Cytometric Response in Multiple Myeloma Cells to Low and Standard Doses of Bortezomib

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 5393-5393
Author(s):  
Nicole Beffermann ◽  
Mauricio Ocqueteau ◽  
Pablo Ramirez ◽  
Mauricio Galleguillos ◽  
Mauricio Sarmiento
Keyword(s):  
Multiple Myeloma ◽  
Flow Cytometry ◽  
Overall Survival ◽  
Plasma Cells ◽  
Conflicts Of Interest ◽  
Light Chains ◽  
Cell Transplant ◽  
Antineoplastic Drugs ◽  
Low Doses ◽  
Number Of Patients

Abstract Bortezomib is a very useful drug in the treatment of multiple myeloma (MM) patients. Used in combination with other antineoplastic drugs it has a well documented impact in progression free survival (PFS) and overall survival (OS) of any age patients, elegible or not for hematopoietic cell transplant. Standard dose (1,3 mg/m2) is used in almost all patients and low dose (0,7-0,8mg/m2) is reserved to patients with kidney disease and neuropathy. However, bortezomib doses used in phase 1 and 2 initial studies were described between 0,7 and 1,3 mg/m2 and were equally effective. The aim of this study was to evaluate the cytometric response of MM naive patients to two different bortezomib doses. We retrospectively analyzed the flow cytometry of fourty eight patients with naive MM treated with VCD scheme (Bortezomib-Cyclophosphamide-Dexametasone), without kidney failure nor neuropathy, of whom 21 received low doses of bortezomib (0.8mg/m2) and 27 standard doses (1.3 mg/m2). Flow cytometry was analyzed at diagnosis and at the end of treatment according to the expression of several clusters of differentiation (CD), establishing the presence of plasmoblastic myeloma clones (>95% of plasma cells with immunophenotype CD19 (-) CD56(+) CD45(-) CD38(+) and restriction of intra cytoplasmatic kappa and or lambda light chains) and normal mature plasma cells (CD19(+) CD56(-) CD45 (+) CD38 (++) with a policlonal expression of kappa/lambda light chains). Cytometric complete response was defined as normalization of the immunophenotype of plasma cells and absence of pathological cells. We found no statistical differences between the 2 groups in flow cytometric response (p>0.1), as shown in figure 1. This retrospective analysis suggests that lower doses of bortezomib could have similar effects in disease control, at least in cytometric response, to standard doses. Further studies should be made to evaluate clinical response and overall survival in patients treated with low doses compared to standard doses of bortezomib. In our country high costs of new generation antineoplastic drugs makes it necessary to find less expensive and equally effective schemes in order to make these well known beneficial treatments available to a greater number of patients. Disclosures No relevant conflicts of interest to declare.

Biclonal Multiple Myeloma with Monoclonal Free IgG3 Heavy Chain and kappa Free Light Chains.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4768-4768
Author(s):  
Alex G. Richter ◽  
Stephen Harding ◽  
Steve Rimmer ◽  
Guy Pratt ◽  
Aarnoud Huissoon ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Western Blot ◽  
Heavy Chain ◽  
Light Chain ◽  
Lymphoproliferative Disorder ◽  
Plasma Cells ◽  
Light Chains ◽  
Cell Transplant ◽  
Free Light Chains ◽  
Night Sweats

Abstract Background: Heavy chain disease (HCD) is a rare lymphoproliferative disorder characterized by a monoclonal heavy chain (HC) unattached to a light chain (LC). IgGHCD or γHCD typically presents as a lymphoproliferative disorder with lymphadenopathy and hepatosplenomegaly. Myeloma has been described associated with γHCD but only with a second intact Ig paraprotein. This report describes a unique presentation of multiple myeloma with monoclonal free γ3HC and kappa free light chains. Case: A 34 year old gentleman presented with mild persistent neutropenia following two episodes of pneumonia, 18 months previously. He admitted to persistent night sweats but no other significant history. Baseline investigations revealed a mild anaemia, neutropenia and a large IgG paraprotein with no associated light chain. Bone marrow aspirate and trephine confirmed myeloma. The patient was treated with cyclophosphamide, thalidomide and dexamethasone and has had a very good partial remission. He is awaiting a sibling allogeneic peripheral blood stem cell transplant. Investigations and results: Serum Electrophoresis confirmed a large IgG paraprotein (23g/l) with no associated light chain in the serum and identified as γ3 subclass by radial immunodiffusion. Western blot showed the γ3HC was truncated with a large deletion. Markedly elevated free kappa (κ) LC (503.58 mg/l [3.30–19.4]) were found in the serum with gross skewing of the kappa/lambda ratio. Urine electrophoresis revealed separate γHC and κ LC paraproteins. Western blot of the fractionated urine protein demonstrated different sized κLC aggregates. Flow cytometry of the marrow aspirate revealed an unusual staining pattern; CD5,19,38,45+ve and CD20,22,23,34,56,138 –ve plasma cells. Cytoplasmic staining revealed 2 distinct populations of plasma cells, the first producing γ3HC and the second only free κLC. Cytogenetics and FISH analysis for 14q, p53 and c-myc abnormalities were normal. Discussion: This is the first description of a Biclonal Myeloma with separate plasma cell populations producing γ3HC and κLC paraproteins. The biclonality confirms the free HC occurs as a result of abnormal synthesis not cleavage. The clinical and immunological findings are clearly different to typical findings in both γ3HCD and Myeloma. HCD has an appalling prognosis and this case is likely to have been ‘smouldering’ for 18 months, evidenced by the 2 pneumonias and persistent night sweats. There is no lymphadenopathy or organomegaly associated with γ3HCD. The immunophenotype of the malignant plasma cells is unique. Other atypical features include frank proteinuria, with a HC in the urine, but normal renal function and no radiological or biochemical evidence of bone involvement. We propose that this unique biclonal myeloma has distinct immunological and clinical features.

Combined Measurement of Plasma Cell Proliferation and Apoptosis in Multiple Myeloma.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 4884-4884
Author(s):  
Jiri Minarik ◽  
Vlastimil Scudla ◽  
Marta Ordeltova ◽  
Tomas Pika ◽  
Jaroslav Bacovsky ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Flow Cytometry ◽  
Cell Proliferation ◽  
Overall Survival ◽  
Plasma Cell ◽  
Poor Prognosis ◽  
Apoptotic Index ◽  
Number Of Patients ◽  
Proliferation And Apoptosis ◽  
Extreme Groups

Abstract Abstract 4884 Background Plasma cell proliferation and apoptosis represent key factors in multiple myeloma (MM) expansion and tumor survival. Both the proliferative and the apoptotic index have been shown to be strong and independent prognosticators in MM. Moreover, they do not correlate with most standard prognostic factors, representing thus different inherent features of the myeloma clone. The aim of the study was to assess the combined measurement of proliferation and apoptosis, giving the best predictive value for long and short MM survivals. Methods We assessed a cohort of 181 patients with newly diagnosed multiple myeloma, treated using conventional chemotherapy (regimens MP, VBMCP, VAD, CyVAD). In all the patients we measured proliferative and apoptotic index at the time of diagnosis, before the start of treatment. The proliferative activity of myeloma plasmocytes in bone marrow aspirate was measured by flow- cytometry using propidium-iodide index (PC-PI), representing the percentage of plasma cells in S-phase of the cell cycle; apoptosis was assessed using flow cytometry using an annexin-V (PC-AI) index. Subsequent statistical analysis of the the Kaplan-Meier curves of overall survival was evaluated using the MATLAB routine. We assessed different PC-PI and PC-AI tresholds with the best separation of groups with good and poor prognosis, using the log rank procedure. Additional measures of performance were obtained looking at the success with which two groups selected using the PC-AI and PC-PI thresholds reflected short term and long term survivors. Results The median follow-up of the group was 25 months (range 1 – 117 months). At the time of analysis, 137 patients had died (76%). Plasma cell proliferative index varied in the range 1.2 – 4.2, with median 2.5; apoptotic index was in the range 1.4 – 24.5 with median 4.3. Patients were divided into 4 groups based on PC-PI and PC-AI thresholds and then keeping only those two groups that demonstrated the worst and best overall survival based on the survival analysis of PC-PI and PC-AI individually. The best discriminating values for patients with poor prognosis (n=20) were PC-PI > 3.0% and PC-AI < 4.75%, and for patients with good prognosis (n = 71) PC-PI ' 3.0% and PC-AI ≥ 4.75%, with median overall survival 8 months versus 40 months respectively, p = 0.0002. The precision of the correct prediction of individual patient prognosis based on this grouping was in the patients with short survival 0.70 and in long survivors 0.58. Due to an imbalanced number of patients especially with high proliferation we were unable to create a single significant combined index across the entire patient group. Moreover, the patients outside the defined ranges substantially influenced the sensitivity and especially the specificity of the test, suggesting that defining of the extreme groups using PC-PI and PC-AI creates a better prognosticator than the assessment of the whole cohort based on a single combined index. Conclusion Plasma cell proliferation and apoptosis reflect different processes of MM kinetics with the growth characteristics on one hand, and survival on the other. Their assessment as single factors provides valuable information about the biology of the clone and contributes to the estimation of overall survival. Their combined measurement, however, significantly separates two extreme groups of patients with different prognosis and may thus become a valuable auxiliary parameter in MM patients stratification. Disclosures No relevant conflicts of interest to declare.

The Prognostic Significance of Polyclonal Bone Marrow Plasma Cells in Patients with Actively Relapsing Multiple Myeloma

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 1194-1194
Author(s):  
Toshi Ghosh ◽  
Wilson I Gonsalves ◽  
Dragan Jevremovic ◽  
S. Vincent Rajkumar ◽  
Michael M. Timm ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Bone Marrow ◽  
Flow Cytometry ◽  
High Risk ◽  
Research Funding ◽  
Plasma Cells ◽  
Labeling Index ◽  
Light Chains ◽  
Fish Cytogenetics ◽  
Kaplan Meier

Abstract Background: Prior studies suggest that the presence of >5% polyclonal plasma cells (pPCs) among total plasma cells (PCs) within the bone marrow (BM) is associated with a longer progression-free survival, higher response rates, and lower frequency of high-risk cytogenetic abnormalities in patients with newly diagnosed multiple myeloma (MM). However, the incidence and prognostic utility of this factor in patients with relapsed and/or refractory MM has not been previously evaluated. Thus, we evaluated the prognostic value of quantifying the percentage of pPCs among the total PCs in the BM of patients with actively relapsing MM. Methods: We evaluated all MM patients with actively relapsing disease (biochemical and/or symptomatic) seen at the Mayo Clinic, Rochester, from 2012 to 2013, who had BM samples evaluated by seven-color multiparametric flow cytometry. All patients had at least 24 months of follow-up from the date of flow evaluation. Cell surface antigens were assessed by direct immunofluorescence antibodies for CD45, CD19, CD38, CD138, cytoplasmic Kappa and Lambda Ig light chains, and DAPI nuclear stain. The flow cytometry data was collected using the Becton Dickinson FACSCanto II instruments that analyzed 150,000 events (cells); this data was then analyzed by multi-parameter analysis using the BD FACS DIVA Software. PCs were selectively analyzed through combinatorial gating using light scatter properties and CD38, CD138, CD19, and CD45. Clonal PCs were separated from pPCs based on the differential expression of CD45, CD19, DAPI (in non-diploid cases), and immunoglobulin light chains. The percentage of pPCs was calculated in total PCs detected. Survival analysis was performed by the Kaplan-Meier method and differences were assessed using the log rank test. Results: There were 180 consecutive patients with actively relapsing MM who had BM biopsies analyzed via flow cytometry as part of their routine clinical evaluation. The median age of this group was 65 years (range: 40 - 87); 52% were male. At the time of this analysis, 104 patients had died, and the 2-year overall survival (OS) rate for the cohort was 58%. The median number of therapies received was 4 (range: 1 - 15). Of these patients, 61% received a prior ASCT, and almost all (99%) received prior regimens containing either immunomodulators or proteasome inhibitors. There were 55 (30%) patients with >5% pPCs among the total PCs in their BM. The median percentage of pPCs among total PCs in these 55 patients was 33% (range: 5 - 99). The median OS for those with >5% pPCs was not reached compared with 22 months for those with <5% pPCs (P = 0.028; Figure 1). Patients with <5% pPCs PCs had a higher likelihood of high-risk FISH cytogenetics compared with the rest of the patients. In a univariate analysis, increasing number of pPCs was associated with an improved OS, while higher labeling index, number of prior therapies, and the presence of high-risk FISH cytogenetics were associated with a worse OS. In a multivariate analysis, only the increasing number of pPCs (P = 0.006), higher labeling index (P = 0.0002) and number of prior therapies (P = 0.003) retained statistical significance. Conclusion: Quantitative estimation of the percentage of pPCs among the total PCs in the BM of patients with actively relapsing MM was determined to be a predictor of worse OS. As such, this parameter is able to identify a group of patients with MM with actively relapsing disease who have a particularly poor outcome. Further studies evaluating its biological significance are warranted. Figure 1 Kaplan-Meier curve comparing OS between patients with ≥5% pPCs and <5% pPCs among the total PCs in their BM. Figure 1. Kaplan-Meier curve comparing OS between patients with ≥5% pPCs and <5% pPCs among the total PCs in their BM. Disclosures Kapoor: Celgene: Research Funding; Amgen: Research Funding; Takeda: Research Funding. Gertz:Prothena Therapeutics: Research Funding; Novartis: Research Funding; Alnylam Pharmaceuticals: Research Funding; Research to Practice: Honoraria, Speakers Bureau; Med Learning Group: Honoraria, Speakers Bureau; Celgene: Honoraria; NCI Frederick: Honoraria; Sandoz Inc: Honoraria; GSK: Honoraria; Ionis: Research Funding; Annexon Biosciences: Research Funding. Kumar:AbbVie: Research Funding; Noxxon Pharma: Consultancy, Research Funding; Celgene: Consultancy, Research Funding; Janssen: Consultancy, Research Funding; Array BioPharma: Consultancy, Research Funding; Sanofi: Consultancy, Research Funding; Onyx: Consultancy, Research Funding; Skyline: Honoraria, Membership on an entity's Board of Directors or advisory committees; Millennium: Consultancy, Research Funding; Kesios: Consultancy; Glycomimetics: Consultancy; BMS: Consultancy.

Multiparameter Flow Cytometry Remission Is the Most Relevant Prognostic Factor for Multiple Myeloma Patients Who Undergo Autologous Stem Cell Transplantation

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 737-737
Author(s):  
Bruno Paiva ◽  
Maria-Belén Vidriales ◽  
Jorge Cerveró ◽  
Gema Mateo ◽  
Jose J. Pérez ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Flow Cytometry ◽  
Stem Cell ◽  
Plasma Cells ◽  
Residual Disease ◽  
Progression Free Survival ◽  
Patient Specific ◽  
Multiparameter Flow Cytometry ◽  
Cell Transplant ◽  
Post Transplant

Abstract Minimal residual disease (MRD) assessment is standard in many hematologic malignancies but is considered investigational in multiple myeloma (MM). We report a prospective analysis of the prognostic importance of MRD detection by multiparameter flow cytometry (MFC) in 295 newly diagnosed MM patients uniformly treated in the GEM2000 protocol (VBMCP/VBAD induction plus autologous stem cell transplant [ASCT]). MRD status by MFC was determined at day 100 post-ASCT. Persistent myelomatous plasma cells (MM-PCs) were detected by MFC in 170 patients (58%), who were considered MRD-positive. Progression-free survival (PFS; median 71 vs 37 months, P < .0001) and overall survival (OS; median not reached vs 89 months, P = .002) were longer in patients who were MRD-negative versus MRD-positive at day 100 post-ASCT, with a 5-year PFS rate of 60% and 22% (P < .0001), respectively. Similar prognostic differentiation was seen in 147 patients who achieved immunofixation (IFx) negative complete response post-ASCT. The 5-year PFS rate was 62% in MRD-negative patients (n=94) versus 30% in MRD-positive patients (n=53; P < .0001), and the respective 5-year OS rates were 87% versus 59% (P = .009). Moreover, MRD− IFx− and MRD− IFx+ patients had significantly longer PFS than MRD+ IFx− patients (median 71, 65, and 37 months, respectively, P = .0002). By multivariate analysis, only MRD status by MFC at day 100 post-ASCT and FISH cytogenetics were identified as independent prognostic factors for PFS, and only MRD status by MFC and age were identified for OS. The relative risks of progression and death among MRD-positive versus MRD-negative patients were 3.64 (P = .002) and 2.02 (P = .02), respectively. Finally, a subgroup of 157 patients in which MRD information was available both pre- and post-ASCT were analyzed. Patients who were MRD-positive both pre- and post-transplant (n=93) had the worst prognosis; patients who were MRD-positive pre-ASCT but improved to MRD-negative post-ASCT (n=48) had an intermediate prognosis, and patients who were MRD-negative both pre- and post-transplant (n=16) had the best prognosis. The 5-year PFS and OS rates in these three prognostic subgroups were 25%, 57%, and 80%, respectively (P = .0001), and 59%, 78%, and 100%, respectively (P = .06). In summary, our results show that MRD evaluation by MFC is a very useful technique to identify patients at different risk of progression. This type of analysis, particularly when performed post-ASCT, may contribute to the design of patient-specific maintenance treatment approaches, as well as the evaluation of the potential benefits of consolidation therapies.

Multiple Myeloma with Secretory and Non-Secretory Biclonal Gammopathy

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 5092-5092
Author(s):  
Namita Vinayek ◽  
Goetz H Kloecker ◽  
Beth C. Riley
Keyword(s):  
Multiple Myeloma ◽  
Plasma Cells ◽  
Conflicts Of Interest ◽  
Bone Biopsy ◽  
M Protein ◽  
Light Chains ◽  
Complete Replacement ◽  
Biclonal Gammopathy ◽  
Kappa Light Chains ◽  
M Spike

Abstract Abstract 5092 Multiple myeloma (MM) accounts for 10 % of hematological malignancies. MM with biclonal gammopathy are rare and seen in 1 % of all MM cases. In 99% of the MM cases, paraproteins are secreted in the serum and/or urine, in the remaining 1% the paraproteins are synthesized but not secreted. Only 2 % of patients are younger than 40 years of age. We report a 36 year old African American female who presented with diffuse lytic lesions. Serum electrophoresis (SPEP) revealed IgG kappa M protein of 3.8 g/dl and a IgG Kappa M spike of 2.6 mg/dl. Free light chains (SFLC) revealed elevated free kappa chains of 500 mg/dl, elevated free lambda chains of 928 mg/dl and K/L ratio of 0.55. Urine protein UPEP showed an M spike of IgG kappa and free kappa light chains. The B2 microglobulin was 26.8 mg/dl. Patient's recent T10 Bone biopsy done at outside hospital had a near complete replacement of the marrow by plasma cells which were biclonal plasma cells by IHC, one kappa restricted and one lambda restricted. Since the IFE showed only a single IgG kappa M protein spike, the lambda plasma clone was apparently non-secretory. Treatment was started with cyclophosphamide, bortezomib and dexamethasone. After 22 days of treatment, the M protein has decreased 1.39 mg/dl. A repeat SFLC also shows further decline in free kappa light chains. As the IFE showed one gamma M spike and bone biopsy had two clonal plasma cells it was concluded that one of the neoplasms is nonsecretory. Although the elevated lambda free light chain in the serum is now does point towards a second plasma cell neoplasm. Biclonal gammopathy is rare and accounts for 1% of all MM cases. To our knowledge, this is the first reported case of a biclonal, secretory and nonsecretory, gammopathy Disclosures: No relevant conflicts of interest to declare.

The Chronic Kidney Disease-Epidemiology Collaboration-Cystatin C (CKD-EPI-CysC) Equation Has an Independent Prognostic Value for Overall Survival in Newly-Diagnosed Patients with Symptomatic Multiple Myeloma: Comparison with 3 Other Equations for GFR Estimation

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 1849-1849
Author(s):  
Meletios A Dimopoulos ◽  
Efstathios Kastritis ◽  
Eirini Katodritou ◽  
Anastasia Pouli ◽  
Eurydiki Michalis ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Overall Survival ◽  
Cystatin C ◽  
Prognostic Value ◽  
Conflicts Of Interest ◽  
Univariate Analysis ◽  
P Value ◽  
Newly Diagnosed ◽  
Number Of Patients ◽  
Ckd Stage

Abstract Abstract 1849 Renal impairment (RI) is a frequent complication in multiple myeloma (MM). The estimation of glomerular filtration rate (GFR) is based on equations that use serum creatinine (sCr) as a marker of RI (i.e. MDRD or CKD-EPI). The IMWG has recommended the use of the MDRD formula for the estimation of GFR in MM patients with stabilized sCr, while the classification of RI is based on the 5 stages of the KDIGO classification (Dimopoulos et al, JCO 2010;28:4976–84). However, the equations based on sCr are imprecise and thus novel markers of renal injury have been used in patients with renal damage, including cystatin-C (CysC). CysC is considered as a more sensitive marker of GFR than sCr. Recently, the CKD-EPI investigators have reported that a combined sCr-CysC (CKD-EPI-sCr-CysC) equation correlated better with GFR than equations based on either of these markers alone (CKD-EPI or CKD-EPI-CysC; Inker et al, NEJM 2012;367:20–9). Although cysC has been reported by our group to be elevated in MM patients, the CKD-EPI equations and their value on MM patients' survival have never been evaluated. Therefore, we studied 220 newly-diagnosed, previously untreated, symptomatic MM patients. The median age was 69 years (range: 36–94 years) and 16% had sCr ≥2 mg/dl. Serum CysC was measured on the BN ProSpec analyser using a latex particle-enhanced nephelometric immunoassay (Dade Behring-Siemens Healthcare Diagnostics, Liederbach, Germany). Serum CysC was increased in MM patients compared to 52 age- and gender-matched controls [median: 1.07 mg/l vs. 0.72 mg/l, p<0.0001]. The median values for eGFR calculated by the MDRD, CKD-EPI, CKD-EPI-CysC and CKD-EPI-sCr-CysC equations were 63.45 ml/min/1.73m2, 68.13 ml/min/1.73m2, 68.11 ml/min/1.73 m2, and 64.87 ml/min/1.73 m2, respectively (p<0.01). Patients were divided in the 5 CKD stages of KDIGO classification, according to eGFR (stage 1: eGFR >90 ml/min/1.73 m2; stage 2: 60–89 ml/min/1.73m2; stage 3: 30–59 ml/min/1.73 m2; stage 4: 15–29 ml/min/1.73 m2; stage 5: <15 ml/min/1.73 m2 or on dialysis). For each studied equation, the number of patients with RI stage 3–5 (i.e. eGFR <60 ml/min/1.732) was 39.5% for MDRD vs. 42.2% for CKD-EPI vs. 43.1% for CKD-EPI-CysC vs. 45% for CKD-EPI-sCr-CysC (p<0.01; see also the table). Concordance for CKD stage allocation for the 4 equations of estimating eGFR was 97% for MDRD vs. CKD-EPI, 60% for MDRD vs. CKD-EPI-CysC and 84% for MDRD vs. CKD-EPI-sCr-CysC. A significant correlation was found between ISS stage and all studied equations (p<0.01 for all). The median overall survival (OS) for all patients was 52 months. In the univariate analysis per CKD stage, the 4 equations could predict for OS (the higher CKD stage had poorer survival) with the following significance: MDRD (p=0.057), CKD-EPI (p=0.01), CKD-EPI-CysC (p<0.0001), and CKD-EPI-sCr-CysC (p=0.006). When we tested the 4 equations as continuous variables, all had prognostic value for OS but the CKD-EPI-CysC had the strongest prognostic value (p<0.0001 and Wald=24.0 vs. p<0.0001 and Wald=19.7 for CKD-EPI-sCr-CysC, p=0.003 and Wald=8.9 for CKD-EPI and p=0.005 and Wald=7.7 for MDRD). In the multivariate analysis, that included ISS stage, LDH ≥300 U/l and eGFR for each different equation (as a continuous variable) only eGFR that included CysC but not sCr (CKD-EPI-CysC and not CKD-EPI-sCr-CysC) had independent significance (p=0.013) along with high LDH (p=0.029). Our data suggest that the CKD-EPI-sCr-CysC equation for the estimation of GFR detects more MM patients with stage 3–5 RI than the MDRD, CKD-EPI or CKD-EPI-CysC equations. However, CKD-EPI-CysC was the only equation that could predict for OS, possibly due to the very strong correlation of CysC with ISS (as myeloma cells produce CysC also). The confirmation of these data will lead to the broader use of equations based on CysC (CKD-EPI-CysC with or without sCr) for the evaluation of RI in patients with MM, as it has been suggested for patients with several other renal disorders. Table. Evaluation of Renal Function Stage by Different Equations CKD stage MDRD equation CKD-EPI equation CKD-EPI-CysC equation CKD-EPI-sCr-CysC equation p-value 1 60 (27%) 57 (26%) 67 (30%) 44 (20%) 2 73 (33%) 70 (32%) 58 (26%) 77 (35% Friedman-test 3 53 (24%) 55 (25%) 57 (26%) 62 (28%) p<0.01 4 21 (9.5%) 25 (11%) 27 (12%) 24 (11%) 5 13 (6%) 13 (6%) 11 (5%) 13 (6%) Disclosures: No relevant conflicts of interest to declare.

Relative Stability Of Plasma Cells Immunophenotype In Multiple Myeloma Over Time

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 1885-1885
Author(s):  
Benoit Tessoulin ◽  
Steven Legouill ◽  
Marion Eveillard ◽  
Nelly Robillard ◽  
Soraya Wuilleme ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Overall Survival ◽  
Plasma Cells ◽  
Conflicts Of Interest ◽  
Survival Rates ◽  
Prospective Trial ◽  
Multiparameter Flow Cytometry ◽  
Single Centre ◽  
Over Time ◽  
Overall Survival Rates

Abstract Emerging evidence indicates that multiple myeloma (MM) can follow a number of evolutionary pathways over a patient’s disease course. Recently, genomic data have shown the coexistence in many patients of several tumour types, which could be either genetically stable, evolving linearly or be constituted of heterogeneous clonal mixtures with predominant clones shifting over time (Keats, Blood 2012;120:1067-1076). Little is known however of immunophenotypic evolution of the myeloma cells (MM cells) during MM progression. In a retrospective single-centre analysis of tumoral immunophenotypes, we examined sequential bone marrow or peripheral blood samples from 51 MM patients. Multiparameter flow cytometry was performed to assess MM phenotype using CD38 and CD138 expression as well as intracytoplasmic light chain usage restriction in most cases. The presence or absence of CD19, CD20, CD27, CD28, CD56 or CD117 on the MM cells’surface was investigated additionally. A median of 4 of the latter antigens was examined per sample. Most MM cells were homogeneous in the expression or absence of expression of the differentiation antigens investigated. Immunophenotype changes were defined as the loss or gain of at least 1 antigen. Whenever partial expression of one or several antigens was observed on MM cells, indicating MM subsets, the presence of subclones was suspected. Samples were obtained at diagnosis and relapse for 33 MM patients, and during consecutive relapses for 18. The total amount of analysed samples was 109, with a median of 2 samples per patient (range: 2-3). A modification of immunophenotype occurred in 22 patients (43%), with one antigen change (68%), two antigens changes (18%) or three antigens changes (14%). Immunophenotypic changes are summarized in table 1. For 20% of the cases or more, this involved CD28 (n=11, balanced gain or loss) or CD27 (n=7, mostly loss). CD56 expression, scarcely negative on the first assessment, was gained at relapse in three cases. Other antigens were expressed in a stable fashion over time. Eighteen (35%) of the patients had at least a subclone on the first immunophenotype, which was lost and/or modified in 14 / 18 patients at time of subsequent relapse. There was no significant impact of immunophenotypic modifications on overall survival rates Similarly, the presence of a subclone at first examination, or the immunophenotypic change of this subclone over time was not associated with differences in overall survival rates. In conclusion, this retrospective single centre study demonstrates that the immunophenotype of MM cells is mostly stable during the course of the disease. When changes occur, they do not seem to affect the clinical course of the patients. These preliminary results need confirmation on a larger and prospective trial. Disclosures: No relevant conflicts of interest to declare.

Prognostic Value Of Quantifying Circulating Plasma Cells By Multiparametric Flow Cytometry In Patients With Relapsed Multiple Myeloma

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 754-754
Author(s):  
Wilson I Gonsalves ◽  
Vinay Gupta ◽  
S. Vincent Rajkumar ◽  
William G Morice ◽  
Michael M Timm ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Flow Cytometry ◽  
Peripheral Blood ◽  
Research Funding ◽  
Prognostic Value ◽  
Plasma Cells ◽  
Light Chains ◽  
Plateau Phase ◽  
Multiparametric Flow Cytometry ◽  
Relapsed Disease

Abstract Background The presence of circulating plasma cells (PCs) in multiple myeloma (MM) is a known poor prognostic marker. Prior studies have utilized a technically challenging slide-based immunofluorescence technique to detect and quantify the presence of circulating PCs which was not widely adapted in clinical practice. More recently, the routine use of flow cytometry has provided the opportunity to quantitatively assess circulating PCs in MM patients with relative ease. We report the prognostic value of quantifying circulating clonal PCs using multiparametric flow cytometry in MM patients with relapsed disease. Methods We evaluated all MM patients seen at the Mayo Clinic, Rochester from 2009 to 2011 with previous or ongoing relapsed disease and who had their peripheral blood samples evaluated by flow cytometry. Each blood sample had its peripheral blood mononuclear cells isolated by ficoll gradient and stained with antibodies to CD45, CD19, CD38, CD138 and cytoplasmic Kappa and Lambda Ig light chains. A six-color multi-parameter flow cytometer (Becton Dickinson FacsCantos II) was used to examine each sample with a target of detecting 150,000 events (cells) that was then analyzed using the Facs Diva Software. Plasma cells were selectively analyzed through combinatorial gating using light scatter properties and CD38, CD138, CD19, and CD45. Normal PC's were then separated from clonal plasma cells based on the differential expression of CD45, CD19 and polytypic Ig light chains. The clonal plasma cells detected were reported out as the number of clonal events/150,000 collected total events. For those samples where less than 150,000 events were gated or examined, the number of final clonal events was adjusted to 150,000 events. Survival analysis was performed by the Kaplan-Meier method and differences in survival assessed using the log rank test. Results There were 647 consecutive patients with a history of treated MM who had a peripheral blood flow cytometry as part of their routine clinical evaluation. The median patient age was 62 years (29-88) and 55% were male. The median time since diagnosis was 12 months (range: 1-363) and the median number of lines of treatment received was 2 (1-11). There were 81 (13%) patients with clonal circulating PCs with a median of 368 cells (4 – 133,464). The 2-year overall survival (OS) for the 81 (13%) patients with any circulating PCs was 17% compared with 65% for those with none (P<0.001). The presence of circulating clonal PCs was associated with high-risk disease by FISH (P <0.001) as well as higher PCLI (P <0.001). We then correlated the presence of circulating clonal PCs with the disease status at flow cytometry assessment. Among the study patients, only 145 (22%) had actively relapsing disease with the remaining 502 (78%) in a plateau including patients in CR. Circulating clonal PCs were more likely to be detected in patients with actively relapsing disease compared with plateau phase (43% vs. 4%, P < 0.001); none of the CR patients and <5% of the remaining plateau phase patients had any detectable circulating clonal PCs. We then restricted the analysis to the actively relapsing patients; the best cutoff predicting 1-year mortality by ROC analysis was 100 events. Based on this, we defined >=100 events as a cutoff for defining the prognostic role of circulating clonal PCs in actively relapsing patients. The median OS for those with >=100 clonal PC events was 12 months compared with not reached for those with <100 events (p<0.001; Figure 1 ). Among patients with actively relapsing disease, >= 100 circulating PCs was associated with a higher ISS stage, plasma cell labeling index and bone marrow PC% compared with those with <100 circulating PCs. In a multivariable model, only LDH > 222 (HR: 2.08, P = 0.045) and >= 100 circulating PCs (HR: 2.48, P = 0.048) were found to adversely affect OS in actively relapsing patients. Conclusion The utilization of flow cytometry to quantify circulating clonal PCs in patients with relapsed MM appears to have significant prognostic relevance. In patients with actively relapsing disease, >= 100 circulating PCs predicted for worse OS with a median of 12 months. Future studies are needed to determine if this would allow an opportunity to develop a more risk adapted approach for patients with relapsed disease. Disclosures: Kumar: Celgene: Consultancy, Research Funding; Millennium: Consultancy, Research Funding; Onyx: Consultancy, Research Funding.

Cerebrospinal Fluid (CSF) Involvement By Plasma Cell Dyscrasias – Single Center Experience

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 5686-5686
Author(s):  
Taiga Nishihori ◽  
Jun Zhou ◽  
Kenneth H. Shain ◽  
Rachid Baz ◽  
Melissa Alsina ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Overall Survival ◽  
Cerebrospinal Fluid ◽  
Plasma Cell ◽  
Plasma Cells ◽  
Cell Transplant ◽  
Cns Involvement ◽  
Color Flow ◽  
Tumor Load ◽  
Plasma Cell Dyscrasias

Abstract Introduction: Central nervous system (CNS) involvement by plasma cell dyscrasias (PCD) is uncommon but poses significant clinical challenges and has a dismal prognosis. Lumbar puncture (LP) is typically performed only for patients with neurologic signs or symptoms and data on patients with CNS involvement are rather scarce. Here, we report a retrospective single institution review of clinicopathological features and treatment outcomes in the setting of cerebrospinal fluid (CSF) involvement by PCD. Methods: We identified consecutive patients with plasma cell disorders who had abnormal cytology or flow cytometry results in the CSF in the Department of Hematopathology database at Moffitt Cancer Center from 1997 to 2014. Cytology slides [Wright-Giemsa (WG) and Papanicolaou (Pap) stained preparations] and the corresponding flow cytometry were reviewed to confirm the diagnosis. Four-color flow cytometry was performed using antibodies against CD38, CD138, CD56, CD117, CD19, and cytoplasmic kappa and lambda light chains, withadditional markers added when necessary. Clinical variables were abstracted from the patient medical records. Overall survival was estimated from the time CSF involvement was identified using the Kaplan-Meier method. Results: Sixty-seven Pap-stained cytology smeas/cytospins and WG stained cytospins from 65 patients who underwent LP for clinical suspicion of CSF involvement were reviewed. Flow cytometry was preformed on 48 cases positive for atypical plasma cells by cytology. Sixteen of 67 (23.9%) were suspicious or diagnostic for PCD (median age of 58 years (range 44 – 75), 56% were male). However, only 4 of 16 cases (25%) were diagnosed as PCD by cytology without additional flow cytometry study. Median tumor load of PCD by flow cytometry was 81% (range 4 - 99%). PCD included 14 patients (88%) with multiple myeloma [MM; 1 patient progressed to secondary plasma cell leukemia (PCL)], 1 with primary PCL, and 1 with Waldenström macroglobulinemia (Neel-Bing syndrome). Of the 14 MM patients, 57% had high-risk disease by cytogenetics/FISH, and immunophenotypes were IgA (50%), IgG (29%), and light chain (21%). All MM patients had Durie-Salmon stage 3 disease. Median number of prior therapies was 2 (1-4), and 44% received stem cell transplant prior to CSF involvement. Median time from diagnosis to CSF involvement was 23 (range, 6 – 78) months. Presenting symptoms included diplopia/vision loss (31%), headache (25%), and leg weakness (cauda equina/cord compression) (19%). Two patients presented with gross orbital involvement and new/enlarging scalp lesions. On radiologic imaging, 5 (31%) had leptomeningeal, 4 (25%) had epi- or extra-dural, and 2 (13%) had dural enhancement/lesions. None had CSF involvement at the time of initial PCD diagnosis. Treatment included intrathecal chemotherapy (methotrexate, cytarabine or triple regimen; 86%), radiation therapy (including whole brain, craniospinal, radiosurgery or involved field; 63%) and systemic chemotherapy (65%). One patient did not receive treatment due to poor performance status. For 5 patients who had repeat CSF analyses, only one had no evidence of disease on cytology but flow cytometry remained positive. Six-month overall survival rate was 20.2% (95% CI 4.4 – 43.5). At the time of data review, only 2 patients were alive. Conclusions: CSF involvement by PCD carries extremely limited prognosis and represents advanced stage of the disease. Patients may be treated with systemic therapy as well as CNS-directed therapy, though the outcomes are dismal. Careful assessment of patients’ neurologic symptoms and low threshold for performing LP is required for early detection of CSF involvement. Application of flow cytometry appears to be a useful tool in the diagnosis of CSF involvement by PCD; improving sensitivity and specificity over cytology alone, particularly when the tumor load is low or cytologically equivocal for atypical plasma cells. Further research is needed to improve the outcomes of these patients. Disclosures No relevant conflicts of interest to declare.

Detection of Circulating Plasma Cells in Multiple Myeloma with Extramedullary Plasmacytoma

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 5328-5328 ◽  
Author(s):  
Jing Wang ◽  
Shuang Geng ◽  
Yuping Zhong ◽  
Mingyi Chen ◽  
Wenming Wang ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Bone Marrow ◽  
Flow Cytometry ◽  
Peripheral Blood ◽  
Disease Surveillance ◽  
Plasma Cells ◽  
Conflicts Of Interest ◽  
Residual Disease ◽  
Bone Marrow Aspiration ◽  
Extramedullary Plasmacytoma

Abstract Objective: To detect the circulating plasma cells (cPCs) in patients of multiple myeloma (MM) with or withoutextramedullary plasmacytoma (EMP). Methods: The 21 patients of MM samples were collected from April 2014 to April 2015. There were 12 males, 9 females, with a median age of 60 (49 to 76 years old). Peripheral blood and bone marrow were examined before treatment or after EMP. Multi parameter flow cytometry (MFC) was used to analyze abnormal plasma cells (tumor cells) in samples of bone marrow and CD138 MACS positive sorting peripheral blood. The antibodies used in the flow cytometry were CD38-APC, CD138-PE, CD81-PE-Cy7, CD45-PacBlue, CD19-Percp-Cy5.5, CD56-mCherry-PE-ef610, CD117-AmCyan, CD16, Zombie-APC-Cy7. Results: In these 21 patients, he ratio of sex is 1.33:1, the median age is 60 (49-76). The immunoglobin type is as follows: IgG κ 7 cases, IgG λ 5 cases, IgG 2 cases, IgA κ 3 cases, IgA λ 2 cases, λ light chain 1 cases. The morphology of bone marrow aspiration showed more than 15% plasma cells and abnormal plasma cells can be seen in bone marrow in cytometry. 9 of 21 patients diagnosed MM with EMP, 2 of them find EMP when initial diagnosis and 7 of them find EMP in the course of disease (6 months to 8 years). The sites of the EMP included head, jaw, chest wall, side of the rib, the soft tissue of the sacral region and the vertebral body and all patients had bone involvement. In 17 patients with complete clinical data, bone marrow and peripheral blood specimens, the cPCs negative rate was 87.5%(7/8) in EMP negative patients, while the cPCs positive rate was 66.7% (6/9) in EMP positive patients, the difference among groups was statistically significant (chi square values: 5.13, p = 0.024). Conclusion: MFC has been widely used in diagnosis and minimal residual disease surveillance of MM, here we established the detection method of cPCs in MM patients, and it is valuable for clinical diagnosis and prognosis judgment. Disclosures No relevant conflicts of interest to declare.

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