bluuebunny/crossref_metadata_2025 huggingface dataset:
Relationship between Serum Concentrations and Urinary Excretions of Monoclonal Free Light Chains (mFLC) Detectable as Bence Jones Proteins (BJP) by Immunofixation Electrophoresis (IFE) in Patients with Multiple Myeloma (MM).
Authors: Nowrousian M.R. , Brandhorst D. , Sammet C. , Kellert M. , Daniels R. , Schuett P. , Poser M. , Mueller S. , Ebeling P. , Welt A. , Buttkereit U. , Opalka B. , Flasshove M. , Seeber S. , Moritz T.
Published: April 2020

Introduction. mFLC are important markers for the diagnosis and monitoring of MM. This study for the first time determines serum concentrations of mFLC which are required to produce renal overflow and BJP in urine detectable by IFE and evaluates the relationship between urinary excretions of mFLC and renal function.

Patients and methods. 378 paired samples of serum and 24-h-urine from 82 patients were evaluated during the course of their disease. Serum FLC concentrations were measured nephelometrically using an automated immunoassay. Urine samples were tested for clonal bands using agarose gel electrophoresis, scanning densitometry and visual checking of electrophoretic gels. BJP were identified by urine IFE.

Results. Among the 378 serum samples, 173 (46%) were normal and 205 (54%) were abnormal for FLC k/l ratios, indicating the presence of mFLC, 98 of kappa and 107 of lambda type. In 98 serum samples with mFLC kappa, 48 (49%) were associated with negative urine IFE analysis and 50 (51%) had positive urine tests. The median serum kappa concentrations were 40 mg/L (range 6–710) for negative urines and 113 mg/L (range 7–39500) for positive urines (p=0.001), indicating an almost threefold greater median value which was approximately six times the upper limit of the reference range (3.3.–19.4 mg/L) for samples with positive urine IFE analysis. In 107 serum samples with mFLC lambda, 70 (65%) were negative in urine and 37 (35%) were positive. The median serum concentrations associated with negative urine IFE tests were 44 mg/L (range 3–561) and were 278 mg/L (range 5–7060) for positive urines (p=0.0001), indicating an almost sixfold difference. This was approximately 2.5-fold greater than for kappa, and approximately 11 times the upper limit of the reference range (5.7–26.3 mg/L) for samples with positive urine IFE analysis. Renal excretions of mFLC, in addition, were determined primarily by serum concentrations for lambda, but by serum concentrations, renal function and, probably, molecular changes for kappa. For both, renal excretions significantly decreased at high serum concentrations combined with renal dysfunction.

Conclusion. Based on these results, relatively high serum concentrations of mFLC are required to produce renal overflow and positive urine IFE tests for BJP. Furthermore, urine excretions of mFLC are determined primarily by serum concentrations, but also by renal function, particularly for kappa.
The Expression and Clinical Significance of Survivin in Multiple Myeloma.
Authors: Li Juan , Zhao Ying , Zhang Dian-Bao , Haung Bei-Hu
Published: April 2020

BACKGROUND & OBJECTIVE:Survivin, a member of the inhibitors of apoptosis protein family (IAPs), is over-expressed in many cancers but not in normal differentiated adult tissues. It is known to be involved in poor prognosis and resistance to chemo- and radiation therapy in many cancers. This study was designed to detect the expression of survivin in normal people, newly diagnosed and relapsed/refractory patients of multiple myeloma (MM), to find out the relationships of its expression with drug resistance and prognosis of MM patients.

METHODS: RT-PCR and Western blot was used to detect mRNA and protein expression of survivin in 29 MM patients (17 newly diagnosed patients and 12 relapsed /refractory patients), 9 cases of normal people and myeloma cell line KM3.

RESULTS: KM3 cell line expessed survivin strongly. The positive rates of survivin mRNA in nomal people, newly diagnosed patients and relapsed/refractory patients were 22.2%, 70.5% and 83.3% respectively, and the expressing levels (survivin/β-actin) were 0.06±0.04, 0.31±0.14 and 0.69±0.24 respectively. On protein level, survivin could not be detected in all of the normal people, while in newly diagnosed patients and relapsed/refractory patients, the positive rate were 41.4% and 58.3%
Falsely Low Serum Free Light Chain Concentration in Patients with Monoclonal Light Chain Diseases.
Authors: Mollee Peter , Tate Jill , Dimeski Goce , Gill Devinder
Published: April 2020

Introduction: Measurement of serum free light chains (FLC) is useful in the diagnosis and monitoring of nonsecretory myeloma, AL amyloidosis (AL) and light-chain myeloma (LCMM). Long-term monitoring of monoclonal FLC disease requires reproducible assay performance. We assessed the FLC assay performance in regard to falsely low FLC values.

Methods: We assessed:

FLC immunoreactivity and, FLC assay recovery on sample dilution.

FLC concentration was measured in serial serum samples collected from patients with AL and LCMM, using a kit assay (The Binding Site Ltd., Birmingham, UK) with the IMMAGETM (Beckman Coulter, Brea, CA) and Hitachi Modular P (Roche Diagnostics) analysers. Monoclonal paraprotein was detected by serum and/or urine protein electrophoresis (Sebia, Issy-les-Moulineaux, France) and immunofixation.

Results:

FLC immunoreactivity. Of 23 patients with LCMM, one patient had discordant Bence Jones protein (BJP) and FLC results. The patient was a 70-year-old male with kappa LCMM where changes in the level of KBJP correlated with the KFLC concentration and the serum kappa to lambda (K/L) FLC ratio in samples measured between 2002 and mid 2004. Thereafter the FLC ratio remained within the reference range despite increasing urine KBJP. Subsequent investigations revealed that the patient’s monoclonal KFLC had failed to react with three different lot numbers of kit reagent. A fourth reagent lot, however, gave elevated KFLC concentration and K/L ratio consistent with disease relapse. A partial response to the patient’s monoclonal KFLC was observed for the same samples using the Modular assay (i.e., KFLC concentration was approximately 50% or less of the IMMAGE value). FLC recovery. In four patients with AL or LCMM the monoclonal FLC was underestimated when using the manufacturer’s recommended 1:10 on-line dilution and gave higher recoveries at 1:20 to 1:110 sample dilution. Values for the monoclonal FLC differed by up to 100% and recovery on dilution did not parallel the standard curve response.

Conclusions:

We report the first case of non-reaction of a monoclonal kappa FLC in a patient with LCMM. Urinary BJP monitoring remains necessary in patients with LCMM. Dilutional non-linearity of FLC suggests that the immunoreactive response of some monoclonal FLC does not parallel that of polyclonal calibration materials. This may result in falsely low FLC results. While the FLC assay represents a major advance for these diseases, clinicians should be aware of the assay’s limitations.
Serum Free Light Chain Measurement Can Replace Urine Electrophoresis in the Detection of B Cell Proliferative Disorders.
Authors: Forsyth Julia M. , Hill Peter G. , Rai Baldeep S. , Mayne Stewart , Mead Graham
Published: April 2020

Retrospective studies have established the high sensitivity of quantitative serum free light chain (SFLC) measurement in the detection of patients with light chain myeloma, nonsecretory myeloma and primary amyloidosis. These disorders account for 15–20% of new diagnoses of B cell proliferative disorders and cannot be confidently excluded by current protocols without electrophoresis of both serum and urine. This prospective study evaluated whether the addition of SFLC to our first line investigations for B cell proliferative disorders detected additional patients with monoclonal gammopathies and if SFLC could replace the requirement for urine for Bence Jones Protein (BJP) analysis. SFLC were added to consecutive requests for serum electrophoresis (SPE) received to investigate patients for possible B-cell proliferative disorders over a 4 month period. SFLC was measured by immunoturbidimetry (The Binding Site, Birmingham, UK) in 923 patients. Subjects with known B-cell disorders were excluded. Concurrent urine samples were received from 370 patients for BJP(40%). The Hydrasis system was used for SPE, urine electrophoresis (UEP) and urine and serum immunofixation (Sebia, Issy-Les-Moulineaux, France.) Kappa/Lambda FLC ratio was abnormal in 71 (7.7%) patients. In 28 of these cases the SPE showed a possible monoclonal band or hypoglobulinaemia and subsequent serum immunofixation confirmed a monoclonal band in 19. In the remaining 43 with negative SPE but abnormal SFLC ratios, lymphoma was diagnosed in 1, multiple myeloma in 2 and monoclonal gammopathy of undetermined significance in 3. Follow-up is continuing in 3 further patients. Of 15 patients with positive urine BJP, 4 patients had normal SFLC ratios but did not have lymphoproliferative disorders requiring treatment. Additional diagnostic information was gained by adding SFLC to SPE for first line investigation of possible B cell disorders. 6 additional patients were identified by abnormal SFLC ratios. Replacing BJP with SFLC ratio would not have missed any significant pathology. In the light of this study we have adopted SPE and SFLC as our first line tests for the investigation of possible B cell disorders. The quality of our diagnostic service has improved; urine samples were received from only 40% of patients being screened and SFLC analysis allows the more confident exclusion of light chain disorders in the absence of a urine sample. The availability of SFLC results when viewing SPE gels assists in the interpretive process and highlights samples requiring serum immunofixation to identify monoclonal bands in the alph-2 or beta globulin regions or light chain monoclonal bands too small for reliable detection by SPE.
IgH Gene Rearrangements Are Strongly Associated with Hyperdiploidy Variant Multiple Myeloma in Korea: A Contradiction to the Pathogenetic Pathway of IgH Tranlocations in Non Hyperdiploidy Variant.
Authors: Moon Hee Won , Kim Tae Young , Kang Seong- Ho , Chi Hyun-Sook , Seo Eul Zu , Park Chan Jeoung , Chun Honggu , Kim Hee Chan , Yoon Sung-Soo , Park Seonyang , Kim Byoung Kook , Cho Han Ik , Lee Dong Soon
Published: April 2020

Recent studies proposed the classification of multiple myeloma (MM) by the pathways involved in the early pathogenesis; nonhyperdiploid variants with a high incidence of IgH translocations and hyperdiploid variants associated with no IgH translocation. Most studies applied cytogenetic study or flow cytometry to define the ploidy. In this study, we combined the cytogenetic results and fluorescent in situ hybridization results to define the ploidy and investigated IgH tranlocation and 13q deletion in relation to the ploidy level on Korean patients with MM. A total of 135 cases diagnosed as MM between 1997 and 2003 from Seoul National University Hospital and the Asan Medical center were enrolled in this study. Conventional cytogenetic studies and FISH studies with different probes specific for the regions containing the genes or chromosomes (RB1, D13S319, D13S25, IgH/FGFR3, IgH/BCL2, IGH dual color, break apart rearrangement probe, IgH/CCND1, 1q, p53, p16, MLL, CEP 7, 11, 12) were performed. Of 135 patients with MM, 62 (45.9%) patients had hyperdiploid karyotype by cytogenetics and FISH. IgH translocations were observed in 37.4% of Korean patients with MM and were more frequent (54.7%) in hyperdiploid variants than in nonhyperdiploid variants (17.4%). Incidence of deletion 13q was 34.7% and also more frequent in hyperdiploid variants (54.2%) than in nonhyperdiploid variants (16.1%). In conclusion, IgH translocations and 13q deletions were not associated with nonhyperdiploid MM and appeared more frequently in hyperdiploid variant in Korean patients with MM.
Characterization of Patients with Multiple Myeloma and t(4;14) Detected by Fluorescence In Situ Hybridization.
Authors: Collado Rosa , Hueso Jose A. , Romaguera Miriam , Mora Carmen , Lopez Aurelio , Benet Carmen , Amigo Victoria , Ferrer Rosa , Navarro Isabel , Ruiz Angeles , Martinez Jose , Pacios Alejandro , Orero Maite , Miguel-Sosa Amparo , Felix Carbonell
Published: April 2020

Multiple myeloma (MM) is a genetically unstable malignancy of postgerminal center B-cells. Almost 40% of intramedullary MM tumors show primary translocations that affect immunoglobulin heavy chain (IgH) gene. These include t(4;14)(p16;q32) which results in the dysregulated expression of two potential oncogenes, MMSET on der(4) and FGFR3 on der(14), and has been associated with poor outcome. The main goals of this study were to determine the incidence and clinical significance of t(4;14) among our MM patients. Therefore, we studied bone marrow specimens from 65 patients with MM by fluorescence in situ hybridization (FISH). All cases were screened for IgH rearrangements, t(4;14), t(11;14), and 13q14 deletions using the locus-specific probes LSI IgH dual color, break apart, LSI IGH/FGFR3, LSI IGH/CCND1, and LSI D13S319 (Vysis). FISH analysis revealed 35 cases (54%) involving IgH locus. 8 patients (12.3%) had a t(4;14), and 13 cases (20%) showed a t(11;14). In the remaining 14 samples (21.5%), IgH rearrangements were observed, but the translocation partner was not one of the loci for we tested. Furthermore, 13q14 deletions were more frequent among patients with t(4;14) than among those with t(11;14) (62.5% vs 30.7% respectively, p=0.03). Regarding clinical parameters, presence of t(4;14) was significantly associated with anemia (mean value: 9.0 g/dl, p=0.049), elevated LDH levels (mean value: 535.4 U/l, p=0.05), Durie III stage (p=0.049), and number of lytic bone lesions >2 (p=0.007). Finally, the survival median of patients with t(4;14) was 23 months vs 48 months for the group without this abnormality. Cases with t(11;14) did not show adverse correlation with survival.

Based on these data, FISH is a successful technique to detect translocations affecting telomeric localization of both chromosomal partner loci. In addition, we confirmed t(4;14) as an important factor of poor prognostic in MM. Its detection is essential to lead a correct evaluation of patients at diagnosis.
Clinical Significance of the Tumor Suppressor p53 Codon 72 Polymorphic Variants in Multiple Myeloma.
Authors: Gonzalez-Paz Natalia C. , Chng Wee Joo , Kumar Shaji , Price-Troska Tammy , Van Wier Scott , Ahmann Gregory , Henderson Kimberly , Gertz Morie , Kyle Robert , Greipp Philip , Fonseca Rafael
Published: April 2020

Background: The p53 tumor suppressor gene has a common sequence polymorphism that results in either proline or arginine at amino-acid position 72. The Pro72 and Arg72 variants have been reported to differ in its functional activity. The Arg72 variant induces apoptosis more efficiently than Pro72 variant by its greater ability to localize to the mitochondria and increase the release of cytochrome c into the cytoplasm. In addition, recent studies have demonstrated that Arg72 variant was a better suppressor of cellular transformation, an activity believed to rely on the pro-apoptotic function of p53. Moreover, the Pro72 variant has been associated with a higher incidence of tumors.

Aim: To evaluate the effect of the polymorphic variants of codon 72 and clinical outcome in patients with plasma cell malignancies.

Patients and Methods: We analyzed 79 patients with plasma cell malignancies seen at Mayo Clinic between 2000 and 2003. Genomic DNA was extracted from CD138+ sorted plasma cell from bone marrow aspirates. PCR amplification of codon 72 and posterior sequencing was used for the analysis.

Results: A total of 79 patients were analyzed. Thirty-four patients (34%) were heterozygous for p53, 57 (59%) homozygous for Arginine and 6 (6%) homozygous for Proline. The distribution among disorders was as follow, 52 patients had MM, 14 patients with SMM and 13 with MGUS. Among patient with MM 25% were Pro/Arg heterozygous, 71 % were Arg/Arg homozygous and 3.8% Pro/Pro homozygous. Survival data among MM patients and genotype were analyzed to Kaplan- Meier method. No significant differences were found among patients who carried homozygous (72R) or heterozygous genotype and overall survival. We exclude the analysis in MGUS and SMM for a low number of patients available.

Conclusions: These findings indicate that codon 72 arginine p53 may not be associated with a prolonged survival in patients with Multiple Myeloma, but further study is needed to assess whether this polymorphism is associated with progression of MGUS to MM, age of onset, clinical manifestations, response to treatment etc.
Hemojuvelin Acts as a Bone Morphogenetic Protein Co-Receptor To Regulate Hepcidin Expression.
Authors: Huang Franklin W. , Babitt Jodie L. , Wrighting Diedra M. , Samad Tarek A. , Xia Yin , Sidis Yisrael , Campagna Jason A. , Chung Raymond T. , Schneyer Alan L. , Woolf Clifford J. , Andrews Nancy C. , Lin Herbert Y.
Published: April 2020

Juvenile hemochromatosis is a severe iron overload disorder resulting from mutations in the hemojuvelin (HJV) gene. To understand its pathogenesis, we developed Hjv−/− mice. Similar to human patients, Hjv−/− animals accumulate excess iron in the liver, pancreas and heart early in life. Tissue macrophages are iron-depleted. Hjv−/− mice express very low levels of hepcidin mRNA and, likely as a consequence, have elevated expression of the iron transporter ferroportin in enterocytes and macrophages. These results suggested that Hjv plays a role in regulating hepcidin expression. Two known Hjv homologs, Rgma and Rgmb, have previously been shown to act as bone morphogenetic protein (BMP) co-receptors. We hypothesized that Hjv regulates hepcidin expression through a BMP signal transduction pathway. We found that Hjv binds radiolabeled BMP, supporting the contention that it is a BMP co-receptor. Transfection of HepG2 cells with Hjv cDNA activated a BMP-responsive reporter construct and augmented its response to exogenous BMP. Both an anti-BMP neutralizing antibody and the natural BMP antagonist Noggin blocked this response, as did co-expressed dominant negative BMP receptor proteins. When cells were transfected with a construct carrying an Hjv mutation known to cause human disease, BMP reporter activation was significantly reduced in the presence and absence of exogenous BMP. Treatment with BMP stimulated hepcidin production in hepatoma cells and activated a reporter construct containing a fragment of the hepcidin promoter. To extend these results, we studied tissues from Hjv−/− mice. BMP signals are transduced through phosphorylation of Smad proteins. We found that Smads 1, 5 and 8 were hypophosphorylated in Hjv−/− liver, consistent with impaired BMP signaling. BMP treatment of wild type and Hjv−/− primary hepatocytes induced hepcidin expression, but induction was blunted in cells from Hjv−/− animals. Taken together, these data suggest that the normal hepatic function of Hjv is to serve as a BMP co-receptor, modulating a signal transduction pathway that culminates in hepcidin expression. [Note - Jodie L. Babitt is the first author of this abstract, but it will be presented by Franklin W. Huang, the second author]
Serum Concentrations of Vitamin B-12 and Alkaline Phosphatase in Newly Diagnosed Multiple Myeloma Patients.
Authors: Elice Francesca , Zangari Maurizio , Barlogie Bart , Burns Michael J. , Tricot Guido J. , Roberti Sara , Rodeghiero Francesco , Hine R. Jean
Published: April 2020

Background: A controversy about a possible link between vitamin B12 deficiency and multiple myeloma (MM) has persisted since the 1970s. Vitamin B12 (B12) deficiency can cause anemia, neuropathy and has been associated with osteoporosis and reduced bone density. Therefore, B12 deficiency might worsen some clinical symptoms of MM.

Methods: We evaluated serum B12 concentrations in 393 untreated MM patients included in the Myeloma Institute Program (October 1998 to March 2005). Total serum B12 was assessed by using ADVIA Centaur immunoassay with chemiluminescent detection; laboratory B12 deficiency was defined as serum B12 levels < 156 mmol/L. Serum alkaline phosphatase (ALP) was determined by a kinetic rate method. Neuropathy was assessed clinically according to NIH guidelines. Correlations of serum B12 concentrations with bone density (radial T- and Z -score) and thalidomide-related neuropathy were investigated. Categorical variables were compared using chi-square and Fisher’s exact test, while correlation coefficient was used for quantitative variables.

Results: The sample was 57% male, median age = 58 years, stage III 70%, median creatinine 1.0 mg/dL, IgG 52%, IgA 24%, and light chain disease 19%. Prevalence of laboratory B12 deficiency was 3.5%, prevalence of functional deficiency (B12 < 241 mmol/L) was 20.4%. No influence of age or MM isotype on B12 levels was observed. Sixty seven percent (264) of the patients received thalidomide in combination with chemotherapy. Levels of B12 did not affect the development of grade ≥ 2 neuropathy after thalidomide treatment (p=0.49). No significant association was observed between serum B12 and the radial T- and Z score at bone densitometry (n= 270). There was a highly significant correlation between serum B-12 and ALP levels (correlation coefficient 0.25, p < 0.0001).

Conclusions: Laboratory B12 deficiency was not common in our series of newly untreated MM patients. B12 levels did not predict the development of thalidomide-related neuropathy, nor are they associated with reduced bone density. The most striking finding of our study was a highly significant correlation between serum B12 and ALP concentrations; clinical significance of this observation requires further evaluation.
Ambulatory Blood Pressure Monitoring in Multiple Myeloma Patients Treated with VAD Protocol.
Authors: Pyszel Angelika , Biedron Monika , Poreba Rafal , Wrobel Tomasz , Mazur Grzegorz , Szuba Andrzej , Kucharski Wojciech , Podgorski Maciej , Kuliczkowski Kazimierz , Andrzejak Ryszard
Published: April 2020

BACKGROUND: Corticosteroids are still a cornerstone in multiple myeloma (MM) therapy, both as a single agent and as a part of many protocols, including VAD regimen (vincristine, adriblastine, dexamethasone). Corticosteroids are known to be involved in blood pressure (BP) regulation and to affect this parameter. The impact of dexamethasone (Dex) administration on BP in MM patients during VAD protocol treatment is poorly documented.

AIM: The purpose of the study was to evaluate the blood pressure changes during Dex administration in patients treated with VAD protocol due to MM.

METHODS: Thirteen patients with MM (7 men and 6 women; mean age 62,45 ± 8,14) were assessed. Primary hypertensive patients (7 persons) were not excluded. They were administered Dex in standard dose of 40 mg (day 1–4, 9–12, 17–21) according to VAD protocol. Blood pressure was assessed by the use of commercially available instruments of Ambulatory Blood Pressure Monitoring (ABPM). The BP recordings lasted 48 hours, were started on the day before the first day of VAD and were obtained every 10 minutes during mornings, every 15 minutes during the rest of days and every 20 minutes during nights. Average of systolic and diastolic blood pressure (SBP/DBP) were estimated for the 2-hour time before Dex and for the 14-hour time in the 2-hour periods after Dex. Minimal and maximal range of SBP/DBP increase and the mean amount of SBP/DBP increase were also determined.

RESULTS: 48-hour BP recordings revealed a significant increase in systolic and diastolic blood pressure after Dex administration in all patients. SBP and DBP began to increase after 3 hours after Dex, then rose continually and reached the peak in the period from 6 to 10 hour after Dex. In comparison to 2-hour period before Dex, in which SBP/DBP amounted 139,63/82,92 ± 23,47/9,38 mmHg, the mean SBP/DBP increase rate was: in the 2–4 hour period after Dex - 146,68/88,15 ± 24,38/10,51 mmHg (p<0,05), in the 6–8 – 148,07/92,66 ± 14,07/9,04 mmHg (ns), in the 8–10 – 147,8/87,99 ± 14,07/9,04 mmHg (p<0,01), in the 10–12 – 143,44/86,12 ± 17,22/9,77 mmHg (p<0,05) and in the 12–14 – 144,47/88,59 ± 17,04/12,54 mmHg (p<0,01). The minimal range of SBP/DBP increase was 10,50/−2,00 mmHg, maximal range of SBP/DBP increase was 35,43/40,00 mmHg and the mean amount of SBP/DBP increase was 20,06/11,6 ± 8,36/14,23 mmHg.

CONCLUSION: Our preliminary study revealed that Dex administration causes an increase in BP in all patients. The mean increase in BP amounted 20,06/11,6 ± 8,36/14,23 mmHg and was similar in all patients, regardless their initial value of BP. So our study demonstrates the need of individualized hypertension treatment with strict control of BP in hypertensive patients when corticosteroid therapy is indicated.
Thalidomide in Relapsed or Refractory Patients with Multiple Myeloma: Monotherapy or Combination Therapy? A Report from Systematic Reviews.
Authors: Glasmacher Axel , Hahn Corinna , Hoffmann Florian , Furkert Kerstin , von Lilienfeld-Toal Marie , Orlopp Katjana , Naumann Ralph , Goldschmidt Hartmut , Schmidt-Wolf Ingo G.H. , Gorschlueter Marcus
Published: April 2020

The activity of thalidomide monotherapy in relapsed or refractory disease is widely accepted but many haematologists have observed better response rates with combination therapies. This communication aims to give an overview on the efficacy of these alternatives.

Methods: Our group has performed two systematic reviews (thalidomide monotherapy: Glasmacher et al., Brit. J. Haematol., 2005; 129 suppl 1: 24; thalidomide and dexamethasone combination therapy: submitted to ASH 2005). Furthermore a search of trials combining thalidomide, cyclophosphamide and dexamethasone (with or without further drugs) was performed. Only trials that included patients with relapsed or refractory multiple myeloma could be evaluated. Response was defined as a reduction of the monoclonal component by more than 50% in the absence of any sign of disease progression. Proportions and 95% confidence intervals were reported (Confidence Interval Analysis, Version 2.1.1, Southampton, UK, 2000).

Results: Thalidomide monotherapy was administered in doses between 50–600 mg/d (median doses). Thalidomide in combination with dexamethasone was given in doses of 100–400 mg/d (median doses), dexamethasone was given 40 mg/d d1–4 every 3–4 weeks in 7 of 8 trials. Thalidomide dosage and combination therapy of trials adding Cy to thalidomide and dexamethasone are shown in Table 1. The response rates of the three different thalidomide applications are demonstrated Table 2.

Conclusion: This compilation of data from phase II trials cannot replace a randomized trial and may be biased. With this limitation it seems that thalidomide combined with dexamethasone (response rate 51%) is more effective than monotherapy (response rate 29%) as the 95% confidence intervals for the response rates do not overlap. Whether a combination of thalidomide, dexamethasone and cyclophosphamide is more effective than thalidomide and dexamethasone alone cannot be decided from the current data.

Table 1: Studies with thalidomide and combination chemotherapy Study Intervention No. of pts. Response (95%CI) Abb.: Cy, cyclophosphamide; Eto, etoposide; T, thalidomide; Dex, dexamethasone; Ida, idarubicin. Values are mg/m2 per course for Cy, Eto, Ida; mg per course for Dex, mg/d for T. Moehler et al. 2003 Cy 1600, Eto 160, T 400, Dex 160 119 55% (46–65) Kropff et al., 2003 Cy 1800, T 400, Dex 240 60 70% (57–81) Dimopoulos et al. 2003 Cy 1500, T 400, Dex 160 43 67% (51–81) Gracia-Sanz et al. 2004 Cy 50 daily, T 800, Dex 160 71 57% (45–67) Glasmacher et al. 2005 Cy 800, Ida 40, T 400, Dex 320 39 57% (41–73) Table 2: Response of different thalidomide combinations Intervention No. of trials No. of pts. Response (95%CI) Abb.: see Table 1 T monotherapy 42 1629 29% (27–32) T + Dex 8 283 51% (45–57) T + Cy + Dex 5 332 60% (55–65)
Anti-Myeloma Effects of a Novel Synthetic Retinoid Am80 (Tamibarotene) through Inhibition of Angiogenesis.
Authors: Sanda Takaomi , Iida Shinsuke , Kuwano Takashi , Ono Mayumi , Ueda Ryuzo
Published: April 2020

In multiple myeloma (MM), the interaction between myeloma cells and bone marrow microenvironment has an important role in the pathogenesis of MM. We first examined the inducing effect of myeloma cells on migration of human umbilical vein vascular endothelial cells (HUVECs). Myeloma cell lines produced varying amounts of vascular endothelial growth factor (VEGF), and migration of HUVECs was induced by co-culture with myeloma cells. We next examined the inhibitory effect of a novel synthetic retinoid Am80 (Tamibarotene) on both myeloma cells and HUVECs. Am80 remarkably inhibited the growth of HUVECs stimulated by VEGF and blocked migration of HUVECs by co-cultured myeloma cells. In addition, VEGF-induced formation of tube-like structures in vitro and neovascularization in mouse corneas were significantly inhibited by Am80. Am80 downregulated both interleukin-6 (IL-6) and IL-6 receptor, and blocked VEGF-induced phosphorylation of VEGF receptor. Furthermore, microarray analysis suggested that Am80 has immunomodulatory effects on myeloma cells. These findings demonstrate that Am80 might be a useful therapeutic agent against MM.
Valproic Acid Induces Non Apoptotic Cell Death in Myeloma Cells and Cell Lines.
Authors: Schwartz Chantal , Palissot Valérie , Brons Rene I. , Leners Bernadette , Bosseler Manon , Dicato Mario A. , Berchem Guy J.
Published: April 2020

Multiple myeloma is, despite the emergence of new treatment strategies in recent years, still a lethal disease in 2005. Over the last several years, significant insights into the dysregulation of various signal transduction pathways of MM have emerged and have evolved the development of new agents. Histone deacetylase inhibitors as valproic acid are compounds that inhibit HDACs enzymes that, in conjunction with histone acetylases, regulate the acetylation state of histones and, by extension, the conformational status of chromatin. VPA is clinically known to be used in treatment of different types of epileptic disease. Effects and signal transduction pathways of cell death have been studied in cells harvested and purified from routine bone marrow aspirates of several patients with multiple myeloma, as well as on myeloma cell lines (OPM2, U266) treated with a physiologic dose range of valproic acid (1–4 mM). We observe significant in vitro toxicity starting at doses of 1 mM for 24h in cell lines as well as in patient cells

using an XTT based cytotoxicity assay. The question we adressed was the mechanisms by which the MM died. Flow cytometric analysis with PI / Annexin V staining does not show apoptosis features, nor do TUNEL staining or DNA fragmentation assays, suggesting non activation of the intrinsic apoptosis pathway. In contrast, cell morphology of treated cells stained with May-Gruenwald-Giemsa staining show increase in multinucleated giant cells. Multinucleation has often been described in cells which die through mitotic catastrophe. Further experiments exploring this hypothesis will be conducted.

Effect of 48h valproic ac on myeloma cell lines Effect of 48h valproic ac on myeloma cell lines Effect of 48h valproic ac on CD138 cells from myeloma patients Effect of 48h valproic ac on CD138 cells from myeloma patients
Treatment Strategy and Prognosis of Myeloma Patients According to ISS Stage at Diagnosis.
Authors: Hatta Yoshihiro , Nishinarita Susumu , Itoh Takeyoshi , Takeuchi Jin , Sawada Umihiko
Published: April 2020

Background: Recently, a new ISS staging system for myeloma has been developed. By analyzing over 10,000 patients, ISS is a simple and useful system to estimate prognosis. However, by ISS, survival time was evaluated only for the time after chemotherapy. The prognosis of untreated patients and survival time after initial diagnosis were not well understood by ISS. Our aim was to evaluate the prognosis and define treatment strategy for newly diagnosed, untreated myeloma patients using ISS at diagnosis. Especially, treatment strategy for ISS stage I was received attention.

Methods: 127 newly diagnosed, untreated myeloma patients referred to Nihon University Itabashi Hospital between 1994 and 2004 were retrospectively analyzed. ISS stage was determined with the laboratory data at diagnosis. Median age of the patients was 62 y (range 30–88). Median follow-up duration was 32.5 months (range 0–171). Transplanted patients were censored at the time of transplantation.

Results: 50% survival duration after initial diagnosis for stage I (n=54), II (n=26), and III (n=47) were 106, 36, and 14.5 months, respectively. For stage I patients given chemotherapy, both 50% survival times after initial diagnosis and after chemotherapy were the same, 85 months, suggesting that stage I patients required chemotherapy were treated early after diagnosis. Fifteen stage I patients have not been treated and all of them are alive. Of these 15 patients, 14 were in Durie-Salmon stage I and one was in Durie-Salmon stage II at diagnosis. 50% progression time from ISS stage I to II in untreated patients was 2 months. For the patients treated at ISS stage I, 50% progression time from stage I to II and I to III were 61 and 155 months, respectively. These data suggest that the progression from stage I to II could be postponed by early chemotherapy. However, overall survival duration of the patients treated at stage I and of those treated after progression to stage II was not different.

Conclusion: We defined treatment strategy for myeloma using ISS at diagnosis. Certain amounts of myeloma patients with Durie-Salmon stage I and ISS stage I concomitantly at diagnosis were not required chemotherapy. Administration of chemotherapy for ISS stage I patients at diagnosis are recommended after they progress to stage II.
A Multicenter Phase I/II Trial Evaluating the Safety and Efficacy of Lenalidomide [Revlimidä (R), CC-5013] in Combination with Doxorubicin and Dexamethasone (RAD) in Patients with Relapsed or Refractory Multiple Myeloma.
Authors: Gerecke C. , Knop S. , Topp M.S. , Kotkiewitz S. , Gollasch H. , Liebisch P. , Hess G. , Watters K. , Einsele H. , Bargou R.C.
Published: April 2020

Introdution:

Lenalidomide (Revlimid™) is Celgene’s lead clinical compound in a new group of drugs called IMiDs, which have immunomodularory properties. The drug has been evaluated in phase-I, II, and III clinical trials for the treatment of multiple myeloma (MM). Lenalidomide shows substantial anti-tumor activity in patients with refractory or relapsed MM and significantly prolongs time to tumor progression (TTP) compared to standard therapy in these patients. Lenalidomide was well tolerated in these trials, the only dose limiting toxicity in a phase-I trial was myelosuppression. In order to further improve therapeutic efficacy and to overcome drug resistance we are currently evaluating Lenalidomide (Revlimid™) in combination with doxorubicin and dexamethasone (RAD) for the treatment of patients with refracrory or relapsed MM in a phase-I/II trial.

Methods:

Patients with relapsed or refractory multiple myeloma recieve a fixed dose of either 10 mg or 15 mg Revlimid Revlimid™ given daily for 21 days (d 1–21) in combination with doxorubicin (adriamycin) and dexamethasone, to be repeated on day 29. Three dose levels of doxorubicin (adramycin) are planned: 4 mg/m2 day 1–4, 6 mg/m2 day 1–4 and 9 mg/m2 day 1–4. 40 mg dexamethasone is given orally day 1–4 and day 17–20 at a fixed dose. 3– 6 cycles are applicated unless severe toxicity or disease progression occurs.

Results:

RAD treatment was well tolerated at dose level 1 and dose level 2. Therefore, current dose escalation is continued. All patients treated at the first two dose levels (6/6) responded to RAD treatment. Further updated results on this trial will be presented.
Salvage Thalidomide Improves Survival of Relapsed Multiple Myeloma Patients When Compared to Other Salvage Chemotherapies: A Retrospective Analysis.
Authors: Bahlis Nizar J. , Savoie Lynn , Chaudhry Ahsan , Valentine Karen , Jenkins Deidre , Brown Christopher , Russell Jim , Stewart Douglas
Published: April 2020

Thalidomide has become one of the standard treatments of multiple myeloma patients either at diagnosis or in relapse and refractory disease. Up to date there are no randomized trials comparing the efficacy of thalidomide to other chemotherapy combinations in relapsed and refractory disease. We have conducted a retrospective analysis comparing the outcome of patients (n=141) younger than 65 years old who received either thalidomide (Group A; n=58) or non-thalidomide containing regimens including dexamethasone, cylophosphamide, melphalan+ prednisone, VAD (Group B; n=83) at first or second relapse post autologous stem cell transplant. The two groups were comparable in respect to age at relapse, Durie Salmon staging, IPS staging, serum beta-2 microglobulin, serum albumin and ECOG performance status. The median time to relapse from the date of initiation of salvage therapy in the thalidomide group was 12.1 months vs 6.5 months in the non-thalidomide group (p < 0.05). The median overall survival from the time of diagnosis was significantly longer in the thalidomide group (median survival = 131 months) vs the non-thalidomide group (median survival 50 months) (p =0.0055) (Figure below). In summary our retrospective analysis supports the superiority of thalidomide in the treatment of younger relapsed and refractory myeloma patients when compared to other previously established salvage chemotherapy regimen.

Figure Figure
The Histone Deacetylase Inhibitor Sodium Valproate Has In Vitro Anti-Neoplastic Activity Against Primary Multiple Myeloma Cells and Cell Lines Both Alone and in Combination with Velcade.
Authors: Basu Supratik , Khanim F. L. , Hayden R. E. , Barkhuinzen A. , MacWhannel A. , Jacob A. , Handa S. , Craddock C. , Drayson M. T.
Published: April 2020

There is growing evidence that histone deacetylase inhibitors (HDAC-I) may have benefit in the treatment of multiple myeloma (MM). We assessed the in vitro responses of the MM cell lines H929, JJN3, KMS11 and U266 to the HDAC-Is trichostatin A (TSA), suberoylanilide hydroxamic acid (SAHA) and sodium valproate (SV). All three HDAC-Is induced dose dependent cell killing and the ranked sensitivity of the lines was remarkably constant with U266 cells being most resistant to all three inhibitors. Real time RT-PCR based analyses of the HDAC expression profiles, identified that the greater resistence of U266 cells was associated with 5–10 fold increased expression of a single HDAC from the sirtuin family of deacetylases (SIRT5). Each HDAC-I displayed different potencies against the cell lines; TSA killed in the range 0.1–1μM, SAHA in the range 1–10μM and SV at the higher doses of 1–10mM. Despite being less potent, the doses of SV are commensurate with those that can be achieved in vivo. Furthermore, we have identified synergy/cooperation between suboptimal SV concentrations and the proteasome inhibitor Velcade (Bortezomib). MTT assays using JJN3 cells, demonstrated that 1mM SV had little effect reducing survival to 89% whereas 0.6 nM velcade reduced cell survival to 42±5.5%. Killing by velcade was not significantly enhanced by the additional presence of 1μM dexamethasone (Dex)(36±3.4%). However, the combination of SV, Dex and Velcade resulted in near total cell killing (4.7±2.6% survival). Collectively, these observations suggested that SV may have value in MM therapy. However, we wished to also investigate the potency of SV against primary MM cells. We have developed an in vitro culture system in which primary MM plasma cells survive for up to 30 days. In this culture system we have observed efficient and selective killing of MM cells by 1mM SV that has been comparable or indeed greater than that observed in response to 1μM SAHA. Thus it appears that primary MM cells are more SV sensitive than MM cell lines. Together our findings indicate that SV may have value in MM therapy and in particular when combined with proteasome inhibitors. Our data also indicate that SIRT5 overexpression may be a resistance factor against such therapies.
Arsenic Trioxide Targets the Interleukin-6 Cascade in Multiple Myeloma by Promoting Lysosomal Degradation of the Interleukin-6 Receptor Complex.
Authors: Cheung Wai Chung , Kwong Yok Lam
Published: April 2020

Introduction. Treatment of multiple myeloma (MM), a B-cell neoplasm characterized by clonal expansion of plasma cells in the bone marrow, remains unsuccessful in a significant proportion of patients, so that innovative strategies are needed. Arsenic trioxide (As2O3) has shown notable efficacy against MM in vitro and in clinical studies. Multiple cellular pathways in MM are targeted by As2O3. As cellular growth of MM cells is interleukin-6 (IL-6) dependent, we investigated if As2O3 also targeted the IL-6 cascade. Materials and methods. The IL-6-dependent MM cell line U266 was used as an in vitro model. Cell growth was measured by MTT assay, and apoptosis by flow cytometry. Protein phosphorylation was studied by Western blotting with specific antibodies. Expression of IL-6 receptor (IL-6R) was investigated by Western blotting and flow cytometry. Gene expression was detected by quantitative polymerase chain reaction (Q-PCR). Results. As2O3 showed a time and dose related inhibition of U266 cellular proliferation by induction of apoptosis. At clinically achievable concentrations (2 – 4 μmol/L), As2O3-induced apoptosis was associated with inhibition of constitutive tyrosine phosphorylation of JAK2 and STAT3, in a time and dose-dependent manner. Furthermore, pre-treatment of U266 cells with As2O3 prevented rescue of phosphorylation of JAK2 and STAT3 by exogenous IL-6, implying that the IL-6 cascade was targeted. Using Western blot analysis, we showed that As2O3 induced a time and dose-dependent down-regulation of both components of the IL-6R complex: IL-6R alpha subunit (IL-6Rα) and gp130 signal transducer. These results were confirmed by flow cytometry, showing that As2O3 treatment led to a down-regulation of surface expression of the IL-6Rα. Interestingly, Q-PCR did not reveal any change in the mRNA levels of the two genes with As2O3 treatment, suggesting that As2O3 downregulated IL-6R complex via a post-transcriptional mechanism. It is known that under physiological conditions, the IL-6R complex is internalized upon ligand binding and is targeted to lysosomes for degradation. Treatment of the U266 cell line with the lysosome inhibitor ammonium chloride totally abrogated As2O3-induced degradation of IL-6Rα and gp130. These results suggested that As2O3 might promote lysosomal degradation of IL-6Rα and gp130 by inducing a ligand-independent internalization of the receptor complex. Conclusion. Our results demonstrated that As2O3 suppressed IL-6-induced JAK/STAT3 signaling pathway in MM cells and this might be, at least partly, mediated by promoting ligand-independent internalization and lysosomal degradation of the IL-6R complex. These results have significant implications on the use of As2O3 in the treatment of patients with MM and other malignancies that are IL-6 dependent.
Exchange of International Staging System for Durie&Salmon Staging System in Therapeutic Strategy for Multiple Myeloma.
Authors: Kurahashi Shingo , Narimatsu Hiroto , Sugimoto Takumi , Sugiura Isamu
Published: April 2020

Introduction: Since multiple myeloma (MM) is not a curative disease and clinical outcome is variable, chemotherapy is started only when patients developed organ impairment or progression of disease. As for clinical staging, Durie&Salmon (DS) system is in use. The International Staging System (ISS) for MM has been recently reported to provide simple and useful prognostic grouping (Greipp et al. 2005). However, its usefulness in therapeutic strategy has not been clearly demonstrated.

Patients and methods: We reviewed medical records of patients with MM, newly diagnosed in Toyohashi Municipal Hospital between May 1997 and April 2004. They were all stratified based on both ISS and DS system.

Results: The median age of 55 patients was 67 years (range; 46–86). M protein isotypes included IgG (n=33), IgA (n=13), BJP (n=6) and IgD (n=1). Fifty-two patients were treated with chemotherapy and 12 of those patients underwent autologous peripheral blood stem cell transplantation. The median follow-up of the patients was 26.8 months (range; 1.4–77.5). Their staging and overall survival (OS) are summarized on the following table. ISS predicted OS more clearly than DS system in our study.

Overall survival based on ISS and DS system ISS DS stage no. of patients OS at 3 yrs no. of patients OS at 3 yrs I 14 1.00 3 0.67 II 22 0.55 20 0.79 III 19 0.25 32 0.40 p-value 0.0102 0.3287

Thirteen of the 14 patients with ISS stage I are alive at median months of 31.5 (10.4–73.8), and only one patient died of secondary malignancy at 44.9 months from diagnosis. The patients with ISS stage I included 86% of DS stage II and III patients, who are usually required treatment.

Conclusions and discussions: ISS could predict clinical outcome more clearly than DS system. The patients’ prognosis was good in ISS stage I although many patients with DS stage II and III was included in this group. We suggests that early treatment to the patients with ISS stage I might not be necessarily required. Further studies are needed to adopt ISS instead of DS system in therapeutic strategy.

Figure Figure
Epidemiology and Features of Multiple Myeloma in a Cohort of 110 Patients from a Central Area of Israel.
Authors: Duek Adrian , Haran Michal , Shvidel Lev , Shtalrid Mordechai , Sigler Erica , Berrebi Alain
Published: April 2020

In the last few years there has been a world wide increase in the incidence of multiple myeloma (MM). Epidemiologic studies have suggested that certain occupational and chemical exposures are likely to increase the risk for MM.

We performed a retrospective clinical and epidemiological study of MM on 110 cases diagnosed during 2000–2005 in our area, which is a rural and agricultural zone of approximately 400000 people. The incidence of MM in our population was 5.5 new cases /100000 people / year. 43% were females (n= 48) and 57 % males (n = 63). 22% of the patients were younger than 60. 58 patients were of European descent (52%), the vast majority of them from the former USSR (including people from near-Chernobyl); 40 from Asia (36%) and 16 patients (14%) from Africa. We separated the population into three categories: non exposed (physician, teacher, housekeeper, secretary, shop assistant, employee, scientific), suspected (painting, cleaning, microbiology, engineer, aviation, building-worker), and exposed: (petroleum, agriculture, plastic manufacture, textile, ammoniac, asphalt, Chernobyl). 65 % did not have any evidence of toxic exposure, in 23% we could document a significant exposure and in 12% there was a possible such exposure. 57% of the patients were in ISS stage I (b2m< 3.5 and albumin > 3.5), 33%- ISS stage II (b2m < 3.5 and albumin< 3.5) and 10% -ISS stage III (b2m > 3.5). Only 13% displayed lytic lesions at the time of survey. Fifty five patients requiring treatment received conventional therapy including APSCT and thalidomide for residual disease. In a cohort of 10 patients thal/dex was given as second line after achievement of a plateau with MP. This oral treatment allows a convenient ambulatory management and does not hamper stem cell harvesting made several months after discontinuation of MP. New clinical trials allowed us to include six patients for treatment with bortezomib.

In conclusion: the high incidence of MM in our area is in accordance with the world wide trend, possibly with a relationship with occupational and environmental risk factors. The heterogeneity of the population of Israel including arabs and new immigrants as well as additional data from others centers of Israel will help us to create a larger data base and to gain a better knowledge of MM in Israel.