High-grade B-cell Neoplasms Research Articles

Antisense oligonucleotide p45Skp-2 suppresses migratory chemotactic and metastasis of oral malignant Burkitt’s lymphoma cell through down-regulation of MTA-1 and induction of E-cadherin mechanism

Introduction: Burkitt’s lymphoma is a high-grade B-cell neoplasm and one of the most aggressive malignancies of lymphoid origins which found mainly in the paediatric population. The treatment options of this tumour are still limited. However, a new strategy for refractory tumour, phosphorothioate oligonucleotide antisense technique has watched with keen interest. This study was aimed to examine the effect of antisense p45Skp-2 (Skp-2 AS) suppressed migratory chemotactic and metastasis of oral malignant Burkitt’s lymphoma (Raji) cell through down-regulation of MTA-1 and E-cadherin. Methods: True experiment laboratory with post-test control group design was confirmed in this study. The efficiency of Skp-2 AS in the suppression of cell chemotactic migration was examined by Boyden chamber assay. To evaluate the inhibition of cell metastasis was conducted by decreasing MTA-1 expression protein. The expressions of MTA-1, E-cadherin and α-tubulin protein were investigated by Western blot analysis. Results: The results revealed that the number of chemotactic migration of Skp-2 AS treated Raji cell was significantly decreased when compared with that of sense p45Skp-2 (Skp-2 S) and scrambled control (SC) cells (P<0.05) followed by decreased expressions of MTA-1 protein and overexpression of E-cadherin. Interestingly, the expression of α-tubulin protein as an internal control was approximately similar in each transfectant cells. Conclusion: p45Skp-2 have an antitumor activity via suppression of migratory chemotactic activity and metastasis on oral Burkitt’s lymphoma cells through down-regulation of MTA-1 and induction of E-cadherin proteins targeting this molecule could represent a promising new therapeutic approach for this type of cancer.

A Rare and Aggressive Lymphoma; Burkitt Lymphoma in the Terminal Ileum

Burkitt Lymphoma (BL) is an aggressive form of high-grade B-cell neoplasm that is rarely identified in the adult population. The World Health Organization (WHO) describes three clinical variants; endemic, sporadic, and immunodeficiency associated.The following case is a rare presentation of Primary small intestine Burkitt lymphoma in an adult male presenting with fatigue, dyspnea on exertion, and unexplained weight loss. We present a case of an 81-year-old male with a past medical history of stage I Hodgkin's lymphoma treated with chemotherapy(may be state chemo regime) approximately 8 years ago, non-insulin-dependent diabetes, hypertension, hyperlipidemia, chronic kidney disease who presented to our a tertiary center with the complaints of increasing weakness, fatigue, and unexplained weight loss. In the emergency department the patient was found to have acute kidney injury and hypercalcemia 14.1mg/dL (Ca 8.4 - 10.3 mg/dL). Due to concern for malignancy with hypercalcemia and the patient's history of Hodgkin's lymphoma a computed tomography (CT) chest, abdomen and pelvis was obtained. Imaging revealed concerning findings of peritoneal carcinomatosis with gastrointestinal malignancy as possible source. Gastroenterology was consulted and an EGD as well as colonoscopy were performed.Colonoscopy revealed diverticulosis in the sigmoid colon and in the descending colon, a malignant-appearing tumor in the distal ileum that was biopsied. Pathology revealed an aggressive B-cell lymphoma with Burkitt like morphology and a germinal center phenotype. The infiltrate was positive for CD 20, CD 10, BCL 2, BCL 6, and MUM1. The Ki 67 proliferation rate was greater than 90%. Overall, the morphology and high proliferation rate were consistent with Burkitt lymphoma. The patient chose to follow up at an alternative facility for his ongoing oncology care. Burkitt lymphoma is a very aggressive form of B-cell lymphoma the sporadic form occurring worldwide with a predilection for children. Clinical symptoms depend on the site of involvement. There is frequently extranodal involvement commonly occurring in the gastrointestinal tract. CT findings can include peritoneal thickening, as well as ascites, and omental involvement. Often times when there is diffuse peritoneal disease, it is usually associated with a focal gastrointestinal mass. Although rare, Burkitt lymphoma should be kept on the differential when investigating CT findings of what appears to be peritoneal carcinomatosis.2530_A Figure 1. peritoneal carcinomatosis2530_B Figure 2. Malignant appearing tumor in the distal ileum

Primary lymphoma of bone, myeloma mimicker: a case report and review of literature

Primary lymphoma of the bone (PLB) primarily arising from the medullary cavity is an extremely rare entity, with only limited number of studies and sporadic cases reported in the literature. The current study presents one case of PLB presented with unique presentation and treated with chemotherapy only. A 66 year old male presented with severe pain in the left hip that persisted for two months. Initial laboratory work up showed anemia, hypercalcemia and increase in free light chains kappa to lambda ratio, a diagnosis of multiple myeloma was suspected, magnetic reso¬nance imaging of the vertebral spine extensive involvement of bone marrow of the vertebrae. An 18F fluorodeoxyglucose positron emission tomography computed tomography (FDG PET CT) scan showed widely scattered moderate to intense uptake in multiple lytic of skeleton, Impression was : Image compatible with extensive multiple myeloma, No extra osseous involvement was found. Bone marrow examination was performed confirmed a diagnosis of extensive bone marrow involvement by High grade B-cell neoplasm with areas of bone marrow necrosis. The patient received six cycles of R-CHOP regimen (Rituximab, cyclo-phosphamide, Anthracycline, vincristine and prednisone) and achieved a complete response, as confirmed by FDG PET CT. At present, the patient is in a good condi¬tion.so this patient presentation mimicking multiple myeloma, which represent diagnostic dilemma. Primary lymphoma of the bone is rare disease entity, there is no strong evidence supporting some diagnostic and therapeutic decisions, and therefore, case presentation of the rare disease may facilitate treatment of similar diseases. A review of the literature to elucidate the clinical, radiological, pathological phenotype and optimal treatment of PLB done.

A Mechanistic Role For Mir-126, a Hematopoietic Stem Cell Microrna, In Acute Leukemias

We have recently shown that miR-126 expression faithfully identifies the engrafting fraction of bone marrow (BM) and cord blood (Gentner et al, Sci Transl Med 2010), in which it regulates hematopoietic stem cell (HSC) pool size by modulating cell cycle progression (Lechman et al., Cell Stem Cell 2012). Antagonizing miR-126 expands HSC by enhancing PI3K/AKT signaling, without causing their exhaustion or transformation.By reconstituting mice with lentivirally transduced BM cells ectopically expressing miR-126, we noted differential effects in HSC and progenitors: while miR-126 overexpressing (126OE) HSC were more quiescent and outcompeted by HSC with physiologic miR-126 expression, progenitor subsets were increased in number and proliferated more upon 126OE. In particular, B lymphopoiesis was enhanced, with the appearance of 126OE-vector marked, CD45(low)CD19+Ig(-) B cell progenitors in the peripheral blood. This abnormal population was transiently observed in all 126OE (n=45), but in none of the control mice (n=30), and BM B cell precursors and progenitors from 126OE mice showed significantly reduced levels of apoptosis, as well as a differentiation block at the immature B cell stage. At least part of this effect was due to interference of miR-126 with p53 activation. Consequentially, 20-30% of 126OE mice (as compared to 0% of control mice) followed over 1 year developed vector-marked, high-grade B-cell neoplasms which resembled diffuse large B-cell lymphoma, lymphoblastic lymphoma or acute lymphoblastic leukemia (ALL). Strikingly, by using a tetracycline-regulated, conditional miR-126OE vector, we observed full regression of the expanded abnormal B cell population after switching miR-126 expression off by doxycycline administration, demonstrating that miR-126 is an oncogenic driver in hematologic malignancies.To establish the relevance of miR-126 in human disease, we studied primary samples from patients with acute leukemia. We measured miR-126 expression in blasts from 12 adult patients with acute lymphoblastic leukemia (ALL). miR-126 was highly expressed in all studied cases of ALL (Phil+: n=7, Phil-: n=5), often surpassing the levels found in normal HSC. Unlike in HSC, miR-126 expression was independent from expression of its host gene EGFL7, suggesting an ALL-specific regulation of the miR-126 locus. By exploiting a lentiviral miR-126 reporter vector, we quantified miR-126 activity with single cell resolution after xenotransplantation of miR-126 reporter vector transduced ALL blasts into NSG mice. In 7 out of 12 ALL cases, we identified distinct subpopulations exhibiting different levels of miR-126 activity, and miR-126 (hi) ALL cells were more frequently contained in the CD34+ cell fraction. Moreover, high miR-126 expression correlated with faster and higher ALL engraftment in NSG mice. In acute myeloid leukemia (AML), miR-126 is significantly enriched in leukemic stem cells (LSC) where it governs quiescence and chemotherapy resistance (see abstract by Lechman et al), and analogous studies in ALL are ongoing. To further substantiate the role of miR-126 in AML, we studied miR-126 activity in 11 primary AML cases (4 AML with CBF, core binding factor, mutations; 7 AML with normal karyotype) using the reporter vector technology and the NSG xenotransplantation model. Globally, we found highest miR-126 activity in CBF-mutated AML. However, there was substantial heterogeneity in subpopulations of each single disease. Significantly higher miR-126 levels were found in CD34+, and particularly in CD34+CD38- AML cells, a fraction that is enriched for LSC. To see whether there is a correlation between miR-126 expression and disease progression, we measured miR-126 levels in sorted blasts from paired diagnosis/relapse (n=6) and diagnosis/chemotherapy-refractory (n=8) patient samples. Strikingly, we found a significant up-regulation of miR-126 in the disease progression sample, suggesting a mechanistic role for miR-126 in AML persistence after chemotherapy.In summary, these data support a clinically relevant role of miR-126 in the pathogenesis, treatment response and progession of acute leukemias.Authors AG and SN contributed equally, and LN and BG contributed equally. Disclosures:No relevant conflicts of interest to declare.

Expression of p16INK4a prevents cancer and promotes aging in lymphocytes

AbstractPrevious authors have suggested that tumor suppressor expression promotes aging while preventing cancer, but direct experimental support for this cancer-aging hypothesis has been elusive. Here, by using somatic, tissue-specific inactivation of the p16INK4a tumor suppressor in murine T- or B-lymphoid progenitors, we report that ablation of p16INK4a can either rescue aging or promote cancer in a lineage-specific manner. Deletion of p16INK4a in the T lineage ameliorated several aging phenotypes, including thymic involution, decreased production of naive T cells, reduction in homeostatic T-cell proliferation, and attenuation of antigen-specific immune responses. Increased T-cell neoplasia was not observed with somatic p16INK4a inactivation in T cells. In contrast, B lineage–specific ablation of p16INK4a was associated with a markedly increased incidence of systemic, high-grade B-cell neoplasms, which limited studies of the effects of somatic p16INK4a ablation on B-cell aging. Together, these data show that expression of p16INK4a can promote aging and prevent cancer in related lymphoid progeny of a common stem cell.

B-cell Lymphomas With Concurrent IGH-BCL2 and MYC Rearrangements Are Aggressive Neoplasms With Clinical and Pathologic Features Distinct From Burkitt Lymphoma and Diffuse Large B-cell Lymphoma

B-cell lymphomas with concurrent IGH-BCL2 and MYC rearrangements, also known as "double-hit" lymphomas (DHL), are rare neoplasms characterized by highly aggressive clinical behavior, complex karyotypes, and a spectrum of pathologic features overlapping with Burkitt lymphoma (BL), diffuse large B-cell lymphoma (DLBCL) and B-lymphoblastic lymphoma/leukemia (B-LBL). The clinical and pathologic spectrum of this rare entity, including comparison to other high-grade B-cell neoplasms, has not been well defined. We conducted a retrospective analysis of clinical and pathologic features of 20 cases of DHL seen at our institution during a 5-year period. In addition, we carried out case-control comparisons of DHL with BL and International Prognostic Index (IPI)-matched DLBCL. The 11 men and 9 women had a median age of 63.5 years (range 32 to 91). Six patients had a history of grade 1 to 2 follicular lymphoma; review of the prior biopsy specimens in 2 of 5 cases revealed blastoid morphology. Eighteen patients had Ann Arbor stage 3 or 4 disease and all had elevated serum lactate dehydrogenase (LDH) levels at presentation. Extranodal disease was present in 17/20 (85%), bone marrow involvement in 10/17 (59%) and central nervous system (CNS) disease in 5/11 (45%). Nineteen patients were treated with combination chemotherapy, of whom 18 received rituximab and 14 received CNS-directed therapy. Fourteen patients (70%) died within 8 months of diagnosis. Median overall survival in the DHL group (4.5 mo) was inferior to both BL (P=0.002) and IPI-matched DLBCL (P=0.04) control patients. Twelve DHL cases (60%) were classified as B-cell lymphoma, unclassifiable, with features intermediate between DLBCL and BL, 7 cases (35%) as DLBCL, not otherwise specified, and 1 case as B-LBL. Distinguishing features from BL included expression of Bcl2 (P<0.0001), Mum1/IRF4 (P=0.006), Ki-67 <95% (P<0.0001), and absence of EBV-EBER (P=0.006). DHL commonly contained the t(8;22) rather than the t(8;14) seen in most BL controls (P=0.001), and exhibited a higher number of chromosomal aberrations (P=0.0009). DHL is a high-grade B-cell neoplasm with a poor prognosis, resistance to multiagent chemotherapy, and clinical and pathologic features distinct from other high-grade B-cell neoplasms. Familiarity with the morphologic and immunophenotypic spectrum of DHL is important in directing testing to detect concurrent IGH-BCL2 and MYC rearrangements when a karyotype is unavailable. The aggressive clinical behavior and combination of genetic abnormalities seen in these cases may warrant categorization as a separate entity in future classifications and call for novel therapeutic approaches.

Primary Burkitt’s Lymphoma Of The Appendix Presenting as Acute Abdomen: A Case Report

Leukemic and lymphomatous involvement of the appendix is rare and even rarer is its presentation as appendicitis. Burkitt's lymphoma is a high grade B-cell neoplasm. Its non-endemic form typically presents as abdominal mass in children. This rapidly growing tumour may cause symptoms due to mass effect or direct involvement of the bowel. Clinical presentations like acute abdomen can be secondary to intestinal obstruction, intussusception or sometimes perforation.We describe here a case of an adult male with an unusual presentation of appendiceal Burkitt's lymphoma mimicking acute cholecystitis or appendicitis.

Optimal primer selection for clonality assessment by polymerase chain reaction analysis III. Intermediate and high-grade B-cell neoplasms

Previous studies have reported that low-grade B-cell lymphoproliferative disorders have variable B-cell monoclonality detection rates by polymerase chain reaction (PCR) analysis. For instance, monoclonal B-cell populations from chronic lymphocytic leukemia/small lymphocytic leukemia and mantle cell lymphoma are most often readily amplified with a single primer pair, whereas follicular lymphomas and plasma cell neoplasms require alternative strategies to approach these higher diagnostic sensitivity standards. Because most published reports have not focused on the variation in PCR B-cell monoclonality detection among subtypes of intermediate and high-grade B-cell neoplasms, additional information is necessary to determine primer selection strategies and identify problematic tumor subtypes within this group. The current investigation, the third part in a series, was aimed at documenting the efficiency of B-cell monoclonality detection by PCR in 71 aggressive B-cell neoplasms of various types using a comprehensive approach. A predetermined panel of primer sets was used in an algorithmic fashion. Specifically, all samples were analyzed with the standard VH-FRIII/JH assay previously shown to have the highest efficiency of monoclonality detection within low-grade B-cell lymphoproliferative disorders. Negative samples were further evaluated with primer sets in the following order until a positive result was observed, or all primer sets were used: (1) bcl-2/JH, (2) VH-FRI family specific/JH, and (3) VH-FRI consensus/JR. Forty-three (61%) of the 71 B-cell neoplasms evaluated with VH-FRIII/JH showed monoclonal B-cell populations. Sequential use of the three reserve primer sets in samples negative with this initial primer pair resulted in an overall improvement in PCR detection from 61% to 82% (58 of 71 specimens) ( P < .001). The VH-FRI family specific assay identified B-cell monoclonality in 11 (73%) of these 15 specimens and was the most productive reserve primer set. Individual categories exhibited the following initial (I) and final (F) PCR detection rates: acute lymphoblastic leukemia/ lymphoblastic lymphoma, 11 specimens (I = 91% to F = 91%); small noncleaved cell lymphoma, 14 specimens (I = 79% to F = 86% [ P > .25]); diffuse large cell lymphoma, 33 specimens (I = 52% to F = 85% [ P < .005]) and large cell, immunoblastic lymphoma, 13 specimens (I = 38% to F = 62% [ P < .01]). The authors have shown that comprehensive PCR analysis is capable of detecting B-cell monoclonality in a significant proportion of samples from each subtype of intermediate and high-grade B-cell neoplasm. The VH-FRIII/JH assay was an adequate initial primer set, but required augmentation with the reserve PCR assays to attenuate the false negative rate and improve diagnostic sensitivity. The B-cell clonality PCR assay is optimally used as a screening tool and when used in this fashion, the more laborious and time-consuming restriction fragment-Southern blot hybridization (RF-SBH) method for IgH gene rearrangement detection may be limited to a relatively small proportion of PCR-negative aggressive B-cell neoplasms.