bluuebunny/crossref_metadata_2025_split huggingface dataset:
Peripheral Blood Cells from Multiple Myeloma Patients Show Increased Osteoclast and Decreased Osteoblast Gene Expression.
Authors: Gordon Melinda S. , Berenson Ariana M. , Drucker Charles B. , Katz Matthew , Lee Hee Jin , Campbell Richard A. , Chen Haiming , Yeh Howard S. , Bonavida Benjamin , Said Jonathan , Berenson James R. 
Published: April 2020

Abstract
Bone resorption leading to osteolytic bone disease is characteristic of multiple myeloma (MM). Recent studies show the presence of bone-resorbing osteoclasts and bone-forming osteoclasts in the circulation, and these cells may correlate with bone disease and change with anti-bone resorptive therapies. We have investigated whether there is an imbalance in the expression of osteoblast and osteoclast genes in the peripheral blood mononuclear cells (PBMCs) from MM patients relative to normal age-matched controls and the effect of bisphosphonate treatment on the expression of these genes. We analyzed the expression of a panel of osteoblast-related (bone alkaline phosphatase [bone AP], bone morphogenic protein 2 [BMP2], collagen I and osteocalcin) and osteoclast-related (b3 integrin, calcitonin, receptor for activation of nuclear factor kappa B [RANK] and tartrate-resistant alkaline phosphatase [TRAP]) genes by semi-quantitative RT-PCR on total RNA isolated from PBMCs obtained following density gradient separation. We demonstrated that the expression of the osteoblast-related gene BMP2 was reduced in eight of nine MM patients when compared with normal donors. In marked contrast, three osteoclast-related genes, b3 integrin, RANK and TRAP, were more highly expressed in all nine MM patients compared to the normal donors; only calcitonin expression was similar to the control subjects. Interestingly, patients receiving bisphosphonate treatment appeared to show increased osteoblast gene expression with higher amounts of bone AP, BMP2 and osteocalcin RNA compared to the patients not receiving anti-bone resorptive therapy. However, there was no alteration in the level of the RNA in any of the four osteoclast genes compared to patients not receiving anti-bone resorptive therapy. We are extending our analysis to a larger panel of MM patients in order to determine the relationship between these circulating cells and bone disease, overall clinical status and change in their levels with anti-bone resorptive therapy. In addition, we are also investigating whether there exist larger and smaller numbers of circulating osteoclasts and osteoblasts, respectively, in MM patients, or whether these circulating cells show alteration of their expression of these genes. Our semi-quantitative RT-PCR results are being correlated with immunohistochemical staining results from osteoblast and osteoclast markers obtained on PBMCs from MM and normal subjects. These studies provide evidence that the number of circulating osteoblasts and osteoclasts is altered in patients with MM, and also may suggest that bisphosphonate therapy may also be associated with changes in these cell populations.
Cycloxygenase-2 (COX-2) Expression Is More in Clonal Plasma Cells.
Authors: Paneesha S. , Leung E. , Borg Anton , Rose Peter , Morris Alan G. 
Published: April 2020

Abstract
Multiple myeloma is characterised by deregulated cytokine network with secretion of inflammatory cytokines. Recent studies have showed the independent poor prognostic value of COX-2 expression in myeloma. We have here studied the COX-2 expression in plasma cells from patients with myeloma, monoclonal gammopathy of undetermined significance (MGUS), Waldenstrom’s Macroglobulinemia (WM) and compared with the COX-2 expression by normal plasma cells.
Methods:
Our study included bone marrow samples from 34 patients (Lymphoma: 15; myeloma: 10; MGUS: 7; WM: 1; Amyloidosis: 1). Mononuclear cells were harvested from the bone marrow samples by density gradient sedimentation using Lymphoprep TM from Axis-Shield, Norway. Mononuclear cells were stained with PE conjugated CD 38 (BD Bioscience) and APC conjugated CD138 (BD Bioscience). FITC conjugated IgG against human COX-2 (Cayman Chemical) was used to study the expression of cycloxygenase-2 following permeabilization with fix and perm kit (Caltag). Flow cytometry was performed on a Becton Dickinson FACS vantage with appropriate isotype controls. The colorectal cell line HT- 29 and human myeloma cell line RPMI 8226 were used as positive controls and the flow results are validated by western blotting and Prostaglandin E2 EIA assay (Cayman Chemical).
Results:
HT-29 cell line showed a peak fluorescence of 58 for COX-2, where as RPMI 8226 cell line revealed peak fluorescence of 141. Median plasma cell count in bone marrow of patients with MGUS was 2 %( range 2–6%). Plasma cells from MGUS patients revealed median peak fluorescence for COX-2 was 37(range: 9–74). Median plasma cell count in bone marrow of patients with myeloma was 25% (range: 10–59%). Median peak fluorescence for COX-2 in plasma cells from patients with myeloma was 24(range: 10–85). Median plasma cell count in bone marrow of patients with lymphoma was 1 %( range1–4%). Median peak fluorescence for COX-2 in plasma cells from patients with lymphoma was 11(range: 2–38).
Conclusions:
Our study reveals COX-2 expression is more in clonal plasma cells as compared to normal plasma cells. Studies are needed to ascertain the role of COX-2 in oncogenesis in myeloma and to discover how COX-2 expression leads to poor outcome in patients with myeloma. This information may lead to the therapeutic role of selective COX-2 inhibitors in the therapy of myeloma.
Synergistic Inhibition of Myeloma Cell Growth by Combinations of Dexamethasone with Other Anti-Myeloma Agents.
Authors: Bakker Frank , Ott Andrea , Richter Kathrin , Ahrens Tanja , Guenther Andreas , Burger Renate , Gramatzki Martin 
Published: April 2020

Abstract
During the past few years, several new agents with anti-myeloma activity have entered the clinic and improved disease outcome. In addition, inhibitors of key signal transduction pathways have shown promising results in preclinical studies. Combining novel and existing therapeutics might be essential to further improve treatment efficacy, while at the same time reducing adverse side effects. In this study, we systematically tested all possible two-fold combinations out of a repertoire of 8 anti-myeloma agents on five human maligant plasma cell lines. We included the conventional therapeutics doxorubicin and dexamethasone, the new therapeutics bortezomib and zoledronate, and the signal transduction inhibitors rapamycin (mTOR), AG490 (JAK/STAT), U0126 (MEK/MAPK) and L744.852 (farnesyltransferase). The anti-myeloma efficacy of each compound combination was tested in an MTS-based cell growth assay. Interestingly, the majority of compound combinations which blocked myeloma cell growth in a synergistic fashion included dexamethasone. The synergistic effect being mainly due to an increased sensitivity of the cell lines towards dexamethasone. In both the L363 and JK-6L cell lines, strong synergistic effects were observed if dexamethasone was combined with the mTOR inhbitor rapamycin, the MEK inhibitor U0126, the farnesyltransferase inhibitor L744.852 or, to a lesser extent, the JAK inhibitor AG490. In the INA-6 cell line, synergistic cell growth inhibition was observed if dexamethasone was combined with the MEK inhibitor U0126. No strong synergisms were found for the U266 and RPMI8226 cell lines. In the L363 cell line, we also observed strong synergistic effects between L744.852 and rapamycin, and between L744.852 and U0126, indicating that a three-fold combination of dexamethasone, L744.852 and rapamycin or U0126 could result in even more potent myeloma cell growth inhibition. Similarly, synergistic cell growth inhibition was seen in the JK-6L cell line when combining rapamycin and U0126, again suggesting that a three-fold combination including these two compounds and dexamethasone could be highly effective. In conclusion, our results offer important insights into the efficacy of combinations of anti-myeloma agents. Systematic in vitro testing could provide a first step towards finding useful combinations of agents to improve clinical outcome.
Stimulation of Erythropoiesis by Thalidomide in Multiple Myeloma Patients: Its Influence on FasL, TRAIL and Their Receptors on Erythroblasts and Plasma Cells.
Authors: Grzasko Norbert , Hus Marek , Soroka-Wojtaszko Maria , Klimek Piotr , Goracy Aneta , Dmoszynska Anna 
Published: April 2020

Abstract
Proteins belonging to the TNF-family, i.e. Fas ligand (FasL), Fas, TRAIL, TRAIL receptor 1 (TRAIL-R1) and TRAIL receptor 2 (TRAIL-R2), are involved in the pathogenesis of anaemia in multiple myeloma patients. Thalidomide (THAL) is successfully used in the treatment of multiple myeloma (MM) and response is usually accompanied by an increase in haemoglobin concentration. In our previous study comprising 234 MM patients treated with THAL we observed that in the group of non-responders more than 50% of patients had improved hemoglobin level after 3 months of treatment. In the present study 29 patients with refractory/relapsed MM were treated with THAL in an initial dose of 100 mg daily, increased to 400 mg. We measured the expression of the erythropoietin receptor (EPOR) and proteins belonging to the TNF-family on erythroblasts as well as the expression of the TNF-family proteins on monoclonal plasmocytes using flow cytometry before and after 3 months of treatment. Erythroblasts and plasma cells were also investigated in short-term cultures of bone marrow mononuclear cells of MM patients. A clinical response was observed in 17 patients (58.6%), but haemoglobin concentration increased in 20 patients (75.9%). An increase in haemoglobin concentration was observed in 12 patients responding to THAL and in 10 patients not responding. The expression of FasL, Fas, TRAIL and TRAIL-R1 decreased significantly during THAL treatment, whereas the expression of EPOR and TRAIL-R2 did not change significantly. In vitro studies revealed that the expression of FasL, TRAIL, TRAIL-R1 and TRAIL-R2 was lower in THAL cultures than in control cultures. The expressions of TNF-like receptors/ligands on monoclonal plasmocytes did not differ significantly between cultures with and without THAL. Altogether our results suggest that THAL may stimulate erythropoiesis and increase haemoglobin level in MM patients and the way in which THAL acts may be connected with a decrease in the expression of TNF-like ligands/receptors on erythroblasts.
Is Survivin a Possible Target Antigen for Cellular Immunotherapy in Multiple Myeloma?.
Authors: Grube Matthias , Moritz Stephanie , Obermann Ellen , Rezvani Katayoun , Mackensen Andreas , Andreesen Reinhard , Holler Ernst 
Published: April 2020

Abstract
Survivin is a member of the inhibitors of apoptosis family and is expressed in different types of malignancies including multiple myeloma. Cytotoxic T cells recognizing survivin epitopes can be elicited in vitro and by vaccination in human individuals. We performed this study to investigate whether survivin peptide specific CD8 T cells occur in patients with multiple myeloma (MM) and in healthy individuals. To detect antigen specific T-cells, we incubated PBMC from HLA-A2.1 positive patients and healthy individuals with antigen-presenting-cells (C1R-A2) pulsed or not pulsed with a recently described, HLA-A2.1 binding survivin derived peptide (Survpep). We used quantitative real time PCR to measure IFN-gamma mRNA expression for the detection of peptide specific CTL in 23 patients and 21 healthy individuals. A peptide stimulation index (SI, test peptide reactivity / no peptide reactivity) of 2.0 or greater was considered a positive response. CD8 T cells recognizing Survpep could be detected in vitro in 9/23 (39%) patients with MM (SI ranging from 2.9 to 83) and 1/21 (5%) healthy individuals (SI 7). Positive responses were confirmed by 4-colour flow cytometry for intracellular IFN-gamma production. T cells producing IFN-gamma were further analyzed for expression of CD45RA and CCR7 to determine phenotypic characteristics in two patients (SI 83 and 63) identifying them as terminally differentiated effector T cells (CD8+, CD45RA+, CCR7−). Additionally, positive expression of survivin antigen in tumor cells was confirmed by imminohistochemical analysis of patients bone marrow specimen. In conclusion we provide for the first time evidence for T cell reactivity against Survivin antigen in patients with MM. Our data suggest (1) the immunogenicity of survivin antigen in multiple myeloma and (2) that survivin may be a useful candidate antigen for cellular immunotherapy of multiple myeloma.
CD8 T cell response (SI) to Survivin Peptide in patients with multiple myeloma (MM, n=23) and healthy individuals (HI, n=21) CD8 T cell response (SI) to Survivin Peptide in patients with multiple myeloma (MM, n=23) and healthy individuals (HI, n=21)
Thrombosis Is Uncommon in Chinese Patients Receiving Thalidomide- Experience in a Regional Hospital in Hong Kong.
Authors: Chan Man-Hong Helen , Lau Sze-Man June , Chen Yi-Tin , Chan Chi-Chung , Chan Cheuk-Hung , Lam Yim-Kwan 
Published: April 2020

Abstract
OBJECTIVE: To determine the incidence of venous thromboembolism and other side effects of thalidomide in Chinese population
Setting: Queen Elizabeth Hospital, Hong Kong SAR.
Subjects and Methods: Patients who had received thalidomide from 2001 to 2005 were evaluated in this retrospective study. Thalidomide was given either as a single agent at a dose of 100–400mg/day (median dose: 200mg; mean dose: 257.3mg) or in a combination with dexamethasone at a dose of 20– 40mg (oral) on days 1–4. A minimum of 4 weeks of duration of thalidomide was recruited for assessment. Toxicities were graded according to WHO criteria.
Results: Of the 55 patients recruited, 41 patients were evaluable for assessing side effects. The diagnosis of our patients included myelodysplastic syndrome (1/41, 2.4%) refractory mantle cell lymphoma (1/41, 2.4%); myelofibrosis (6/41, 14.6%); and multiple myeloma (33/41, 80.5%). Nine (22%) patients received thalidomide as first line therapy. Thirty-two (78%) patients received thalidomide as 2nd line therapy. Thirty-four (82.9%) patients used thalidomide as single agent. Seven (17.1%) patients used thalidomide as combination therapy. Duration of thalidomide treatment ranged from 4 weeks to 219.3 weeks (Mean 29.4 weeks, median: 20 weeks ). The most commonly observed toxicities were constipation (51.2%), somolence (24.4%), peripheral neuropathy (22%). Only one (2.4%) patient who suffered from myeloma complicated by Miller Fisher syndrome and tetraplegia developed deep vein thrombosis after 4 weeks treatment with thalidomide (300mg/day) plus oral dexamethasone.
Conclusion: Chinese patients in our center tolerated thalidomide well. The low incidence of venous thrombosis observed suggests that anticoagulant prophylaxis may not be warranted in Chinese patiens.
Immunomodulatory Drug Lenalidomide (CC-5013, IMiD3) Augments Anti-CD40 SGN-40-Induced Cytotoxicity in Human Multiple Myeloma: Clinical Implications.
Authors: Tai Yu-Tzu , Catley Laurence , Li Xian-Feng , Coffey Rory , Breitkreutz Iris , Bae Jooeun , Podar Klaus , Hideshima Teru , Chauhan Dharminder , Schlossman Robert , Richardson Paul , Treon Steven P. , Grewal2 Iqbal S. , Munshi Nikhil C. , Anderson Kenneth C. 
Published: April 2020

Abstract
SGN-40, a humanized immoglobulin G1 (IgG1) anti-CD40 monoclonal antibody, mediates cytotoxicity against human multiple myeloma (MM) cells via suppression of IL-6-induced proliferative and antiapoptotic effects, as well as antibody-dependent cell-mediated cytotoxicity (ADCC). We here studied the clinical significance of an immunomodulatory drug lenalidomide on SGN-40-induced cytotoxicity against CD138+CD40+ MM lines and patient MM cells. Pretreatment with lenalidomide sensitized MM cells to SGN-40-induced cell death. Combined lenalidomide and SGN-40 significantly induced MM apoptosis, evidenced by enhanced cleavage of caspase 3/8/PARP and increased subG0 cells, compared with either single agent at the same doses. Pretreatment of effector cells with lenalidomide augmented SGN-40-induced MM cell lysis, associated with an increased number of CD56+CD3− NK cells expressing CD16 and LFA-1. Importantly, pretreatment with lenalidomide or lenalidomide and SGN-40 markedly enhanced NK-cell-mediated lysis of autologous patient MM cells by SGN-40. Lenalidomide also upregulated CD40L on CD56+CD3− NK cells, facilitating IL-2-mediated activation of NK cells. In addition, lenalidomide induced the CD56dim NK subset, which are more potent mediators of ADCC against target MM cells than the CD56bright NK subset. Finally, pretreatment of both effector and target MM cells with lenalidomide markedly enhanced SGN-40-mediated ADCC against CD40-expressing MM cells. These studies therefore demonstrate that the addition of lenalidomide to SGN-40 enhances cytotoxicity against MM cells, providing the framework for combined lenalidomide and SGN-40 in a new treatment paradigm to both target MM cells directly and induce immune effectors against MM.
Didox Induced Apoptosis Occurs by Inhibiting DNA Synthesis and Repair Via Down-Regulation of Ribonucleotide Reductase M1 in Multiple Myeloma (MM).
Authors: Raje Noopur , Kumar Shaji , Hideshima Teru , Ishitsuka Kenji , Yasui Hiroshi , Shiraishi Norihiko , Kiziltepe Tanyel , Chhetri Shweta , Munshi Nikhil C. , Anderson Kenneth C. 
Published: April 2020

Abstract
Ribonucleotide reductase is the rate-limiting enzyme of deoxynucleoside triphosphate synthesis and is therefore an excellent target for cancer chemotherapy. Inhibition of ribonucleotide reductase results in inhibition of DNA synthesis and has anti-neoplastic effects. Several ribonucleotide reductase inhibitors (RRIs) are already in clinical practice, including Hydroxyurea and Gemcitabine. Here we examine the anti-MM activity of a novel RRI, Didox. Our data shows that Didox has potent cytotoxicity against MM cells in 48-hour cultures. Apoptosis induced in MM cell lines by Didox (200mmol/l), evidenced by increased sub-G1 cell fraction, is caspase dependent, confirmed by blocking experiments with Z-VAD-FMK. Unlike other RRIs which mainly target the pyrimidine metabolism pathway (hydroxyurea and gemcitabine), didox targets both the purine and pyrimidine metabolism pathways in MM, as demonstrated by transcriptional profiling using the Affymetrix U133A 2.0 gene chip. Specifically, our data shows a ≥ 2 fold downregulation of genes in these pathways as early as 12 hours after exposure to Didox. Importantly, ribonucleotide reductase (RR) M1 component transcript was downregulated, associated with downregulation of this enzyme by Western Blot analysis, and inhibition of DNA synthesis. Furthermore, genes involved in DNA repair mechanisms were also downregulated after exposure to Didox, demonstrated by heirachial clustering of gene chip data and further validation by western blotting. Specifically RAD 51 homologue, which is involved in nucleotide excision repair and recombination repair, was downregulated in MM cells both transcriptionally as well as at the protein level. This was accompanied by downregulation of the BCL family protein including Bcl-2, Bclxl, and XIAP both at the transcriptional and post transcriptional levels. Since Didox acts on MM cells by inhibiting DNA synthesis and repair, combination studies with melphalan, an alkylating agent commonly used in MM, were next performed. Our data shows a strong in vitro synergism, with combination indices of &lt;0.7 by the Chou Talahay method. When combined with DNA damaging agents like doxorubicin, additive cytotoxicity was noted. These studies therefore provide the preclinical rationale for evaluation of Didox, alone and in combination with alkylating agents and anthracyclines, to improve patient outcome in MM.
Engaging the NKG2D Receptor with a Novel Bispecific Protein (ULBP2-antiCD138) Activates NK Cells and Has Potent Antitumor Activity Against Human Multiple Myeloma.
Authors: von Strandmann Elke Pogge , Hallek Michael , Engert Andreas 
Published: April 2020

Abstract
NK cells, a component of the innate immune system, attack virus-infected and malignant cells without prior antigen stimulation, mediate cellular cytotoxicity and produce cytokines such as interferon gamma (IFN-gamma) upon stimulation. There is growing evidence that NK cells also participate directly in adaptive immune responses, mainly by cross-talk with dendritic cells. One key factor responsible for the activation of innate and adaptive immune responses is NKG2D, a stimulatory receptor expressed on natural killer cells that binds to cellular ligands on malignant cells. Therefore we designed a recombinant NK receptor ligand (ULBP2) fused to an antibody (BB4) detecting the tumor antigen CD138 which is overexpressed on a variety of malignancies including multiple myeloma (MM). The major findings were that (1) ULBP2-BB4 bound both NK cells and tumor cells, (2) triggered NK-mediated cell lysis of CD138+ malignant cell lines and primary MM cells in the allogenic and autologous setting, (3) activated IFN-gamma secretion of NK cells exposed to immobilized protein, and (4) the co-therapy with ULBP-BB4 and human peripheral blood lymphocytes abrogated the tumor growth in a nude mouse model with subcutaneously growing MM cells. This is the first report on the design, expression, purification and functional pre-clinical investigation of a recombinant NKG2D ligand. The results suggest not only a potential clinical use of this novel construct in patients with MM, but might also offer an innovative therapy approach which is based on NKG2D engagement transferable to other malignancies.