ABSTRACT
A recently discovered, enhanced Ge diffusion mechanism along the oxidizing interface of Si/SiGe nanostructures has enabled the formation of single-crystal Si nanowires and quantum dots embedded in a defect-free, single-crystal SiGe matrix. Here, we report oxidation studies of Si/SiGe nanofins aimed at gaining a better understanding of this novel diffusion mechanism. A superlattice of alternating Si/Si0.7Ge0.3 layers was grown and patterned into fins. After oxidation of the fins, the rate of Ge diffusion down the Si/SiO2 interface was measured through the analysis of HAADF-STEM images. The activation energy for the diffusion of Ge down the sidewall was found to be 1.1 eV, which is less than one-quarter of the activation energy previously reported for Ge diffusion in bulk Si. Through a combination of experiments and DFT calculations, we propose that the redistribution of Ge occurs by diffusion along the Si/SiO2 interface followed by a reintroduction into substitutional positions in the crystalline Si.
ABSTRACT
BACKGROUND: The objective of the current study was to establish a process for validating immunohistochemistry (IHC) protocols for use on the Cellient cell block (CCB) system. METHODS: Thirty antibodies were initially tested on CCBs using IHC protocols previously validated on formalin-fixed, paraffin-embedded tissue (FFPE). Cytology samples were split to generate thrombin cell blocks (TCB) and CCBs. IHC was performed in parallel. Antibody immunoreactivity was scored, and concordance or discordance in immunoreactivity between the TCBs and CCBs for each sample was determined. Criteria for validation of an antibody were defined as concordant staining in expected positive and negative cells, in at least 5 samples each, and concordance in at least 90% of the samples total. Antibodies that failed initial validation were retested after alterations in IHC conditions. RESULTS: Thirteen of the 30 antibodies (43%) did not meet initial validation criteria. Of those, 8 antibodies (calretinin, clusters of differentiation [CD] 3, CD20, CDX2, cytokeratin 20, estrogen receptor, MOC-31, and p16) were optimized for CCBs and subsequently validated. Despite several alterations in conditions, 3 antibodies (Ber-EP4, D2-40, and paired box gene 8 [PAX8]) were not successfully validated. CONCLUSIONS: Nearly one-half of the antibodies tested in the current study failed initial validation using IHC conditions that were established in the study laboratory for FFPE material. Although some antibodies subsequently met validation criteria after optimization of conditions, a few continued to demonstrate inadequate immunoreactivity. These findings emphasize the importance of validating IHC protocols for methanol-fixed tissue before clinical use and suggest that optimization for alcohol fixation may be needed to obtain adequate immunoreactivity on CCBs.
Subject(s)
Antibodies/immunology , Cytodiagnosis/methods , Immunohistochemistry/methods , Research Personnel , Female , Humans , Male , Neoplasms/pathology , Sensitivity and Specificity , Specimen Handling , Tissue Embedding , Tissue Fixation/methodsABSTRACT
Peripheral T-cell lymphomas (PTCLs) are generally aggressive non-Hodgkin lymphomas with poor overall survival rates following standard therapy. One-third of PTCLs express interferon regulatory factor-4 (IRF4), a tightly regulated transcription factor involved in lymphocyte growth and differentiation. IRF4 drives tumor growth in several lymphoid malignancies and has been proposed as a candidate therapeutic target. Because direct IRF4 inhibitors are not clinically available, we sought to characterize the mechanism by which IRF4 expression is regulated in PTCLs. We demonstrated that IRF4 is constitutively expressed in PTCL cells and drives Myc expression and proliferation. Using an inhibitor screen, we identified nuclear factor κB (NF-κB) as a candidate regulator of IRF4 expression and cell proliferation. We then demonstrated that the NF-κB subunits p52 and RelB were transcriptional activators of IRF4. Further analysis showed that activation of CD30 promotes p52 and RelB activity and subsequent IRF4 expression. Finally, we showed that IRF4 transcriptionally regulates CD30 expression. Taken together, these data demonstrate a novel positive feedback loop involving CD30, NF-κB, and IRF4; further evidence for this mechanism was demonstrated in human PTCL tissue samples. Accordingly, NF-κB inhibitors may represent a clinical means to disrupt this feedback loop in IRF4-positive PTCLs.
Subject(s)
Interferon Regulatory Factors/genetics , Ki-1 Antigen/metabolism , Lymphoma, T-Cell, Peripheral/genetics , Lymphoma, T-Cell, Peripheral/metabolism , NF-kappa B/metabolism , Adult , Aged , Cell Line, Tumor , Cell Proliferation , DNA Copy Number Variations , Female , Gene Expression Regulation, Neoplastic , Genes, myc , Germ Cells/metabolism , Humans , Male , Middle Aged , Models, Biological , Polymorphism, Genetic , Transcription, GeneticABSTRACT
Anaplastic large cell lymphoma (ALCL) is one of the most common T-cell non-Hodgkin lymphomas and has 2 main subtypes: an anaplastic lymphoma kinase (ALK)-positive subtype characterized by ALK gene rearrangements and an ALK-negative subtype that is poorly understood. We recently identified recurrent rearrangements of the DUSP22 locus on 6p25.3 in both primary cutaneous and systemic ALK-negative ALCLs. This study aimed to determine the relationship between these rearrangements and expression of the chemokine receptor gene, CCR8. CCR8 has skin-homing properties and has been suggested to play a role in limiting extracutaneous spread of primary cutaneous ALCLs. However, overexpression of CCR8 has also been reported in systemic ALK-negative ALCLs. As available antibodies for CCR8 have shown lack of specificity, we examined CCR8 expression using quantitative real-time PCR in frozen tissue and RNA in situ hybridization (ISH) in paraffin tissue. Both approaches showed higher CCR8 expression in ALCLs with DUSP22 rearrangements than in nonrearranged cases (PCR: 19.5-fold increase, P=0.01; ISH: 3.3-fold increase, P=0.0008). CCR8 expression was not associated with cutaneous presentation, cutaneous biopsy site, or cutaneous involvement during the disease course. These findings suggest that CCR8 expression in ALCL is more closely related to the presence of DUSP22 rearrangements than to cutaneous involvement and that the function of CCR8 may extend beyond its skin-homing properties in this disease. This study also underscores the utility of RNA-ISH as a paraffin-based method for investigating gene expression when reliable antibodies for immunohistochemical analysis are not available.
Subject(s)
Dual-Specificity Phosphatases/genetics , Lymphoma, Large-Cell, Anaplastic/genetics , Mitogen-Activated Protein Kinase Phosphatases/genetics , Receptors, CCR8/genetics , Adolescent , Adult , Aged , Aged, 80 and over , Child , Female , Humans , Immunohistochemistry , In Situ Hybridization, Fluorescence , Male , Middle Aged , Real-Time Polymerase Chain Reaction , Young AdultABSTRACT
Anaplastic lymphoma kinase (ALK)-negative anaplastic large cell lymphoma (ALCL) is a CD30-positive T-cell non-Hodgkin lymphoma that morphologically resembles ALK-positive ALCL but lacks chromosomal rearrangements of the ALK gene. The genetic and clinical heterogeneity of ALK-negative ALCL has not been delineated. We performed immunohistochemistry and fluorescence in situ hybridization on 73 ALK-negative ALCLs and 32 ALK-positive ALCLs and evaluated the associations among pathology, genetics, and clinical outcome. Chromosomal rearrangements of DUSP22 and TP63 were identified in 30% and 8% of ALK-negative ALCLs, respectively. These rearrangements were mutually exclusive and were absent in ALK-positive ALCLs. Five-year overall survival rates were 85% for ALK-positive ALCLs, 90% for DUSP22-rearranged ALCLs, 17% for TP63-rearranged ALCLs, and 42% for cases lacking all 3 genetic markers (P < .0001). Hazard ratios for death in these 4 groups after adjusting for International Prognostic Index and age were 1.0 (reference group), 0.58, 8.63, and 4.16, respectively (P = 7.10 × 10(-5)). These results were similar when restricted to patients receiving anthracycline-based chemotherapy, as well as to patients not receiving stem cell transplantation. Thus, ALK-negative ALCL is a genetically heterogeneous disease with widely disparate outcomes following standard therapy. DUSP22 and TP63 rearrangements may serve as predictive biomarkers to help guide patient management.
Subject(s)
Lymphoma, Large-Cell, Anaplastic/genetics , Lymphoma, Large-Cell, Anaplastic/metabolism , Receptor Protein-Tyrosine Kinases/genetics , Receptor Protein-Tyrosine Kinases/metabolism , Adolescent , Adult , Aged , Aged, 80 and over , Anaplastic Lymphoma Kinase , Biomarkers, Tumor/genetics , Biomarkers, Tumor/metabolism , Child , Dual-Specificity Phosphatases/genetics , Female , Gene Rearrangement , Humans , Immunohistochemistry , In Situ Hybridization, Fluorescence , Interferon Regulatory Factors/genetics , Kaplan-Meier Estimate , Lymphoma, Large-Cell, Anaplastic/pathology , Male , Middle Aged , Mitogen-Activated Protein Kinase Phosphatases/genetics , Prognosis , Transcription Factors/genetics , Tumor Suppressor Proteins/genetics , Young AdultABSTRACT
Using laser microdissection and mass spectrometry (MS)-based proteomics, we subtyped amyloid deposits from 130 cases of hepatic amyloidosis. Although we confirmed that immunoglobulin light chain amyloidosis was the most frequent cause of hepatic amyloidosis, leukocyte cell-derived chemotaxin 2 (LECT2) amyloidosis (ALect2) accounted for 25% of cases. This novel finding was associated with Hispanic ancestry, incidental discovery of amyloid in liver specimens sampled for other unrelated conditions, and a characteristic pattern of hepatic amyloid deposition. Although ALect2 patients had a common LECT2 polymorphism, pathogenic mutations were not discovered, suggesting that constitutive or compensatory LECT2 overexpression led to ALect2 deposition. These findings indicate that ALect2 is a common cause of hepatic amyloidosis in the population of the United States, and subtyping hepatic amyloid deposits by an accurate analytic method such as MS is required for optimal clinical management of hepatic amyloidosis patients and to avoid incorrect and unnecessarily toxic therapies.
Subject(s)
Amyloidosis/metabolism , Intercellular Signaling Peptides and Proteins/metabolism , Liver Diseases/metabolism , Liver Diseases/pathology , Adult , Aged , Amyloidosis/diagnosis , Female , Humans , Liver/metabolism , Liver/pathology , Liver Diseases/diagnosis , Male , Mass Spectrometry , Microdissection , Middle Aged , Prospective Studies , Proteomics/methods , United StatesABSTRACT
The traditional model of hematopoiesis is based on unidirectional maturation of hematopoietic precursors into lineage-committed cells. However, recent studies indicate that mature B lymphocytes may demonstrate significant lineage plasticity. We and others have reported transdifferentiation of follicular lymphomas (FLs) into clonally related histiocytic/dendritic cell neoplasms. Here, we describe 2 patients with FL who developed clonally related Langerhans cell neoplasms. The first was a 52-year-old man diagnosed with FL, grade 1. He received immunochemotherapy and had stable disease for 8 years. He then developed increasing lymphadenopathy, and lymph node biopsy showed Langerhans cell sarcoma with no evidence of FL. The second patient was a 77-year-old woman who presented with lymphadenopathy, an abdominal mass, and pulmonary nodules. Lymph node biopsy showed both Langerhans cell histiocytosis and minimal involvement by FL, grade 1. In each case, a combination of immunoglobulin gene rearrangement and fluorescence in situ hybridization studies provided evidence to support a clonal relationship between the FL and Langerhans cell neoplasm. These cases provide striking examples of neoplastic transdifferentiation and expand the spectrum of lesions clonally identical to otherwise typical FL. Awareness of this phenomenon may aid in diagnosis when histologically dissimilar tumors arise synchronously or metachronously in patients with lymphoma.
Subject(s)
Cell Transdifferentiation , Langerhans Cell Sarcoma/pathology , Langerhans Cells/pathology , Lymphoma, Follicular/pathology , Neoplasms, Second Primary/pathology , Aged , Clone Cells , Female , Flow Cytometry , Gene Rearrangement , Humans , Immunoglobulin Heavy Chains/genetics , Immunophenotyping , In Situ Hybridization, Fluorescence , Langerhans Cell Sarcoma/genetics , Langerhans Cell Sarcoma/metabolism , Langerhans Cells/metabolism , Lymphoma, Follicular/genetics , Lymphoma, Follicular/metabolism , Male , Middle Aged , Neoplasms, Second Primary/genetics , Neoplasms, Second Primary/metabolism , Proto-Oncogene Proteins B-raf/genetics , Proto-Oncogene Proteins B-raf/metabolismABSTRACT
STAT3 regulates cell growth by up-regulating downstream targets, such as Myc. The frequency of phosphorylated STAT3 (pSTAT3) and Myc expression and their prognostic relevance is unknown within diffuse large B-cell lymphoma (DLBCL) germinal center B-cell (GCB) and non-GCB subtypes. pSTAT3 and Myc were studied by immunohistochemistry (IHC) on tumors from 40 DLBCL patients uniformly treated on a clinical trial of epratuzumab/rituximab-CHOP. A total of 35% of cases were pSTAT3-positive, and pSTAT3 positivity was more frequent in the non-GCB (P = .06) type but did not correlate with event-free survival (EFS). Myc expression was observed in 50% of cases and was more frequent in non-GCB type (P = .07). Myc-positive cases had inferior EFS in all patients, including the GCB and pSTAT3-positive cases, were more likely to express Myc (P = .06). Myc translocations involving the major breakpoint regions were found in 10% (3 of 29) of cases, and all 3 cases were GCB and had an inferior EFS (P = .09). pSTAT3, but not Myc expression, was correlated with elevated pretreatment serum cytokines, such as IL-10 (P = .05), G-CSF (P = .03), and TNF-α (P = .04). pSTAT3 IHC in DLBCL tumors has the potential to identify patients for STAT3 pathway-directed therapy; Myc IHC is a potential marker for inferior EFS in GCB patients.
Subject(s)
Antibodies, Monoclonal, Humanized/administration & dosage , Antineoplastic Combined Chemotherapy Protocols/administration & dosage , Lymphoma, Large B-Cell, Diffuse/diagnosis , Lymphoma, Large B-Cell, Diffuse/drug therapy , Proto-Oncogene Proteins c-myc/physiology , STAT3 Transcription Factor/physiology , Aged , Aged, 80 and over , Antibodies, Monoclonal, Murine-Derived/administration & dosage , Biomarkers, Pharmacological/metabolism , Biomarkers, Tumor/metabolism , Biomarkers, Tumor/physiology , Cyclophosphamide/administration & dosage , Disease-Free Survival , Doxorubicin/administration & dosage , Humans , Lymphoma, Large B-Cell, Diffuse/metabolism , Middle Aged , Phosphorylation , Prednisone/administration & dosage , Prognosis , Protein Kinases/metabolism , Proto-Oncogene Proteins c-myc/metabolism , Rituximab , STAT3 Transcription Factor/metabolism , Treatment Outcome , Validation Studies as Topic , Vincristine/administration & dosageABSTRACT
Peripheral T-cell lymphomas (PTCLs) are aggressive malignancies of mature T lymphocytes with 5-year overall survival rates of only â¼ 35%. Improvement in outcomes has been stymied by poor understanding of the genetics and molecular pathogenesis of PTCL, with a resulting paucity of molecular targets for therapy. We developed bioinformatic tools to identify chromosomal rearrangements using genome-wide, next-generation sequencing analysis of mate-pair DNA libraries and applied these tools to 16 PTCL patient tissue samples and 6 PTCL cell lines. Thirteen recurrent abnormalities were identified, of which 5 involved p53-related genes (TP53, TP63, CDKN2A, WWOX, and ANKRD11). Among these abnormalities were novel TP63 rearrangements encoding fusion proteins homologous to ΔNp63, a dominant-negative p63 isoform that inhibits the p53 pathway. TP63 rearrangements were seen in 11 (5.8%) of 190 PTCLs and were associated with inferior overall survival; they also were detected in 2 (1.2%) of 164 diffuse large B-cell lymphomas. As TP53 mutations are rare in PTCL compared with other malignancies, our findings suggest that a constellation of alternate genetic abnormalities may contribute to disruption of p53-associated tumor suppressor function in PTCL.
Subject(s)
Gene Rearrangement , Lymphoma, T-Cell, Peripheral/genetics , Mutation , Transcription Factors/genetics , Tumor Suppressor Protein p53/genetics , Tumor Suppressor Proteins/genetics , Cell Line, Tumor , Cyclin-Dependent Kinase Inhibitor p16/chemistry , Cyclin-Dependent Kinase Inhibitor p16/genetics , Cyclin-Dependent Kinase Inhibitor p16/metabolism , DNA Mutational Analysis , Female , Genome-Wide Association Study , Humans , Lymphoma, Large B-Cell, Diffuse/genetics , Lymphoma, Large B-Cell, Diffuse/metabolism , Lymphoma, Large B-Cell, Diffuse/mortality , Lymphoma, Large B-Cell, Diffuse/pathology , Lymphoma, T-Cell, Peripheral/metabolism , Lymphoma, T-Cell, Peripheral/mortality , Lymphoma, T-Cell, Peripheral/pathology , Male , Mutant Proteins/chemistry , Mutant Proteins/metabolism , Oligonucleotide Array Sequence Analysis , Oncogene Proteins, Fusion/chemistry , Oncogene Proteins, Fusion/genetics , Oncogene Proteins, Fusion/metabolism , Oxidoreductases/chemistry , Oxidoreductases/genetics , Oxidoreductases/metabolism , Repressor Proteins/chemistry , Repressor Proteins/genetics , Repressor Proteins/metabolism , Sequence Homology, Nucleic Acid , Transcription Factors/chemistry , Transcription Factors/metabolism , Tumor Suppressor Protein p53/chemistry , Tumor Suppressor Protein p53/metabolism , Tumor Suppressor Proteins/chemistry , Tumor Suppressor Proteins/metabolism , United States , WW Domain-Containing OxidoreductaseABSTRACT
CD30-positive T-cell lymphoproliferative disorders are classified as cutaneous (primary cutaneous anaplastic large cell lymphoma and lymphomatoid papulosis) or systemic. As extent of disease dictates prognosis and treatment, patients with skin involvement need clinical staging to determine whether systemic lymphoma also is present. Similar processes may involve mucosal sites of the head and neck, constituting a spectrum that includes both neoplasms and reactive conditions (eg, traumatic ulcerative granuloma with stromal eosinophilia). However, no standard classification exists for mucosal CD30-positive T-cell lymphoproliferations. To improve our understanding of these processes, we identified 15 such patients and examined clinical presentation, treatment and outcome, morphology, phenotype using immunohistochemistry, and genetics using gene rearrangement studies and fluorescence in situ hybridization. The 15 patients (11 M, 4 F; mean age, 57 years) had disease involving the oral cavity/lip/tongue (9), orbit/conjunctiva (3) or nasal cavity/sinuses (3). Of 14 patients with staging data, 7 had mucosal disease only; 2 had mucocutaneous disease; and 5 had systemic anaplastic large cell lymphoma. Patients with mucosal or mucocutaneous disease only had a favorable prognosis and none developed systemic spread (follow-up, 4-93 months). Three of five patients with systemic disease died of lymphoma after 1-48 months. Morphologic and phenotypic features were similar regardless of extent of disease. One anaplastic lymphoma kinase-positive case was associated with systemic disease. Two cases had rearrangements of the DUSP22-IRF4 locus on chromosome 6p25.3, seen most frequently in primary cutaneous anaplastic large cell lymphoma. Our findings suggest mucosal CD30-positive T-cell lymphoproliferations share features with cutaneous CD30-positive T-cell lymphoproliferative disorders, and require clinical staging for stratification into primary and secondary types. Primary cases have clinicopathologic features closer to primary cutaneous disease than to systemic anaplastic large cell lymphoma, including indolent clinical behavior. Understanding the spectrum of mucosal CD30-positive T-cell lymphoproliferations is important to avoid possible overtreatment resulting from a diagnosis of overt T-cell lymphoma.
Subject(s)
Head and Neck Neoplasms/classification , Head and Neck Neoplasms/pathology , Lymphoma, T-Cell/classification , Lymphoma, T-Cell/pathology , Mucous Membrane/pathology , Adolescent , Adult , Aged , Aged, 80 and over , Child , Child, Preschool , Female , Head and Neck Neoplasms/metabolism , Humans , Immunohistochemistry , In Situ Hybridization, Fluorescence , Ki-1 Antigen/metabolism , Lymphoma, T-Cell/metabolism , Lymphoma, T-Cell, Cutaneous/pathology , Male , Middle Aged , Mucous Membrane/metabolism , Neoplasm Staging , Phenotype , Young AdultABSTRACT
Lymphoplasmacytic lymphomas and marginal zone lymphomas of nodal, extra-nodal and splenic types account for 10% of non-Hodgkin lymphomas. They are similar at the cell differentiation level, sometimes making difficult to distinguish them from other indolent non-Hodgkin lymphomas. To better characterize their genetic basis, we performed array-based comparative genomic hybridization in 101 marginal zone lymphomas (46 MALT, 35 splenic and 20 nodal marginal zone lymphomas) and 13 lymphoplasmacytic lymphomas. Overall, 90% exhibited copy-number abnormalities. Lymphoplasmacytic lymphomas demonstrated the most complex karyotype (median=7 copy-number abnormalities), followed by MALT (4), nodal (3.5) and splenic marginal zone lymphomas (3). A comparative analysis exposed a group of copy-number abnormalities shared by several or all the entities with few disease-specific abnormalities. Gain of chromosomes 3, 12 and 18 and loss of 6q23-q24 (TNFAIP3) were identified in all entities. Losses of 13q14.3 (MIRN15A-MIRN16-1) and 17p13.3-p12 (TP53) were found in lymphoplasmacytic and splenic marginal zone lymphomas; loss of 11q21-q22 (ATM) was found in nodal, splenic marginal zone and lymphoplasmacytic lymphomas and loss of 7q32.1-q33 was found in MALT, splenic and lymphoplasmacytic lymphomas. Abnormalities affecting the nuclear factor kappa B pathway were observed in 70% of MALT and lymphoplasmacytic lymphomas and 30% of splenic and nodal marginal zone lymphomas, suggesting distinct roles of this pathway in the pathogenesis/progression of these subtypes. Elucidation of the genetic alterations contributing to the pathogenesis of these lymphomas may guide to design-specific therapeutic approaches.
Subject(s)
Lymph Nodes/pathology , Lymphoma, B-Cell, Marginal Zone/pathology , Waldenstrom Macroglobulinemia/pathology , Chromosome Aberrations , DNA, Neoplasm/analysis , Gene Expression Profiling , Gene Expression Regulation, Neoplastic , Genomics , Humans , In Situ Hybridization, Fluorescence , Lymphoma, B-Cell, Marginal Zone/genetics , Waldenstrom Macroglobulinemia/geneticsABSTRACT
Most lymphomas arising in the ovary are diffuse large B-cell or Burkitt lymphomas. Follicular lymphomas of the ovary are rare, and their clinicopathologic features are incompletely understood. We investigated clinical, morphologic, immunophenotypic, and genetic features of follicular lymphomas initially diagnosed in the ovary in 16 women. We performed immunohistochemistry for CD20, CD3, CD10, BCL6, IRF4/MUM1, and BCL2 and interphase fluorescence in situ hybridization for translocations involving BCL2, BCL6, and IRF4. Clustering was performed on the basis of clinicopathologic and genetic variables using Cluster 3.0. Ages ranged from 42 to 73 years. Unsupervised hierarchical clustering analysis revealed 2 distinct groups of patients. One group (group A) included 7 cases, which were negative or weakly positive for BCL2 protein and lacked IGH@/BCL2 translocations (6 cases evaluable). Four patients were of grade 3A, and 3 patients were of grade 2. Four had stage IE disease with unilateral ovarian involvement (subgroup A(1)), and the stage was unknown in 3 patients (subgroup A(2)). The second group (group B) included 9 cases of low histologic grade, which strongly expressed BCL2 protein and had IGH@/BCL2 translocations (7 cases evaluable). Six patients had stage III/IV disease, 2 had stage II disease, and the stage was unknown in 1 patient. Although follow-up intervals were limited, patients with higher-grade/low-stage disease tended to have favorable outcomes. These data indicate 2 distinct types of follicular lymphomas presenting in the ovary: a low-grade/high-stage/BCL2-positive group and a higher-grade/low-stage/BCL2-negative group. This latter group likely has distinct biological and immunologic characteristics, as has been suggested for higher-grade/low-stage follicular lymphomas presenting at other extranodal sites.
Subject(s)
Lymphoma, Follicular/diagnosis , Ovarian Neoplasms/diagnosis , Adult , Aged , Biomarkers, Tumor/analysis , Cluster Analysis , Female , Gene Expression Regulation, Neoplastic , Humans , Immunohistochemistry , Immunophenotyping , In Situ Hybridization, Fluorescence , Lymphoma, Follicular/chemistry , Lymphoma, Follicular/classification , Lymphoma, Follicular/genetics , Lymphoma, Follicular/pathology , Middle Aged , Minnesota , Neoplasm Grading , Neoplasm Staging , Ovarian Neoplasms/chemistry , Ovarian Neoplasms/classification , Ovarian Neoplasms/genetics , Ovarian Neoplasms/pathology , Prognosis , Time Factors , Translocation, GeneticABSTRACT
The BCL-6 proto-oncogene encodes a transcriptional repressor protein. Among normal tissues, BCL-6 expression is confined to germinal center B-cells and a subpopulation of T-helper cells. Little is known about BCL-6 expression in mesenchymal tissues. We examined a series of solitary fibrous tumor (SFT) and other mesenchymal tumors for BCL-6 expression. Immunohistochemistry for BCL-6 was performed on 64 mesenchymal tumors [26 SFT (19 benign/uncertain, 7 malignant), 6 synovial sarcomas (SS), 5 gastrointestinal stromal tumors (GIST), 5 malignant peripheral nerve sheath tumors (MPNST), 5 leiomyosarcomas (LMS), 9 leiomyomas (LM) 4 desmoid tumors (DT), 4 perineuriomas (PN)]. Nuclear immunoreactivity was considered positive. Six BCL-6 positive SFT were also tested for BCL-6 gene rearrangement/amplification by FISH. Nuclear expression of BCL-6 was seen in 13/26 SFT, 5/5 LMS, 1/9 LM, 5/6 SS, 1/5 GIST, 1/5 MPNST, 1/4 PN, and 0/5 DT. BCL-6 expression was significantly more frequent in malignant (6/7) as compared with benign/uncertain SFT (6/19) (p=0.02) and in LMS (5/5) as compared with LM (1/9) (p=0.003). FISH for BCL-6 rearrangement/amplification was negative in all tested cases. We have observed BCL-6 expression in 50% or more of SFT, SS, and LMS, and in a lesser percentage of LM, GIST, MPNST and PN. Significantly more frequent expression of BCL-6 in malignant compared with benign/uncertain SFT and in LMS compared with LM suggests abnormalities in the BCL-6 signaling pathway may contribute to malignant transformation in at least some mesenchymal tumors. It is unlikely that BCL-6 expression in mesenchymal tumors is due to BCL-6 gene amplification or rearrangement.
Subject(s)
Biomarkers, Tumor/analysis , Biomarkers, Tumor/genetics , DNA-Binding Proteins/analysis , DNA-Binding Proteins/genetics , Immunohistochemistry , In Situ Hybridization, Fluorescence , Mesoderm/chemistry , Neoplasms, Connective Tissue/chemistry , Neoplasms, Connective Tissue/genetics , Cell Transformation, Neoplastic/chemistry , Cell Transformation, Neoplastic/genetics , Gene Amplification , Gene Rearrangement , Humans , Mesoderm/pathology , Minnesota , Neoplasms, Connective Tissue/pathology , Proto-Oncogene Mas , Proto-Oncogene Proteins c-bcl-6 , Solitary Fibrous Tumors/chemistry , Solitary Fibrous Tumors/geneticsABSTRACT
Distinguishing mantle cell lymphoma (MCL), from low-grade B-cell lymphoma is important because MCL is clinically more aggressive and is treated differently. Though most MCL overexpress cyclinD1 (CCND1) and have a t(11;14)(q13;q32), MCL that are negative for CCND1 exist. Some have translocations involving cyclinD2 (CCND2) and either the immunoglobulin heavy chain or kappa light chain locus. We present a CD5-positive, CCND1-negative B-cell lymphoma with a novel translocation involving CCND2 and the immunoglobulin lambda (IGL) gene. A 64-year-old male underwent resection of a polypoid mass of the ileum. Histology showed atypical, medium-sized lymphoid cells positive for CD20, CD5, CD43, and CCND2 by immunohistochemistry, and negative for CCND1, CCND3, and p27. Fluorescence in situ hybridization was negative for CCND1 abnormalities, but demonstrated a CCND2/IGL fusion. Clinical workup revealed stage IV disease. Current diagnostic criteria are insufficient for subclassifying this case, highlighting the need for additional studies on CCND2-translocated B-cell lymphomas to guide therapy appropriately.
Subject(s)
CD5 Antigens/biosynthesis , Cyclin D2/genetics , Immunoglobulin lambda-Chains/genetics , Lymphoma, B-Cell/genetics , Lymphoma, Mantle-Cell/genetics , Bone Marrow Neoplasms/diagnosis , Bone Marrow Neoplasms/genetics , Bone Marrow Neoplasms/metabolism , Chromosomes, Human, Pair 12 , Chromosomes, Human, Pair 22 , Cyclin D1/metabolism , Diagnosis, Differential , Gastrointestinal Neoplasms/diagnosis , Gastrointestinal Neoplasms/genetics , Gastrointestinal Neoplasms/metabolism , Humans , Immunohistochemistry , In Situ Hybridization, Fluorescence , Lymphoma, B-Cell/diagnosis , Lymphoma, B-Cell/metabolism , Lymphoma, Mantle-Cell/diagnosis , Lymphoma, Mantle-Cell/metabolism , Male , Middle Aged , Translocation, GeneticABSTRACT
Acute bilineal leukemias are rare and are commonly associated with t(9;22) and MLL abnormalities. Herein, we report a pediatric case of bilineal T/myeloid acute leukemia associated with del (9q)(q13q22) and TLX3/BCL11B fusion due to the cryptic t(5;14)(q35;32). FISH studies confirmed the TLX3/BCL11B fusion in both the myeloid and lymphoid blasts, while the 9q deletion was restricted to the lymphoid component. Optimal therapy for such patients remains controversial and it is not clear if they should be treated with ALL or AML-based chemotherapeutic regimens. Our patient has been in extended remission following ALL-based chemotherapy and a matched unrelated cord blood transplant. Inc.
Subject(s)
Chromosomes, Human, Pair 9/genetics , Homeodomain Proteins/genetics , Leukemia, Biphenotypic, Acute/therapy , Leukemia, Myeloid, Acute/therapy , Leukemia, T-Cell/therapy , Oncogene Proteins, Fusion/genetics , Repressor Proteins/genetics , Tumor Suppressor Proteins/genetics , Antineoplastic Agents/therapeutic use , Bone Marrow Transplantation , Child , Chromosomes, Human, Pair 22/genetics , Combined Modality Therapy , Humans , In Situ Hybridization, Fluorescence , Leukemia, Biphenotypic, Acute/diagnosis , Leukemia, Biphenotypic, Acute/genetics , Leukemia, Myeloid, Acute/diagnosis , Leukemia, Myeloid, Acute/genetics , Leukemia, T-Cell/diagnosis , Leukemia, T-Cell/genetics , Male , RNA, Messenger/genetics , Remission Induction , Reverse Transcriptase Polymerase Chain Reaction , Sequence Deletion , Translocation, Genetic/genetics , Treatment OutcomeABSTRACT
Current pathologic criteria cannot reliably distinguish cutaneous anaplastic large cell lymphoma from other CD30-positive T-cell lymphoproliferative disorders (lymphomatoid papulosis, systemic anaplastic large cell lymphoma with skin involvement, and transformed mycosis fungoides). We previously reported IRF4 (interferon regulatory factor-4) translocations in cutaneous anaplastic large cell lymphomas. Here, we investigated the clinical utility of detecting IRF4 translocations in skin biopsies. We performed fluorescence in situ hybridization (FISH) for IRF4 in 204 biopsies involved by T-cell lymphoproliferative disorders from 182 patients at three institutions. In all, 9 of 45 (20%) cutaneous anaplastic large cell lymphomas and 1 of 32 (3%) cases of lymphomatoid papulosis with informative results demonstrated an IRF4 translocation. Remaining informative cases were negative for a translocation (7 systemic anaplastic large cell lymphomas; 44 cases of mycosis fungoides/Sézary syndrome (13 transformed); 24 peripheral T-cell lymphomas, not otherwise specified; 12 CD4-positive small/medium-sized pleomorphic T-cell lymphomas; 5 extranodal NK/T-cell lymphomas, nasal type; 4 gamma-delta T-cell lymphomas; and 5 other uncommon T-cell lymphoproliferative disorders). Among all cutaneous T-cell lymphoproliferative disorders, FISH for IRF4 had a specificity and positive predictive value for cutaneous anaplastic large cell lymphoma of 99 and 90%, respectively (P=0.00002, Fisher's exact test). Among anaplastic large cell lymphomas, lymphomatoid papulosis, and transformed mycosis fungoides, specificity and positive predictive value were 98 and 90%, respectively (P=0.005). FISH abnormalities other than translocations and IRF4 protein expression were seen in 13 and 65% of cases, respectively, but were nonspecific with regard to T-cell lymphoproliferative disorder subtype. Our findings support the clinical utility of FISH for IRF4 in the differential diagnosis of T-cell lymphoproliferative disorders in skin biopsies, with detection of a translocation favoring cutaneous anaplastic large cell lymphoma. Like all FISH studies, IRF4 testing must be interpreted in the context of morphology, phenotype, and clinical features.
Subject(s)
Biomarkers, Tumor/genetics , Interferon Regulatory Factors/genetics , Lymphoma, Primary Cutaneous Anaplastic Large Cell/genetics , Lymphoproliferative Disorders/genetics , Skin Neoplasms/genetics , Translocation, Genetic , Adolescent , Adult , Aged , Aged, 80 and over , Biomarkers, Tumor/analysis , Biopsy , Child , Child, Preschool , Diagnosis, Differential , Female , Humans , Immunohistochemistry , In Situ Hybridization, Fluorescence , Lymphoma, Large-Cell, Anaplastic/genetics , Lymphoma, Primary Cutaneous Anaplastic Large Cell/chemistry , Lymphoma, Primary Cutaneous Anaplastic Large Cell/pathology , Lymphomatoid Papulosis/genetics , Lymphoproliferative Disorders/metabolism , Lymphoproliferative Disorders/pathology , Male , Middle Aged , Mycosis Fungoides/genetics , Predictive Value of Tests , Sensitivity and Specificity , Skin Neoplasms/chemistry , Skin Neoplasms/pathology , United States , Young AdultABSTRACT
The genetics of peripheral T-cell lymphomas are poorly understood. The most well-characterized abnormalities are translocations involving ALK, occurring in approximately half of anaplastic large cell lymphomas (ALCLs). To gain insight into the genetics of ALCLs lacking ALK translocations, we combined mate-pair DNA library construction, massively parallel ("Next Generation") sequencing, and a novel bioinformatic algorithm. We identified a balanced translocation disrupting the DUSP22 phosphatase gene on 6p25.3 and adjoining the FRA7H fragile site on 7q32.3 in a systemic ALK-negative ALCL. Using fluorescence in situ hybridization, we demonstrated that the t(6;7)(p25.3;q32.3) was recurrent in ALK-negative ALCLs. Furthermore, t(6;7)(p25.3;q32.3) was associated with down-regulation of DUSP22 and up-regulation of MIR29 microRNAs on 7q32.3. These findings represent the first recurrent translocation reported in ALK-negative ALCL and highlight the utility of massively parallel genomic sequencing to discover novel translocations in lymphoma and other cancers.
Subject(s)
Chromosomes, Human, Pair 6/genetics , Chromosomes, Human, Pair 7/genetics , Lymphoma, Large-Cell, Anaplastic/genetics , Translocation, Genetic , Adult , Aged , Aged, 80 and over , Anaplastic Lymphoma Kinase , Base Sequence , Chromosome Breakage , Chromosome Breakpoints , Dual-Specificity Phosphatases/genetics , Female , Humans , In Situ Hybridization, Fluorescence , Lymphoma, Large-Cell, Anaplastic/pathology , Male , MicroRNAs/genetics , Middle Aged , Mitogen-Activated Protein Kinase Phosphatases/genetics , Molecular Sequence Data , Protein-Tyrosine Kinases/genetics , Receptor Protein-Tyrosine Kinases , Sequence Analysis, DNA/methodsABSTRACT
Diffuse large B-cell lymphoma (DLBCL) can be separated for prognostic purposes using gene expression profiling (GEP) into 2 subgroups: germinal center B-cell (GCB) and activated B-cell phenotypes. However, GEP is impractical for routine clinical use, and immunophenotyping is an imperfect surrogate. Therefore, we studied the relationship between expression of the purported germinal center marker LMO2 and the presence of IGH-BCL2 fusions, BCL6 translocations, and LMO2 translocations. In addition, we investigated the usefulness of LMO2 expression as a marker of GCB subtype in DLBCL. Immunohistochemical and fluorescence in situ hybridization studies were successfully performed on 101 cases of de novo DLBCL that had been incorporated into a tissue microarray. There was a statistically significant association between IGH-BCL2 fusion and LMO2 protein expression (P = .02) but not between BCL6 translocations and LMO2 expression. LMO2 translocations were not identified. Although uncommon, all cases that had both IGH-BCL2 fusion and BCL6 translocations expressed LMO2. The findings suggest LMO2 as a potential marker for the GCB phenotype.
Subject(s)
Biomarkers, Tumor/analysis , DNA-Binding Proteins/biosynthesis , Genes, Immunoglobulin Heavy Chain/genetics , Genes, bcl-2/genetics , Lymphoma, Large B-Cell, Diffuse/genetics , Metalloproteins/biosynthesis , Proto-Oncogene Proteins c-bcl-6/genetics , Adaptor Proteins, Signal Transducing , Chromosomes, Human, Pair 14/genetics , Chromosomes, Human, Pair 16/genetics , Gene Expression , Gene Expression Profiling , Humans , Immunohistochemistry , In Situ Hybridization, Fluorescence , LIM Domain Proteins , Lymphoma, Large B-Cell, Diffuse/metabolism , Oncogene Proteins, Fusion , Proto-Oncogene Proteins , Tissue Array Analysis , Translocation, GeneticABSTRACT
Cell lineage is the major criterion by which lymphomas are classified. Immunohistochemistry has greatly facilitated lymphoma diagnosis by detecting expression of lineage-associated antigens. However, loss or aberrant expression of these antigens may present diagnostic challenges. Anaplastic large cell lymphoma is a T-cell lymphoma that shows morphologic and phenotypic overlap with classical Hodgkin's lymphoma, which is a tumor of B-cell derivation. Staining for the B-cell transcription factor, paired box 5 (PAX5), has been suggested to be helpful in this differential, as it is positive in most classical Hodgkin's lymphomas, but absent in anaplastic large cell lymphomas. In this study we report four systemic T-cell anaplastic large cell lymphomas that were positive for PAX5 by immunohistochemistry, with weak staining intensity similar to that observed in classical Hodgkin's lymphoma. All diagnoses were confirmed by a combination of morphologic, phenotypic, and molecular criteria. Three cases were anaplastic lymphoma kinase (ALK) negative and one was ALK positive. PAX5 immunohistochemistry was negative in 198 additional peripheral T-cell lymphomas, including 66 anaplastic large cell lymphomas. Unexpectedly, although PAX5 translocations were absent, all evaluable PAX5-positive anaplastic large cell lymphomas showed extra copies of the PAX5 gene locus by fluorescence in situ hybridization (FISH). In contrast, only 4% of PAX5-negative peripheral T-cell lymphomas had extra copies of PAX5. We conclude that aberrant expression of PAX5 occurs rarely in T-cell anaplastic large cell lymphomas, and may be associated with extra copies of the PAX5 gene. PAX5-positive lymphomas with morphologic features overlapping different lymphoma types should be evaluated with an extensive immunohistochemical panel and/or molecular studies to avoid diagnostic errors that could lead to inappropriate treatment. As PAX5 overexpression causes T-cell neoplasms in experimental models, PAX5 may have contributed to lymphomagenesis in our cases.