bluuebunny/crossref_metadata_2025_split huggingface dataset:
Evidence for Non-Random Immunoglobulin VH Gene and Light Chain Isotype Usage in Follicular Non-Hodgkin’s Lymphoma.
Authors: Yohn Christopher B. , Van Beveren Charles P. , Mu Xi Y. , Shier Peter , Silverman Gregg J. , Gold Daniel P. 
Published: October 2019

Abstract
Background: Antibody diversity is generated by recombination of individual immunoglobulin (Ig) gene segments and subsequent somatic diversification driven by antigen recognition. In the repertoire of expressed B cell receptors (BCR) among normal peripheral B cells, variable heavy (VH) gene segments are not equally represented. The ratio of kappa to lambda light chain usage is also skewed; the normal κ/λ is 1.5. Investigation of the BCR repertoire may provide clues to the genesis of B cell malignancies, as suggested in chronic lymphocytic leukemia (CLL) and mantle cell lymphoma (MCL). BCR V gene identification during the production of recombinant antibodies used in our ongoing PhII and PhIII FavId® (idiotype/KLH) immunotherapy studies has enabled us to analyze V gene usage from 475 B cell follicular lymphoma (FL) tissue samples. This study reports the results of VH gene and κ/λ gene expression in this FL sample collection.
Methods: Ig heavy chain (HC) and light chain (LC) isotypes from B cell FL samples were identified by flow cytometry. VH and VL regions were sequenced from gene specific cDNA libraries prepared from these samples. VH gene usage and κ/λ ratios were compared to frequencies determined for normal peripheral B cells isolated from six healthy volunteers as well as published reports for normal peripheral B cells and other B cell malignancies.
Results: Compared to VH gene family usage determined for normal B cells, VH3 usage is higher (68% vs. 42%), VH1 usage is lower (7.8% vs. 22%) and VH4 usage is equivalent (22% vs. 26%) in our cohort of FL patients while VH2, 5, 6 and 7 are infrequently used in both populations. Usage of the VH3 genes within FL derived sequences also depends upon isotype, in that this gene family is preferentially associated with the IgM HC isotype relative to IgG (76% and 57% respectively). Additionally, the combined usage of the specific genes VH3-23 and VH3-48 in our patient collection accounts for over 29% of all VH genes - compared to 9% among normal B cells. These VH gene usages also differ from reports of VH gene expression among CLL and MCL patients. With respect to LC usage, VH3 isolates are associated with a normal κ/λ ratio of 1.6 while VH4 gene isolates are preferentially associated with λ light chains with a κ/λ ratio of 0.9. Finally, FL B cells expressing the IgM HC isotype preferentially co-express κ light chains (κ/λ ratio of 2.4) while IgG expressing cells preferentially utilize λ chains (κ/λ ratio of 0.6).
Conclusions: Non-random V gene and LC expression among patients with FL is noted. These distortions in Ig gene expression suggest that lymphomagenesis in FL may be associated with B cell stimulation by common antigens. A program to investigate the epitopes recognized by FL derived BCRs via binding of recombinant FL derived antibodies to protein arrays containing common auto-antigens is currently underway.
Transmission of a Follicular Lymphoma by Allogeneic Bone Marrow Tranplantation — Evidence to Support the Existence of a Lymphoma Progenitor Cell.
Authors: Hart Jason , Turner A. Robert , Larratt Loree M. , Russell James A. , Franko Bevin , Frantz Christine , Paonessa Teresa , Mansoor Adnan , Lai Raymond 
Published: October 2019

Abstract
Background: Lymphomagenesis is believed to depend on clonal proliferation of neoplastic progenitor cells. For follicular lymphoma, the neoplastic progenitors are likely derived from the early B-cells, but their exact nature remains elusive. We report a case of transmission of a follicular lymphoma by allogeneic bone marrow transplantation. Follicular lymphoma developed in the donor, and subsequently in the recipient, years after the BMT. Testing supports a common progenitor cell for both the malignancies.
Methods: Sequencing of the amplifiable translocation (14;18) BCL2/IgH fusion gene product and microsatellite analysis using a commercially available human identity testing kit were used to establish the biological relatedness of the donor and recipient lymphomas. The monoclonal IgH gene rearrangements from these two lymphomas were further examined for their somatic hypermutational status.
Results: Microsatellite analysis confirmed the recipient’s lymphoma was donor cell-derived. Analysis of the monoclonal IgH gene rearrangements revealed post-germinal center cell phenotype in both tumors. The VH regions utilized were different in the donor and recipient lymphoma, demonstrating two distinct clonal populations. Despite this difference, the sequences of the translocation (14;18) fusion gene products (BCL2/IgH) from the two lymphomas were identical confirming a common progenitor cell for both malignancies.
Conclusion: Our findings strongly suggest the same neoplastic progenitor cells are responsible for the lymphomagenesis in both the donor and recipient. These findings support the concept of a lymphoma progenitor cell.
A Vascular Targeted Pan PI-3 Kinase Inhibitor, SF1126 with Activity Against Multiple Myeloma In Vivo.
Authors: De Pradip , Peng Qiong , Dey Nandini , McDermitt Breanne , Peng Xiaodong , Garlich Joseph , Lonial Sagar , Durden Donald L. 
Published: October 2019

Abstract
Background: Considerable evidence suggests an important role for the PI-3 kinase and AKT signaling pathways in survival and chemoresistance in multiple myeloma (MM) and other lymphoid malignancies. Our group and others have demonstrated that downregulation of p-AKT with combination therapy (bortezomib + lonafarnib; David et al, Blood, 2005) is a surrogate marker for myeloma apoptosis. It has been demonstrated that the compound, LY294002 has significant pan PI-3 kinase inhibitory properties but is not suitable for clinical use due to PK issues. SF1126 is a novel RGD targeted derivative of LY294002 that has been shown to have activity in a number of different tumor models. Herein, we evaluated the activity of SF1126 against the MM.1S and MM.1R MM cell lines in vitro and in vivo for sensitivity to PI-3 kinase inhibition. The results demonstrate that MM.1S and MM.1R tumor cell growth is sensitive to SF1126 with IC50 of 7.5 and 10.8 uM, respectively. The effects of SF1126 on MM.1R signaling in vitro was examined with profound inhibition of HIF1a induction under hypoxia, the suppression of phosphorylation states of MDM2, ERK and RS6 kinase. The IC50 for inhibition of p-AKT in MM.1S and MM.1R cells was determined to be 2.4 and 2.8 uM, respectively. SF1126 treatment (50 mg/kg/dose sc given every other day) inhibited MM.1R tumor growth in nude mouse xenografts 95% as compared to untreated controls on day 38 (p < .01). Microvessel density analysis of MM.1R tumor tissue demonstrated that SF1126 had significant antiangiogenic activity in vivo.
Conclusion: The results provide preclinical data to support SF1126 as a clinically viable antiangiogenic, pan PI-3 kinase inhibitor for Phase I clinical trials in the treatment of multiple myeloma. Further studies in primary myeloma cells and in combination with conventional agents will be presented.
A Phase II Study of Denileukin Diftitox (Ontak®) with CHOP Chemotherapy in Patients with Newly-Diagnosed Aggressive T-Cell Lymphomas, the CONCEPT Trial: Interim Analysis.
Authors: Foss Francine M. , Sjak-Shie Nelida , Goy Andre , Advani Ranjana , Jacobsen Eric , Acosta Mark 
Published: October 2019

Abstract
Denileukin diftitox, a genetically engineered fusion protein comprised of the enzymatically active domain of diphtheria toxin and the full length sequence of human interleukin-2 (IL-2) that targets malignancies expressing the IL-2 receptor, is approved for cutaneous T-cell lymphoma and has demonstrated a 48% objective response rate in relapsed/refractory PTCL. Because denileukin diftitox is not myelosuppressive and induces apoptosis by inhibition of protein synthesis, a mechanism of action non-cross-resistant with chemotherapy, we evaluated the efficacy and safety of the combination of denileukin diftitox plus CHOP in newly diagnosed aggressive T-cell lymphomas. In this multicenter Phase II study, patients (pts) with aggressive T-cell histologies according to REAL classification were eligible. Pts were required to have absolute neutrophil count ≥1000, platelets ≥50,000, adequate renal and hepatic function with serum albumin ≥3.0 g/dL, and LVEF ≥50%. Denileukin diftitox was given by IV infusion at 18 mcg/kg/day on Days 1 and 2 of each cycle, followed by CHOP chemotherapy on Day 3, and G-CSF support starting Day 4 of each cycle. Cycles were administered every 3 weeks for up to 6 cycles. Restaging evaluation of response is performed after every 2 cycles. The study objective for efficacy is to achieve an overall response rate (CR+PR) of 70%. Twenty-one patients (76% PTCLnos) have been enrolled to date, with a median age of 56 (range 40–80), 10 male, and 11 female. Six patients are not evaluable for response; one discontinued therapy during Cycle 1 due to a hypersensitivity reaction, and 5 are too early to evaluate (1 cycle of therapy given). For the 15 evaluable patients, the overall response rate is 87% with 9 CR or CRu (60%), 4 PR (27%) and 1 each for stable and progressive disease (see Table below). Four of 13 responders (31%) have progressed, with duration of response ranging from 1.5 to 5.9 months. Toxicities have generally been grade 1–2, were transient, and caused few treatment delays. The most common adverse events have been edema (27%), elevated transaminases (20%), and hypoalbuminemia (14%). Only two Grade 4 toxicities were noted (one each of neutropenia and deniluekin diftitox infusion-related allergic reaction). This is the first trial to report on the use of ONTAK in combination with chemotherapy. The novel combination of denileukin diftitox and CHOP appears to be clinically active, and is well tolerated with no unexpected, new, or overlapping toxicities. Pt accrual is ongoing.
PTCL subtype No. of Patients No. evaluable Responses ORR PTCLnos 16 12 4 CR 4 CRu, 3 PR, 1 PD 92% Angioimmunoblastic 2 1 1 PR 50% Enteric T-cell 2 2 1 CR, 1 SD 50% Subcutaneous panniculitic 1 0 n/a n/a
Cutaneous T Cell Lymphoma: Responses in Phase 1 Trial of Combination Therapy with Liposomal Doxorubicin, Bortezomib, and Gemcitabine.
Authors: Dhillon Navneet , Bakkannagari Srinivasa , Ng Chaan , Lim Joann , Duvic Madeline , Kurzrock Razelle 
Published: October 2019

Abstract
BACKGROUND: There is preclinical data suggesting pro-apoptotic synergy between the proteosome inhibitor bortezomib, doxorubucin and gemcitabine. Therefore, we are currently conducting a Phase I dose finding trial of this combination. As a part of this trial, high response rates with excellent tolerance were noted in advanced cutaneous T- cell lymphoma, and are presented here.
METHODS: A 3+ 3 design was used. There were two cohorts of patients: < 65 and ≥65 years of age. The older cohort was dose escalated after the younger cohort demonstrated safety at a dose level. Initial dose escalations were in bortezomib alone, followed by gemcitabine, and then by liposomal doxorubicin.
RESULTS: There were 33 patients who were treated on this trial to date. We are reporting the results of 7 patients with cutaneous T cell lymphoma. These patients had been heavily pretreated with a median of 6 prior therapies (range 1–12). All six of six evaluable patients (100%) achieved a partial remission (PR). Some of these responses are ongoing and the patients are still improving. The most common toxicities were grade 2 thrombocytopenia (50% of patients), grade 1–2 fatigue (44% patients), grade1–2 mucositis (14% patients), and myelosuppression (12% patients). Four patients needed hospitalization early in the course for sepsis. All of these patients had advanced disease with ulcerating skin lesions. Dose limiting toxicity has not been seen.
CONCLUSIONS: The above combination is safe up to the doses tested so far- (dose level 5 = doxil 24mg/m2 on day1, gemcitabine 800 mg/m2 on days 1 and 8 and bortezomib 1.0 mg/m2 on days 1,4,8,11) and has activity in heavily pretreated, advanced cutaneous T-Cell lymphoma.
Responses in Cutaneous T Cell Lymphoma Age Sex Stage Number of prior therapies Dose Level Response Duration of therapy in months 71 F IV 12 1 PR 4 32 F IV 6 1 PR 3 (sent to BMT) 61 F IV 8 2 Lost to follow up after 2 doses 72 M IV 2 2 PR 7+ 73 M IV B 6 2 PR 4+ 53 M II B 1 5 PR 7+ 60 F IV 5 5 PR 8+
Atiprimod Inhibits Growth of Mantle Cell Lymphoma In Vitro and In Vivo and Induces Apoptosis Via Activation of the Mitochondrial Pathway.
Authors: Wang Michael , Zhang Liang , Han Xiaohong , Yang Jin , Qian Jianfei , Wang Siqing , Hong Sungyoul , Zhang Xiang , Zhou Yuhong , Romaguera Jorge , Yi Qing 
Published: October 2019

Abstract
Mantle cell lymphoma (MCL) has poor outcome and is a therapeutic challenge. Atiprimod, a cationic amphiphilic compound, inhibits the proliferation of most human tumor cell lines. Our study was undertaken to evaluate the therapeutic efficacy of Atiprimod on MCL cells and elucidate the mechanism by which it induces cell apoptosis. Four human MCL cell lines, SP53, MINO, Grant 519, and Jeko-1; freshly isolated primary MCL cells from three MCL patients; and normal peripheral blood mononuclear cells (PBMCs) from five healthy donors were treated with Atiprimod. A 3H-thymidine incorporation assay showed that Atiprimod not only inhibited the growth of the MCL cell lines SP53, MINO, Grant 519, and Jeko-1, but also freshly isolated patient MCL cells in a dose-dependent manner. Interestingly, Atiprimod also inhibited the proliferation of PHA-, PMA/ionomycin-, or anti-CD3 mAb/anti-CD28 mAb-activated, but not resting PBMCs from healthy donors. Flow cytometry analysis with fluorescence-labeled Annexin V and propidium iodide showed that Atiprimod induced apoptosis in both the cell lines and primary MCL cells in time-dependent and dose-dependent manners. Notably, Atiprimod did not induce apoptosis of normal T cells and B cells. Atiprimod was also effective and therapeutic in a MCL mouse model established in severe combined immunodeficient (SCID) mice. SP53 cells (5 × 106) were inoculated subcutaneously into the right leg of SCID mice. Three weeks later, after palpable tumors developed, mice were treated intraperitoneally with either vehicle alone (PBS) or Atiprimod (25 mg/kg per day) for 6 consecutive days. Tumor growth was significantly inhibited after Atiprimod treatment compared with vehicle control, and the survival time of tumor-bearing mice was significantly prolonged in the treatment group. Western blot analysis showed that apoptosis of MCL cells was induced through the activation of caspase-9, caspase-3, and PARP pathway. Furthermore, the expression of phosphorylated Bcl-2, Bax, Bad, Bax-xL, and cytochrome C was increased in Atiprimod-treated MCL cells. Pretreatment of cells with caspase-9 inhibitor (z-LEHD) or pan-caspase inhibitor (z-VAD) but not caspase-8 inhibitor (z-IETD) completely blocked Atiprimod-induced apoptosis. In conclusion, Atiprimod inhibits growth and induces apoptosis of MCL cells in vitro and in vivo. Cell apoptosis was induced via activation of the mitochondrial signaling pathway. The therapeutic efficacy of Atiprimod on MCL cells in the mouse model supports the use of Atiprimod as a potential agent in MCL chemotherapy.
Inhibition of the Pan-Bcl-2 Family by the Small Molecule GX15-070 Induces Apoptosis in Mantle Cell Lymphoma (MCL) Cells and Enhances the Activity of Two Proteasome Inhibitors (NPI-0052 and Bortezomib), and Doxorubicin Chemotherapy.
Authors: Yazbeck Victor Y. , Georgakis Georgios V. , Li Yang , McConkey David , Andreeff Michael , Younes Anas 
Published: October 2019

Abstract
Mantle cell lymphoma (MCL) accounts for 6–8% of all non-Hodgkin lymphomas (NHLs). It is an aggressive lymphoma with a poor prognosis--it is generally considered incurable with conventional treatments, and median survival is 3–4 years with a 10-year survival of only 10–15%. There is no accepted standard of care and effective treatments are greatly needed. Bcl-2 family proteins are important regulators of the intrinsic apoptotic pathway and are involved in oncogenesis and chemoresistance of a variety of tumor types, including lymphoma. Antiapoptotic proteins of the Bcl-2 family are overexpressed in mantle cell lymphoma (MCL) cells and may be responsible, in part, for drug resistance. GX15-070 is a small-molecule antagonist of the BH3-binding groove of the Bcl-2 family of proteins, and is currently in Phase I clinical trials. Consequently, we determined the activity of GX15-070 in 3 MCL cell lines (Jeko-1, Mino, and SP53). Cell viability was determined by MTS assay, apoptosis by Annexin-V binding and FACS analysis, and molecular changes by western blot. GX15-070 induced apoptosis in all three MCL cell lines in a dose and time-dependent manner. In the SP53 cell line, GX15-070 decreased MCL-1 and Bak levels, increased Bax and cleaved caspase 3. Furthermore, GX15-070 activated both the extrinsic and intrinsic apoptotic pathway as evident by cleavage of caspase 8, 9, and Bid. Both bortezomib and the novel proteasome inhibitor NPI-0052 induced single agent antiproliferative activity in MCL. GX15-070 enhanced the effect of both proteasome inhibitors. Additionally, GX15-070 showed an additive effect with doxorubicin. These studies suggest that GX15-070 may have a therapeutic value in MCL either alone or in combination with proteasome inhibitors or chemotherapy.
Defined Bone Marrow Niche Components Mediate the In Vitro Resistance of Acute Myeloid Leukaemic Stem and Progenitor Cells (LSPC) to Cytosine Arabinoside.
Authors: Mony Ullas , Jawad Mays , Russell Nigel H. , Pallis Monica 
Published: October 2019

Abstract
The majority of Acute Myeloid Leukaemia (AML) patients respond to remission-induction chemotherapy, but many patients will suffer disease relapse following induction therapy due to the presence of minimum residual disease (MRD) cells in the bone marrow. Relapse is underpinned by outgrowth of leukaemic stem and progenitor cells (LSPC). We aimed to establish a methodology to distinguish normal from leukaemic SPC, to optimise maintenance of the LSPC phenotype in 48 hour culture and to quantify viable LSPC following culture with and without drugs. The CD34+ CD38− CD123+ high phenotype was used to distinguish LSPC. CD123 fluorescence intensity was measured using fluorescence standards. Miltenyi CD 133 and CD34 coated beads were compared as enrichment strategies. The concentration of LSPC at the start of 48h culture ranged from 0.2 – 16 × 104/ml (median 3.4 × 104/ml). We compared various culture conditions aimed at maintaining these cells in culture without differentiation (i.e. without loss of phenotype), including serum, immobilized fibronectin, SCF, IL-3, IL-6, GM-CSF and angiopoietin-1. We used two flow cytometric assays in parallel for analysis of LSPC survival: in the first assay, viable bulk cells were enumerated using the dye 7-AAD along with an internal standard for cell counting. In the second assay, cells were labelled with CD34FITC, CD123PE, 7-AAD and CD38APC in order to quantify LSPC as a percentage of viable cells. We found that serum-free culture medium, fibronectin-coated wells and a cocktail of IL-3, IL-6, SCF, and angiopoietin-1 was the most successful at maintaining the concentration of CD34+ CD38− CD123+ cells in culture for 48h (median 0% change), although there was considerable variation between samples. We then examined the effect of these niche conditions on the sensitivity of LSPC to cytosine arabinoside (Ara-C). Our data indicates that the survival of LSPC treated with 500 ng/ml Ara-C was 50 per cent of untreated cell survival in the presence of fibronectin and cytokine support (n=9), but 33% per cent of untreated cells survived without this support. Moreover, under these conditions LSPC were less sensitive to Ara-C (median 50% survival, n=10) than unselected cells (median 26% survival). We have defined a system for assessing the in vitro chemosensitivity of LSPC from AML samples to anti-leukaemic agents and have validated the assay by showing that phenotypically-identified LSPC as well as niche components fibronectin and cytokines mediate in vitro resistance to cytosine arabinoside.
Novel Markers for the Isolation of Primary Bone Marrow Derived MSC with Multi-Lineage Differentiation Capacity.
Authors: Buhring Hans-Jorg , Battula Venkata L. , Treml Sabrina , Kanz Lothar , Vogel Wichard 
Published: October 2019

Abstract
We have previously described a novel culture protocol to grow MSC from bone marrow (BM) and non-amniotic placenta (PL) with an immature phenotype and multi-lineage differentiation capacity (1). Using the low affinity nerve growth factor receptor (CD271) (2) as a key marker for isolation of MSC derived from femur shaft bone marrow cells (BM-MSC) of patients undergoing a total hip replacement, we could identify two CD271+ distinct populations: CD271dull and CD271bright cells. Two-color flow cytometer analysis revealed that only the CD271bright population coexpressed the mesenchymal markers CD10, CD13, CD73, and CD105, but was negative for CD45. CD271dull cells were positive for CD45 and HLA-DR but negative for the other markers. To analyze the mesenchymal stem cell potential, colony-forming-unit fibroblast (CFU-F) assays were performed. Not surprisingly, the CFU-F were exclusively detected in the CD271bright but not in the CD271dull fraction. By screening a battery of antibodies against known and unknown antigens, we identified several reagents that selectively detected the CD271brightCD45- population but no other bone marrow cells. These markers included the PDGF-RB (CD140b), the embryonic stem cell marker TRA-1-49, the clustered markers HER-2/erbB2 (CD340), the recently described W8B2 antigen (3), as well as the cell surface antigens defined by the antibodies W1C3, W3D5, W4A5, W5C4, W5C5, W7C6, 9A3, 58B1, F9-3C2F1, and HEK-3D6. In conclusion we identified several novel markers for the prospective isolation and characterization of BM-MSC.