bluuebunny/crossref_metadata_2025_split huggingface dataset:
IgA and IgG Hypogammaglobulinemia Is a Constitutive Feature in Most Waldenstrom’s Macroglobulinemia Patients and May Be Related to Mutations Associated with Common Variable Immunodeficiency Disorder (CVID)
Authors: Treon Steven , Hunter Zachary , Ciccarelli Bryan , Zhou Yangsheng , Hanzis Christina , Manning Robert , Ioakimidis Leukothea , Xu Lian , Patterson Christopher , Morra Massimo 
Published: October 2019

Abstract
Background: Hypogammaglobulinemia of the “uninvolved” immunoglobulins is commonly observed in Waldenstrom’s macroglobulinemia (WM), and has often been attributed to disease-related suppression. However, there is a paucity of information related to the pathogenesis of hypogammaglobulinemia in these disorders.
Methods: We evaluated the incidence of IgA and IgG hypogammaglobulinemia in 207 patients with WM, and addressed the impact of therapy and response on IgA and IgG levels for 93 of these patients who required subsequent treatment. We also performed extensive sequence analysis of the promoter, all exonic, and flanking intronic regions from peripheral blood of 19 untreated WM patients who demonstrated IgA and/or IgG hypogammaglobulinemia for 8 genes often observed in common variable immunodeficiency disorders (CVID) and B cell deficiency i.e. AICDA, BTK, CD40, CD154, NEMO(IkBkG), TACI, SH2D1A, UNG.
Results: At baseline, 120/207 (58.0%), 131/207 (63.3%), and 102/196 (49.3%) patients had abnormally low levels of serum IgG (<700 mg/dL), IgA (<70 mg/dL), or both, respectively. No correlation between baseline bone marrow disease involvement and immunoglobulin levels was observed. With a median follow-up of 12 months following completion of therapy, IgA and/or IgG levels remained abnormally low for 92.1% and 87.3% of responding patients, respectively, including those who achieved a complete remission. Sequence analysis of the NEMO and CVID gene panel demonstrated intronic variation at position c.1056-6T>C (n=2) and a hemizygous missense mutation at codon 113 for NEMO (n=1), and a heterozygous missense mutation at codon 142 in UNG (n=1).
Conclusions: The results of these studies demonstrate that IgA and IgG hypogammaglobulinemia is a constitutive feature of patients with WM, which neither correlates with, nor is impacted by disease burden, despite therapeutic intervention and response. The results also suggest that patients with WM may harbor sequence mutations, which may be a contributor to the pathogenesis and/or morbidity of WM.
Early Response Using Serum Immunoglobulin Free Light Chain (sFLC) Measurement Predicts Response to Therapy in Waldenstrom Macroglobulinemia (WM)
Authors: Leleu Xavier , Roccaro Aldo M. , Leduc Renee , Poulain Stephanie , Moreau Anne-Sophie , Gay Julie , Dulery Remy , Richardson Paul G. , Ghobrial Irene M. 
Published: October 2019

Abstract
Background. Measurement of serum M-spike is used to determine response to therapy and treatment free survival in Waldenstrom Macroglobulinemia (WM). However, there are many limitations to the use of IgM M-spike, and new sensitive markers are needed to determine early response or progression in these patients. We have previously shown that involved serum free light chain (sFLC) accurately diagnosed patients with WM, and correlated with markers of poor prognosis, specifically beta- 2 microglobulin (B2M). In this study, we sought to determine the role of levels of involved in the response to therapy in patients with WM treated on clinical trials, and compare it to the response observed using the traditional M-spike measurement.
Methods. We prospectively studied involved sFLC in 61 WM patients enrolled on 2 clinical trials, at diagnosis (N=8) and with relapse/refractory disease (N=53). Patients were treated on one of two clinical trials: either perifosine (N=30; given 150mg oral daily) or combination of bortezomib and rituximab (N=30; given IV bortezomib 1.6mg/m2 at days 1, 8, 15 q 28 days × 6 cycles and rituxan 375 mg/m2 at days 1, 8, 15, 22 on cycles 1 and 4). Response was assessed after cycle 2, confirmed on 2 consecutive measurements, and included minor response (MR), partial response (PR) or complete remission (CR).
Results. The baseline characteristics of the patients were as follows: the median age was 65 years (44–83), male/female ratio 2.38, serum B2M 3.0mg/L (1.00–7.30), hemoglobin 11.0g/dL (7.00–14.90), platelet count 216 ×109/mm3 (46–563), serum M-spike 2.24g/L (0.41–4.62), and involved sFLC 95.2mg/L (4.92–868). The WM International staging system (WM-ISS) showed that 28% of patients had low ISS, 31% intermediate, and 41% high ISS scores. The overall response rate for the entire cohort was 62.3%, assessed by M-spike measurement (MR=23, PR=13, nCR=2). The overall response rate using involved sFLC level was 72.1% (MR=14, PR=29, and CR=1), (p<0.001). We then investigated the role of sFLC in predicting early response to therapy in patients treated with bortezomib and rituximab. We studied the level of sFLC weekly for the first month of therapy. Interestingly, early measurement of sFLC in the first month of therapy significantly predicted response; 93% of the patients who obtained at least an MR within the first 3 weeks of therapy by sFLC demonstrated a response on further follow up using serum M-spike (p=0.024).
Conclusion. In this study, we identified that involved sFLC is a new and reliable marker for monitoring response to therapy in WM disease, especially to determine early response in these patients.
A Prognostic Role for Absolute Lymphocyte Counts in Pediatric Non- Hodgkin’s Lymphoma
Authors: Levy Alejandro G. , Sabella-River Romina , Ghisoli Maurizio L. , Nunez Cesar A. , Anderson Pete M. , Palla Shana L , De Angulo Guillermo , Zweidler-McKay Patrick A. 
Published: October 2019

Abstract
A role for the immune system during treatment for cancer therapy has not been widely established. However, we have previously reported that lymphocyte counts during chemotherapy independently predict outcome in pediatric patients with acute lymphoblastic leukemia (ALL), acute myelogenous leukemia, and Ewing’s sarcoma. Others have reported similar observations in adults with AML, follicular lymphoma and diffuse large B cell lymphoma. This led us to evaluate the prognostic ability of lymphocyte counts in pediatric patients with non-Hodgkin’s lymphoma (NHL). Seventy-six consecutive patients with de novo NHL, age 1–21 years, who were treated at M. D. Anderson Cancer Center were reviewed. We analyzed absolute lymphocyte counts (ALC) at diagnosis and days 7, 15, 21 and 28 of induction chemotherapy. We found that similar to children with ALL, lymphocyte counts in young patients with NHL during induction chemotherapy are independently prognostic for both overall survival and relapse-free survival. For example, children with low ALC on day 7 (ALC-7) of induction had poor RFS and OS. Specifically, patients with ALC-7 <600 cells/μL had a 2-year RFS of 56% versus 95% for those with ALC-7 >600 cells/mcL (HR=4.4, p=0.003). Strikingly, the 19% (12/63) of children with low ALC on both day 7 and 15 (<600 cells/mcL) had a 2-year RFS of 23% compared with 90% for those with either an ALC-7 or -15 >600/mcL (HR=8, p=0.0004). This effect was so dramatic that when ALC-7 of treatment was evaluated as a continuous variable, each cell/mcL increase in ALC was protective for OS decreasing the hazard ratio by 0.002 (p=0.04–0.05). This predicts that an increased ALC of only 250 cells/mcL would yield an HR of 0.5. Importantly, in multivariate analysis with disease stage (I+II versus III+IV), ALC-7<600 remained significantly prognostic for OS and RFS (HRs 6.2–12.2, p=0.006–0.018). Interestingly, lymphocyte counts prior to starting treatment also were prognostic with ALC<500–1000/mcL predicting poor OS (HRs 3.4–4, p=0.04–0.05).
These data reveal that low lymphocyte counts prior to and during induction chemotherapy in children with NHL are significant independent predictors of poor prognosis. These findings support a potential role for the immune system in the treatment of pediatric NHL which should be explored. Outcome differences of 39% are highly clinically relevant and suggest that ALC during induction should be evaluated prospectively and may contribute to future risk stratification of pediatric NHL.
Figure Figure
From Signalosome to Enhanceosome: The Translocation of CD40 from Membrane to Nucleus Involves Multiple Mechanisms
Authors: Zhou Hai-Jun , Tamayo Archie , Pham Lan , Lin-Lee Yen-Chiu , Ford Richard J. 
Published: October 2019

Abstract
CD40 plays important roles in the proliferation, survival and differentiation of lymphocytes. Constitutively active CD40 recruits TRAFs and IKKs within the lipid rafts to form a signalosome that mediates pivotal downstream proliferation and survival mechanisms involving NF-kB. Recently, we have reported that nuclear localization of CD40, through its interaction with c-Rel, promotes growth, cell cycle progression and survival in large B cell lymphoma. Our studies have opened a new paradigm in the functional role of CD40 in non-Hodgkin lymphomas of B cell origin (NHL-B). However, the mechanism about how CD40 enters nuclear still remains elusive. In this study, we show that CD40 ligation enhances its nuclear accumulation with activation of c-Rel in both normal B-lymphocytes and B cell lymphoma cells with cell fractionation assay and con-focal microscopy. Over-expression of c-Rel in B cell lymphoma cells drives CD40 into cell nucleus. We hypothesize that the route CD40 enters nucleus may involve endosome-endoplasmic reticulum-nuclear pore complex. Indeed, further studies show CD40 co-localizes with endosome marker-EEA1 and endoplasmic reticulum marker-Sec61. Furthermore, our co-immunoprecipitation assay has demonstrated CD40 interacts with Sec61. CD40 also co-localizes and immuno-precipitates with nuclear pore complex (NPC) proteins-NUP62 in normal B-lymphocytes and B lymphoma cells, which suggests NPC proteins may facilitate the nuclear translocation of CD40 protein. Overall, our study suggests that translocation of CD40 into cell nucleus involves multiple pathways. Blocking nuclear localization may modulate the function of CD40 in lymphoma cells; which could provide a new-targeted therapeutic approach for lymphoma therapy.
Dissecting Proto-Oncogenic PIM Serine/Threonine Kinases in FLT3-ITDInduced Leukemogenesis: PIM1 Regulates CXCL12/CXCR4-Mediated Homing and Migration
Authors: Gasser Christelle , Grundler Rebekka , Brault Laurent , Bullock Alec , Dechow Tobias , Nawijn Martjin , Woetzel Sabine , Ehret Sabine , Berridge Georgina , Spoo Anke , Dierks Christine , Biondi Andrea , Knapp Stefan , Duyster Justus , Schwaller Juerg 
Published: October 2019

Abstract
Previous work has shown that FLT3-ITD mediated leukemogenesis is associated with increased expression of PIM1 and PIM2 serine/threonine kinases. Here we show that retroviral expression of FLT3-ITD could not compensate impaired clonogenic in vitro growth of PIM1−/− bone marrow cells. Induction of a lethal myelo- and lymphoproliferative disorder by FLT3-ITD in vivo was independent of PIM2, but rather unexpectedly, lethally irradiated recipients could not be reconstituted with FLT3-ITD expressing bone marrow cells lacking PIM1. Transplants of CSFE-labeled PIM1−/− cells revealed an impaired homing capacity to bone marrow and spleen. Expression of lower surface CXCR4 levels (while maintaining normal total CXCR4 levels) in PIM1−/− bone marrow cells was associated with significantly reduced migration towards a CXCL12 gradient and impaired CXCL12-mediated intracellular Ca2+ release. Using siRNA-mediated knockdown, a small molecule PIM inhibitor, expression of a dominant-negative acting PIM1 mutant or re-expression of PIM1 in knockout cells, we observed that PIM1 activity was critical for CXCR4 surface expression. In vitro kinase assays and masspectrometric analysis further revealed that PIM1 directly phosphorylated serine 339 located in the CXCR4 intracellular domain known to be essential for proper receptor recycling. Interestingly, in leukemic blasts from acute myeloid leukemia (AML) patients, we found an association of increased PIM1 expression and high-level of surface CXCR4. In addition, treatment of the cells with a small molecule PIM inhibitor resulted in decreased surface CXCR4 expression in some patients. Our work suggests that PIM1 exerts its oncogenic activity not only by supporting proliferation and survival but also by regulation of cell homing and migration through direct modification of the CXCL12/CXCR4 axis. As CXCR4 is a key mediator of cancer stem cell homing and metastasis, targeting of PIM1 may offer new therapeutic avenues against tumor progression and relapse.
The C-Terminus of the CBFβ-SMMHC Fusion Protein Is Required for the Myeloblastic Transformation of inv16 Leukemia
Authors: Kamikubo Yasuhiko , Garrett-Beal Lisa , Kirby Martha , Liu Pu Paul 
Published: October 2019

Abstract
Inv(16)(p13q22) is found in almost all human acute myeloid leukemia (AML) subtype M4Eo cases and forms a fusion gene CBFB-MYH11, which encodes a fusion protein CBFβ–SMMHC. CBFβ forms a heterodimeric transcription factor with RUNX1 and both are required for embryonic hematopoiesis, while SMMHC is the smooth muscle myosin heavy chain. Using knock-in mouse strategy, we previously demonstrated that Cbfb+/MYH11 F1 embryos have severe blockage of definitive hematopoiesis and die from CNS hemorrhage. The phenotype was similar to that of embryos with homozygous deletion of Cbfb or Runx1, suggesting that Cbfb-MYH11 dominantly represses Runx1/CBFb function. We further demonstrated that Cbfb-MYH11 is necessary but not sufficient for leukemia to develop, as the Cbfb+/MYH11 mice required additional mutations for leukemogenesis. Several hypotheses have been proposed, based on in vitro studies, to explain how CBFβ-SMMHC dominantly inhibits RUNX1/CBFβ. The C-terminal region of CBFβ-SMMHC is responsible for multimerization and also interacts with corepressors; thus this region might be critical for RUNX1 repression. To determine the importance of this multimerization domain in vivo, we generated knock-in mice expressing CBFβ-SMMHC with a 95 aa C-terminal deletion (CBFβ-SMMHCdC95), which truncates this multimerization/repression domain. CBFβ-SMMHCdC95 expressing F1 heterozygous embryos (Cbfb+/MYH11dC95) developed normally with no hematopoietic defects and no hemorrhage. Hematopoiesis was normal in the adult Cbfb+/MYH11dC95 mice except for mild increase of mature neutrophils and minor T cell developmental defects in the first year. However, the mice proceeded to a lethal myeloproliferative disorder (MPD) during their second year of life. There was a significant increase of Mac1/Gr1 double positive cells in the peripheral blood and the spleen, which were negative for the stem cell/progenitor cell marker, c-kit. Morphologically the erythrocytes and neutrophils were dysplastic, and the mice developed severe splenomegaly. ENU treatment of the Cbfb+/MYH11dC95 mice accelerated the development of the MPD phenotype but could not induce overt, transplantable, myeloid leukemia. These data suggest that the multimerizatin domain of SMMHC is important for both hematopoiesis blockage and leukemogenesis, especially the blastic transformation of inv16 leukemia.
The Role of Hepcidin in Anemia of Cancer
Authors: Rivera Seth , Kim Airie , Ganz Tomas 
Published: October 2019

Abstract
Anemia is a common occurrence in a wide variety of malignancies. When the anemia is not due to bleeding, bone marrow infiltration or chemotherapy, it is termed anemia of cancer (AC). AC is presumed to be a form of anemia of inflammation (AI, aka anemia of chronic disease). By limiting the availability of iron absorption from diet and release from stores, hepcidin causes AI. To test whether hepcidin contributes to AC, we created two syngeneic mouse models of lung cancer (TC-1 and LLC) and a syngeneic mouse model of melanoma (B16-F10). In all three models, significant anemia developed (Hgb 10.9±2.0, 9.1±2.0, 7.8±2.0 for TC-1, LLC, and B16-F10 vs. 13.7±1.2 for controls; p<0.001 for all compared to controls). There was no evidence of bleeding in the tumors or intraperitoneally. The anemia was microcytic in TC-1 mice but not LLC or B16-F10. Hepcidin mRNA (hepatic production measured by qRT-PCR) was increased in TC-1 mice but not LLC or B16-F10. To test whether hepcidin was necessary for the development of anemia in the TC-1 model, we generated tumors in hepcidin knockout mice. Hgb did not fall significantly in hepcidin knockout mice with TC-1 induced tumors (Hgb 14.5±2.4 vs. 15.5±2.2, p=0.3).
CONCLUSIONS: Hepcidin is necessary for the development of AC when hepcidin production is induced by the tumor and microcytosis is present. However, AC is likely a heterogenous disease; experiments are underway to determine whether hepcidin plays a role in cancers that do not upregulate hepcidin production or cause microcytosis.
Inhibition of Hamp1 Expression by TMPRSS6, GDF15, PIAS4 and SMAD6 Utilize Several Target Sites in the Hamp1 Promoter
Authors: Truksa Jaroslav , Lee Pauline , Beutler Ernest 
Published: October 2019

Abstract
Hepcidin, the key regulator of iron homeostasis, is up-regulated by iron excess, bone morphogenetic proteins (BMPs) and inflammatory cytokines and down-regulated by hypoxia and anemia. Known positive regulators at the level of transcription factors include SMAD1/4, STAT3 and CEBPα. In this study, we focused on negative regulators of hepcidin regulation:
Matriptase II/TMPRSS6 (Transmembrane serine protease 6, a recently identified negative regulator in which disruption leads to anemia in mice as well in humans); Protein inhibitor of activated STATs no. 4 (PIAS4); Growth and differentiation factor 15 (GDF15, a potential erythroid negative regulator); and SMAD6 (Mothers against decapentaplegic homolog 6, an inhibitory SMAD blocking the SMAD/BMP pathway).
All tested inhibitors significantly decreased expression of the luciferase reporter under the control of 2.5 Kb murine Hamp1 promoter with GDF15 and PIAS4 Hamp1 specific since none of the inhibitors were able to reduce expression of the luciferase reporter under the control of the murine Hamp2 promoter. Inhibition of the luciferase reporter under the control of the 2.5 Kb murine Hamp1 promoter by SMAD6, unlike TMPRSS6, PIAS4 and GDF15, did not require liver specific transcription factors since the inhibition could also be observed in transfected HEK293T cells. GDF15, PIAS4, TMPRSS6 and SMAD6 all reduced basal level expression of the luciferase reporter under the control of the 2.5 Kb murine Hamp1 promoter as well as the total level of reporter expression induced by IL-6 and BMP-4. Nevertheless, GDF15 did not affect responsiveness (fold induction) to IL-6 and BMP-4. PIAS4 and TMPRSS6 inhibited responsiveness to IL-6 but had little effect on responsiveness to BMP-4. In contrast, SMAD6 did not affect responsiveness to IL-6 but reduced responsiveness to BMP-4. Deletion of the −140 bp −260 bp region of the murine Hamp1 or double deletion of the BMP-RE1 and BMP-RE2 motifs severely reduced the ability of all inhibitors to reduce reporter expression. Deletion of the STAT site abrogated PIAS4 inhibition while deletion of either BMP-RE1 or BMP-RE2 motifs alone partially reduced inhibition by TMPRSS6 and SMAD6. We conclude that there are several independent pathways that inhibit hepcidin expression.
cIAP1-Dependent TRAF2 Degradation Is Required for Macrophage Differentiation and Function
Authors: Dupoux Alban , Cartier Jessy , Marivin Arthur , Cathelin Séverine , Filomenko Rodolphe , Solary Eric , Dubrez-Daloz Laurence 
Published: October 2019

Abstract
TRAF2 (Tumor necrosis factor (TNF) Receptor associated factor) is an adaptor molecule involved in TNF family membrane receptor signalling pathway. We demonstrate that TRAF2 is required for macrophage differentiation. Its silencing prevents Iκ-Bα degradation, NF-κB p65 nuclear translocation and the differentiation process. Then, TRAF2 is degraded by the proteasome system in the cytoplasm. Inhibition or silencing of the E3 ubiquitine ligase cIAP1 from the IAP (Inhibitor of apoptosis protein) family of proteins, prevents the differentiation-associated TRAF2-degradation. In the presence of proteasome inhibitor, TRAF2 colocalizes with cIAP1 in a golgi-associated compartment, suggesting that cIAP-1-mediated TRAF2 degradation occurred in this subcellular compartment. An overexpression of TRAF2 or silencing of cIAP1 prevents the outcome of differentiation as revealed by the expression of differentiation markers at the cell surface, and inhibits the secretion of pro-inflammatory cytokines such as interleukin-8 and MCP-1 by fully differentiated-macrophages in response to CD40 stimulation. Our data suggest that TRAF2 expression and its subsequent cIAP1-dependent degradation is required for the differentiation of monocytes into macrophages and allows the macrophages to be fully functional.
Conditional T-Cell Activation for Tumor Under Hypoxia
Authors: Ang Sonny O , Olivares Simon , Shpall Elizabeth J. , Lee Dean A. , Champlin Richard , Yotnda Patricia , Gottschalk Stephen , Cooper Laurence 
Published: October 2019

Abstract
Solid tumors are characterized by cell clusters experiencing chronic and intermittent hypoxia due to haphazard growth. Importantly, highly migratory tumorigenic stem cells are localized within hypoxic niches, shielded by hypoxia-induced metabolites that inhibit tumor-infiltrating T cells, restricting their survival, cytotoxicity, and beneficial immunomodulatory cytokine responses. To eliminate bulky solid tumors including stem cells, T cells must overcome physiological changes distinctive for the tumor microenvironment, such as hypoxia, depleted nutrient levels, and low extracellular pH. To exploit the hypoxic tumor microenivornment, we introduce a new approach that exploits hypoxia as a condition for T-cell activation. We demonstrate a chimeric antigen receptor (CAR), which is specific for CD19 and capable of activating T cells through chimeric CD28 and/or CD3-zeta signaling endodomain, that can be conditionally expresed in a strictly oxygen-sensitive manner. Using the Sleeping Beauty transposon/transposase system, we can achieve stable, persistent CAR transgene expression without the expense and complexity of retroviral production. Cell surface expression of our CAR is high at 1% O and not detectable at 20% O (Figure). Anticipating the circulatory nature of T cells in vivo, our oxygen-sensitive CAR is designed to deactivate upon exiting from hypoxic tumor microenvironment to minimize deleterious off-target effects. When supplemented with one or more survival factors and/or homing receptors, this approach to CAR expression has the advantage of
transforming hypoxia from an adverse factor to a triggering mechanism, efficiency, and high levels of transgene expression.
Figure: Flow cytometry expression of CAR under normoxia and hypoxia. Figure:. Flow cytometry expression of CAR under normoxia and hypoxia.