Congenital B cell lymphocytosis explained by novel germline CARD11 mutations

Nuclear factor-κB (NF-κB) controls genes involved in normal lymphocyte functions, but constitutive NF-κB activation is often associated with B cell malignancy. Using high-throughput whole transcriptome sequencing, we investigated a unique family with hereditary polyclonal B cell lymphocytosis. We found a novel germline heterozygous missense mutation (E127G) in affected patients in the gene encoding CARD11, a scaffolding protein required for antigen receptor (AgR)–induced NF-κB activation in both B and T lymphocytes. We subsequently identified a second germline mutation (G116S) in an unrelated, phenotypically similar patient, confirming mutations in CARD11 drive disease. Like somatic, gain-of-function CARD11 mutations described in B cell lymphoma, these germline CARD11 mutants spontaneously aggregate and drive constitutive NF-κB activation. However, these CARD11 mutants rendered patient T cells less responsive to AgR-induced activation. By reexamining this rare genetic disorder first reported four decades ago, our findings provide new insight into why activating CARD11 mutations may induce B cell expansion and preferentially predispose to B cell malignancy without dramatically perturbing T cell homeostasis.

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The Emergence of Clonally Related Hodgkin Lymphoma Following B-Cell Malignancies Treated with Rituximab.

Blood ◽

10.1182/blood.v114.22.3664.3664 ◽

2009 ◽

Vol 114 (22) ◽

pp. 3664-3664

Author(s):

Catherine H Burton ◽

Sheila J.M. zsnO'Connor ◽

Roger G Owen ◽

Andrew S Jack

Keyword(s):

Follicular Lymphoma ◽

B Cell ◽

Hodgkin Lymphoma ◽

B Cell Lymphoma ◽

Tumour Cells ◽

Classical Hodgkin Lymphoma ◽

Composite Lymphoma ◽

Original Diagnosis ◽

B Cell Malignancy ◽

Cell Malignancy

Abstract Abstract 3664 Poster Board III-600 Classical Hodgkin lymphoma in association with another type of B-cell malignancy is a well recognised entity. In at least some of these composite lymphoma cases, a clonal relationship between the lymphoma sub-types can be demonstrated by the presence of common cytogenetic abnormalities or mutational patterns within the IGH locus. There is little known of possible mechanisms of clonal divergence in these cases. It is possible that treatment or the tumour micro-environment could, in some circumstances, favour the outgrowth of Reed Sternberg cells. These cells having lost many central phenotypic characteristics of mature B-cells may have an advantage over clonal tumour cells that remain under the control of normal regulatory pathways. In such a model, treatment with rituximab could add selective pressure favouring the development of a composite lymphoma. We have recently identified three male patients who relapsed with classical Hodgkin lymphoma after treatment for another form of B-cell malignancy. Patient 1, aged 76 years, had stage 3, asymptomatic follicular lymphoma, and was treated with rituximab alone. Eight months from the original diagnosis he developed new submental lymphadenopathy. Patient 2, aged 49 years, had stage four, symptomatic follicular lymphoma, treated with R-CVP. He developed biopsy proven high grade transformation at the end of his treatment and proceeded to two cycles of R-DHAP. At the end of treatment, twelve months from the original diagnosis, he developed inguinal lymphadenopathy. Patient 3, aged 70 years, had stage four, diffuse large B-cell lymphoma, treated at presentation with R-CHOP. Two years after initial diagnosis, he developed cervical lymphadenopathy. In all three presenting cases, CD20 was strongly expressed on the lymphoma cells and a t(14;18) was identified in the biopsy. In all three relapsed cases the biopsy showed morphologically typical classical Hodgkin lymphoma with CD30/IRF4 co-expression and absence of Oct2, Bob1 and CD20 expression within the Reed Sternberg cells. A t(14;18) was detected by FISH in the Reed Sternberg cells, demonstrating clonal identity with the underlying lymphoma. There was no evidence of the preceding lymphoma and complete absence of a normal B-cell population in the biopsy. Epstein Barr Virus was not detectable by EBER in situ hybridisation. All patients have been treated for Hodgkin lymphoma. Patient 1 has subsequently died, patient 2 is awaiting a stem cell transplant and patient 3 is currently receiving treatment. Treatment with rituximab is associated with the development of CD20 negative phenotypic change. In some cases this may be due to the selection of tumour cells that have epigenetically silenced the expression of CD20. Epigenetic silencing of key transcription factors may also be the mechanism of loss of the B-cell phenotype, including CD20, in Reed Sternberg cells. The relatively frequent occurrence of composite lymphoma suggests that this may occur regularly with neoplastic B-cell populations and the outgrowth of these cells would be strongly favoured by rituximab treatment. The cases presented here highlight the need for a more systematic approach to the collection of data from patients with relapsed lymphoma. An association between rituximab treatment and the development of composite lymphoma has practical consequence but may also provide a unique insight into the pathogenesis of Hodgkin lymphoma. Disclosures: No relevant conflicts of interest to declare.

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Association of Renal Cell Carcinoma and B-Cell Hematological Malignancy

Blood ◽

10.1182/blood.v120.21.5086.5086 ◽

2012 ◽

Vol 120 (21) ◽

pp. 5086-5086 ◽

Cited By ~ 1

Author(s):

Rangaswamy Chintapatla ◽

Leticia Varella ◽

Peter Wiernik ◽

Valerie Rusciano ◽

Janice P. Dutcher

Keyword(s):

Acute Myeloid Leukemia ◽

B Cell ◽

Renal Cell ◽

Myeloid Leukemia ◽

Family Members ◽

B Cell Lymphoma ◽

B Cell Malignancies ◽

B Cell Malignancy ◽

Acute Myeloid ◽

Cell Malignancy

Abstract Abstract 5086 Purpose of the study: We observed an increased frequency of hematologic malignancy (HM) in patients and family members of patients with renal cell cancer (RCC) and sought to characterize the association further in terms of frequency and characteristics of HM, and the importance of such an association. Methods: We performed a chart review of our data base of approximately 700 RCC patients seen by us from 2004 to the present in an effort to determine the frequency of HM in patients and in the families of patients diagnosed with RCC. Results: Of the 700 charts reviewed, both HM and RCC occurred in 19 individuals. [11 males and 8 females]. HM diagnosis included acute myeloid leukemia in 1 patient, Hodgkin's lymphoma (HL) in 4 pts, non-Hodgkin lymphoma (NHL) in 7 pts, (3 small cell and 4 large B-cell lymphoma), chronic lymphocytic leukemia (CLL) in 2 pts and hairy cell leukemia (HCL), monoclonal gammopathy of undetermined significance (MGUS) myelodysplasia (MDS) in one patient each. A family history of HM was found in 71 relatives involving 56 families of patients with RCC. Of these, 48/71 cases of HM were in first degree relatives and 18/71 were in second degree relatives. The most common HMs were lymphoma and leukemia: 24 NHL, 9 HL, 6 lymphoma not further specified (NOS), 11 CLL, 1 acute myeloid leukemia, 5 acute leukemia NOS and 6 leukemia NOS. Other HM observed once were multiple myeloma, Waldenström's macroglobulinemia, chronic myeloid leukemia, myelofibrosis and polycythemia vera. In addition, 2 family members had blood cancers that were NOS. Thus, of 77 patients/family members with known HM diagnosis 94% were B-cell malignancies. Clear cell histology was the most common subtype of RCC, and all subtypes of RCC occurred in the study population with expected frequency. RCC and HM occurred in the same patient in this study more frequently at 2. 7% than would be expected from a SEER database. In that database the observed to expected (O/E) ratio of NHL and RCC was 1. 86 to 2. 07% [Kunthur et al. Am J Hematol 2006; 81:271–80]. Conclusions: Increased incidence of B-cell malignancy has been reported in individuals with RCC [Dutcher et al. Proc Am Fed Clin Res, Eastern Division, April 2011]. Wiernik et al. [Cancer J 2000] reported a similar increase was noted between adenocarcinoma of the breast and B cell malignancies in same individual and mouse mammary tumor virus was proposed a potential causative agent. There is a preponderance of B-cell malignancy in both the individuals and in the families of the patients with RCC noted in this study. The etiology of this association between RCC and HM is unclear and suggests a common etiopathogenesis for RCC and B-cell tumors, or a familial immunologic defect that facilitates both malignancies. We plan to further explore the relationship of HM to RCC. Disclosures: No relevant conflicts of interest to declare.

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The Eµ-Myc/TCL1 Transgenic Mouse As a New Aggressive B-Cell Malignancy Model Suitable for Preclinical Therapeutics Testing

Blood ◽

10.1182/blood.v126.23.2752.2752 ◽

2015 ◽

Vol 126 (23) ◽

pp. 2752-2752

Author(s):

Kerry A. Rogers ◽

Dalia El-Gamal ◽

Harrington K. Bonnie ◽

Hing A. Zachary ◽

Goettl M. Virginia ◽

...

Keyword(s):

B Cell ◽

Cell Transformation ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Large Cell ◽

Aggressive Lymphoma ◽

Targeted Therapeutics ◽

B Cell Malignancy ◽

Large Cell Transformation ◽

Cell Malignancy

Abstract Background: Aggressive B-cell lymphomas occurring in the setting of Chronic Lymphocytic Leukemia (CLL) as a large cell transformation are an important clinical problem, and improved mouse models to test novel and targeted therapeutics are needed. The Eµ-Myc mouse overexpresses c-Myc gene which is placed under control of the Myc promoter and lymphoid-specific IgH enhancer (Eµ), resulting in c-Myc overexpression and spontaneous B-cell lymphoma development. The Eµ-Myc mice have been used in drug development, however malignancy develops at variable ages and with differing genetics and response to therapeutic agents, making drug studies difficult. The Eµ-TCL1 transgenic mouse overexpresses the human TCL1 oncogene, under the control of the B-cell specific IgVH promoter and Eµ enhancer. Mice develop a spontaneous mature B-cell leukemia after a long latency period and represent a well-established model of human CLL. We crossed the Eµ-Myc and TCL1 mice to create a new model of aggressive B-cell lymphoma to test novel therapeutics that would be more homogeneous than the Eµ-Myc model. Methods: Eµ-Myc and TCL1 mice on C57BL/6 background strain were crossed and resulting genotypes were verified by PCR. All mice were kept in standard pathogen-free housing until death or removal from study. All mice born within a 24 month time period were followed for survival. At the time of death, spleen, lymph nodes, bone marrow, and liver were fixed in 10% neutral buffered formalin, embedded in paraffin, sectioned at 4uM, and stained with hematoxylin and eosin for histologic evaluation. For engraftment studies, adoptive cell transfer was done by injecting fresh splenic white blood cells (1e7 cells, >75% CD5+/CD19+) into the tail vein of C57BL/6 wild type recipient mice. In the drug treatment experiments ibrutinib (~30mg/kg/day) was administered in drinking water and KPT-8602 (15mg/kg) by daily oral gavage. Survival was shown using Kaplan-Meier curves and survival curves compared statistically using a log-rank test. Results: The Eµ-Myc/TCL1 mice had significantly worse survival (median 45.5, range 29-74 days) compared to Eµ-Myc (median 118, range 42-520 days) or TCL1 (median 359.5, range 188-531 days) mice (p=<0.0001). Survival curves are show in Figure 1. Pathologic examination revealed aggressive B-cell histology similar to Burkitt lymphoma in both the Eµ-Myc mice and Eµ-Myc/TCL1 mice, whereas the TCL1 mice showed mature B-cell morphology. Infiltration of the bone marrow of Eµ-Myc and Eµ-Myc/TCL1 mice was extensive in comparison with TCL1 mice. In all three mouse genotypes the normal architecture in the spleen was effaced by neoplastic cells. Adoptive transfer of Eµ-Myc/TCL1 splenic lymphocytes produced similar pathologic findings to the Eµ-Myc/TCL1 transgenic mice with less severe splenic enlargement in the engrafted mice. Median survival of the engrafted mice was short (38 days). There was a differential response to targeted therapeutics between the groups. Ibrutinib, an irreversible inhibitor of Bruton tyrosine kinase (BTK) previously demonstrated to improve survival in TCL1 mice, had no survival advantage over vehicle treatment in either Eµ-Myc or Eµ-Myc/TCL1 mice (p=0.876 and P=0.83 respectively). KPT-8602, a second generation selective inhibitor of exportin 1 (XPO1) was tested in Eµ-Myc/TCL1 engrafted mice, and demonstrated a significant improvement in survival compared to vehicle alone (p=<0.0001). A first generation agent (selinexor) targeting XPO1 is currently in human clinical trials for aggressive lymphomas, including large cell transformation of CLL. Conclusions: Mice with both c-Myc and TCL1 transgenes develop an aggressive B-cell malignancy and have decreased survival compared to Eµ-Myc or TCL1 mice. The histology of the disease is similar to that of Eµ-Myc mice, but with shorter survival with less variability. The resulting mouse model has differential responses to targeted therapeutics, with a poor response to ibrutinib more similar to aggressive lymphoma than CLL. The malignant B cells can be adoptively transferred into immunocompetent mice for experimental drug treatment studies. These qualities make Eµ-Myc/TCL1 mice a useful tool to test new therapies for aggressive lymphoma where there is an unmet clinical need. Work is currently underway to further define the genetic characteristics of this mouse in comparison to human lymphoma sub-types and responsiveness to targeted agents. Disclosures Byrd: Acerta Pharma BV: Research Funding.

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Ph Chromosome and B-Cell Malignancy-Associated Chromosomal Aberrations in Non-Leukaemic Immunoblastic B-Cell Lymphoma

Acta Haematologica ◽

10.1159/000206202 ◽

1985 ◽

Vol 74 (3) ◽

pp. 171-174 ◽

Cited By ~ 8

Author(s):

Gunnar Juliusson ◽

Kristina Friberg ◽

Gösta Gahrton

Keyword(s):

B Cell ◽

Chromosomal Aberrations ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

B Cell Malignancy ◽

Ph Chromosome ◽

Cell Malignancy

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Anti-Infectious Pathogen Specificity of Purified Monoclonal Immunoglobulins from Patients with MGUS and Multiple Myeloma

Blood ◽

10.1182/blood.v128.22.4413.4413 ◽

2016 ◽

Vol 128 (22) ◽

pp. 4413-4413

Author(s):

Adrien Bosseboeuf ◽

Delphine Feron ◽

Jean Harb ◽

Cathy Charlier ◽

Anne Tallet ◽

...

Keyword(s):

Multiple Myeloma ◽

B Cell ◽

Western Blotting ◽

B Cell Lymphoma ◽

Solid Cancer ◽

Varicella Zoster ◽

B Cell Malignancy ◽

H Pylori ◽

Cell Malignancy ◽

Hsv 1

Abstract Background: Multiple myeloma (MM) is characterized by the accumulation of malignant plasma cells, which secrete >30g/L of monoclonal immunoglobulin (mc Ig). MM is always preceded by the usually asymptomatic stage called "monoclonal gammapathy of undetermined significance" (MGUS). Presently the detection of a mc Ig does not lead to analysis of its specificity of antigen (Ag) recognition. Yet certain infectious pathogens are associated with the development of B-cell malignancy: for instance, Epstein-Barr virus (EBV) and Burkitt lymphoma, Helicobacter pylori (H. pylori) and MALT lymphoma. Objective: Our aim was to determine the frequency of recognition by purified mc Ig from MGUS and MM patients, of infectious pathogens associated with chronic infection and B-cell malignancy. Methods: We used the "Multiplex Infectious Antigen Array" (MIAA)test set up by the team to study the specificity of purified mc IgG against 8 infectious pathogens: hepatitis C virus (HCV), EBV, H. pylori, varicella zoster virus (VZV), cytomegalovirus (CMV), herpesvirus simplex 1 (HSV-1), HSV-2, and Toxoplasma gondii. Second or third methods were used to verify the specificity of mc IgG (western blotting, ELISA, or another multiplex array). Sialylation of mc Ig, which reflects chronic inflammation, was also studied, via ELISAs and Western blotting, for 98 MGUS and MM patients and 43 controls. Results: The specificity of purified mc IgG was analyzed for 291 patients (60 MGUS, including 4 associated with a B-cell lymphoma; 108 MM; for the 123 other patients the diagnosis of MGUS or MM was not available). Purified mc IgG were found to be specific for one of the 8 infectious pathogens of the MIAA test for 17.5% of patients (51/291: 25 MGUS, 1 SMM, and 25 MM). Purified mc Ig specifically recognized EBV (n=27, including 8 MGUS, 1 SMM and 18 MM ; the Ag recognized were EBNA-1, 25 cases; and VCA, 2 cases); HCV (10 cases: 9 MGUS and 1 MM) ; HSV-1 (7 cases: 5 MGUS including 3 associated with a B-cell malignancy, and 2 MM) ; H. pylori (5 cases: 2 MGUS including 1 associated with a B-cell malignancy, and 3 MM); VZV (1 MM); and CMV (1 MGUS). The Ag most frequently recognized by purified mc IgG was EBV EBNA-1; MM patients with EBNA-1-specific mc IgG represented 10% of the MM in this series. Consistent with a pro-inflammatory effect, for 81.6% of MGUS and MM patients (80/98), the purified mc IgG was found to be hyposialylated. Moreover, purified polyclonal Ig were also hyposialylated for 37% of MM (compared to <2.3% of the polyclonal Ig from healthy donors or MGUS). Conclusion: At least 6 infectious pathogens (EBV, HCV, H. pylori, HSV, VZV, CMV) may be involved in the pathogenesis of MGUS and MM with mc IgG. Importantly, EBV, HCV, H. pylori and HSV are known as oncogenic pathogens associated with solid cancer and/or B-cell malignancy. Finally, for 37% of the MM patients examined, both the polyclonal and mc Ig were found to be hyposialylated, which suggests that inflammation existed prior to clonality and MM. Disclosures Garderet: Takeda: Consultancy; Amgen: Consultancy; BMS: Consultancy, Honoraria; Novartis: Consultancy.

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