bluuebunny/crossref_metadata_2025_split huggingface dataset:
Flow Cytometric Disease Monitoring in Patients with Multiple Myeloma Undergoing Autologous Stem Cell Transplantation: A Retrospective Study.
Authors: Liu Hong , Yuan Constance M. , Braylan Raul C. , Chang Myron N. , Wingard John R. , Moreb Jan S. 
Published: April 2020

Abstract
The persistence of abnormal neoplastic plasma cells (APC) detectable in the bone marrow by flow cytometry at more than 3 months after autografting for multiple myeloma (MM) has been reported to predict early disease progression. In this study, we retrospectively reviewed the flow cytometric data from bone marrow aspirates of MM patients before and after autologous stem cell transplantation (ASCT). Light scatter properties and CD38 expression were used to identify plasma cells, and CD19/CD45/CD56 further distinguished normal plasma cells (NPC) from APC. Conventional response criteria (Blade criteria) and survival data were also collected. Forty-seven (47) patients treated with the same conditioning regimen were screened. Median follow up from ASCT was 19 months. After ASCT, 66% (31/47) patients achieved complete remission (CR)/very good partial remission (VGPR), as compared to only 36% (17/47) prior to ASCT. In 39 patients with data before and after ASCT, all 39 (100%) had a detectable abnormal plasma cell population identified phenotypically by flow cytometry prior to ASCT. Of these patient, 18/39 (46%) had greater than 30 APC and these patients had significantly shorter PFS independent of other covariates (1-sided P=0.036, 2 sided P=0.072, logrank). Twenty-six out of 39 patients (67%) also had detectable NPC. Following ASCT, the number of patients with detectable NPC increased to 35/39 (89%), while 3/39 (8%) had no detectable NPC and 1/39 (3%) had neither NPC or APC. The proportion of APC decreased significantly after transplant (81% prior to transplant vs. 59% post-transplant, P=0.008, 2 tailed t-test). Patients with a APC to NPC ratio &lt; 1 post transplant has higher PFS rate at 2 year (54%) when compared to patients with higher APC/NPC ratio (29% PFS at 2 year), however, the difference is not statistically significant. In addition to the presence of APC, the ratio of APC to NPC, age, beta-2 microglobulin levels, and the presence of normal immunoglobulin levels were analyzed. Patients who achieved CR/VGPR after transplant had significantly longer PFS (23 months vs. 11 months, P=0.03). All other covariates were not found to be significant. Because only 10 deaths were observed, covariate analysis for OS was not feasible. In conclusion, the recovery of NPC after ASCT is seen in a substantial propotion of patients with a trend towards better PFS in patients with low APC/NPC ratio. On the other hand, the presence of a significant population of APC (&gt; 30) prior to transplant appears to correlate with poorer PFS in MM patients.
Genotype-Phenotype Associations in Patients with Severe Congenital Neutropenia.
Authors: Rosenberg Philip S. , Stein Steven , Rodger Elin , Bolyard Audrey Anna , Bonilla Mary Ann , Dror Yigal , Kannourakis George , Newburger Peter E. , Boxer Laurence A. , Alter Blanche P. , Dale David C. 
Published: April 2020

Abstract
BACKGROUND: G-CSF therapy has reduced sepsis mortality in patients with severe congenital neutropenia (SCN), revealing a syndromic predisposition to myelodysplastic syndrome and acute myeloid leukemia (MDS/AML). The MDS/AML risk appears to be higher in SCN patients who are less-responsive to therapy; however, the genetic determinants of susceptibility remain to be elucidated.
METHODS: We studied 82 North American and Australian patients with SCN on long-term G-CSF enrolled in the Severe Chronic Neutropenia International Registry. These patients had prospective follow-up, sufficient banked DNA for genotyping, and validated clinical data.
RESULTS: Fifty-two patients (63%) were positive for germline mutations in neutrophil elastase (ELA2); the remaining 30 patients (37%) had no detectable mutations. Prior to G-CSF therapy, blood cell counts were significantly different between ELA2 positive and negative patients. Compared with ELA2 positive patients, ELA2 negative patients had significantly higher median absolute neutrophil count (ANC) values (158 versus 53 cells/uL, P=0.02), significantly lower monocyte and eosinophil counts (P&lt;0.001), and significantly lower platelet counts (P=0.002). ELA2 negative patients were maintained on significantly lower doses of G-CSF (5.2 versus 9.9 ug/kg/day, P=0.02), consistent with higher baseline ANC values. However, ELA2 negative patients were significantly less responsive to G-CSF therapy. Compared with ELA2 positive patients, on therapy, ELA2 negative patients had significantly smaller median increases in ANC values (1200 versus 2700 cells/uL, P=0.02); the difference remained significant after controlling for individual G-CSF dose (P=0.04). ELA2 negative patients had 5 MDS/AML events in 132 person-years, versus 8 MDS/AML events in 373 person-years among ELA2 positive patients. Using the Cox proportional hazards model, the hazard of MDS/AML was 3.1-fold higher in ELA2 negative patients compared with ELA2 positive patients. This association was of borderline statistical significance (P=0.08). No patient in either group died of sepsis. We found 34 distinct mutations in 52 ELA2 positive patients. The mutation sites were not clustered in the 8 ELA2 positive patients who developed MDS/AML. We found no associations of phenotype or outcome with specific ELA2 mutations; however, the numbers were limited.
CONCLUSIONS: SCN patients without mutations in ELA2 constitute a clinically distinct subgroup. ELA2 negative patients are significantly less responsive to G-CSF therapy than ELA2 positive patients, and they may be at substantially higher risk of leukemia. Additional studies are needed to confirm the latter association and identify the causative gene or genes.
Comparison of Myeloma Cells Killing Efficiency by Geldanamycin and Its Analogs.
Authors: Zebzda Anna , Jurczyszyn Artur , Czogala Wojciech , Lesko Ewa , Gozdzik Jolanta , Skotnicki Aleksander B. , Majka Marcin 
Published: April 2020

Abstract
Multiple Myeloma (MM) is the second most commonly diagnosed hematologic malignancy and still an incurable disease with dismal prognosis and average survival of 3 – 4 years. None of conventional therapies gives total cure or long survival, so it is urgently necessary to find new treatment strategies. Heat shock protein 90 (HSP90) is a potentially good therapeutic target, because it interacts with diverse and wide panel of client proteins that are important for proliferation and survival of tumor cells. Geldanamycin (GA) and its analog 17-allylaminogeldamycin (17AAG) are HSP90 inhibitors and were shown to block various tumor cells growth including MM cells. 17AAG has been already tested in clinical trials and showed some antitumor properties. There are, however, other inhibitors belonging to geldanamycin family such as: 17DMAG, 17DMAP and 17AEP which have not been tested against multiple myeloma cell lines and primary samples.
In this study we looked at the influence of GA and its analogs on proliferation and apoptosis of MM cells. We found that GA at 100 nM concentration inhibited growth of MM cells by 80%, 17AEP by 75%, 17DMAG and 17DMAP by 70% and 17AAG by 30%. Primary MM cells were more resistant and at 100 nM concentration we observed 30% inhibition with GA and 17DMAG. At the same time 17AAG did not effectively inhibit proliferation of primary MM cells.
We also analyzed influence of 100 nM concentration of GA and analogs on the apoptosis of MM primary cells and cell lines after 6 h and 24 h of incubation. Percentage of Anexin V and propidium iodine double positive geldanamycines treated primary MM cells was similar to the control after 6 h but change after 24 h. In comparison to control, number of apoptotic cells increased by: 175% for 17DMAG, 140% for GA, 80% for 17DMAP and 17AEP, no changes in number of apoptotic cells were observed for 17AAG. Similar observation was made for MM cell lines and increase in number of Anexin V positive cells was noticed. Again, percentage of Anexin V positive cells did not increase after treatment with 17AAG and remained at the control level. We also observed, similarly to others, substantial block in G1 and G2/M phase of cell cycle in cells treated with GA and its analogs. In conclusion, besides to 17AAG other HSP90 inhibitors could be considered as potential therapeutics for treatment of MM. Particularly, based on our data, we suggest that 17DMAG and 17AEP are new HSP90 inhibitors that should be tested in a clinic because they exert strong MM cells growth inhibition.
Bone Marrow Adipocytes Influence Multiple Myeloma Development by Secretion of Different Growth Factors and Chemokines.
Authors: Caers Jo , Belaid Zakia , Deleu Sarah , Defresne Marie Paule , Vanderkerken Karin 
Published: April 2020

Abstract
In multiple myeloma (MM) monoclonal plasma cells accumulate in the bone marrow (BM) where they interact with neighboring cells to receive supportive signals. Since BM adipocytes are known to participate in normal and malignant hematopoiesis, we presumed interactions between BM adipocytes and myeloma cells. Using an adipocytic cell line and primary isolated BM adipocytes, we performed functional assays indicating that adipocytes support MM cell development by affecting proliferation, apoptosis, cell migration and adhesion. We subsequently analyzed the secretion of different cytokines by BM adipocytes and analyzed the contribution of a specific adipokine, leptin, to these processes. We confirmed the expression of leptin by adipocytes and leptin receptor by myeloma cells. Immunostaining showed strong presence of leptin receptor on MM cells in 13 of 28 patients tested. This expression was not correlated to disease stage. On cell proliferation, leptin acted as a moderate growth factor. We conclude that, next to the earlier described interactions within the BM, adipocytes influence MM cell behavior by affecting proliferation, apoptosis and migration and that part of these effects may be contributed to leptin.
Over Expression of YY1 and RKIP in Multiple Myeloma (Cell Lines and MM Tissues).
Authors: Baritaki Stavroula , Huerta-Yepez Sara , Katsman Alina , Yeung Kam C. , Chatterjee Devasis , Chen Haiming , Berenson James , Bonavida Benjamin 
Published: April 2020

Abstract
There is a need to identify underlying molecular mechanisms and gene products (targets for intervention and biomarkers) involved in MM chemo- or immuno-resistance. Recent findings from our laboratory have reported high expression of the transcription factor Yin Yang 1 (YY1) in several tumor types and have demonstrated its fundamental role in tumor cell chemo/immuno-resistance (Gordon, S., et al., Oncogene25:1125, 2006). In contrast, low levels of the Raf kinase inhibitor protein (RKIP) has been shown to play a pivotal role in the negative regulation of cell survival signaling pathways, and has been considered a metastasis tumor suppressor. The overall objective of our studies is to examine whether MM cell lines and tissues derived from MM patients present deregulated expression patterns of RKIP and YY1, and to demonstrate the roles of YY1 and RKIP in MM pathogenesis and response to various therapies. In the present study, the myeloma cell lines RPMI-8226 and MM-1S were screened for RKIP and YY1 expression at the protein and m-RNA levels. In addition, fresh tissues from MM patients were also examined for RKIP and YY1 expression by IHC and their patterns compared with those observed in normal bone marrow cells. The findings demonstrate that both MM cell lines express remarkably higher levels of RKIP protein and transcripts compared to other tumor cell lines examined (lymphomas and prostate cancer), as assessed by western, RT-PCR and IHC. There was significantly higher expression of RKIP in MM patient’s samples compared to normal bone marrow cells. YY1 protein and m-RNA levels were detected in the MM cell lines; however, they were lower compared to Ramos and PC-3 cells. YY1 expression in MM tissues was significantly elevated compared to normal bone marrow cells. RKIP was basically present in the cytoplasm while YY1 was mainly accumulated in the nucleus. In contrast, normal bone marrow cells presented primarily cytoplasmic YY1 and RKIP expression. The YY1 expressing population was the CD38+/CD138− cell subset, which has been reported to be more susceptible to apoptosis (Mitsiadis, S.C., et al., Blood 98:795, 2001). Overall, the present findings support the potential involvement of two new gene products, such as YY1 and RKIP, in MM pathogenesis and their implication in regulating MM resistance to conventional therapeutics. Moreover, these gene products suggest their potential use as new therapeutic targets and/or biomarkers in multiple myeloma. Future studies with large cohorts of tissues will elucidate the importance of the above findings and will give new insights in the unexpected RKIP over expression and activity in multiple myeloma.
Evaluation of a 5-Color Flow Cytometric Technique for Immunophenotyping and Quantitation of Plasma Cell Myeloma in Post-Therapy Bone Marrows.
Authors: Isaacson Tove , Jakubowiak Andrzej , Stoolman Lloyd , Kota Usha , Finn William , Ross Charles 
Published: April 2020

Abstract
Multiparameter flow cytometry is a useful tool for comprehensive immunophenotyping of plasma cell myeloma, and has been proposed as a sensitive method for the evaluation of minimal residual disease in patients following treatment. This study aimed to assess the value of flow cytometry in quantitation of residual disease, in comparison to routine morphologic examination of first-pull bone marrow aspirate smears, in myeloma patients post-therapy. Heparinized bone marrow aspirates were obtained from 27 treated patients with plasma cell myeloma. Cells were prepared for 5-color flow cytometric analysis within 24-hours of specimen draw. Surface membrane staining with anti-CD19, CD20, CD38, CD45, CD56, and CD138 was followed by ammonium chloride lysis of red cells. Fixed and permeabilized cells were analyzed for cytoplasmic light chains to confirm clonality. Data were acquired using an FC500 flow cytometer (Beckman-Coulter), analyzed with CXP software with plasma cells isolated based on bright CD38+ or CD138+ expression. A median of 97,639 cellular events (range 14,279 to 262,508) were collected per analysis. Flow cytometric enumeration of plasma cells was compared to 500-cell differential counts of Wright-Giemsa-stained first-pull aspirate smears from the same cases. The median plasma cell count as determined by flow cytometry was 0.5% (range 0–7.9%). The median plasma cell count estimated by morphologic review was 8.0% (range 0–84.4%). Flow cytometry underestimated the plasma cell content in all but one case. Clonal plasma cells expressed CD38 and CD138 in all cases; 87.5% (21/24) coexpressed CD56, 25% (6/24) coexpressed CD45, and 4.2% (1/24) coexpressed CD19. None was positive for CD20. Although detection of minimal residual disease after therapy for acute leukemia is routinely achieved by flow cytometric analysis, successful quantitation of minimal residual disease in treated myeloma patients using flow cytometry remains limited as it usually underestimates the plasma cell content of bone marrow samples compared to routine morphology of first-pull aspirates. We have observed that this holds true for both pre-treatment and post-treatment specimens. Causes for the discrepancy may include hemodilution of second-pull aspirates used for flow cytometry, fragility and loss of plasma cells during preparation for flow cytometry, and incomplete disaggregation of plasma cells from bone marrow spicules. With improved outcome of treatments, better and more reliable methods of detection of minimal residual disease are needed for optimal prognostic stratification. We are currently validating alternative methods, which may offer more sensitivity while at the same time allow more objectivity, for assessing the amount of minimal residual disease in myeloma patients.
Immunoglobulin-Like Transcript 2 (ILT2) Is Not Differentially Expressed in MGUS and Myeloma, but Appears To Be Downregulated at an Earlier Stage of Plasma Cell Disease.
Authors: Schreder Martin , Huebl Wolfgang , Koch Gudrun , Strasser-Weippl Kathrin , Zojer Niklas , Ludwig Heinz 
Published: April 2020

Abstract
Background: Immunoglobulin-like transcript 2 (ILT2/CD85j) belongs to the Ig superfamily and has homology to the killer cell inhibitory receptors (KIRs). It is expressed on natural killer (NK) cells, monocytes, macrophages, dendritic cells and (naive) B lymphocytes. A differential expression of ILT2 was described for monoclonal gammopathy of undetermined significance (MGUS) and multiple myeloma (MM). Gene expression profiling studies found ILT2 to be downregulated 8.26 fold in myeloma as compared to MGUS, being the most differentially expressed gene between these two subsets. However, as RNA from CD138+ cells was used in this analysis, a varying percentage of normal, non-malignant plasma cells will impact on the results, especially in MGUS cases.
Aims: We aimed to delineate ILT2 expression in different plasma cell subsets (normal compared to monoclonal cells) in MGUS and MM and the eventual prognostic impact of a differential expression level.
Methods: ILT2 expression was measured by flow cytometry using a PE-conjugated antibody (clone HP-F1, Beckman Coulter) in a series of 30 MGUS patients and 91 myeloma patients. Phenotypically normal and malignant plasma cells were defined by differential expression of CD38, CD45, CD19 and CD56. Expression levels are given as mean fluorescence intensity (MFI) after correction for background staining.
Results: ILT2 was not differentially expressed in monoclonal plasma cells from patients with MGUS (MFI median 112.0, range 13.5–274.4) and myeloma (MFI median 96.6, range 0.4–454.5). In contrast, monoclonal cells from MGUS and MM showed a significantly lower expression of ILT2 as compared to phenotypically normal plasma cells in the majority of samples (p=0.007). Results were confirmed by quantitative real time PCR studies in 25 MM patients showing a linear correlation of ILT2 mRNA levels with the intensity of ILT2 protein expression. ILT2 levels did not vary with state of disease and showed no correlation with clinical parameters or prognosis in our series of myeloma patients.
Conclusions: In the majority of patients with monoclonal plasma cell disorders, ILT2 seems to be downregulated at an early stage of disease, i.e. upon transformation from a normal plasma cell to the MGUS/MM stage. The expression level of ILT2 in monoclonal plasma cells is neither correlated with the state of disease (MGUS versus newly diagnosed myeloma versus advanced disease) nor to prognosis of myeloma patients or other clinical parameters.
Induction of Cyclin D1 by HGF, IGF-1 and IL-6 in a Human Myeloma Cell Line with a t(11:14) Translocation.
Authors: Hov Håkon , Våtsveen Thea K. , Waage Anders , Sundan Anders , Borset Magne 
Published: April 2020

Abstract
Almost all patients with multiple myeloma have dysregulation of cyclin D1 -2 or -3. The t(11:14) translocation juxtapose the CCND1 locus coding for the cyclin D1 protein to the IgH locus. This leads to upregulation of the cyclin D1 gene, and thus to increased proliferative capacity. Despite the presence of IgH translocations, the myeloma cells are highly dependent of the bone marrow microenvironment for malignant growth. Whether the expression of cyclin Ds is additionally regulated by the microenvironment needs to be further examined. Here we report a myeloma cell line, INA-6, with a t(11;14) translocation in which expression of cyclin D1 is regulated by several cytokines. INA-6 cells were analysed by interphase FISH with probes matching the CCND1 gene (coding for the cyclin D1 protein) and the Ig heavy chain locus. We found co-localisation of the two probes indicating a translocation between chromosome 14 and chromosome 11. We detected a total of five signals for CCND1, indicating the occurrence of three extra copies of chromosome 11. This was confirmed by an analysis with a single probe detecting the CCND1 locus. We further analysed the content of cyclin D1 protein by western blotting after the cells had been washed and grown in serum free media with or without cytokines for 24 hours. INA-6 constitutively expressed cyclin D1. HGF, IL-6 and IGF-1 further enhanced the expression of cyclin D1. In combination with IL-6, HGF had a synergistic effect on cyclin D1 expression. The constitutive cyclin D1 expression is probably caused by the translocation between chromosome 11 and 14. However the increase in number of cyclin D1 genes not co-localized to the IgH chain can also contribute to the basal level of cyclin D1 expression in this cell line as well as post-transcriptional regulation. The increase in cyclin D1 expression after cytokine stimulation can be explained by an increase in transcription from the normal CCND1 copy or the other extra copies. One intriguing possibility is that the translocation itself makes the cell susceptible to cytokine regulation of cyclin D1. At least IGF-1 and IL-6 are known for their ability to enhance immunoglobulin production and may therefore regulate cyclin D1 through the IgH promotor in the t(11:14) translocation.
The different ways to induce cyclin D1 protein in this cell line is a functional example of the collaboration between the dysregulated myeloma cell and the components of its microenvironment in the bone marrow in regulating a key component of cell cycling and a possible oncogene in multiple myeloma.
The Sesquiterpene Lactone Parthenolide Induces Apoptosis and Overcomes the Protective Effect of the Bone Marrow Microenvironment in Human Multiple Myeloma Cells.
Authors: Suvannasankha Attaya , Crean Colin D. , Shanmugam Rajasubramaniam , Abonour Rafat , Boswell H. Scott , Nakshatri Harikrishna 
Published: April 2020

Abstract
Parthenolide is a sesquiterpene lactone, a new class of antitumor agents which target nuclear factor kappa B (NFkB). Although the activation of NFkB has been implicated in the growth and drug resistance of multiple myeloma (MM) cells, the efficacy of parthenolide in MM is unknown. Here we show that parthenolide inhibits the NFkB DNA binding in MM cells in a time- and dose-dependent fashion. Parthenolide inhibits the proliferation of MM cell lines, as well as tumor cells from chemotherapy-resistant MM patients; however, at the effective doses it is not toxic to peripheral blood mononuclear cells or bone marrow stromal cells. Neither exogenous interleukin-6 (IL-6) nor insulin-like growth factor 1 (IGF-1) overcome the parthenolide-induced cytotoxicity. In addition, parthenolide inhibits the proliferation of MM cells adherent to bone marrow stromal cells. Parthenolide has a minimal effect on cell cycle progression but strongly induces apoptosis. Parthenolide activates caspases 8 and 3 more strongly than caspase 9, resulting in the cleavage of poly (ADP)-ribose polymerase (PARP) and XIAP. Z-VAD-FMK partially blunts the PARP cleavage but does not protect MM cells against apoptosis. Parthenolide induces p21 accumulation, while the levels of Bax, IAP2, cyclin D, Bcl-X and p53 or the phosphorylation status of JNK, AKT and ERK are unaffected. In addition, parthenolide augments dexamethasone-induced cytotoxicity. Our study therefore provides the rationale for the future clinical development of parthenolide, or its combination with dexamethasone, in MM therapy.