microRNA sequencing for biomarker detection in the diagnosis, classification and prognosis of Diffuse Large B Cell Lymphoma.

Despite being considered a single disease, Diffuse Large B Cell Lymphoma (DLBCL) presents with variable backgrounds, which results in heterogeneous outcomes among patients, with 40% of them still having primary refractory disease or relapse. Thus, novel biomarkers are needed. In addition, multiple factors regarding its pathogenesis remain unclear. In this context, recent investigations point to the relevance of microRNAs (miRNAs) in cancer. However, regarding DLBCL, there is inconsistency in the data reported. Therefore, in this work, the main goals were to determine a miRNA set with utility as biomarkers for DLBCL diagnosis, classification, prognosis and treatment response, as well as to decipher the mechanism of action of deregulated miRNAs in the origin of the disease. We analyzed miRNA expression in a cohort of 78 DLBCL patients and 17 controls using small RNA sequencing and performed a miRNA-mRNA interaction network analysis. This way, we were able to define new miRNA expression signatures for diagnosis, classification, treatment response and prognosis, and we identified plausible mechanisms of action by which deregulated miRNAs could be involved in DLBCL pathogenesis. In summary, our study remarks that miRNAs could play an important role in DLBCL.

SOX9 Triggers Different Epithelial to Mesenchymal Transition States to Promote Pancreatic Cancer Progression.

BACKGROUND: Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal cancers mainly due to spatial obstacles to complete resection, early metastasis and therapy resistance. The molecular events accompanying PDAC progression remain poorly understood. SOX9 is required for maintaining the pancreatic ductal identity and it is involved in the initiation of pancreatic cancer. In addition, SOX9 is a transcription factor linked to stem cell activity and is commonly overexpressed in solid cancers. It cooperates with Snail/Slug to induce epithelial-mesenchymal transition (EMT) during neural development and in diseases such as organ fibrosis or different types of cancer. METHODS: We investigated the roles of SOX9 in pancreatic tumor cell plasticity, metastatic dissemination and chemoresistance using pancreatic cancer cell lines as well as mouse embryo fibroblasts. In addition, we characterized the clinical relevance of SOX9 in pancreatic cancer using human biopsies. RESULTS: Gain- and loss-of-function of SOX9 in PDAC cells revealed that high levels of SOX9 increased migration and invasion, and promoted EMT and metastatic dissemination, whilst SOX9 silencing resulted in metastasis inhibition, along with a phenotypic reversion to epithelial features and loss of stemness potential. In both contexts, EMT factors were not altered. Moreover, high levels of SOX9 promoted resistance to gemcitabine. In contrast, overexpression of SOX9 was sufficient to promote metastatic potential in K-Ras transformed MEFs, triggering EMT associated with Snail/Slug activity. In clinical samples, SOX9 expression was analyzed in 198 PDAC cases by immunohistochemistry and in 53 patient derived xenografts (PDXs). SOX9 was overexpressed in primary adenocarcinomas and particularly in metastases. Notably, SOX9 expression correlated with high vimentin and low E-cadherin expression. CONCLUSIONS: Our results indicate that SOX9 facilitates PDAC progression and metastasis by triggering stemness and EMT.

Endothelial cell loss rate after penetrating keratoplasty: optical versus therapeutic grafts.

Background: This study aimed to compare the rate of endothelial cell loss (ECL) after penetrating keratoplasty (PKP) for optical versus therapeutic grafts at 3-, 6-, and 12-month postoperatively. Furthermore, the study aimed to investigate postoperative graft viability and the rate of graft rejection during the first year of follow-up for both indications. Methods: This was a prospective, observational, comparative study that included patients who sought medical advice at the cornea outpatient clinic of Ain Shams University Hospitals, Cairo, Egypt. The study recruited 60 patients: group 1 included 30 transplanted corneas of 30 patients who underwent optical PKP for various indications, while group 2 included 30 transplanted corneas of 30 patients who underwent therapeutic PKP for unhealed, resistant infectious keratitis. Specular microscopy was performed for all patients at the 3-, 6-, and 12-month follow-up visits using Nidek CEM-530 specular microscopy. Postoperative clinical examinations were performed at the same follow-up visits to detect graft rejection. Results: There were no statistically significant differences between the groups concerning the postoperative timing of graft clarity or the rate of ECL at 3- and 6-months postoperatively; however, the rate of ECL was significantly greater in group 2 than in group 1 at 12-months postoperatively (P = 0.03), although the difference was small from a clinical point of view. Moreover, there was no statistically significant difference between the groups in terms of the graft rejection rate. Conclusions: Therapeutic PKP results were comparable to optical PKP with respect to graft viability, the rate of ECL, and the rate of graft rejection 1 year after grafting.

Identification of Recurrent Mutations in the microRNA-Binding Sites of B-Cell Lymphoma-Associated Genes in Follicular Lymphoma.

Follicular lymphoma (FL) is a common indolent B-cell lymphoma that can transform into the more aggressive transformed FL (tFL). However, the molecular process driving this transformation is uncertain. In this work, we aimed to identify microRNA (miRNA)-binding sites recurrently mutated in follicular lymphoma patients, as well as in transformed FL patients. Using whole-genome sequencing data from FL tumors, we discovered 544 mutations located in bioinformatically predicted microRNA-binding sites. We then studied these specific regions using targeted sequencing in a cohort of 55 FL patients, found 16 recurrent mutations, and identified a further 69 variants. After filtering for QC, we identified 21 genes with mutated miRNA-binding sites that were also enriched for B-cell-associated genes by Gene Ontology. Over 40% of mutations identified in these genes were present exclusively in tFL patients. We validated the predicted miRNA-binding sites of five of the genes by luciferase assay and demonstrated that the identified mutations in BCL2 and EZH2 genes impaired the binding efficiency of miR-5008 and miR-144 and regulated the endogenous levels of messenger RNA (mRNA).

Distinct molecular profile of IRF4-rearranged large B-cell lymphoma.

Pediatric large B-cell lymphomas (LBCLs) share morphological and phenotypic features with adult types but have better prognosis. The higher frequency of some subtypes such as LBCL with IRF4 rearrangement (LBCL-IRF4) in children suggests that some age-related biological differences may exist. To characterize the genetic and molecular heterogeneity of these tumors, we studied 31 diffuse LBCLs (DLBCLs), not otherwise specified (NOS); 20 LBCL-IRF4 cases; and 12 cases of high-grade B-cell lymphoma (HGBCL), NOS in patients ≤25 years using an integrated approach, including targeted gene sequencing, copy-number arrays, and gene expression profiling. Each subgroup displayed different molecular profiles. LBCL-IRF4 had frequent mutations in IRF4 and NF-κB pathway genes (CARD11, CD79B, and MYD88), losses of 17p13 and gains of chromosome 7, 11q12.3-q25, whereas DLBCL, NOS was predominantly of germinal center B-cell (GCB) subtype and carried gene mutations similar to the adult counterpart (eg, SOCS1 and KMT2D), gains of 2p16/REL, and losses of 19p13/CD70. A subset of HGBCL, NOS displayed recurrent alterations of Burkitt lymphoma-related genes such as MYC, ID3, and DDX3X and homozygous deletions of 9p21/CDKN2A, whereas other cases were genetically closer to GCB DLBCL. Factors related to unfavorable outcome were age >18 years; activated B-cell (ABC) DLBCL profile, HGBCL, NOS, high genetic complexity, 1q21-q44 gains, 2p16/REL gains/amplifications, 19p13/CD70 homozygous deletions, and TP53 and MYC mutations. In conclusion, these findings further unravel the molecular heterogeneity of pediatric and young adult LBCL, improve the classification of this group of tumors, and provide new parameters for risk stratification.

Precise Immunodetection of PTEN Protein in Human Neoplasia.

PTEN is a major tumor-suppressor protein whose expression and biological activity are frequently diminished in sporadic or inherited cancers. PTEN gene deletion or loss-of-function mutations favor tumor cell growth and are commonly found in clinical practice. In addition, diminished PTEN protein expression is also frequently observed in tumor samples from cancer patients in the absence of PTEN gene alterations. This makes PTEN protein levels a potential biomarker parameter in clinical oncology, which can guide therapeutic decisions. The specific detection of PTEN protein can be achieved by using highly defined anti-PTEN monoclonal antibodies (mAbs), characterized with precision in terms of sensitivity for the detection technique, specificity for PTEN binding, and constraints of epitope recognition. This is especially relevant taking into consideration that PTEN is highly targeted by mutations and posttranslational modifications, and different PTEN protein isoforms exist. The precise characterization of anti-PTEN mAb reactivity is an important step in the validation of these reagents as diagnostic and prognostic tools in clinical oncology, including their routine use in analytical immunohistochemistry (IHC). Here, we review the current status on the use of well-defined anti-PTEN mAbs for PTEN immunodetection in the clinical context and discuss their potential usefulness and limitations for a more precise cancer diagnosis and patient benefit.

Metástasis en hueso malar de tumor neuroendocrino oncocítico pulmonar / Malar bone metastasis of oncocytic neuroendocrine tumour of the lung

Las metástasis en hueso malar de tumores sólidos infraclaviculares son excepcionales. En aquellos de estirpe neuroendocrina, se debe considerar la expresión de receptores de somatostatina mediante diferentes técnicas de medicina nuclear para realizar terapias dirigidas. Se presenta un caso clínico de un varón de 66 años con neoplasia pulmonar con diferenciación oncocítica, cuyo debut es una metástasis malar de lenta evolución. Es tratado con lanreotido y ácido zolendrónico con enfermedad estable al año de seguimiento

Malar bone metastases of solid infraclavicular tumours are exceptional. Expression of somatostatin receptors should be considered in neuroendocrine strains, in order to carry out targeted therapies. We report a clinical case of a 66-year-old man with a tumour of the lung with oncocytic features, which debut is a malar metastasis of slow evolution. He is treated with Lanreotide and Zolendronic acid with stable disease at 1-year follow-up

Precise definition of PTEN C-terminal epitopes and its implications in clinical oncology.

Anti-PTEN monoclonal antibodies (mAb) are arising as important tools for immunohistochemistry (IHC) and protein quantification routine analysis in clinical oncology. Although an effort has been made to document the reliability of tumor tissue section immunostaining by anti-PTEN mAb, and to standardize their IHC use in research and in the clinical practice, the precise topological and biochemical definition of the epitope recognized by each mAb has been conventionally overlooked. In this study, six commercial anti-PTEN mAb have been validated and characterized for sensitivity and specificity by IHC and FISH, using a set of prostate and urothelial bladder tumor specimens, and by immunoblot, using PTEN positive and PTEN negative human cell lines. Immunoblot precise epitope mapping, performed using recombinant PTEN variants and mutations, revealed that all mAb recognized linear epitopes of 6-11 amino acid length at the PTEN C-terminus. Tumor-associated or disease-associated mutations at the PTEN C-terminus did not affect subcellular localization or PIP3 phosphatase activity of PTEN in cells, although resulted in specific loss of reactivity for some mAb. Furthermore, specific mimicking-phosphorylation mutations at the PTEN C-terminal region also abolished binding of specific mAb. Our study adds new evidence on the relevance of a precise epitope mapping in the validation of anti-PTEN mAb for their use in the clinics. This will be substantial to provide a more accurate diagnosis in clinical oncology based on PTEN protein expression in tumors and biological fluids.

Mutational profiling can identify laryngeal dysplasia at risk of progression to invasive carcinoma.

Early diagnosis of laryngeal squamous cell carcinoma (LSCC) at the stage of dysplasia could greatly improve the outcome of affected patients. For the first time we compared the mutational landscape of non-progressing dysplasia (NPD; n = 42) with progressing dysplasia (PD; n = 24), along with patient-matched LSCC biopsies; a total of 90 samples. Using targeted next-generation sequencing identified non-synonymous mutations in six genes (PIK3CA, FGFR3, TP53, JAK3, MET, FBXW7), and mutations were validated by Sanger sequencing and/or qPCR. Analysis was extended in silico to 530 head and neck (HNSCC) cases using TCGA data. Mutations in PIK3CA and FGFR3 were detected in PD and LSCC cases, as well as other HNSCC cases, but absent in NPD cases. In contrast, mutations in JAK3, MET and FBXW7 were found in NPD cases but not PD, LSCC or other HNSCC cases. TP53 was the most frequently mutated gene in both PD and NPD cases. With the exception of R248W, mutations were mutually exclusive. Moreover, five of seven PD mutations were located in motif H2 of p53, whereas none of the NPD mutations were. In summary, we propose that the mutational profile of laryngeal dysplasia has utility for the early detection of patients at risk of progression.

DNMT1 is predictive of survival and associated with Ki-67 expression in R-CHOP-treated diffuse large B-cell lymphomas.

DNMT1 is a target of approved anti-cancer drugs including decitabine. However, the prognostic value of DNMT1 protein expression in R-CHOP-treated diffuse large B-cell lymphomas (DLBCLs) remains unexplored. Here we showed that DNMT1 was expressed in the majority of DLBCL cases (n = 209/230, 90.9%) with higher expression in germinal centre B-cell-like (GCB)-DLBCL subtype. Low and negative DNMT1 expression (20% cut-off, n = 33/230, 14.3%) was predictive of worse overall survival (OS; p < 0.001) and progression-free survival (PFS; p < 0.001). Nonetheless, of the 209 DNMT1 positive patients, 33% and 42% did not achieve 5-year OS and PFS, respectively, indicating that DNMT1 positive patients showed considerably heterogeneous outcomes. Moreover, DNMT1 was frequently expressed in mitotic cells and significantly correlated with Ki-67 or BCL6 expression (r = 0.60 or 0.44, respectively; p < 0.001). We demonstrate that DNMT1 is predictive of DLBCL patients' survival, and suggest that DNMT1 could be a DLBCL therapeutic target due to its significant association with Ki-67.

[Renal Cell Carcinoma metastases in the maxillofacial area: Case series.] / Metástasis en región maxilofacial de carcinoma renal: serie de casos.

Renal cell carcinoma is an unpredictable malignancy. Sometimes, metastases are the disease debut. On the other hand, metastases could present years after treatment of the primary tumor. Four clinical cases of atypical metastases in the head and neck location are presented: parotid gland, mandibular bone, attached molar gingiva and masticator space. Physiopathology, clinics, histology and management of metastatic renal cell carcinoma at those anatomical regions are reviewed.

Metástasis en región maxilofacial de carcinoma renal: serie de casos / Renal Cell Carcinoma metastases in the maxillofacial area: Case series

El carcinoma renal es un tumor de evolución imprevisible. A veces las metástasis son el debut de la enfermedad, mientras que en otras, éstas se manifiestan años tras el tratamiento del primario. Se presentan 4 casos clínicos de metástasis atípicas de carcinoma renal en región de cabeza y cuello: glándula parótida, hueso mandibular, encía adherida molar y espacio masticador. Se revisa la fisiopatología, clínica, histología y manejo del cáncer renal metastásico en esas localizaciones

Renal cell carcinoma is an unpredictable malignancy. Sometimes, metastases are the disease debut. On the other hand, metastases could present years after treatment of the primary tumor. Four clinical cases of atypical metastases in the head and neck location are presented: parotid gland, mandibular bone, attached molar gingiva and masticator space. Physiopathology, clinics, histology and management of metastatic renal cell carcinoma at those anatomical regions are reviewed

Low HIP1R mRNA and protein expression are associated with worse survival in diffuse large B-cell lymphoma patients treated with R-CHOP.

Huntingtin-interacting protein 1-related (HIP1R) is an endocytic protein involved in receptor trafficking, including regulating cell surface expression of receptor tyrosine kinases. We have previously shown that low HIP1R protein expression was associated with poorer survival in diffuse large B-cell lymphoma (DLBCL) patients from Denmark treated with R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, prednisone). In this multicenter study, we extend these findings and validate the prognostic and subtyping utility of HIP1R expression at both transcript and protein level. Using data mining on three independent transcriptomic datasets of DLBCL, HIP1R transcript was preferentially expressed in germinal center B-cell (GCB)-like DLBCL subtype (P<0.01 in all three datasets), and lower expression was correlated with worse overall survival (OS; P<0.01) and progression-free survival (PFS; P<0.05) in a microarray-profiled DLBCL dataset. At the protein level examined by immunohistochemistry, HIP1R expression at 30% cut-off was associated with GCB-DLBCL molecular subtype (P=0.0004; n=42), and predictive of OS (P=0.0006) and PFS (P=0.0230) in de novo DLBCL patients treated with R-CHOP (n=73). Cases with high FOXP1 and low HIP1R expression frequency (FOXP1(hi)/HIP1R(lo) phenotype) exhibited poorer OS (P=0.0038) and PFS (P=0.0134). Multivariate analysis showed that HIP1R<30% or FOXP1(hi)/HIP1R(lo) subgroup of patients exhibited inferior OS and PFS (P<0.05) independently of the International Prognostic Index. We conclude that HIP1R expression is strongly indicative of survival when utilized on its own or in combination with FOXP1, and the molecule is potentially applicable for subtyping of DLBCL cases.

MYC protein expression is associated with poor prognosis in primary diffuse large B-cell lymphoma of the central nervous system.

MYC and BCL2 gene translocations and protein expression have recently demonstrated to be of prognostic significance in systemic diffuse large B-cell lymphoma (DLBCL). However, their role in primary central nervous system DLBCL (CNS-DLBCL) prognosis has been scarcely analyzed. We studied the immunophenotype, the status of the MYC, BCL2, and BCL6 genes and the clinical features of a series of 42 CNS-DLBCL and evaluated their prognostic significance. We found high MYC protein expression in 43% of cases, and this was associated with lower overall survival (OS). Cases with concurrent expression of MYC and BCL2 showed a lower OS, although the difference did not reach statistical significance. Translocations involving the MYC or BCL2 genes were not detected. The BCL6 gene was frequently translocated, but was unrelated to survival. We conclude that MYC protein expression detected by immunohistochemistry identifies a CNS-DLBCL subset with worse prognosis and may contribute to a more accurate risk stratification of CNS-DLBCL patients.

Should PET/CT be implemented in the routine imaging work-up of locally advanced head and neck squamous cell carcinoma? A prospective analysis.

PURPOSE: The objective of this study was to determine the incremental staging information provided by positron emission tomography/computed tomography (PET/CT) and its impact on management plans in patients with untreated stage III-IV head and neck squamous cell carcinoma (HNSCC). METHODS: We prospectively studied, between September 2011 and February 2013, 84 consecutive patients [median age 63.5 years (39-84); 73 men] with histologically confirmed HNSCC. First, based on a conventional work-up (physical examination, CT imaging of the head, neck and chest), the multidisciplinary Head and Neck Tumour Board documented the TNM stage and a management plan for each patient, outlining the modalities to be used, including surgery, radiation therapy (RT), chemotherapy or a combination. After release of the PET/CT results, new TNM staging and management plans were agreed on by the multidisciplinary Tumour Board. Any changes in stage or intended management due to the PET/CT findings were then analysed. The impact on patient management was classified as: low (treatment modality, delivery and intent unchanged), moderate (change within the same treatment modality: type of surgery, radiation technique/dose) or high (change in treatment intent and/or treatment modality → curative to palliative, or surgery to chemoradiation or detection of unknown primary tumour or a synchronous second primary tumour). TNM stage was validated by histopathological analysis, additional imaging or follow-up. Accuracy of the conventional and PET/CT-based staging was compared using McNemar's test. RESULTS: Conventional and PET/CT stages were discordant in 32/84 (38 %) cases: the T stage in 2/32 (6.2 %), the N stage in 21/32 (65.7 %) and the M stage 9/32 (28.1 %). Patient management was altered in 22/84 (26 %) patients, with a moderate impact in 8 (9.5 %) patients and high impact in 14 (16.6 %) patients. PET/CT TNM classification was significantly more accurate (92.5 vs 73.7 %) than conventional staging with a p value < 0.001 (McNemar's test). CONCLUSION: PET/CT should be implemented in the routine imaging work-up of stage III-IV HNSCC.

Metaplasia escamosa descamativa queratinizante (colesteatoma) del uréter / Keratinizing desquamative squamous metaplasia (cholesteatoma) of the ureter

La metaplasia escamosa descamativa queratinizante del tracto urinario es una enfermedad poco frecuente que puede simular un proceso neoplásico. Se presenta un ejemplo típico de esta entidad en una paciente de 71 años de edad con una larga historia de cólicos de repetición. El estudio radiológico mostró una lesión sospechosa, ocupante de espacio, en el muñón ureteral de una nefrectomía previa, y el examen histológico mostró metaplasia escamosa del urotelio y abundante descamación córnea en la luz formando masas irregulares(AU)

Keratinizing desquamative squamous metaplasia of the urinary tract is a rare condition that may mimic neoplasia. We report a characteristic case occurring in a 71-year old woman with a long previous history of recurrent nephritic colic. Imaging studies showed a suspicious mass in the ureteral stump of a previous nephrectomy. Histology revealed squamous metaplasia of the urothelium with abundant keratinizing material in the lumen forming irregular masses(AU)

La médula renal normal y patológica / The normal and abnormal renal medulla

Antecedentes: La médula renal constituye un intrincadosistema de túbulos, vasos sanguíneos e intersticio pococonocido por la mayor parte de los patólogos generales.Métodos: Amplia revisión de la literatura sobre la médularenal y de archivo de toda su patología. Resultados: Se pormenorizandatos de interés para el patólogo sobre el desarrollonormal y patológico, la anatomía microscópica, histologíae inmunohistoquímica, la fisiología, la patología dela diferenciación medular (displasia renal multiquística,enfermedades poliquísticas renales autosómicas dominantey recesiva, enfermedad quística medular) la patología inflamatoria(pielonefritis xantogranulomatosa, malacoplaquia),las displasias, y las neoplasias (oncocitoma, tumor oncocíticoatípico, carcinoma renal de células cromófobas, carcinomade los túbulos colectores, carcinoma urotelial, otros carcinomas,fibroma renomedular y tumores metastáticos) deesta topografía. Conclusiones: El conocimiento compendiadode la génesis, del funcionamiento y de la patologíamedular renal, tanto del desarrollo, como inflamatoria yneoplásica, redundará en un mayor interés por esta zona delriñón que habitualmente pasa desapercibida para el patólogoasistencial(AU)

Background: The renal medulla is composed of a complexsystem of tubules, blood vessels and interstitium,which the general pathologist often are unfamiliar with.Methods: An in depth review of the literature and of materialfrom our archives related to the pathology of the renalmedulla was made. Results: Interesting data on normal andabnormal development, microscopic anatomy, histology,immunohistochemistry, physiology, and pathology, includingrenal medulla differentiation disorders (multicysticrenal dysplasia, adult and infantile type polycystic diseases,cystic medullary disease), inflammatory diseases (xanthogranulomatouspyelonephritis, malakoplakia) and neoplasias(oncocytoma, atypical oncocytic tumour, chromophoberenal cell carcinoma, collecting duct carcinoma, urothelialcarcinoma, other carcinomas, renomedullary fibroma andmetastatic tumours), was reviewed. Conclusions: A comprehensiveknowledge of the genesis, function, and pathologyof the renal medulla would result in a greater interestbeing taken in an area often ignored by pathologists(AU)

Prostatic remnants in mature cystic teratoma of the ovary.

Mature cystic teratomas of the ovary containing prostatic remnants are reported in 2 women aged 31 and 20 years. Both cases showed the expected histology of mature teratomas with a mixture of ecto- and endodermal structures lying in a fibrous stroma. In both cases, the foci of prostate tissue were composed of typical prostatic glands arranged in acinar structures. One case displayed a transitional cell-lined duct resembling the urethra. Prostate glands showed intense positive immunostaining with prostatic specific antigen and prostatic acidic phosphatase. Focal images suggesting high-grade prostatic intraepithelial neoplasia were detected in 1 case. The literature on this unusual finding in these common tumors is reviewed and commented on.