico 90 minutos, 54% de pacientes transfundidos con una media de 0,89 concentrados por paciente sin diferencias estadisticamente significativas entre aquellos en lo que se colocó y tapón y los que no; 63% de los pacientes deambulan si ayuda en el pstoperatorio. Conclusiones: el sistema Gamma 3 proporciona resultados fiables y reproducible en cualquier variedad de fractura trocanterea y nos parece un sistema eficaz y válido para el tratamiento de este tipo de fracturas.Objectives: to study the results of Gamma 3 for the treatment of trochanteric fractures. Material and Methods: 73 patients older than 60 years old with trochanteric fracture tre- ated using Gamma 3 nailing during year 2005. Follow-up 8-18 months. Mean age 80 years old, 40% preoperative walking without help, 50% ASA III and IV. Results: surgical time 90 minutes, 54% of the patients need blood transfusion with 0,89 concentrates per patients mean. No difference was found in blood transfusion with the use of end cup. 63% walking wit- hout help of with cane postoperatively. Conclusions: Gamma 3 nail is a valid system for the treatment of all types of tro- chanteric fractures

Aplasia parcial congénita de arco posterior de atlas.

Los defectos de arco posterior de C1 son anomalías raras pero bien documentadas en la bibliografía, la mayoría de los casos son asintomáticos pero existen casos publicados de mielopatía y tetraparesia transitoria asociadas a defectos de arco posterior de atlas. Se presentan dos casos: una mujer de 31 años que presentó cérvico-braquialgia secundaria a accidente de tráfico en la que se detectó aplasia parcial de arco posterior de atlas que un año después continúa con dolor y un paciente de 28 años que sufrió accidente de tráfico en la que se detectó aplasia parcial de arco posterior de atlas. Se realiza una revisión bibliográfica.Partial aplasia of posterior arch of the atlas are rare but well documented anomalies, most cases are asymp- tomatic but cases of myelopaty and quadraparesis associa- ted to defects of posterior arch of the atlas have been publis- hed in the past. Case 1: female, 31 years old, suffered traffic accident with neck and upper limb pain. Partial aplasia of posterior arch to the atlas was detected in X-Rays, at the moment the patient continues with neck pain. Case 2: female 28 years old who referred neck pain after traffic accident, partial aplasia of posterior arch of the atlas was detected. Bibliographic review

Identification Of A Germline F692L Drug Resistance Variant In Cis With Flt3-ITD In Knock-In Mice

Letter to the Editor.-- Dovey, Oliver M. et al.Internal tandem duplication (ITD) mutations in the juxtamembrane domain of the fms-like tyrosine kinase 3 (FLT3) gene occur in approximately one quarter of cases of acute myeloid leukemia (AML), are associated with constitutive activation of the kinase and confer a poor prognosis.BC is funded by the >China Scholarship Council> for his visiting studies in UK. AM is funded by the Kay Kendall Leukaemia Fund project grant. CG was funded by a Bloodwise Clinical Research Training Fellowship. IV is funded by Spanish Ministerio de Economía y Competitividad subprograma Ramón y Cajal. We thank Servicio Santander Supercomputación for their support. OMD, JLC and GSV are funded by a Wellcome Trust Senior Fellowship in Clinical Science (WT095663MA) and this work was also funded by the Wellcome Trust Sanger InstitutePeer Reviewe

Identification Of A Germline F692L Drug Resistance Variant In Cis With Flt3-ITD In Knock-In Mice

Letter to the Editor.-- Dovey, Oliver M. et al.Internal tandem duplication (ITD) mutations in the juxtamembrane domain of the fms-like tyrosine kinase 3 (FLT3) gene occur in approximately one quarter of cases of acute myeloid leukemia (AML), are associated with constitutive activation of the kinase and confer a poor prognosis.BC is funded by the >China Scholarship Council> for his visiting studies in UK. AM is funded by the Kay Kendall Leukaemia Fund project grant. CG was funded by a Bloodwise Clinical Research Training Fellowship. IV is funded by Spanish Ministerio de Economía y Competitividad subprograma Ramón y Cajal. We thank Servicio Santander Supercomputación for their support. OMD, JLC and GSV are funded by a Wellcome Trust Senior Fellowship in Clinical Science (WT095663MA) and this work was also funded by the Wellcome Trust Sanger InstitutePeer Reviewe

Evaluation of Toll-like-receptor gene family variants as prognostic biomarkers in rheumatoid arthritis

Rheumatoid arthritis (RA) is a systemic autoimmune disease whose main feature is persistent joint inflammation. Toll-like receptors (TLRs) play critical roles in the activation of innate and adaptive immune responses, and influence the activity of NF?B, a key player in chronic inflammation. We aimed at investigating the association of TLR allelic variants with susceptibility and severity of RA through a systematic, high-throughput, analysis of TLR genes. All coding exons and flanking regions of nine members of the TLR family (TLR1-9) were analyzed in 66 patients with RA and 30 healthy controls by next generation sequencing. We focussed on three single allelic variants, N248S in TLR1, Q11L in TLR7 and M1V in TLR8 based on the allelic frequencies in both patient and control populations, the predicted impact on protein function and the novelty in RA research. Analysis of these selected variants in a larger cohort of 402 patients with RA and in 208 controls revealed no association with susceptibility. However, the M1V allele was associated with a lower need for disease-modifying antirheumatic drugs (DMARDs) (p =0.008) and biologic treatments (p =0.021). Functional studies showed that the M1V variant leads to a reduced production of inflammatory cytokines, IL-1?, IL-6 and TNF?, in response to TLR8 agonists. Thus, the presence of this variant confers a significant protective effect on disease severity. These results show for the first time the association between the M1V variant of TLR8 and reduced disease severity in RA, which could have prognostic value for these patients.This work was supported by the Instituto de Salud Carlos III (ISCIII), Spanish Ministry of Science and Innovation grant PI11/02012, and grant RD12/0036/0022 from Red Temática de Investigación Cooperativa en Cáncer, Sociedad Española de Reumatología grant FER13/13 and Instituto de Investigación Valdecilla (IDIVAL) grant APG-03. I.V. is funded by programa Ramón y Cajal, Ministerio de Economia y Competitividad, Spain

Analysis pipelines for cancer genome sequencing in mice

Mouse models of human cancer have transformed our ability to link genetics, molecular mechanisms and phenotypes. Both reverse and forward genetics in mice are currently gaining momentum through advances in next-generation sequencing (NGS). Methodologies to analyze sequencing data were, however, developed for humans and hence do not account for species-specific differences in genome structures and experimental setups. Here, we describe standardized computational pipelines specifically tailored to the analysis of mouse genomic data. We present novel tools and workflows for the detection of different alteration types, including single-nucleotide variants (SNVs), small insertions and deletions (indels), copy-number variations (CNVs), loss of heterozygosity (LOH) and complex rearrangements, such as in chromothripsis. Workflows have been extensively validated and cross-compared using multiple methodologies. We also give step-by-step guidance on the execution of individual analysis types, provide advice on data interpretation and make the complete code available online. The protocol takes 2?7 d, depending on the desired analyses.D.S. is supported by the European Research Council (Consolidator Grant 648521) and the Deutsche Forschungsgemeinschaft (SA1374/4-2; SFB 1321). I.V. is supported by the European Research Council (Starting Grant INTRAHETEROSEQ) and the Spanish Goverment (SAF2016-76758-R). R.R. is supported by the European Research Council (Consolidator Grants PACA-MET and MSCA-ITN-ETN PRECODE), the Deutsche Forschungsgemeinschaft (DFG RA1629/2-1; SFB1243; SFB1321; SFB1335), the German Cancer Consortium Joint Funding Program, and the Deutsche Krebshilfe (70112480)

A functional variant of TLR10 modifies the activity of NFkB and may help predict a worse prognosis in patients with rheumatoid arthritis

Background: Toll-like receptor (TLR) family members are key players in inflammation. TLR10 has been poorly studied in chronic inflammatory disorders, and its clinical relevance in rheumatoid arthritis (RA) is as yet unknown. We aimed at identifying TLR10 variants within all coding regions of the gene in patients with RA as well as studying their functional and clinical significance. Methods: TLR10 gene variants were studied by performing sequencing of 66 patients with RA and 30 control subjects. A selected variant, I473T, was then analyzed in 1654 patients and 1702 healthy control subjects. The capacity of this TLR10 variant to modify the transcriptional activity of nuclear factor kappa-light-chain-enhancer of activated B cells (NFkB) was determined by using a luciferase reporter assay and analyzing the expression of NFkB target genes by quantitative polymerase chain reaction. Differences between groups were analyzed by using the Mann-Whitney U test and the unpaired two-tailed Student’s t test. Results: We detected ten missense variants in the TLR10 gene and focused on the I473T substitution based on allele frequencies and the predicted functional impact. I473T variant is not associated with susceptibility to RA, but it significantly correlates with erosive disease in patients seropositive for antibodies to citrullinated protein antigens (p = 0.017 in the total cohort and p = 0.0049 in female patients) and with a lower response to infliximab treatment as measured by the change in Disease Activity Score in 28 joints (p = 0.012) and by the European League Against Rheumatism criteria (p = 0.049). Functional studies showed that TLR10 reduced activation of the NFkB inflammatory pathway in hematopoietic cells, whereas the I473T variant lacked this inhibitory capacity. Consistently, after exposure to infliximab, cells expressing the I437T variant showed higher NFkB activity than cells carrying wild-type TLR10. Conclusions: A TLR10 allelic variant, I473T, has impaired NFkB inhibitory activity and is highly associated with disease severity and low response to infliximab in patients with RA

Characterization of gene mutations and copy number changes in acute myeloid leukemia using a rapid target enrichment protocol

Prognostic stratification is critical for making therapeutic decisions and maximizing survival of patients with acute myeloid leukemia. Advances in the genomics of acute myeloid leukemia have identified several recurrent gene mutations whose prognostic impact is being deciphered. We used HaloPlex target enrichment and Illumina-based next generation sequencing to study 24 recurrently mutated genes in 42 samples of acute myeloid leukemia with a normal karyotype. Read depth varied between and within genes for the same sample, but was predictable and highly consistent across samples. Consequently, we were able to detect copy number changes, such as an interstitial deletion of BCOR, three MLL partial tandem duplications, and a novel KRAS amplification. With regards to coding mutations, we identified likely oncogenic variants in 41 of 42 samples. NPM1 mutations were the most frequent, followed by FLT3, DNMT3A and TET2. NPM1 and FLT3 indels were reported with good efficiency. We also showed that DNMT3A mutations can persist post-chemotherapy and in 2 cases studied at diagnosis and relapse, we were able to delineate the dynamics of tumor evolution and give insights into order of acquisition of variants. HaloPlex is a quick and reliable target enrichment method that can aid diagnosis and prognostic stratification of acute myeloid leukemia patients.This project was funded by the Wellcome Trust. NB is a fellow of the European Hematology Association and was supported by the Academy of Medical Sciences. EP is a European Hematology Association Advanced Research Fellow. GV is a Wellcome Trust Senior Fellow in Clinical Science. IV is funded by Spanish Ministerio de Economía y Competitividad subprograma Ramón y Cajal