bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Phase II Study of Velcade (V), Doxil (D) in Combination with Low-Dose Thalidomide (T) as Salvage Therapy for Patients (pts) with Relapsed (rel) or Refractory (ref) Multiple Myeloma (MM) and Waldenstorm Macroglobulinemia (WM): Encouraging Preliminary Results.
Authors: Chanan-Khan Asher Alban , Miller Kena C. , McCarthy Philip , DiMiceli Laurie A. , Yu Jihnee , Bernstein Zale P. , Czuczman Myron S. 
Published: April 2020

Abstract
In MM, tumor microenvironment (ME) plays an important role in disease progression, dissemination, and development of resistance to therapy. Pts with rel/ref MM have limited treatment options. Therefore, targeting the ME simultaneously with malignant cells may be an effective way to overcome resistance in pts with rel/ref MM. Orlowski et al recently reported synergistic activity of V+D in patients with hematologic malignancies. A phase II trial initiated at our institute is exploring this approach, targeting the MM cell as well as its ME, using a combination of Velcade (V), Doxil (D) and low-dose thalidomide (T) as salvage therapy for pts with rel/ref MM. Pts with rel/ref disease are eligible for this study. V is given at 1.3mg/m2 (D1,4,15,18) and D at 20mg/m2 (D1,15) every 4 weeks with daily T (200 mg) for 4–6 cycles. SWOG criterion was used to evaluate response. Low-dose coumadin (1–2 mg po qd) was used for prevention of venous thromboembolism (VTE). Eighteen pts (7M, 11F; median age 56, range 44–80 yrs; 16MM, 2 WM) have been enrolled to date. All pts had Stage III disease, median b2M of 4.8 (nl range: 0–2.15) and median prior therapies 2 (range 1–7). Prior therapies included stem cell transplant in (46%), T (54%), adriamycin(A) (70%) and steroids (92%). Thirteen pts have completed at least 1 cyc and are available for toxicity and response evaluation. One pt died of sepsis prior to completing 1 cyc and 1 pt with PR was taken of study for non-compliance after 1 cyc. ORR was 100% (13/13) with 5PR and 8MR. All pts are currently continued on therapy. Toxicity: 2 pts developed Gr. II plantar-palmar erythrodysthesia (PPE) and 1 had Gr. III cellulitis. No VTE was noted. VDT is a highly active salvage regimen in pts with rel/refr MM. Responses were noted despite prior failure of steroids, T or A. It is well tolerated without any significant non-hematologic Gr. III/IV toxicity. VTE does not appear to be a problem. Updated results of this first cohort of pts will be presented at the annual ASH meeting.
Discriminative HMGA2 Isoform Expression in CD15+ Granulocytic Cells in Myeloid Metaplasia with Myelofibrosis (MMM).
Authors: Pierre-Louis Olivier , Andrieux Joris , Desterke Christophe , Lippert Eric , Praloran Vincent , Demory Jean-Loup , Le Bousse-Kerdiles Marie-Caroline , Bilhou-Nabera Chrystele 
Published: April 2020

Abstract
MMM is a myeloproliferative disorder characterized by extramedullary hematopoiesis and reactive myelofibrosis. Recently, HMGA2 dysregulation has been demonstrated in 2 MMM patients showing 12q15 rearrangement and confirmed in 25 consecutive MMM patients without cytogenetic abnormalities (Andrieux, 2004). HMGA2 proteins belong to the high mobility group A (HMGA) family of architectural transcription factors regulating the expression of several genes. As MMM is a clonal disorder of CD34+ hematopoietic progenitors, we analyzed HMGA2 expression in peripheral blood sub-populations of 5 MMM patients and 7 healthy donors to determine in which sub-population HMGA2 was dysregulated.
RNA was extracted from peripheral blood mononuclear cells (PBMC) and CD15+ granulocytic cells (PBCD15+) separated through Ficoll centrifugation or from immunomagnetically selected circulating CD34+ cells (PBCD34+). Real-time quantitative PCR (RQ-PCR) using Taqman technology was performed on cDNA. As different isoforms were described in malignancies, we used two primer sets : the first one allowing the amplification of all HMGA2 isoforms (exon 1 to 3) (HMGA2 1–3), the second one allowing the amplification of the full length HMGA2 isoform (exon 1 to 5)(HMGA2 1–5).
In healthy donors and in MMM, PBMC HMGA2 expression levels were heterogeneous, depending of the cellular sub-population purity. HMGA2 1–3 or HMGA2 1–5 were both expressed in MMM and normal PBCD34+ cells, but with a higher expression level for HMGA2 1–3 as compared to HMGA2 1–5. Furthermore, both HMGA2 1–3 and HMGA2 1–5 expression levels were significantly increased in PBCD34+ MMM patients (p&lt;10−6) compared to healthy donors. In MMM, HMGA2 expression level was significantly increased (p&lt;10−5) in PBCD15+ as compared to PBCD34+. Moreover, PBCD15+ HMGA2 1–3 expression level was significantly higher in MMM patients compared to PBCD15+ from healthy donors (p&lt;10−7). A persistence of HMGA2 1–5 expression was only observed in MMM PBCD15+ but was undetectable neither in normal PB neutrophils (purity&gt;98%) nor in PB neutrophils from other myeloproliferative disorders (Polycythemia Vera and Essential Thrombocythemia).
To determine if HMGA2 level was modified during hematopoietic differentiation, we quantified HMGA2 1–3 and 1–5 isoform expression on purified healthy donor PB CD34+ and MMM CD34+ before and after culture with specific lineage growth factors (12 day-culture). Primary results showed that both HMGA2 isoform expression levels were higher during granulocytic differentiation. Our results demonstrate that HMGA2 1–5 isoform is discriminately overexpressed in MMM PBCD15+. The persistence of this HMGA2 full length expression in MMM myeloid lineage could be considered as a marker of the disease.
NFκB Activation in Primary Mediastinal Large B-Cell Lymphoma: Nuclear Localization of c-REL and Coordinate Upregulation of NFκB Target Genes.
Authors: Feuerhake Friedrich , Kutok Jeffery L. , Monti Stefano , Cattoretti Giorgio , Kurtin Paul , Pinkus Geraldine S. , de Leval Laurence , Harris Nancy L. , Savage Kerry J. , Habermann Thomas M. , Dalla-Favera Riccardo , Golub Todd R. , Aster Jon C. , Shipp Margaret A. 
Published: April 2020

Abstract
Primary mediastinal large B-cell lymphoma (MLBCL) is a clinically distinct entity that typically presents as localized, sclerotic disease in young, female patients. We previously characterized the transcriptional profiles of MLBCLs and identified important shared features with a clinically related disorder, classical Hodgkin lymphoma (cHL) (Blood 102:3871, 2003). Given the documented role of the NFkB survival pathway in Hodgkin Reed-Sternberg cells, we previously assessed NFkB activation in MLBCL by determining the subcellular location of the c-REL subunit of the NFkB heterodimer with a 2-color immunofluoresence assay. In a small pilot MLBCL series, c-REL was localized to the nucleus in the majority of examined cases, consistent with NFkB activation. In the current study, we evaluated c-REL subcellular localization in an additional series of MLBCLs and DLBCLs using a broadly applicable immunoperoxidase method. 100% of MLBCLs exhibited nuclear c-REL staining whereas DLBCL c-REL subcellular localization was more variable. Thereafter, we analyzed the transcription profiles of the 34 MLBCLs and 176 DLBCLs for coordinate expression of NFkB target genes, using literature-curated NFkB target gene lists from three independent sources and gene set enrichment analysis (GSEA). MLBCL signatures exhibited significant enrichment of 2 of the 3 NFkB target gene sets. In addition, 32 NFkB target genes from the combined set were significantly more abundant in MLBCLs than DLBCLs (&gt; 30% more abundant and &gt; 99th percentile in permutation analysis). Similar results were obtained in an independent series of MLBCLs and DLBCLs with available gene expression profiles (J. Exp. Med. 198:851, 2003). To assess the role of c-REL amplification in NFkB activation in our lymphoma series, we compared c-REL amplification, c-REL subcellular localization and coordinate expression of the identified NFkB target genes and classified the DLBCLs according to putative cell of origin. The majority of c-REL amplifications (67%) were found in DLBCLs of germinal center (GC) subtype, consistent with the observation that c-REL is part of the described GC signature. However, most (71%) of the examined GC DLBCLs had cytoplasmic c-REL expression and the GC DLBCLs did not have increased expression of NFkB target genes. Taken together with the MLBCL analyses, these studies indicate that: 1) NFkB is consistently activated in MLBCL; 2) c-REL amplification is not closely associated with NFkB activation in large cell lymphomas (LCLs); and 3) NFkB activation in LCL subtypes does not require amplification of the c-REL locus.
Expression of HMGA2 in PB Leukocytes and Purified CD34+ Cells from Normal Individuals and Patients with Myelofibrosis and Myeloid Metaplasia.
Authors: Andrieux Joris R. , Bilhou-Nabera Chrystèle , Demory Jean Loup , Pierre-Louis Olivier , Dupriez Brigitte , Grardel Nathalie , Charpentier Agnès , Praloran Vincent , Bauters Francis , Laï Jean Luc , Le Bousse-Kerdilès Marie-Caroline 
Published: April 2020

Abstract
Background: Myelofibrosis with myeloïd metaplasia (MMM) is a rare myeloproliferative disorder which associates clonal proliferation of multipotent hematopoietic progenitors and reactive fibrosis. Deregulation and overexpression of HMGA2 has recently been demonstrated in MMM. Using quantitative RT-PCR on blood mononuclear cells from 27 patients, we found various levels of expression. It could be hypothesized that HMGA2 expression reflected the variable increase in circulating CD34+ progenitors in MMM though we failed to establish significant correlation between the level of expression and the blood enumeration of CD34+ progenitors.
Material and Methods: We studied 24 patients diagnosed as primary MMM according to the set of criteria recently updated through an Italian Consensus Conference. Six individuals free from hematologic malignancy were used as controls. We also studied-purified CD34+ cells: 8 samples were isolated from the PB of MMM patients; given the very low concentrations of progenitors in normal PB, 2 samples pooling 8 healthy donors each were also prepared. In addition, 7 CD34+ cell samples were purified from the BM of consenting subjects collected during surgery for hip replacement.
Results: We assayed HMGA2 expression in purified CD34+ populations from MMM patients and controls: both expressed HMGA2 but markedly more the CD34+ cells from the patients. We also evidenced transcripts in their CD15+ granulocytic cells suggesting that the enhanced expression of the gene affects the whole pathological clone. HMGA2, not expressed in normal blood cells, is involved in benign solid tumors of mesenchymal origin.
Conclusion: Our data support the hypothesis that its reactivation in the clonal progenitors contribute to the pathogenesis of MMM.
HLA-Cw6 Alloreactive CTL Recognizing HLA-CW6+ Myeloma Cells but Not Non-Myeloma HLA-Cw6+ Cells Are Peptide Dependent.
Authors: Geffroy Alexandrine , Moreau-Aubry Agnes , Bataille Regis , Pellat-Deceunynck Catherine 
Published: April 2020

Abstract
We investigated the generation of myeloma-specific cytotoxic T lymphocytes (CTL) from unrelated normal donors non-HLA matched with the myeloma cell line SBN. The aim was to obtain alloreactive CTL specific of peptides to identify genes preferentially expressed by to myeloma cells. After coculture of SBN with PBL of unrelated donor, the T-cell line obtained was cloned and each CTL was assessed against SBN and SB-EBV (B-EBV cell line obtained by infecting B cells of SB patient with EBV) simultaneously. Only the CTL recognizing SBN but not SB-EBV were kept for further study. Among more than 200 clones screened, we isolated two different CTL CD8+ recognizing SBN only (60% of specific lysis at effector: target ratio 5). Their cytotoxicity was blocked by antibodies against HLA-I molecules and more precisely by mAb against HLA-B/Cw molecules. Both CTL recognized also other MM cell lines but only those matched with SBN for HLA-Cw0602 (XG6, BCN) suggesting an HLA-Cw6 recognition. Furthermore, antiserum directed against HLA-Cw6 abrogated the recognition of SBN, XG6 and BCN. Both CTL were not cytotoxic against BC-EBV nor against 3 other B-EBV cell lines derived from HLA-Cw0602 donors. This unreactivity was not restricted to B-EBV cell lines since PBL from 2 normal HLA-Cw0602 donors were also not recognized (cytotoxicity, TNF secretion). We directly measured cell surface HLA-Cw6 expression of both B-EBV and myeloma cells with a scFv directed against HLA-Cw6 and we showed that HLA-Cw6 was more expressed by B-EBV cells as compared to myeloma cells (2fold). So, the lack of recognition of B-EBV cells was not related to HLA-Cw6 expression level. We also looked for KIR expression by the CTL since KIR molecules could have blocked B-EBV recognition that expressed more HLA-Cw6 molecules as compared to myeloma cells. No KIR molecules (p58.1, p58.2, CD94) were found to be expressed by the CTL.
To investigate whether the recognition of myeloma cells was peptide dependent, we performed two experiments i.e., acid elution and cold target competition. Acid elution (glycine 0.3M, pH=2.5) abrogated myeloma recognition and cold target competition showed that B-EBV cells were not recognized by the CTL. These data suggest that the CTL recognize a peptide or a set of peptides restricted to myeloma cells or that the CTL recognition is activated by tumor cells only. Anyway, such alloreactive CTL could be helpful for GVL reaction in allotransplanted patients with myeloma.
Periocular MALT Lymphomas (Conjunctival/Orbital): Complete Responses with Antibiotic Therapy.
Authors: Coleman Morton , Leonard John P. , Decter Jules , Furman Richard R. , Niesvizky Ruben , Rollins Indira , Abramson David H. 
Published: April 2020

Abstract
There is mounting epidemiologic, histologic, experimental and therapeutic evidence that MALT (mucosa-associated lymphoid tissue) lymphomas may be related to chronic infections. While H. pylori infection-initiated MALT lymphomas of the stomach are now well documented, only recently has evidence for infectious sources of MALT lymphomas of the intestine (Campylobacter), skin (Borrelia), and the eye (Chlamydia) been reported. Chlamydia psittaci has been implicated in ocular MALT lymphoma in 2 reports (Ferreri et al, JNCI, 96(8):586–594, 2004 and Yeung et al, Cornea, 23:71–75, 2004). In these reports, only 2 of 4 evaluable patients treated with doxycycline were noted to have objective responses. We retrospectively reviewed data on 4 patients who have now undergone antibiotic therapy for subconjunctival MALT lymphomas, three of whom are evaluable for response to treatment. Complete evaluations for systemic disease (including CT scans of neck, chest, abdomen, pelvis, PET scans, and bone marrow examination) were all negative. Serologic tests for antibodies to Chlamydia psittaci were negative in all 4 individuals. Patients were 58, 74, 76 and 79 years of age with 2 males and 2 females. Treatment of one patient was with a PrevpacR (lansoprazole 30 mg capsule, amoxicillin 500 mg capsules, and clarithromycin 500 mg tablets BID) x 14 days, while three received doxycycline 100 mg daily for one month. Of the three patients evaluable to date for response, all demonstrated prompt and sustained responses (2 CR and 1 PR) with followup of 38 months, 12 months, and 7 months respectively. Definitive evidence for a particular infectious organism was not demonstrated, but the prompt and sustained response to antibiotics is consistent with an infectious origin. While further evaluation and followup is warranted in additional patients, we conclude that a trial of anti-microbial therapy should be considered for patients diagnosed with ocular MALT lymphoma.
A Novel N-Oxide Drug, AQ4N, Has In Vitro Activity in Lymphoma and Leukemia Cell Lines and Selectively Targets Lymphocytes and Lymphatic Tissues in Vivo.
Authors: Cleland Jeffrey L. , Wong Alvin , Alters Susan E. , Harris Peter A. , Dunk Chris R. , Curd John G. , Capizzi Robert L. , Henner William D. 
Published: April 2020

Abstract
An ideal treatment for lymphoma and leukemia is the use of highly selective compounds to eliminate diseased cells with minimal systemic toxicity to normal tissues (cf. imatinib mesylate; Gleevec). AQ4N (1,4 bis[[2-(dimethylamino)ethylamino}-5,8-hydroxyanthracene-9,10-dione bis N-oxide) is designed to have little or no toxicity until selectively activated by bioreduction in hypoxic cells to AQ4 (reduced AQ4N), a highly potent DNA topoisomerase II inhibitor. In a series of studies, AQ4 has been shown to have potent cytotoxicity on lymphoma and leukemia cell lines in vitro and AQ4N has selective activity in lymphatic tissues in vivo. The IC50 of AQ4, was 0.63, 12.0, 90.5 and 150 nM in Namalwa, Daudi, Ramos, and Raji human lymphoma cell lines and 1.0, 6.0, and 20 nM in HL-60, KG1a and K562 human leukemia cell lines. On several of the tumor lines the activity of AQ4 was more potent than doxorubicin (i.e. IC50 for Dox was 20.3 nM on Namalwa). AQ4N also had anti-proliferative activity at μM levels indicating a potential mechanism for activation by these cell lines. In repeat dose toxicology studies of AQ4N in pigmented rats and cynomolgus monkeys, the maximum tolerated doses (MTD; rats: 20 mg/kg/wk x 6; monkeys 6 mg/kg/wk x 6) resulted in lymphoid tissue atrophy. A decrease in lymphocyte levels and atrophy of the spleen, thymus, and mandibular and mesenteric lymph nodes were observed at terminal sacrifice of the animals. In contrast, there was an absence of myelosuppression and only mild neutropenia and minor bone marrow atrophy at the MTD. Administration of radiolabeled AQ4N (14C-benzene) to pigmented rats and cynomolgus monkeys indicated persistence of AQ4N radioactivity in lymphoid tissues for several weeks after a single dose (rats: 20 mg/kg (130–140 μCi/kg); monkeys: 10 mg/kg (135 μCi/kg)). For example, in rats the half-life of radioactive AQ4N in the spleen was 538 hrs with 0.9 μg AQ4N/g tissue (spleen) remaining one week after dosing. Monkeys demonstrated a similar effect with 76.5–86.8 μg AQ4N/g tissue observed in the spleen one week after treatment. Other tissues contained significantly less radioactive AQ4N with the exception of the liver (67.9–78.6 μg AQ4N/g tissue) and adrenal cortex (78.7–86.6 μg AQ4N/g tissue). While some hypertrophy and eosinophila was observed in the adrenal glands, liver toxicity was not observed at the MTD in the repeat dose cynomolgus monkey toxicology study. Overall, these initial findings indicate that AQ4N is active in vitro against human lymphoma and leukemia cell lines and selectively targets lymphoid tissues in vivo suggesting the potential benefit of AQ4N in the treatment of lymphoproliferative diseases.
Analysis of the Effect of the Novel Histone Deacetylase Inhibitor, Depsipeptide (FK228/FR901228) on Mantle Cell Lymphoma.
Authors: Paner Gladell P. , Alkan Serhan 
Published: April 2020

Abstract
Background: Mantle cell lymphoma (MCL) comprises approximately 3–5% of the non-Hodgkin’s lymphomas and the vast majority of these patients are incurable. Histone deacetylases (HDAC) inhibitors are a new class of chemotherapeutic drugs that have been shown to inhibit cell proliferation and induce apoptosis in a variety of malignancies. Depsipeptide (FK228/FR901228) is a novel inhibitor of class I HDAC and was shown to be effective in patients with cutaneous T-cell lymphoma.
Design: A MCL cell line, JeKo-1 was treated with varying concentrations of depsipeptide and analyzed for evidence of apoptosis. Apoptosis was analyzed by flow cytometry (Annexin and 7AAD staining) and by microscopic examination. The effect on the status of H3 histone acetylation and changes in the expression of the Bcl-2 family proteins (Bcl-2, Bcl-xl and Mcl-1) following treatment with depsipeptide were investigated by flow cytometric evaluation. Requirement for de novo protein synthesis and activation of JNK in depsipeptide-induced apoptosis was analyzed by treatment with cycloheximide (CHX) and JNK inhibitor (SP600125) prior to addition of depsipeptide respectively. The effect of depsipeptide in 5 primary MCL patient samples was then analyzed for morphologic evidence of apoptosis.
Results: Depsipeptide (0.25 mM/L; 80%apoptosis) induced marked apoptosis in JeKo-1 cells and this was shown to be a dose dependent fashion following 12 hours incubation. There was increase in H3 acetylation, after treatment with depsipeptide. Bcl-2, Bcl-xl and Mcl-1 showed decreased expression in the depsipeptide treated samples. Treatment with CHX (100ng/ml and 500ng/ml) and SP600125 (20ml/L) had no effect to the ability of depsipeptide-induced apoptosis. Primary MCL cells of 5 patients also showed marked apoptosis with depsipeptide treatment.
Conclusion: Depsipeptide causes increased H3 acetylation and induces apoptosis in MCL cell line and primary MCL cells. The mitochondrial apoptotic pathway is possibly altered by the depsipeptide treatment causing apoptosis, by down-regulation of Bcl-2, Bcl-xl and Mcl-1 proteins. Based on the recent phase I clinical trial demonstrating clinical response in cutaneous T cell lymphoma, our study supports the potential clinical utilization of depsipeptide in patients with MCL.
Immunotargetting of the Wilms’ Tumor WT1 Antigen for Dendritic Cell and B-Cell-Based Vaccination of Leukemia.
Authors: Berneman Zwi N. , Van Driessche Ann , Ponsaerts Peter , Gao Liquan , Stauss Hans J. , Schroyens Wilfried A. , Van de Velde Ann , Gadisseur Alain , Lenjou Marc , Nijs Griet , Van Bockstaele Dirk R. , Van Tendeloo Viggo F. 
Published: April 2020

Abstract
The Wilms’ tumor antigen (WT1) is overexpressed in almost all leukemias and in several solid tumors. Overexpression of WT1 blocks the normal differentiation and enhances proliferation of hematopoietic progenitor cells. WT1 is also used in the detection of minimal residual disease. Using WT1-specific MHC class I tetramers, we were able to detect ex vivo low numbers of WT1-specific CD8+ T cells in the peripheral blood or bone marrow of leukemia patients, but not of healthy donors. In one particular donor we could detect up to 24% WT1 tetramer positive cells at the time of diagnosis. WT1 tetramer positive cells were present in all types of leukemia, except for CLL, and also in patients with MDS. Because WT1 plays an important role in leukemogenesis, it could serve as an antigenic target for dendritic cell-based immunotherapy. We used the mRNA electroporation strategy that allows presentation of multiple WT1 epitopes by MHC class I molecules, irrespective of the HLA haplotype. Monocyte-derived DC (Mo-DC) were electroporated with in vitro transcribed WT1 mRNA. RT-PCR and Western blot analysis showed that WT1 RNA and protein, respectively, was present for up to 5 days in WT1-electroporated DC, but not in mock- or EGFP mRNA-electroporated Mo-DC. Importantly, using a CD8+ T cell clone that secretes IFN-gamma upon recognizing the HLA-A2 immunodominant WT1126–134 epitope, we showed that WT1 mRNA-electroporated Mo-DC processed the WT1 protein via the MHC class I pathway and presented the WT1 epitope to the T cells in an HLA- and antigen-specific manner. Since Mo-DCs are a non-expandable source of antigen-presenting cells, we also used proliferating CD40-activated B (CD40-B) cells as inducers for WT1-specific T cell immunity. CD40-B cells were expanded to high numbers from a limited amount of peripheral blood and subsequently electroporated with WT1 mRNA. In T cell clone activation experiments, WT1 mRNA-electroporated CD40-B cells were as efficient as Mo-DC in presenting the WT1 epitope in a MHC class I-restricted manner. Based on these results, we are currently focusing on the in vitro (re)activation of autologous WT1-specific cytotoxic T cells of leukemia patients using WT1-loaded autologous Mo-DC or CD40-B cells and on the immunological parameters to break immune tolerance against the WT1 tumor self antigen.