Hevylite™ Assays Detect a Hidden Immunoparesis Associated with Adverse Biology in Myeloma Precursor Disease: A Prospective Clinical Study

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 5065-5065
Author(s):  
Rene Costello ◽  
Karina Espana ◽  
Neha Korde ◽  
Mary Kwok ◽  
Adriana Zingone ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Bone Marrow ◽  
Prospective Study ◽  
Heavy Chain ◽  
Light Chain ◽  
Conflicts Of Interest ◽  
Prospective Clinical Study ◽  
Biological Features ◽  
Increased Risk ◽  
M Spike

Abstract Abstract 5065 Recent studies have found immunoparesis (suppression of an uninvolved heavy-chain immunoglobulin (Ig) isotype; e.g. low quantitative IgA and/or IgM levels in a patient with an IgG M-spike) to be an independent risk factor for transformation from smoldering multiple myeloma (SMM) to multiple myeloma (MM). About 70–80% of SMM patients have evidence of immunoparesis. Among remaining 20–30% of SMM patients without immunoparesis, it is currently unknown whether there could be a hidden immunoparesis, reflected in suppression of the uninvolved light chain counter part of the same heavy-chain Ig isotype (e.g. IgG kappa in a patient with an IgG lambda M-spike), which in turn may be associated with adverse biology. A total of 50 SMM patients were enrolled in the first analysis of this prospective study (target: 124 SMM patients). At baseline, a bone marrow core biopsy/aspirate was conducted, and comprehensive immunohistochemistry and flow cytometry analyses were done to confirm the diagnosis and to generate risk profile data for individual patients. For each patient we obtained peripheral blood at baseline. Using serum, we performed the following clinical analyses: serum protein electrophoresis (SPEP); immunofixation electrophoresis (IFE); serum free light chains (sFLC); and quantitative Ig levels for IgG, IgA, and IgM. Using the Hevylite™ assay on a SPAplus (Specialty Protein Analyzer) platform, for each patient, we measured the heavy-light chain (HLC) protein pairs for the complete pair of the patient's heavy-chain Ig isotype (e.g. IgG kappa and IgG lambda for a patient with an IgG M-spike). Consistent with the literature; overall, we found 32/50 (64%) SMM patients to have immunoparesis of an uninvolved heavy-chain Ig isotype; 31 (97%) of these patients also had suppression of the uninvolved HLC counterpart. Among remaining SMM patients without immunoparesis of an uninvolved heavy-chain Ig isotype, using the Hevylite™ assay we found 8/18 (44%) to have a hidden immunoparesis of the uninvolved HLC counterpart. Compared to SMM patients without any type of Ig suppression, adverse biological features were profound in patients with a hidden immunoparesis of the uninvolved HLC counterpart. For example, the FLC-ratios were more skewed (mean: 14.2 vs. 2.5), the distribution of abnormal/normal plasma cells was more pronounced (90.1% vs. 80.3%), the concentration of the M-spike was higher (1.4 g/dL vs. 1.0 g/dL), and the plasma cell percentage in the bone marrow was higher (15.1% vs. 11.9%). In conclusion, using the Hevylite™ assay we identified, for the first time, among SMM patients without immunoparesis, a hidden immunoparesis of the uninvolved HLC counterpart, associated with adverse biological features. Our ongoing prospective study will evaluate whether immunoparesis of the uninvolved HLC counterpart is associated with an increased risk of progression to MM. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Uninvolved Immunoglobulin Suppression Determined By "Hevylite" ASSAY. Strongly Predicts Evolution of Smoldering Myeloma (SMM) to Symptomatic Multiple Myeloma (MM)

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 5584-5584
Author(s):  
Paraskevi Papaioannou ◽  
Annita Ioanna Gkioka ◽  
Kallirroi Tsalimalma ◽  
Aspasia Koudouna ◽  
Anastasia Kopsaftopoulou ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Plasma Cell ◽  
Light Chain ◽  
Conflicts Of Interest ◽  
Statistical Significance ◽  
Rapid Evolution ◽  
Great Majority ◽  
Bone Marrow Infiltration ◽  
Prognostic Information ◽  
Increased Risk

Abstract SMM may have an indolent course and remain stable for years; however, some patients evolve to MM requiring treatment within two years or less. To timely recognize these patients, three biomarkers (FLCR>100, ≥1 osteolysis and ≥60% plasma cell bone marrow infiltration) were recently established to discriminate asymptomatic MM patients at increased risk of rapid evolution and therefore, patients displaying the aforementioned biomarkers are considered symptomatic and receive immediate treatment. Despite the usage of the new IMWG definition criteria, there are still patients evolving rapidly and the notion of High risk SMM remains subjective among scientific groups. We herein studied wherever immunoglobulin (Ig) Heavy/Light Chain (HLC) measurements with the "Hevylite" assay that measures separately HLC-IgA, -G, -M-kappa or lambda, could add prognostic information on SMM evolution, with special attention to the significance of polyclonal Ig suppression. This method allows exact quantification of the amount of pure monoclonal fraction but also the degree of suppression of uninvolved polyclonal Igs. Patients and Methods: We studied 79 SMM patients of whom 30 were men and 49 women, with median age 65 (31-84) Ig type was IgG in 64 patients (81%) and IgA in the rest (19%). None of the patients had a free light chain ratio (FLCR) of more than 100, bone disease or more than 60% plasma cell bone marrow infiltration, in accordance to the last IMWG definition criteria. Patients were regularly followed (each 3 months) since SMM diagnosis and for a long period of time (median 98 months) during which 15 patients evolved to MM. "HevyLite" assay measurements were performed by nephelometry according to the manufacturer's instructions in frozen sera sample drawn at the time of diagnosis. We then calculated in all patients the ratio of involved/uninvolved Ig (HLCR); we also scored on a 5 points scale the eventuality of uninvolved Ig kappa or lambda below normal limits, that are, according to the manufacturer's testing on healthy individuals, 3608, 2023, 300, 312, 267, and 185 mg/L for IgG-kappa, IgG-lambda, IgA-kappa, IgA-lambda, IgM-kappa and IgM-lambda respectively. Results' relationship with SMM evolution were analyzed by standard methods using the SPSS v22.0. software and p values below 0,05 were considered statistically significant. Survival curved were drawn according to Kaplan-Mayer method. Results: Monoclonal Ig type was IgG-kappa, IgG-lambda, IgA-kappa, IgA-lambda in 61%, 19%, 11% and 9% of SMM patients respectively. Uninvolved IgG-kappa, IgG-lambda, IgA-kappa, IgA-lambda, IgM-kappa, IgM-lambda ranged from 625 to 14300 mg/L, 495 to 12700 mg/L , 214 to 2930 mg/L, 38 to 2580 mg/L, 19 to 1300 mg/L, 51 to 950 mg/L respectively. Median HLCR was 7 (range 0,24-707). Patients with HLCR above median tended to evolve faster than the others (p=0,1-not reaching statistical significance). Thirty-five patients had no one uninvolved Ig class depressed (score 0), 28 had 1, 9 had 2, 5 had 3 and 2 had 4; No score 5 was observed. Time to evolution strongly correlated to high score uninvolved Ig suppression (p< 0,00001). A simplified score separating 3 groups, the first with 0 Ig suppression, the 2nd with 1 or 2 and the 3rd with 3 or 4 proved to be equally potent in separating 3 risk-groups of SMM patients at risk of evolution to myeloma (p< 0,00001) and the great majority of the patients in the 3rd group evolved within two years (figure). In conclusion, the presence of decreased uninvolved Ig levels as determined by "Hevylite" constitutes a powerful prognostic marker of SMM evolution to MM. Figure. Figure. Disclosures No relevant conflicts of interest to declare.

The Natural History of Smoldering (Asymptomatic) Multiple Myeloma.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3396-3396 ◽  
Author(s):  
Robert Kyle ◽  
Ellen Remstein ◽  
Terry Therneau ◽  
Angela Dispenzieri ◽  
Paul Kurtin ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Bone Marrow ◽  
Plasma Cell ◽  
Light Chain ◽  
Plasma Cells ◽  
M Protein ◽  
Cell Content ◽  
Bone Marrow Plasma ◽  
M Spike

Abstract Smoldering multiple myeloma (SMM) is characterized by a serum M protein ≥ 3g/dL and/or 10% or more of plasma cells in the bone marrow. However, the definition is not standardized, and it is not known whether both serum M protein levels and bone marrow plasma cell counts are necessary for diagnosis or if one parameter is sufficient. We reviewed the medical records and bone marrows of all patients from Mayo Clinic seen within 30 days of recognition of an IgG or IgA M protein ≥ 3g/dL or a bone marrow containing ≥ 10% plasma cells from 1970 to 1995. This allows for a minimum potential follow-up of 10 years. Patients with end-organ damage at baseline from plasma cell proliferation, including active multiple myeloma (MM) and primary amyloidosis (AL) and those who had received chemotherapy were excluded. A differential of the bone marrow aspirate coupled with the bone marrow biopsy morphology and immunohistochemistry using antibodies directed against CD138, MUM-1 and Cyclin D1 were evaluated in every case in order to estimate the plasma cell content. In all, 301 patients fulfilled either of the criteria for SMM. Their median age was 64 years and only 3% were less than 40 years of age; 60% were male. The median hemoglobin value was 12.9 g/dL; 7% were less than 10 g/dL, but the anemia was unrelated to plasma cell proliferation. IgG accounted for 75%, IgA 22%, and biclonal proteins were found in 3%. The serum light-chain was κ in 67% and λ in 33%. The median serum M spike was 2.9 g/dL; 11% were at least 4.0 g/dL. Uninvolved serum immunoglobulins were reduced in 81%; only 1 immunoglobulin was reduced in 31% and both were decreased in 50%. The urine contained a monoclonal κ protein in 36% and λ in 18% and 46% were negative. The median size of the urine M spike was 0.04 g/24h; only 5 (3%) were &gt; 1 g/24h. The median bone marrow plasma cell content was 15 – 19%; 10% had less than 10% plasma cells, while 10% had at least 50% plasma cells in the bone marrow. Cyclin D-1 was expressed in 17%. Patients were categorized into 3 groups: Group 1, serum M protein ≥ 3g/dL and bone marrow containing ≥ 10% plasma cells (n= 113, 38%); Group 2, bone marrow plasma cells ≥ 10% but serum M protein &lt; 3g/dL (n= 158, 52%); Group 3, serum M protein ≥ 3g/dL but bone marrow plasma cells &lt; 10% (n= 30, 10%). During 2,204 cumulative years of follow-up 85% died (median follow-up of those still living 10.8 years), 155 (51%) developed MM, while 7 (2%) developed AL. The overall rate of progression at 10 years was 62%; median time to progression was 5.5 yrs. The median time to progression was 2.4, 9.2, and 19 years in groups 1, 2, and 3 respectively; correspondingly at 10 years, progression occurred in 76%, 59%, and 32% respectively. Significant risk factors for progression with univariate analysis were serum M spike ≥ 4g/dL (p &lt; 0.001), presence of IgA (p = 0.003), presence of urine light chain (p = 0.006), presence of λ urinary light chain (p = 0.002), bone marrow plasma cells ≥ 20% (p &lt; 0.001) and reduction of uninvolved immunoglobulins (p &lt; 0.001). The hemoglobin value, gender, serum albumin, and expression of cyclin D-1 were not of prognostic importance. On multivariate analysis, the percentage of bone marrow plasma cells was the only significant factor predicting progression to MM or AL.

Monitoring Minimum Residual Disease In Multiple Myeloma Patients By LC-MS/MS

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 3152-3152
Author(s):  
H. Robert Bergen ◽  
David L Murray ◽  
Diane F. Jelinek ◽  
Renee C. Tschumper ◽  
David R Barnidge ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Bone Marrow ◽  
Bone Marrow Biopsy ◽  
Light Chain ◽  
Plasma Cells ◽  
Residual Disease ◽  
Patent Application ◽  
Sds Page ◽  
Minimum Residual ◽  
M Spike

Abstract Proteomics with great sensitivity and specificity effectively analyzes proteins by prior tryptic digestion and subsequent analysis by LC-MS/MS of the tryptic digest. We have utilized this approach in the development of a LC-MS/MS method to characterize minimum residual disease in multiple myeloma. The abundant antibodies produced by multiple myeloma plasma cells are identical and appear as a spike (M-spike) upon protein electrophoresis. The M-spike and histological examination of the bone marrow constitute part of the myeloma diagnostic repertoire. Upon treatment, myeloma cells and the antibodies they produce are reduced in number and amount and become increasingly hard to detect. The bone marrow biopsy serves as the “gold standard” test for clinical remission. We have developed a sensitive test for the presence of the monoclonal antibody produced by the plasma cells which may serve as a substitute for invasive bone marrow biopsy. We have focused on tryptic peptides comprising the variable CDR regions of the Ig light chains that are unique to each patients antibody clone. Utilizing 2-5 µL of patient plasma/serum from the initial M-spike sample we have utilized SDS-PAGE to yield a crudely purified immunoglobulin light chain. The light chain band is isolated, reduced, alkylated and trypsin digested with subsequent LC-MS/MS analysis to identify CDR specific tryptic peptide(s). These can be identified as variable peaks (elution time and mass) in a base peak ion chromatogram where constant region peptides of either lambda or kappa light chains (clone dependent) serve as “internal standards” for identifying the CDR tryptic peptides. The peptide’s mass and its corresponding MS/MS spectra are unique to this CDR tryptic peptide and this patient’s clone. The unique diagnostic peptide is isolated in subsequent samples by immunoaffinity purification of the target kappa/lambda clone, SDS-PAGE separation and LC-MS/MS analysis of the light chain gel band. An extracted ion chromatogram is generated based upon the CDR peptides identified in the initial analysis of the M-spike sample. In patients in clinical remission the presence of a significant signal from the targeted peptides indicates that targeting a CDR peptide from the M-spike protein is more sensitive than currently available diagnostic tools including immunofixation. Comparison of this test to the gold standard bone marrow biopsy will be examined. Disclosures: Barnidge: Mayo Foundation: provisional patent application for technology, provisional patent application for technology Patents & Royalties.

Patient With IgG Heavy Chain Disease Presents With Parathyroid Hormone-Related Peptide Mediated Hypercalcemia

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 5344-5344
Author(s):  
Damien Hansra ◽  
Victoria Sujoy ◽  
Ikpatt Offiong ◽  
Chris Wunsch ◽  
James E. Hoffman
Keyword(s):  
Multiple Myeloma ◽  
Bone Marrow ◽  
Renal Failure ◽  
Renal Function ◽  
Bone Marrow Biopsy ◽  
Heavy Chain ◽  
Bone Destruction ◽  
Light Chains ◽  
M Spike ◽  
Heavy Chain Disease

Abstract Background Multiple myeloma is associated with excessive tumor-induced, osteoclast-mediated bone destruction. Hypercalcemia remains the most frequent metabolic complication of myeloma in patients, and excessive osteolysis plays a major contributory role in its pathogenesis. Hypercalcemia caused by increased blood levels of PTHrP have been found in patients with solid tumors and are uncommon in patients with hematologic malignancies including multiple myeloma. Here we present a unique case of a rare variant of multiple myeloma, IgG heavy chain disease, presenting with PTHrP associated hypercalcemia. Methods A retrospective chart review of one patient from 2011-2013 including clinical history, laboratory data, imaging, and pathology was performed. Results A 60 year old female with past medical history of stage IIA Hodgkin's lymphoma diagnosed in 1992, treated with total nodal radiation with recurrence in 1997 treated with 6 cycles of ABVD achieving complete remission. The patient was found to have hypercalcemia (11.0 mg/dL) and renal failure (1.29 mg/dL) in March 2011. Hypercalcemia workup revealed suppressed PTH (<3 pg/ml), elevated ionized calcium (6.1 mg/dL) elevated parathyroid related peptide (38pg/ml), normal vitamin D and ACE levels. Serum protein electrophoresis (SPEP) showed M-spike of 0.9 g/dL. Immunofixation electrophoresis (IFE) demonstrated an IgG monoclonal immunoglobulin without a corresponding light chain (figure 1). Free serum kappa and lamda light chains were within normal limits. Serum IgG was elevated (4678 mg/dL), normal IgA (127 mg/dL) and low IgM (39mg/dL). Also, Beta-2 microglobulin was elevated (14.3 mg/L). The patient was seen by hematology for monoclonal gammopathy and hypercalcemia and a solid tumor work up was recommended given that the most common the mechanism of hypercalcemia for multiple myeloma is osteoclast-driven and not PTHrP related. CT chest/abdomen/pelvis, bone scan, bone survey, mammography, pelvis ultrasound were negative. Bone marrow biopsy was performed November 2011 showing 10% plasma cells (PC) by CD138, non clonal by kappa/lambda. Patient was placed on zolendonic acid for hypercalcemia and her renal function and hypercalcemia continued to worsen over a period of months. A kidney biopsy was performed in April, 2012 and revealed acute and chronic tubulointerstitial nephritis with secondary glomerulosclerosis and mild interstitial fibrosis and tubular atrophy suggestive of sarcoidosis and the patient was placed on a course of prednisone with transient improvement in calcium and renal function. The patient presented in emergency room in April 2013 with altered mental status. She was found to have hypercalcemia (12.0 mg/dL), renal failure (1.55 mg/dL),). SPEP revealed M-spike 0.64 g/dl. IFE displayed a broad band of IgG heavy chain, without associated light chains and severe depression of the non-monoclonal IgG. Serum immunoglobulins demonstrated elevated IgG (2110 mg/dL), normal IgA (46 mg/dL) and decreased IgM (<21 mg/dL). Bone marrow biopsy showed 5% PCs (figure 2), non clonal by kappa/lambda- but exclusive for IgG by IHC, without any staining for IgA or IgM (figure 3). Cytogenetics were normal. Based on the constellation of findings and similarity to prior workup the patient was diagnosed with IgG heavy chain disease and therapy with cytoxan, dexamethasone, bortezomib was initiated. Calcium levels improved and she has recovered clinically; she is currently completing her second cycle of therapy. We plan to follow PTHrP levels along with routine paraprotein assessments. Conclusions PTHrP mediated hypercalcemia is rare in patients with multiple myeloma and the main mechanism of hypercalcemia in multiple myeloma is osteoclast-mediated bone destruction. This is the first reported case of IgG heavy chain disease presenting with PTHrP related hypercalcemia. Another critical point related to this case, is that kappa/lambda staining, the primary mechanism of determining plasma cell clonality, is not useful in exclusively heavy chain disease- and can, in such cases, obscure the diagnosis of this plasma cell malignancy. Disclosures: No relevant conflicts of interest to declare.

scholarly journals CRISPR-Mediated Loss of Immunoglobulin Heavy Chain in Multiple Myeloma Cell Line Results in Metabolic Pathway Alterations

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 1885-1885
Author(s):  
Denise K Walters ◽  
Renee C Tschumper ◽  
Dominique B Hoelzinger ◽  
Sophia J Quinton ◽  
Xiaojian Shi ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Cell Line ◽  
Cell Lines ◽  
Heavy Chain ◽  
Light Chain ◽  
Conflicts Of Interest ◽  
Dna Demethylation ◽  
Growth Patterns ◽  
Amino Acid Synthesis ◽  
Significant Difference

Abstract Aberrant over production of monoclonal immunoglobulin is a classic feature of the plasma cell (PC) malignancy multiple myeloma (MM). At diagnosis and more frequently upon relapse, some MM patients present with increased production of monoclonal free light chain (FLC) without a corresponding increase in intact immunoglobulin. This phenomenon is commonly referred to as light chain escape (LCE). As previously described (Walters et al. 2018, Experimental Hematology, Vol 57 p42-49), we have established a monoclonal IgA kappa MM cell line, designated as MC-B11/14, from the bone marrow (BM) aspirate of a 46 year old man diagnosed with MM. This cell line is non-hyperdiploid and possesses an 11;14 chromosomal translocation. Given our long standing interest in immunoglobulin function and secretion, we used CRISPR technology to knock out IgA heavy chain production in an effort to create a model of LCE. Successful knockout (KO) of IgA expression was demonstrated by immunofluorescence, flow cytometry, and western blot. As expected, secretion of intact IgA was undetectable in the MC-B11/14IgA- cells. Notably, no significant difference in morphology, phenotypic markers, or growth patterns was observed between the MC-B11/14WT and MC-B11/14IgA- cells. Examination of drug sensitivity between the MC-B11/14WT and MC-B11/14IgA- cells revealed that the response to pomalidomide treatment was similar between the MC-B11/14WT and MC-B11/14IgA- cells; however, the MC-B11/14IgA- cells were found to be more resistant to treatment with bortezomib. Given that increased bortezomib resistance was the only significant difference detected between the MC-B11/14WT and MC-B11/14IgA- cell lines in our initial study, we decided to take a metabolomics approach to examine whether loss of IgA heavy chain expression may alter cellular metabolism. Metabolomics or the profiling of small molecules is a powerful tool to characterize the metabolome of cells/biological systems and has proven to be useful in identifying alterations that occur in malignancy. In this regard, we employed pathway-specific targeted LC-MS/MS protocols in order to examine over 330 metabolites in 35 metabolic pathways in MC-B11/14WT and MC-B11/14IgA- methanolmetabolite extracts using an Agilent 1290 UPLC-6490 QQQ-MS liquid chromatography mass-spectrometer. Of the 330 metabolites examined, 28 were found to be significantly more abundant in the MC-B11/14IgA- cells compared to MC-B11/14 WT cells while 79 were found to be significantly less abundant. In general, the majority of significant metabolic differences between the two cell lines were found to involve metabolites from various amino acid synthesis pathways. More specifically, the MC-B11/14 WT cell line was found to possess significantly more citraconic acid, anthranilic acid, and homovanillic acid. By contrast, further analysis revealed that the MC-B11/14IgA- cell line was found to possess an 11.5 fold increase of the metabolite 2-hydroxyglutarate (2-HG). 2-HG has previously been shown to act as a competitive antagonist of α-ketoglutarate (α-KG), which results in the inhibition of α-KG-dependent dioxygenases including the JmjC domain-containing histone demethylases (KDMs) and the ten-eleven translocation (TET) family of DNA hydroxylases. Notably, these proteins are known to play a role in histone and DNA demethylation, respectively. Taken together, these data suggest that MM cells that have undergone LCE may also possess an accumulation of 2-HG which could lead to altered patterns of histone and DNA methylation. Given that MM patients who relapse with LCE typically have a poorer prognosis, further investigation of metabolomics in light-chain only expressing cell lines and patients who have relapsed with LCE is warranted. Disclosures No relevant conflicts of interest to declare.

scholarly journals Multiple Myeloma Cell Defects Erythropoiesis and Results Anemia Via High Level of CCL3 in Bone Marrow Microenvironment

Blood ◽  
2020 ◽  
Vol 136 (Supplement 1) ◽  
pp. 55-55
Author(s):  
Lanting Liu ◽  
Zhen Yu ◽  
Hui Cheng ◽  
Weiwei Sui ◽  
Shuhui Deng ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Bone Marrow ◽  
Mouse Model ◽  
Conflicts Of Interest ◽  
Erythroid Differentiation ◽  
Myeloma Cell ◽  
Cell Infiltration ◽  
Prognostic Impact ◽  
Multiple Myeloma Cell ◽  
Increased Risk

Background: Anemia is the most common complication of myeloma, and is associated with worse clinical outcomes including diminished quality of life, and increased risk of morbidity and mortality. Bone marrow (BM) is the main location of the growth and differentiation of HSPCs. Although marrow replacement by myeloma cell is widely considered a mechanistic rationale for myeloma-related anemia, however the molecular mechanism has not been fully understood. Methods: The clinical characteristics and the prognostic impact of myeloma-related anemia were investigated in patients with multiple myeloma. The erythroid differentiation defects of HSPCs were examined by flow cytometry and colony-formation assay. RNA-seq were conducted to clarify the different gene expression and the signaling pathway in HSPCs. Relative protein levels were determined by immunoblotting and relative gene levels were determined by quantitative real-time PCR. 5TGM1 murine myeloma mouse model was utilized. Results: The data of our large cohort of 1,363 myeloma patients demonstrated that 84.4% of patients had anemia (Hgb&lt;120 g/L), more than 50% of patients had moderate (Hgb=90-120 g/L) or severe anemia (Hgb=60-90 g/L) at the time of diagnosis. Anemia positively correlated with myeloma cell infiltration in the BM and worse outcome of patient. Patient sample and mouse model indicated that the quantity and the erythroid differentiation of HSPCs were affected by myeloma cell infiltration. The number of HPCs was significantly declined in the BM of myeloma, and negatively correlated with the quantity of tumor cells. The master regulator of erythropoiesis GATA1 and KLF1, was obviously down-regulated in myeloma HSPCs cells. However, the gene of chemokine CCL3 was significantly upregulated. Elevated CCL3 in the BM plasma of myeloma further inhibited erythropoiesis of HSPCs through activation of CCL3/CCR1/p38 signaling, and suppressed the expression of GATA1. CCR1 antagonist treatment effectively inhibits CCL3 activity and recovered the expression of GATA1 and rescued the erythropoiesis of HSPCs. Conclusions: myeloma cell infiltration causes the elevation of CCL3 in microenvironment that suppresses the erythropoiesis of HSPCs and results in anemia by downregulating the level of GATA1 of HSPCs. Thus, our study indicates targeting CCL3 would be a potential strategy against anemia and improvement the survival of myeloma patient. Disclosures No relevant conflicts of interest to declare.

Tissue Factor Is Frequently Expressed in Multiple Myeloma Cells.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 2132-2132 ◽  
Author(s):  
Sameer Gupta ◽  
Tanisha R Hayes ◽  
Yuchuan Liu ◽  
Matthew T Hurford ◽  
Charalambos Solomides ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Cell Line ◽  
Tissue Factor ◽  
Cell Lines ◽  
Light Chain ◽  
Conflicts Of Interest ◽  
Mrna Levels ◽  
Bone Marrow Biopsies ◽  
Increased Risk ◽  
Mcf 7

Abstract Abstract 2132 Poster Board II-109 Tissue factor (TF) is a 47 kDa transmembrane glycoprotein that initiates blood coagulation when complexed with factor VIIa. TF is constitutively expressed in a variety of tumor cells and has been shown to have a role in cellular signaling, angiogenesis and solid tumor progression. However, the role of TF in the hematologic malignancies is poorly defined. Multiple myeloma (MM) is associated with an increased risk of venous thromboembolic disease. However, whether increased TF expression contributes to the hypercoagulable state associated with MM remains controversial. In this study, we determined the expression of TF on archived bone marrow biopsies and plasmacytomas, and human MM cell lines. Immunohistochemical staining of TF was carried out on paraffin-embedded specimens from eighteen advanced stage MM patients. Staining for TF expression was scored as 0 (null), 1+ (weak), 2+ (moderate) and 3+ (strong). TF expression for the MM cell lines (U266B1, MM1.RL and MM1.S) was carried out by semi-quantitative real time RT-PCR. TF mRNA levels were normalized to 18S ribosomal mRNA levels. The TF: 18S ratios were then compared to that of a low TF expressing human breast cancer cell line cell line, MCF-7. Paraprotein distribution in the MM patient specimens was: IgG kappa (7), IgG lambda (5), IgA kappa (3), lambda light chain (2) kappa light chain (1). Overall, TF expression was observed in 10/18 (56%) of the patient specimens. Six specimens stained 1+, and two each stained 2+ and 3+. Staining was mainly cytoplasmic and did not correlate with the type of secreted paraprotein. TF expression (relative to MCF-7) was 17.3, 1.97 and 0.77 for the U266B1, MM1.RL and MM1.S cell lines, respectively. Results from these studies suggest that TF is frequently expressed in MM cells and might contribute to the hypercoagulability associated with this disease. In addition, TF may play a role in MM cell progression. Disclosures: No relevant conflicts of interest to declare.

Associations Of VEGF and VEGFR2 Polymorphisms With Increased Risk and Aggressiveness Of Multiple Myeloma

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 1886-1886
Author(s):  
Angelo Borsarelli Carvalho Brito ◽  
Leisa Lopes Aguiar ◽  
Gislaine Borba Oliveira ◽  
Gustavo Jacob Lourenco ◽  
Carmino Antonio De Souza ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Bone Marrow ◽  
Vascular Endothelial Growth Factor ◽  
Growth Factor ◽  
Clinical Features ◽  
Conflicts Of Interest ◽  
Vascular Endothelial ◽  
Healthy Humans ◽  
Increased Risk ◽  
Endothelial Growth Factor

Abstract Background Increased angiogenesis (AG) has been demonstrated in the bone marrow microenvironment in multiple myeloma (MM), suggesting its potential pathophysiologic role in the disease. The most important mediators of AG during tumor development are the vascular endothelial growth factor (VEGF) and the vascular endothelial growth factor receptor 2 (VEGFR2). Both factors are encoded by polymorphic genes and, therefore, their levels or functions are variable in healthy humans. It is already known that allele C of the VEGF 2578C/A (rs699947), allele G of the 1154G/A (rs1570360) and allele C of the 634 G/C (rs2010963) are related to higher concentration of serum VEGF compared to the remaining alleles. It is also established that the G allele of the VEGFR2 1192G/A (rs2305948) has higher binding efficiency and the allele C of the VEGFR2 604T/C (rs2071559) has lower transcription activity. Since the roles of these genetic polymorphisms in the risk and clinical manifestation of MM are still unknown, these were the aims of the present study. Material and methods Genomic DNA from peripheral blood of 192 consecutive MM patients and 202 age and race-matched controls was analyzed by real-time polymerase chain reaction for genotyping of the above mentioned polymorphisms. The differences between groups were analyzed by the logistic regression model. Power analysis (PA) was used to verify the effect of sample size on the results obtained in the study. Results The VEGF 2578CC genotype alone or combined with VEGF 634GG and VEGFR2 1192GG was higher in patients than in controls (47.4% versus 36.8%, P=0.01, PA: 88%; 44.3% versus 20.0%, P=0.001, PA: 99%; 71.3% versus 58.9%, P=0.01, PA: 81%; respectively). Carriers of these genotypes had a 1.89, 5.52 and 2.56 increased risks for MM than those with the remaining genotypes, respectively. Also, the VEGF 634GG genotype combined with VEGF 2578CC and VEGF 1154GG genotypes, with VEGF 2578CC and VEGFR2 1192GG genotypes and with VEGF 1154GG and VEGFR2 604TT genotypes were higher in patients than in controls (43.3% versus 32.5%, P=0.006, PA: 96%; 66.7% versus 32.0%, P=0.004, PA: 96%; 30.5% versus 12.5%, P=0.001, PA: 99%, respectively). Carriers of these genotypes had a 4.91, 10.97 and 14.10 increased risks for MM than those with the remaining genotypes, respectively. When only patients were analyzed, SNPs on the AG pathway were associated with clinical features. The VEGFR2 1192GG genotype, alone or combined with VEGF 2578CC and VEGF 1154GG, was related with lower counts of plasma cells in the bone marrow (80.4% versus 64.4%, P=0.03; PA: 52.5%; 80.5% versus 60.0%, P=0.02, PA: 44.4%; 85.1% versus 67.8%, P=0.01; PA: 42.8%, respectively). The frequency of the VEGFR2 604TT genotype was higher in patients with tumors of II and III Durie & Salmon stages than in those with tumors of stage I (29.9% versus 9.1%, P=0.02; PA: 75.5%). The VEGF 634GG genotype, alone or combined with VEGFR2 1192GG, was related to lower rates of renal failure (53.2% versus 35.3%, P=0.02; PA: 59.3%; 76.7% versus 53.8%; P=0.02, PA: 59.1%, respectively). Conclusions The data present, for the first time, preliminary evidence that inherited abnormalities of AG pathways, specifically SNPs on VEGF and VEGFR2, alone or combined, alter the risk for MM and clinical features of the disease. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Multiple Myeloma with Suspected Non-Secretory Type

2020 ◽  
Vol 27 (1) ◽  
Author(s):  
Annisa Ginar Indrarsi ◽  
Usi Sukorini
Keyword(s):  
Multiple Myeloma ◽  
Bone Marrow ◽  
Monoclonal Gammopathy ◽  
Plasma Cells ◽  
Immunoglobulin A ◽  
Protein Electrophoresis ◽  
Palpebral Conjunctiva ◽  
General Weakness ◽  
M Spike ◽  
G Ratio

Multiple Myeloma (MM) is a hematological malignancy characterized by clonal plasma cell in bone marrow that produceabnormal globulin, which resulted in monoclonal gammopathy. Multiple Myeloma Non-Secretory (MMNS) is a very rareform of multiple myeloma with monoclonal plasmocytic proliferation in bone marrow supported by clinical manifestationand radiological findings. However, plasma cells fail to secrete immunoglobulin. A 44-year-old female came to SardjitoGeneral Hospital with main complaints of weakness and back pain. General weakness and pale palpebral conjunctiva were6 observed (+/+), liver and spleen were not palpable. Blood test results were as follows: Hb 3.0 g/dL, RBC 1.07 x 10 / μL, WBC3 3 562 x 10 /μL, PLT 114 x 10 /μL, A/G ratio 1.07, BUN 51.5 mg/dL, creatinine 4.62 mg/dL, and calcium 3.1 mmol/L. Skeletalsurvey suggested a multiple osteolytic. Protein electrophoresis revealed hypogammaglobulinemia with no M-spike. Therewere 66% of plasma cells in bone marrow. Patient was diagnosed by MMNS. Diagnosis MMNS can be established if clonalplasmacytes is accompanied with renal insufficiency and hypercalcemia. However, monoclonal gammopathy was not foundin serum protein electrophoresis. A case reported of 44-year-old female diagnosed as MMNS with 'punched out' multipleosteolytic, increased plasma cells in bone marrow without evidence of paraprotein in circulation proved by low A/G ratio andnegative M-spike.

The Certainty of a Post-Bone Marrow Diagnosis: A Study of the Yield of Bone Marrow Biopsies in a Community Hospital Setting

Blood ◽  
2020 ◽  
Vol 136 (Supplement 1) ◽  
pp. 34-35
Author(s):  
Manasi M. Godbole ◽  
Peter A. Kouides
Keyword(s):  
Multiple Myeloma ◽  
Bone Marrow ◽  
Bone Marrow Biopsy ◽  
Fever Of Unknown Origin ◽  
Conflicts Of Interest ◽  
Diagnostic Yield ◽  
Hospital Setting ◽  
Unknown Origin ◽  
Nutritional Deficiencies ◽  
Bone Marrow Biopsies

Introduction: Most studies on the diagnostic yield of bone marrow biopsy including the one by Hot et al. have focused on the yield of bone marrow biopsies in diagnosing the source of fever of unknown origin. However, there have not been any studies performed to our knowledge looking at overall practice patterns and yield of bone marrow biopsies for diagnoses other than fever of unknown origin. We aim to determine the most common indications for performing bone marrow biopsies in a community-based teaching hospital as well as the yield of the biopsies in patients with specified and unspecified pre-test indications to estimate the rate of uncertain post-test diagnoses. Methods: We performed a retrospective data collection study at Rochester General Hospital, NY. A comprehensive search was conducted in our electronic medical data to identify all patients who underwent bone marrow biopsies over a 5 year period from January 2011 - December 2016 for indications other than fever of unknown origin. Patient data including demographics, pre-bone marrow biopsy diagnosis and post-bone marrow diagnosis was obtained. All patients above the age of 18 who underwent bone marrow biopsy for indications other than fever of unknown origin or follow up treatment of a hematological malignancy were included. Results: A total of 223 biopsies were performed. The median age was 59 years (age range- 23-95). One hundred and sixteen patients were male and 107 were female. The most common indications for performing bone marrow biopsy were evaluation of the following possible conditions: multiple myeloma (n=54), myelodysplastic syndrome [MDS] (n=47), lymphoma (n=28) and leukemia (n=18) as well as non-specific indications such as pancytopenia (n=40), anemia (n=22) and thrombocytopenia (n=11). The proportion of cases confirmed by bone marrow biopsy was 45/54 (83%) with the pre-marrow diagnosis of multiple myeloma, 34/47 cases (72%) with the pre-marrow diagnosis of MDS, 15/18 (83%) with the pre-marrow diagnosis of leukemia and 13/28 (46%) in those with the pre-marrow diagnosis of rule out lymphoma. Thirteen cases (18%) with possible MDS had post-bone marrow diagnoses of leukemia, anemia of chronic disease, myelofibrosis or medication-related changes. Five out of twenty two cases (23%) for anemia and 3/11 cases (27%) for thrombocytopenia without otherwise specified pre-bone marrow etiology had uncertain diagnosis after bone marrow biopsy. Conclusion: In about a fifth of patients necessitating a bone marrow, the diagnosis is discordant and can be surprising. It is also worth reporting that in these discordant results, non-hematological causes such as medications, anemia due to chronic diseases or conditions such as cirrhosis or splenomegaly from other etiologies were among the final diagnoses. Interestingly, 20% of the patients with unspecified pre-bone marrow diagnoses such as anemia or thrombocytopenia in our study had an unclear post-bone marrow diagnosis despite undergoing bone marrow biopsy. Our findings are a reminder that the bone marrow exam does not always lead to a definitive diagnosis and the need by exclusion to include in the differential non-hematological etiologies such as nutritional deficiencies, chronic kidney disease or autoimmune disorders. Disclosures No relevant conflicts of interest to declare.

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