bluuebunny/crossref_metadata_2025_split huggingface dataset:
PPARβ-Mediated Suppression of the Growth and Survival in Human Myeloma Cells Conteracting NF-kB Activity.
Authors: Otsuyama Ken-ichiro , Ma Zi , Liu Shangqin , Abroun Saeid , Asaoku Hideki , Kawano Michio M. 
Published: April 2020

Abstract
PPAR(peroxisomal proliferators-activated receptor)β is considered to be involved in the lipid metabolism and regulating the inflammatory response and in human myeloma cells PPARβis predominantly expressed among these PPARs. However, it remains to be clarified what is the functional role of PPARβ in myeloma cells. In order to identify what are the ligands for PPARβ, we constructed the PPREβ-luciferase reporter gene and performed the reporter assay in Hela cells. Since we have already identified several reagents (DHEA (dehydroepiandrosterone), DHEA-S, baicalein, baicalin, dexamethasone) that suppressed the growth or survival of human myeloma cells (Cancer Res65:2269,2005; Blood105:3312,2005), we analyzed whether these reagents augmented the expression of reporter gene. DHEA-S, baicalein and dexamethasone showed the possibility of their PPARβ binding. Furthermore, these reagents could induce or upregulate the expression of PPREβ-target genes such as ILK and COX2 in myeloma cell lines. We’ve already confirmed that DHEA-S inhibited the proliferation and survival of primary myeloma cells as well as myeloma cell lines, and downregulated the activity of NF-kB, also baicalein showed the growth suppression through the downregulation of NF-kB. Therefore, it is possible that PPARβ-binding reagents such as DHEA-S and baicalein could counteract NF-kB activity to suppress the growth or survival in myeloma cells, while the exact mechanism is under investigation.
Characterization of Oncogene Dysregulation in Multiple Myeloma by Combined FISH and RT-PCR Analyses.
Authors: Qian Shenxian 
Published: April 2020

Abstract
Chromosomal translocations involving the immunoglobulin heavy chain (IGH) locus and various partner loci frequently are associated with multiple myeloma (MM). We investigated the expression profiles of the FGFR3/MMSET, CCND1, CCND3, MAF genes, which are involved in t(4;14)(p16.3;q32), t(11;14)(q13;q32), t(6;14)(p21;q32), and t(14;16)(q32;q23), respectively. The analysis was performed by RT-PCR from purified plasma cell populations from 57 MMs and we compared the results with the presence of translocations as assessed by dual-color FISH. A t(4;14) was found in 11MMs, t(11;14) in 9 MMs, t(6;14) in 5 MM, and t(14;16) in 4 MMs. In all cases, the translocations were associated with the spiked expression of target genes. Furthermore, gene expression profiling enabled the identification of putative translocations causing dysregulation of CCND1 (1 MM) and MAF (1 MM) without any apparent involvement of immunoglobulin loci. Notably, all of the translocations were mutually exclusive. IGH-MMSET hybrid transcripts were found in 10 of the 57 (17.7%) MM samples. There was complete concordance between the findings of RT-PCR and FISH analyses of the MM samples, with 19.2% (11/57) t(4; 14) detected by FISH. Samples were separated further into three major groups based on the size of the RT-PCR product. The 1064bp, 438bp, and 275bp of IGH-MMSET were found in 7, 2, and in 1 sample, respectively. We then screened all 57 MM samples for the expression of FGFR3 using RT-PCR, with primers amplifying the 283bp fragments. Specific transcripts were detected in 11 (19.2%) samples that validate the t(4; 14) from cytogenetic studies. In the remaining 46 MM patients without t(4; 14), and 10 normal bone marrow controls, the FGFR3 amplified transcript was barely detectable. Only one patient sample without t(4; 14) revealed detectable levels of FGFR3 expression. Thus, RT-PCR assay for FGFR3 expression can detect all cases with evident or cryptic t(4; 14) translocation (P&lt; 0.01). Using the primers corresponding to 7–10 exon in 11 cases of MM patients with overexpression of FGFR3, we directly sequenced the FGFR3 cDNA fragments amplified by PCR. Polymorphism (GGC&gt;GGT) was detected in nine of the 11 patients. This polymorphism was tightly associated with higher expression of FGFR3. No FGFR3 mutations were found in the remaining 2 MM patients with overexpression of FGFR3. Our data indicate that RT-PCR is a sensitive and reliable method for the detection of FGFR3 and IGH-MMSET. Translocation t(4; 14) in MM detected by FISH can be validated by RT-PCR method. We examined our result by the Chi-Square test and revealed 90% sensitivity and 100% specificity. The Youden Index remains 0.9. This rapid and reliable detection of FGFR3 and IGH-MMSET overexpression may have practical clinical utility in the analysis and monitoring of the disease in MM patients with t(4; 14). Our data suggest the importance of using combined molecular cytogenetic and gene expression approaches to detect genetic aberrations in MM.
Circulating Endothelial Cells (CEC) and Angiogenesis-Related Factors in Multiple Myeloma (MM).
Authors: Jobe Dean A. , Asp Krista E. , Callister Steven M. , Cole Craig E. , Meyer Lori A. , Mathiason Michelle A. , Bottner Wayne A. , Farnen John P. , Go Ronald S. 
Published: April 2020

Abstract
Background: The profile of CEC can potentially be a biomarker of angiogenesis. As angiogenesis is increased in patients with MM, we studied the CEC and their relationships to angiogenesis-related factors in the blood and disease activity.
Methods: We prospectively quantitated (CD45-/CD146+/CD31+) and characterized (apoptotic [annexin stain], activated [CD106]) the CEC in the whole blood of 20 patients with MM (7 untreated [4 newly diagnosed, 3 smoldering], 9 receiving therapy, and 4 treated previously) using four-color flow cytometry and compared the findings to those from 20 normal volunteers. In addition, we determined the associations between CEC and disease activity and the plasma levels of several angiogenesis-related factors (vascular endothelial growth factor [VEGF], fibroblast growth factor-2 [FGF-2], hepatocyte growth factor [HGF], and thrombospondin-1 [TSP-1]) pertinent to MM.
Results: The median level of CEC was higher (P&lt;0.001) in MM patients (124.8 cells/mL [range, 15–572] versus 21.9 cells/mL [range, 4–66]). However, only 6 (30%) MM patients had levels that were greater than 3 standard deviations from the mean of normal. In general, more apoptotic CEC were present in MM patients (41.5% [range, 0–100] vs 25.9% [range, 0–57.1]; P=0.028), while activated CEC were similar in both groups. There were no significant associations between CEC levels and disease stage, disease status (progressive versus stable/responding), or serum monoclonal protein levels. MM patients had higher levels of HGF (1159 pg/mL [range, 628–5401] vs 524.5 pg/mL [range, 260–938]; P&lt;0.001) but lower levels of TSP-1 (43.9 mcg/mL [range, 3.9–134.1] vs 66.5 mcg/mL [range, 6.4–214.4]; P=0.021), and FGF-2 (20 pg/mL [range, 0–52] vs 34.5 pg/mL [range, 8–82]; P=0.018) in the plasma. In the subgroup of MM patients with elevated CEC, VEGF level was significantly higher than control (211 pg/mL [range, 13–1156] vs 61 pg/mL [0–401]; P=0.012).
Conclusions: Increased levels of CEC can be detected in some patients with MM. In these patients, measurement of CEC may be useful as a biomarker of angiogenesis. As we did not find a significant association between CEC and disease activity, additional studies are necessary to determine the clinical significance of increased CEC in MM.
CD 138 Immunostaining of Bone Marrow Trephine Specimens Is the Most Sensitive Method for Quantifying Marrow Involvement in Patients with Plasma Cell Dyscrasias.
Authors: Ng Ashley P. , Wei Andrew , Bhurani Dinesh , Chappell Peter , Feleppa Frank , Juneja Surender 
Published: April 2020

Abstract
AIM In this study plasma cell quantitation comparing four modalities comprising morphology of the aspirate, flow cytometry, HEtrephine section examination and bone marrow immunohistology was undertaken to determine the most sensitive technique.
METHODS 70 patients with a plasma-cell dyscrasia were retrospectively analysed. Plasma-cell quantitation was performed on Romanowsky-stained aspirate slides by a 200-cell differential. Flow cytometric quantitation of plasma-cell burden was performed by identifying CD38/138 co-expressing cells. Trephine specimens were analysed after HEand CD138 staining. Statistical analysis was performed using a two-tailed paired t-test.
RESULTS See Table 1.
DISCUSSION Significant discrepancy was noted between the four methods for determining plasma-cell infiltration. CD138 immunohistochemical staining of paraffin embedded trephine specimens was the most sensitive modality. Flow-cytometric analysis underestimated the degree of marrow infiltration. Morphology of the aspirate sample may also underestimate plasma-cell burden as sampling may be affected by patchy marrow infiltration as well hypoplastic, fibrotic marrows or clotted specimens. Quantification of plasma cells based on HEstained trephine specimens is less sensitive, and is dependent on the technical quality of the specimen and observer experience. Immunostaining has the advantages of improved plasma-cell identification compared to HEsections and avoids the problems inherent in the analysis of aspirate samples. In conclusion, CD138 immunostaining is the most sensitive method for quantifying plasma-cell burden in the bone marrow.
Two tailed Paired t-test analysis of Quantitation of Plasma Cell burden by various modalities Flow Cytometry Aspirate Hematoxylin & Eosin Immunostaining Mean % of plasma cells 5.6 18.9 23.6 29.7 Paired t-test (two tailed) Mean Difference compared to CD138 immunostaining −24.12 −11 −6.14 - 95% C.I. −30.5 to −17.8 −15.8 to −6.1 −8.34 to −3.9 - p-value p&lt;0.0001 p&lt;0.0001 p&lt;0.0001 -
CD66 Expression on Malignant and Normal Plasma Cells: A Potential Target for Therapy.
Authors: Richardson Deborah , Hodges Elizabeth , Mani Adnan , Orchard Kim 
Published: April 2020

Abstract
CD66, a member of the carcinoembryonic antigen family, is known to be expressed on cells of myeloid and monocytic origin and has also been demonstrated on blasts from patients with B-lineage acute lymphoblastic leukaemia. An analysis of CD66 expression has been undertaken in bone marrow samples from patients with plasma cell disorders. Diagnostic bone marrow aspirate samples from 53 patients with multiple myeloma or monoclonal gammopathy were examined by multiparametric flow cytometry in order to demonstrate and quantitate plasma cells. Samples were analysed initially using a primary screening panel of antibodies including a combination of antiCD19 fluoroscein isothiocyanate (FITC)(Pharminogen), antiCD5 phycoerythrin (PE)(in-house), antiCD45 peridin chlorophyll protein (PerCP)(Becton Dickinson) and antiCD38 allophycocyanin (APC)(Phar). Samples shown to contain a CD45 negative, CD19 negative, CD38 positive population, consistent with the presence of plasma cells were then examined with a myeloma panel, including antiCD66. Samples from 41 patients were analysed using antiCD66 naked antibody (TheraPharm, GmBH) double-layered with sheep anti-mouse IgG F(ab’)2 fragment conjugated to FITC, antiCD138 PE, antiCD38 APC and antiCD45 PerCP. The correlation between CD38/CD138 and CD38/CD66 dual expression was 0.997. A further 13 samples were analysed using a commercial antiCD66 antibody conjugated to FITC (Dako). Again the correlation between CD38/CD138 and CD38/CD66 dual expression was 0.990. All patients apart from one, with morphologically detectable plasma cells, co-expressed CD66 with CD38. One patient with highly plasmablastic morphology did not express either CD38/138 or CD38/66 on plasma cells.The bone marrow of a normal individual was also examined and found to contain plasma cells which co-expressed CD19, CD138, CD38 and CD66. Plasma cells express CD66 in almost all patients with plasma cell disorders and expression has also been demonstrated on plasma cells in a normal individual. CD66 is therefore an attractive target for immunotherapy. A clinical trial using anti-CD66 targeted radiotherapy as part of the conditioning regimen for patients undergoing autologous or allogeneic stem cell transplantation, as therapy for multiple myeloma, is currently being conducted at our institution.
The Immunomodulatory Action of Thalidomide in Patients with Multiple Myeloma Involves a Clonal Expansion of Late-Differentiated Cytotoxic Effector Cells.
Authors: Brown Ross D. , Murray Allan , Sze Daniel M. , Spencer Andrew , Ho Phoebe Joy , Gibson John , Joshua Douglas E. 
Published: April 2020

Abstract
Although thalidomide has been reported to increase T cell activity and NK cell cytotoxicity in the blood of patients with multiple myeloma (MM) it is not known whether thalidomide stimulates tumour-specific T cells or causes a general lymphocytosis. We investigated the immunodulatory effects of thalidomide by studying peripheral blood samples from 64 patients in the Australian ALLG MM6 trial, a multicentre randomised phase III study of low-dose thalidomide, prednisolone and Zometa versus prednisolone and Zometa used as post-autologous stem cell transplant (ASCT) maintenance therapy in patients with MM. We have previously demonstrated that TCR Vβ clonal expansions are present in approximately 50% of patients with MM and that their presence correlates with a good prognosis. Sequencing has confirmed that these cells are a clonal expansion of CD3+CD8+CD57+CD28−CD27− late-differentiated cytotoxic effector cells. Flow cytometric analysis of TCR Vβ expansions (24 families) was performed on blood samples from 64 patients enrolled in the MM6 trial both prior to and 12 months post transplant. Prior to transplant, 59% of the 64 patients had TCR Vβ expansions. After thalidomide, T cell expansions were found in a further 8/34 patients compared with 3/30 in the control, no thalidomide group. In the thalidomide group, 70% of the patients had an expansion of more than one T cell clone compared with 32% in the control group (chi2=28.8; p&lt;0.01). In the thalidomide treated patients there was a total of 32 new clones compared with 22 in the patients who did not receive thalidomide. There was a trend for an increase in NK and NKT cells following thalidomide. Further patients will be studied and the prognostic significance will be evaluated but these preliminary studies provide further evidence that thalidomide has an immunomodulatory action in patients with MM and that this is due, at least in part, to a clonal expansion of late-differentiated cytotoxic effector cells.
BP1 Expression in a Myeloma Cell Line: Modulation by Arsenic Trioxide and Ascorbic Acid.
Authors: Do Khanh , Berg Patricia , Mincheff Milcho , Ascensao Joao L. 
Published: April 2020

Abstract
BP1 belongs to a family of homeobox genes which are not expressed in normal bone marrow cells but are seen in multiple leukemic cell lineages including erythroid, myeloid, and T- and B-cell lineages, and whose modulation is important in differentiation and oncogenic transformation (Shen et al. EMBO J 1992). Multiple Myeloma (MM) is a clonal B-cell neoplasm marked by proliferation of plasma cells within bone marrow. BP1 expression has not previously been demonstrated in MM cell lineages. Recent unpublished data demonstrated elevated BP1 expression in the bone marrow of Acute Promyelocytic Leukemia (APL) patients and its down regulation with the differentiating agent ATRA. With recent data demonstrating arsenic trioxide (As2O3) induced remission in chemorefractory APL and MM (Grad et al. Blood 2001), we hypothesized BP1 may share a role in this response conversion.
In the current study, we treated ARH77 cells - a multiple myeloma cell line, with 0.25 – 3 uM As2O3 and adjuvant ascorbic acid (AA) for 24–96hrs; AA had been previously documented to potentiate As2O3-mediated cell death in multiple myeloma cells (Grad et al. Blood 2001). Viability, cell numbers and mRNA expression were determined at each time point. The % of viable cells at 72 and 96 hrs decreased to less than 10% of the control in cultures flasks containing 2 and 3uM; 0.25 uM had minimal effect in this assay. We then used 0.5 and 1uM, which caused growth arrest compared to controls at each interval. 1 uM caused a decrease of 12% viability at 72 hrs. A significant reduction in the expression of BP1 was seen in these cultures when balanced with beta-actin levels. Myeloma cells are marked by a myriad of antiapoptotic signaling mechanisms, which account for their acquired resistance to current chemotherapy. A better understanding of signaling pathways and genetic events may provide potential therapeutic targets and aid in correlation between genetic abnormalities and clinical outcomes.
Type I MAGE Proteins: Novel Markers of Proliferation in Multiple Myeloma Progenitor Cells.
Authors: Cho Hearn J. , Caballero Otavia , Jungbluth Achim A. , DiLiberto Maurizio , Niesvizky Ruben , Old Lloyd J. , Chen-Kiang Selina 
Published: April 2020

Abstract
CT7 (MAGE-C1) and MAGE-A3, two members of the type I MAGE family of Cancer-Testis antigens, are commonly expressed at both the mRNA and protein levels in primary tumor specimens from multiple myeloma patients. In previous analyses, tumors with higher percentages of type I MAGE-expressing cells had a positive correlation with abnormally elevated plasma cell proliferation. These data support the hypothesis that type I MAGE proteins are molecular markers of proliferating myeloma progenitor cells (the so-called “myeloma stem cell”) and may play a role in the pathobiology of this disease. To test this hypothesis, we examined the expression of type I MAGE in proliferating myeloma cells by flow cytometry. Human multiple myeloma cell lines U266, RPMI-8226, and KMS-11 were co-cultured for 12 or 24 hours with the nucleoside analog bromodeoxyuridine (BrdU), then fixed, permeabilized, and stained with CT7-33, a monoclonal antibody (mAb) to CT7, or M3H67 (to MAGE-A3), followed by a phycoerythrin (PE)-conjugated secondary mAb. The cells were then treated with DNAse and stained with a fluoroisothiocyanate (FITC)-conjugated mAb against BrdU. Proliferating cells that incorporated BrdU into their DNA exhibited high FITC fluorescence. For mAb M3H67, dual color analysis of this population showed that greater than 99% demonstrated a significant shift in PE fluorescence in all three of these cell lines as measured by Mean Fluorescence Index (MFI= geometric mean fluorescence [specific primary antibody]/mean fluorescence [no primary antibody], table 1). For CT7-33 mAb, greater than 85% demonstrated a shift in two of three lines (U266 and KMS-11), but not in RPMI-8226. For all three of these cell lines, dual color analysis of the BrdU-low population demonstrated less than 65% staining with either type I MAGE mAb. Interestingly, RT-PCR with CT7-specific primers of total RNA from RPMI-8226 revealed a product of lower molecular weight than expected, suggesting that a gene deletion occurred in this cell line possibly resulting in a stop codon, decreased translation, or decreased protein stability. This PCR product is being sequenced to determine the nature of the deletion. These results demonstrate that type I MAGE proteins are expressed in proliferating myeloma cells and are molecular markers of this population. These data suggest that novel therapeutics such as vaccines that target type I MAGE may preferentially eliminate the cycling myeloma cells, resulting in long-term cures.
Table 1. Type I MAGE expression in proliferating (BrdU+) myeloma cells Cell line MFI CT7-33 MFI M3H67 U266 65.8 62.5 KMS-11 9.9 48.4 RPMI-8226 3.1 12.3
Lovastatin and Thalidomide Have an Synergic Effect on the Rate of Multiple Myeloma Cell Apoptosis in Short Term Cell Cultures.
Authors: Podhorecka Monika , Klimek Piotr , Grzasko Norbert , Dmoszynska Anna 
Published: April 2020

Abstract
Multiple myeloma is characterized by an accumulation of plasma cells in the bone marrow. Despite many therapeutic regimens introduced recently, the prognosis for patients suffering from treatment-resistant or relapsing multiple myeloma is still very poor. Thus, there is an urgent medical need for novel innovative drugs. Thalidomide is successfully used in myeloma patients being reported to induce apoptosis or G1 growth arrest of plasma cells, regulate microvessel density and cytokine secretion. Statins, largely used for the treatment of hypercholesterolemia, seem to be promising drug in multiple myeloma also. High dose of lovastatin has been shown to have antiproliferative effect by inhibition of malignant cell proliferation and inducing programmed cell death. The aim of this study was the assessment of multiple myeloma cells apoptosis induced by mixture of lovastatin and thalidomide in short-term cell cultures. We analyzed plasmocytes of bone marrow samples obtained from 10 patients with treatment-resistant or relapsing multiple myeloma. To assess apoptosis we used Annexin V and propidium iodide binding. We also examined the regulation of BCL-2 and BAX protein expression in the population of CD138+ plasmocytes. The cells were analyzed with use of flow cytometry technique. The experiments were done before and after 72 hours of cell culture. We observed an increase of apoptotic cell number in all cultures supplemented with analyzed drugs in comparison to 0 h culture and to 72 h control. The percentage of Annexin V positive cells in culture with lovastatin and thalidomide mixture was significantly higher in comparison to culture with lovastatin or thalidomide alone (the mean percentages were 33.40 versus 27.04 and 26.49, respectively, p&lt;0.05). The BCL-2/BAX ratio was lower in cell cultures supplemented with mixture of lovastatin and thalidomide (mean ratio 0.95) in comparison to cultures supplemented with lovastatin or thalidomide alone (mean ratio 1.25 and 1.17, p=0.06 and 0.05, respectively) indicating the tendency to apoptosis induction in analyzed cells. Basing on these results we can conclude that lovastatin and thalidomide may have an synergic effect on the rate of multiple myeloma cell apoptosis and may act together on BCL-2 and BAX regulation. Thus, further research should establish both the precise mechanism of this synergic action of statins and thalidomide and the new therapeutic option for myeloma patients.