Decoding the molecular heterogeneity of pediatric monomorphic post-solid organ transplant lymphoproliferative disorders.

Posttransplant lymphoproliferative disorders (PTLDs) represent a broad spectrum of lymphoid proliferations, frequently associated with Epstein-Barr virus (EBV) infection. The molecular profile of pediatric monomorphic PTLDs (mPTLDs) has not been elucidated, and it is unknown whether they display similar genetic features as their counterpart in adult and immunocompetent (IMC) pediatric patients. In this study, we investigated 31 cases of pediatric mPTLD after solid organ transplantation, including 24 diffuse large B-cell lymphomas (DLBCLs), mostly classified as activated B cell, and 7 cases of Burkitt lymphoma (BL), 93% of which were EBV positive. We performed an integrated molecular approach, including fluorescence in situ hybridization, targeted gene sequencing, and copy number (CN) arrays. Overall, PTLD-BL carried mutations in MYC, ID3, DDX3X, ARID1A, or CCND3 resembling IMC-BL, higher mutational burden than PTLD-DLBCL, and lesser CN alterations than IMC-BL. PTLD-DLBCL showed a very heterogeneous genomic profile with fewer mutations and CN alterations than IMC-DLBCL. Epigenetic modifiers and genes of the Notch pathway were the most recurrently mutated in PTLD-DLBCL (both 28%). Mutations in cell cycle and Notch pathways correlated with a worse outcome. All 7 patients with PTLD-BL were alive after treatment with pediatric B-cell non-Hodgkin lymphoma protocols, whereas 54% of patients with DLBCL were cured with immunosuppression reduction, rituximab, and/or low-dose chemotherapy. These findings highlight the low complexity of pediatric PTLD-DLBCL, their good response to low-intensity treatment, and the shared pathogenesis between PTLD-BL and EBV-positive IMC-BL. We also suggest new potential parameters that could help in the diagnosis and the design of better therapeutic strategies for these patients.

Diverse mutations and structural variations contribute to Notch signaling deregulation in paediatric T-cell lymphoblastic lymphoma.

BACKGROUND: T-cell lymphoblastic lymphoma (T-LBL) is an aggressive neoplasm closely related to T-cell acute lymphoblastic leukaemia (T-ALL). Despite their similarities, and contrary to T-ALL, studies on paediatric T-LBL are scarce and, therefore, its molecular landscape has not yet been fully elucidated. Thus, the aims of this study were to characterize the genetic and molecular heterogeneity of paediatric T-LBL and to evaluate novel molecular markers differentiating this entity from T-ALL. PROCEDURE: Thirty-three paediatric T-LBL patients were analyzed using an integrated approach, including targeted next-generation sequencing, RNA-sequencing transcriptome analysis and copy-number arrays. RESULTS: Copy number and mutational analyses allowed the detection of recurrent homozygous deletions of 9p/CDKN2A (78%), trisomy 20 (19%) and gains of 17q24-q25 (16%), as well as frequent mutations of NOTCH1 (62%), followed by the BCL11B (23%), WT1 (19%) and FBXW7, PHF6 and RPL10 genes (15%, respectively). This genetic profile did not differ from that described in T-ALL in terms of mutation incidence and global genomic complexity level, but unveiled virtually exclusive 17q25 gains and trisomy 20 in T-LBL. Additionally, we identified novel gene fusions in paediatric T-LBL, including NOTCH1-IKZF2, RNGTT-SNAP91 and DDX3X-MLLT10, the last being the only one previously described in T-ALL. Moreover, clinical correlations highlighted the presence of Notch pathway alterations as a factor related to favourable outcome. CONCLUSIONS: In summary, the genomic landscape of paediatric T-LBL is similar to that observed in T-ALL, and Notch signaling pathway deregulation remains the cornerstone in its pathogenesis, including not only mutations but fusion genes targeting NOTCH1.

Relevance of dynamic studies with magnetic resonance enterography in Crohn's disease.

INTRODUCTION: A proper quantification of the inflammatory activity in Crohn's disease (CD) lesions is needed to establish the appropriate management for each patient. The aim of this study is to evaluate the inflammatory activity of affected segments in small bowel lesions using dynamic studies of magnetic resonance enterography (MRE) in patients undergoing surgery, and their correlation with the level of inflammation and histological fibrosis of the surgical piece. METHODS: A prospective, consecutive, observational, clinical study was conducted that included all the patients with small bowel CD that underwent surgery in this center between March 2011 and September 2013. Diagnosis was established according to Lennard-Jones criteria and the Montreal classification. All the patients underwent MRE within three months before surgery, using a routine protocol involving Liver Acquisition with Volume Acceleration-Extended Volume (LAVA-XV) sequence for the dynamic studies before intravenous administering of gadolinium and 30, 70, 120, and 420s after administering this. The results allowed the designing of graphics with different uptake patterns. The Chiorean classification was used in the histological analysis, as well as a modified version published previously by this study group. RESULTS: A total of 28 patients with 47 lesions were analyzed. There was a significant correlation between both curve patterns, including the modified Chiorean classification (P<0.0001) as well as the level of inflammation (P<0.0001) and fibrosis (P<0.002). Inflammatory patterns of dynamic studies are related to histological findings with 80.9% accuracy (sensitivity=75.7%; specificity=100%). CONCLUSION: There is a high correlation between dynamic enhancement studies and the level of inflammatory activity. MRE is a suitable tool to differentiate between inflammatory and fibrotic lesions, making it useful to decide the appropriate management of each patient.

Presentation and Long-term Outcome of High-grade Osteosarcoma: A Single-institution Experience.

PURPOSE: To evaluate clinicopathologic characteristics, prognostic factors, and treatment outcome of pediatric/adolescent high-grade osteosarcoma patients. METHODS/PATIENTS: Retrospective evaluation of patients 21 years of age or younger with newly diagnosed high-grade osteosarcoma treated in a single institution. Effects of variables on event-free survival and overall survival (OS) were determined by using Kaplan-Meier survival analysis. Variables found to be significant were evaluated with multivariable Cox regression analysis. RESULTS: Seventy-seven patients diagnosed between January 1985 and December 2011 were included. Median follow-up time was 11.0 years (range, 1.6 to 26.4 y). Event-free survival at 5 and 10 years was 38%±11% and 38%±11%, respectively. OS at 5 and 10 years was 51%±12% and 45%±12%, respectively. Metastatic disease, prolonged time interval to resumption of chemotherapy, lower tumor necrosis rate, and lack of achievement of complete response at the end of first-line chemotherapy treatment were associated with inferior OS probabilities in univariate analysis. Upon multivariate analysis, only achievement of complete response at the end of first-line chemotherapy and tumor necrosis rate retained independent prognostic significance. CONCLUSIONS: Prognostic factors and long-term survival are similar to those previously described. Reduction of global time interval to resumption of chemotherapy as well as a more specific and validated definition of pulmonary metastases at diagnosis are needed.

[Clinicopathological diagnosis of blastic plasmacytoid dendritic cell neoplasm: Report of three cases]. / Diagnóstico clínico-patológico de la neoplasia de células dendríticas plasmocitoides blásticas: reporte de 3 casos.

Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is a rare disease with a poor prognosis. It frequently affects the skin; indeed, dermal lesions may be the first clinical manifestation. We report three cases of BPDCN where the patients presented with skin lesions and describe the clinical, histopathological and immunohistochemical findings, its molecular characteristics and metastatic work-up. One of the patients remains in a clinical trial with IMGN632, a molecule directed against CD123, while the other two patients died after different therapeutic regimens. BPDCN is a complex diagnostic challenge which, together with its poor prognosis, requires close clinical-pathological cooperation in order to accelerate its diagnosis and offer early therapeutic alternatives with drugs directed against specific molecular targets.

Germline variant in Ctcf links mental retardation to Wilms tumor predisposition.

CTCF germline mutations have been related to MRD21. We report the first bilateral Wilms tumor suffered by a MRD21 patient carrying an unreported CTCF missense variant in a zinc finger domain of CTCF protein. We found that germline heterozygous variant I446K became homozygous in the tumor due to a loss of heterozygosity rearrangement affecting the whole q arm on chromosome 16. Our findings propose CTCF I446K variant as a link between MRD21 and Wilms tumor predisposition.

Next-Generation Sequencing Identifies Potential Actionable Targets in Paediatric Sarcomas.

Background: Bone and soft-tissue sarcomas represent 13% of all paediatric malignancies. International contributions to introduce next-generation sequencing (NGS) approaches into clinical application are currently developing. We present the results from the Precision Medicine program for children with sarcomas at a reference centre. Results: Samples of 70 paediatric sarcomas were processed for histopathological analysis, reverse transcriptase polymerase chain reaction (RT-PCR) and next-generation sequencing (NGS) with a consensus gene panel. Pathogenic alterations were reported and, if existing, targeted recommendations were translated to the clinic. Seventy paediatric patients with sarcomas from 10 centres were studied. Median age was 11.5 years (range 1-18). Twenty-two (31%) had at least one pathogenic alteration by NGS. Thirty pathogenic mutations in 18 different genes were detected amongst the 22 patients. The most frequent alterations were found in TP53, followed by FGFR4 and CTNNB1. Combining all biological studies, 18 actionable variants were detected and six patients received targeted treatment observing a disease control rate of 78%. Extrapolating the results to the whole cohort, 23% of the patients would obtain clinical benefit from this approach. Conclusions: Paediatric sarcomas have a different genomic landscape when compared to adult cohorts. Incorporating NGS targets into paediatric sarcomas' therapy is feasible and allows personalized treatments with clinical benefit in the relapse setting.

Retinoblastoma and mosaic 13q deletion: a case report.

BACKGROUND: Patients with 13q-syndrome are at risk of retinoblastoma when the RB1 gene, located in the chromosomal band 13q14.2, is deleted. This syndrome is frequently associated with congenital malformations and developmental delay, although these signs could be mild. Mosaic 13q-deletion patients have been previously reported in the literature; their phenotype is variable, and they may not be recognized. CASE PRESENTATION: Retinoblastoma diagnosed in a child with 13q-mosaicism confirmed in blood, oral mucosa, healthy retina and retinoblastoma. A second RB1 hit is present exclusively in the retinoblastoma sample (RB1 c.958C>T p.Arg320Ter). Other detected molecular events in retinoblastoma are 6p12.3pter gain and 6q25.3qter loss. Clinical examination is unremarkable except for clinodactyly of the right fifth finger. DISCUSSION AND CONCLUSIONS: We describe a case of mosaic 13q deletion syndrome affected by retinoblastoma. Molecular data obtained from the tumor analysis are similar to previous data available about this malignancy. High clinical suspicion is essential for an adequate diagnosis of mosaic cases.

Testicular tumours in children: an approach to diagnosis and management with pathologic correlation.

Testicular tumours are rare in children. Painless scrotal mass is the most frequent clinical presentation. Tumoural markers (alpha-fetoprotein, beta-human gonadotropin chorionic) and hormone levels (testosterone) contribute to the diagnosis and management of a testicular mass in boys. Ultrasonography is the best imaging modality to study testicular tumours. A benign tumour is suggested when ultrasonography shows a mainly cystic component, well-defined borders, echogenic rim or normal to increased echogenicity lesion when compared to the healthy testicular parenchyma. Malignant tumour is suspected when ultrasonography shows inhomogeneous, hypoechoic, not well-circumscribed or diffuse infiltration lesion. However, these ultrasonographic findings may overlap. Colour Doppler, power Doppler, elastography and contrast-enhanced ultrasonography are useful complementary methods to characterise the focal testicular lesions. Chest computerised tomography and abdominopelvic magnetic resonance are necessary to establish the extension in case of malignant proved tumours.Benign tumours are more frequent in prepuberal boys and malignant tumours in pubertal boys. Mature teratoma prepubertal-type is the most common histologic type. Testicular sparing surgery is the choice in benign tumours. Radical inguinal orchiectomy is indicated in malignant tumours. Prognostic is excellent.The purpose of our study is to show an approach to the diagnosis and management of the most frequent testicular tumours in children according to clinical manifestations, imaging findings and tumour markers levels based on histologically confirmed tumours in our hospital.

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.

Diagnóstico clínico-patológico de la neoplasia de células dendríticas plasmocitoides blásticas: reporte de 3 casos / Clinicopathological diagnosis of blastic plasmacytoid dendritic cell neoplasm: Report of three cases

La neoplasia de células dendríticas plasmocitoides blásticas (NCDPB) es una enfermedad de baja incidencia y muy mal pronóstico, que con gran frecuencia afecta a la piel, pudiendo ser el primer signo clínico de la enfermedad. Se presentan 3 casos en los que la primera manifestación de la enfermedad fueron lesiones cutáneas. Se describe el cuadro clínico, los hallazgos histopatológicos e inmunohistoquímicos, así como los estudios de extensión y las características moleculares de las 3 neoplasias. Uno de los pacientes permanece en un ensayo clínico con IMGN632, una molécula dirigida contra CD123, mientras que los otros 2 pacientes fallecieron tras distintos regímenes terapéuticos. La NCDPB es una entidad de diagnóstico complejo. Esto, unido a su mal pronóstico, obligan a una comunicación clínico-patológica estrecha que acelere su diagnóstico y ofrezca alternativas terapéuticas precoces con fármacos dirigidos contra dianas moleculares específicas de esta entidad. (AU)

Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is a rare disease with a poor prognosis. It frequently affects the skin; indeed, dermal lesions may be the first clinical manifestation. We report three cases of BPDCN where the patients presented with skin lesions and describe the clinical, histopathological and immunohistochemical findings, its molecular characteristics and metastatic work-up. One of the patients remains in a clinical trial with IMGN632, a molecule directed against CD123, while the other two patients died after different therapeutic regimens. BPDCN is a complex diagnostic challenge which, together with its poor prognosis, requires close clinical-pathological cooperation in order to accelerate its diagnosis and offer early therapeutic alternatives with drugs directed against specific molecular targets. (AU)

Diagnóstico clínico-patológico de la neoplasia de células dendríticas plasmocitoides blásticas: reporte de 3 casos / Clinicopathological diagnosis of blastic plasmacytoid dendritic cell neoplasm: Report of three cases

La neoplasia de células dendríticas plasmocitoides blásticas (NCDPB) es una enfermedad de baja incidencia y muy mal pronóstico, que con gran frecuencia afecta a la piel, pudiendo ser el primer signo clínico de la enfermedad. Se presentan 3 casos en los que la primera manifestación de la enfermedad fueron lesiones cutáneas. Se describe el cuadro clínico, los hallazgos histopatológicos e inmunohistoquímicos, así como los estudios de extensión y las características moleculares de las 3 neoplasias. Uno de los pacientes permanece en un ensayo clínico con IMGN632, una molécula dirigida contra CD123, mientras que los otros 2 pacientes fallecieron tras distintos regímenes terapéuticos. La NCDPB es una entidad de diagnóstico complejo. Esto, unido a su mal pronóstico, obligan a una comunicación clínico-patológica estrecha que acelere su diagnóstico y ofrezca alternativas terapéuticas precoces con fármacos dirigidos contra dianas moleculares específicas de esta entidad. (AU)

Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is a rare disease with a poor prognosis. It frequently affects the skin; indeed, dermal lesions may be the first clinical manifestation. We report three cases of BPDCN where the patients presented with skin lesions and describe the clinical, histopathological and immunohistochemical findings, its molecular characteristics and metastatic work-up. One of the patients remains in a clinical trial with IMGN632, a molecule directed against CD123, while the other two patients died after different therapeutic regimens. BPDCN is a complex diagnostic challenge which, together with its poor prognosis, requires close clinical-pathological cooperation in order to accelerate its diagnosis and offer early therapeutic alternatives with drugs directed against specific molecular targets. (AU)

Relevance of dynamic studies with magnetic resonance enterography in Crohn's disease / Relevancia de los estudios dinámicos en Resonancia Entérica en la Enfermedad de Crohn

INTRODUCTION: A proper quantification of the inflammatory activity in Crohn's disease (CD) lesions is needed to establish the appropriate management for each patient. The aim of this study is to evaluate the inflammatory activity of affected segments in small bowel lesions using dynamic studies of magnetic resonance enterography (MRE) in patients undergoing surgery, and their correlation with the level of inflammation and histological fibrosis of the surgical piece. METHODS: A prospective, consecutive, observational, clinical study was conducted that included all the patients with small bowel CD that underwent surgery in this center between March 2011 and September 2013. Diagnosis was established according to Lennard-Jones criteria and the Montreal classification. All the patients underwent MRE within three months before surgery, using a routine protocol involving Liver Acquisition with Volume Acceleration-Extended Volume (LAVA-XV) sequence for the dynamic studies before intravenous administering of gadolinium and 30, 70, 120, and 420s after administering this. The results allowed the designing of graphics with different uptake patterns. The Chiorean classification was used in the histological analysis, as well as a modified version published previously by this study group. RESULTS: A total of 28 patients with 47 lesions were analyzed. There was a significant correlation between both curve patterns, including the modified Chiorean classification (p < 0.0001) as well as the level of inflammation (p < 0.0001) and fibrosis (p < 0.002). Inflammatory patterns of dynamic studies are related to histological findings with 80.9% accuracy (sensitivity=75.7%; specificity=100%). CONCLUSION: There is a high correlation between dynamic enhancement studies and the level of inflammatory activity. MRE is a suitable tool to differentiate between inflammatory and fibrotic lesions, making it useful to decide the appropriate management of each patient

INTRODUCCIÓN: Se necesita una cuantificación adecuada de la actividad inflamatoria en las lesiones de la enfermedad de Crohn (EC) para establecer el tratamiento adecuado para cada paciente. El objetivo de este estudio es evaluar la actividad inflamatoria de los segmentos afectados en las lesiones del intestino delgado mediante estudios dinámicos de enterografía por resonancia magnética (ERM). MÉTODOS: Estudio prospectivo, consecutivo, observacional y clínico, que incluye a todos los pacientes con EC del intestino delgado que se sometieron a cirugía en nuestro centro entre marzo de 2011 y septiembre de 2013. El diagnóstico se estableció de acuerdo con los criterios de Lennard-Jones y la clasificación de Montreal. Todos los pacientes se sometieron a una ERM dentro de los 3 meses previos a la cirugía, aplicando el protocolo de rutina y secuencias preestablecidas. Para el estudio dinámico se empleó la secuencia Adquisición hepática con aceleración de volumen-Volumen extendido (LAVA-XV), antes de la administración intravenosa (IV) de gadolinio, y 30, 70, 120 y 420s después de esta administración. Los resultados permiten diseñar gráficos con diferentes patrones de captación. En el análisis histológico se utilizó la clasificación de Chiorean, así como una versión modificada creada por nuestro grupo de estudio. RESULTADOS: En total se analizaron 28 pacientes con 47 lesiones. Se detectó una correlación significativa entre ambos patrones de curva, incluyendo la clasificación de Chiorean modificada (p < 0,0001), así como el grado inflamatorio (p < 0,0001) y de fibrosis (p < 0,002). Los patrones inflamatorios de los estudios dinámicos se relacionaron con los hallazgos histológicos con una precisión del 80,9% (S=75,7%; E=100%). CONCLUSIÓN: Existe una alta correlación entre los estudios dinámicos y el grado de actividad inflamatoria. La ERM constituye una herramienta adecuada para diferenciar entre lesiones inflamatorias y fibróticas, siendo útil para colaborar en la decisión terapéutica