Factor In Multiple Myeloma Research Articles

Epha3 acts as proangiogenic factor in multiple myeloma.

This study investigates the role of ephrin receptor A3 (EphA3) in the angiogenesis of Multiple Myeloma (MM) and the effects of a selective target of EphA3 by a specific monoclonal antibody on primary bone marrow endothelial cells (ECs) of MM patients.EphA3 mRNA and protein were evaluated in ECs of MM patients (MMECs), in ECs of patients with monoclonal gammopathies of undetermined significance (MGECs) and in ECs of healthy subjects (control ECs). The effects of EphA3 targeting by mRNA silencing (siRNA) or by the anti EphA3 antibody on the angiogenesis were evaluated. We found that EphA3 is highly expressed in MMECs compared to the other EC types. Loss of function of EphA3 by siRNA significantly inhibited the ability of MMECs to adhere to fibronectin, to migrate and to form tube like structures in vitro, without affecting cell proliferation or viability. In addition, gene expression profiling showed that knockdown of EphA3 down modulated some molecules that regulate adhesion, migration and invasion processes. Interestingly, EphA3 targeting by an anti EphA3 antibody reduced all the MMEC angiogenesis-related functions in vitro. In conclusion, our findings suggest that EphA3 plays an important role in MM angiogenesis.

The Evolution of Prognostic Factors in Multiple Myeloma.

Multiple myeloma (MM) is a heterogeneous hematologic malignancy involving the proliferation of plasma cells derived by different genetic events contributing to the development, progression, and prognosis of this disease. Despite improvement in treatment strategies of MM over the last decade, the disease remains incurable. All efforts are currently focused on understanding the prognostic markers of the disease hoping to incorporate the new therapeutic modalities to convert the disease into curable one. We present this comprehensive review to summarize the current standard prognostic markers used in MM along with novel techniques that are still in development and highlight their implications in current clinical practice.

Prognostic Impact of Molecular Response Assessed By Next-Generation Sequencing in a Large Cohort of Multiple Myeloma Patients

INTRODUCTIONMinimal residual disease (MRD) assessment is an essential prognosis factor in multiple myeloma (MM). In this way different high-sensitivity quantification methods are being developed and improved using molecular or flow-cytometry approaches. Currently, the only method available to study molecular response by NGS in MM is clonoSEQ; this is offered as external service by Adaptative Technologies. We have optimized a simplified method to quantify molecular response using NGS. In this work, we compare results when quantifying molecular response by our simplified NGS method or by clonoSEQ. Also we study the prognostic impact of molecular response assessed by NGS techniques in a large series of 181 cases.METHODSWe evaluated the molecular response in 71 patients included in GEM2010MAS65 clinical trial by our simplified method, and in 110 patients included in GEM2005 clinical trial by clonoSEQ. Immunoglobulin clonotype quantification by our simplified method: DNA was amplified in accordance with the terms of BIOMED-2 Concerted Action BMH4-CT98-3936. Standard kits were used to prepare libraries (Life Technologies or New England Biolabs), and sequencing was performed with Ion Torrent™ PGM, Ion S5, or MiSeq systems. The resulting FASTQ were analyzed with specific bioinformatic applications in order to identify and quantify clonal specific sequences (clonotype) present in every proband.RESULTSMRD negativity showed significant longer Progression Free Survival (PFS), regardless of the method employed (median 34 months for local method and 32 months for clonoSEQ method in MRD+ patients (MRD>10-5) vs median not reached and 81 months in MRD- patients, respectively, p=0.0001). Likewise, significant differences were observed in terms of Overall Survival (OS) between patients with MRD+ (median 81 months for local method vs 50 months for clonoSEQ method) and MRD- patients (median not reached for both methods), p=0.014. No significant differences were observed in OS and PFS between both local and clonoSEQ NGS techniques (Figure 1).Then, we performed a global analysis with all patients to assess the potential of molecular response evaluated by any NGS method to predict survival regardless of treatment, Molecular response by SEQ was achieved in 43 cases out of 181 (23.7%). Median PFS were 34 vs 80 months for MRD+ and MRD- patients, respectively (p<0.0001, HR=2.8). Median OS were not reached in MRD- patients vs 81 months in MRD+ patients (p=0.004, HR=2.78) (Figure 2).Regression analysis did not show association between any biological variable and MRD- achievement by NGS. When analyzing MRD- patients, no significant differences in PFS or OS were detected between high or standard-risk cytogenetics cases, comparing to differences observed in overall series or in the MRD+ patients group.CONCLUSIONMolecular response assessed by NGS is able to identify patients with higher risk, independently of the treatment administered and the method employed. MRD negativity achievement by NGS is an independent risk factor to other clinical and biological variables. Our simplified NGS method offers molecular response information with prognostic value similar to standardized molecular techniques. [Display omitted] [Display omitted] DisclosuresPaiva:Celgene: Honoraria, Research Funding; Janssen: Honoraria; Takeda: Honoraria, Research Funding; Sanofi: Consultancy, Research Funding; EngMab: Research Funding; Amgen: Honoraria; Binding Site: Research Funding. Martínez-López:Novartis: Honoraria, Speakers Bureau.

Abstract 3085: The BET family bromodomain inhibitor ABBV-075 targets multiple pathogenesis factors in multiple myeloma and exhibits robust in vivo efficacies as a single agent and in combination with bortezomib

Abstract Small molecule inhibitors of the bromodomain and extraterminal domain (BET) proteins represent a potential new class of cancer therapeutic agents. ABBV-075 is a potent and selective BET family bromodomain inhibitor that was recently advanced to Phase 1 studies. Here we report the in vitro and in vivo characterization of ABBV-075 in preclinical models of multiple myeloma. ABBV-075 exhibited significant anti-proliferative activities and triggered robust apoptosis across many multiple myeloma cell lines, including cell lines that were engineered to become resistant to bortezomib. As reported for other BET inhibitors, ABBV-075 potently inhibited super-enhancer regulated potential cancer drivers such as Myc, IRF8 etc. In addition to directly impacting multiple myeloma cell growth and survival, ABBV-075 also diminished stroma cell derived IL-6 secretion and prevented HUVEC cell proliferation in vitro. Given the importance of angiogenesis and the survival factor IL-6 for multiple myeloma pathogenesis, ABBV-075 could potentially produce significant therapeutic benefits in myeloma by simultaneously targeting multiple critical pathogenesis mechanisms for this disease. ABBV-075 caused significant tumor growth delay in the OPM2 flank tumor model as a monotherapy. Combining ABBV-075 with Bortezomib provided further therapeutic benefit compared with the treatment of either of the two agents individually. Taken together, our results suggest that ABBV-075 could be a promising option to explore for the treatment of multiple myeloma. Citation Format: Tamar Uziel, Xiaoyu Lin, Emily Faivre, Aparna Sarthy, Daniel H. Albert, Leiming Li, Denise Wilcox, Xiaoli Huang, Terry Magoc, Lloyd Lam, Steven W. Elmore, Keith McDaniel, Warren Kati, Yu Shen. The BET family bromodomain inhibitor ABBV-075 targets multiple pathogenesis factors in multiple myeloma and exhibits robust in vivo efficacies as a single agent and in combination with bortezomib. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3085.

Abstract 1762: Body mass index shows etiologic heterogeneity for risk of diffuse large B-cell lymphoma (DLBCL) subtype defined by cell-of-origin

Abstract Background. DLBCL is the most common non-Hodgkin lymphoma (NHL) subtype in western countries and is known to be clinically heterogeneous. Gene expression profiling has identified two major, biologically distinctive DLBCL subtypes on the basis of their cell-of-origin (COO): the germinal center B-cell (GCB) - characterized by BCL2 rearrangement and C-REL amplification, and activated B-cell (ABC) - characterized by constitutive activation of the NF-kB pathway. Recently, the InterLymph Subtypes Project identified medical history, lifestyle, and family history risk factors for DLBCL using a large pooled analysis of case-control studies. In this study we evaluated these same risk factors for etiologic heterogeneity as defined by DLBCL COO. Methods. For this analysis we used a clinic-based study of newly diagnosed NHL cases and frequency matched controls, enrolled from 2002-2012, with 474 DLBCL cases and 2203 controls. COO was determined clinically using the Hans algorithm, which is based on immunohistochemistry markers using formalin-fixed, paraffin-embedded tumor tissue. Amongst DLBCL cases 107 were ABC, 207 were GCB, and 160 were missing subtype (tissue unavailable). Risk factor data, including body mass index (BMI), smoking, alcohol use, any allergy, asthma, blood transfusion, diabetes, rheumatoid arthritis, sun exposure, lived on a farm and family history of hematologic malignancy, were collected from self-administered questionnaires. Polytomous logistic regression was used to estimate odds ratios (ORs) and 95% confidence intervals for ABC and GCB subtypes, adjusted for age and sex. Results. The mean age of cases was 60.4 years with 53% male; the mean age of controls was 61.6 years and 53% male. We identified a strong positive association for BMI with risk of ABC but not GBC DLBCL (p-heterogeneity 0.025). Compared to BMI 18.5-24.9 kg/m2, those with a BMI of 25.0-29.9 (OR = 1.65; 0.93-2.93) or 30+ (OR = 2.61; 1.48-4.62) were at increased risk of ABC DLBCL. There was no association for a BMI of 25.0-29.9 (OR = 0.91; 0.63-1.31) or 30+ (OR = 1.21; 0.83-1.76) with GCB DLBCL. There was no evidence of significant heterogeneity between ABC and GCB DLBCL for the other risk factors that we evaluated. Further adjustment of the BMI association for educational level, smoking, and alcohol only slightly attenuated these findings. Conclusions. We observed a strong positive association of BMI with ABC but not GCB DLBCL. In contrast, other evaluated risk factors showed no heterogeneity by COO. While BMI has not been clearly associated with NHL overall, there has been evidence for an association with DLBCL. The etiologic heterogeneity we observed for ABC DLBCL is notable as BMI is also a strong risk factor for multiple myeloma, another post-germinal center B-cell malignancy. While requiring replication, our findings suggest BMI may influence B-cell neoplasia through effect on post-germinal center pathways. Citation Format: Matthew K. Breitenstein, Megan M. O’Byrne, Andrew L. Feldman, Carrie A. Thompson, William R. Macon, Grzegorz S. Nowakowski, Stephen M. Ansell, Susan L. Slager, Thomas M. Habermann, James R. Cerhan. Body mass index shows etiologic heterogeneity for risk of diffuse large B-cell lymphoma (DLBCL) subtype defined by cell-of-origin. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1762.

Potential crosstalk of the interleukin-6-heme oxygenase-1-dependent mechanism involved in resistance to lenalidomide in multiple myeloma cells.

Interleukin (IL)-6 is one of the most important survival factors in multiple myeloma (MM), and determines the poor prognosis of MM. IL-6 mainly has a paracrine bone marrow stromal cell origin and an autocrine MM cell origin. As an enzyme having cytoprotective effects, heme oxygenase-1 (HO-1) promotes the growth and drug resistance of various malignant tumors. HO-1 expression levels in bone marrow CD138(+) cells of MM patients were significantly higher than those in healthy donors, and positively correlated with both serum IL-6 and intracellular IL-6 mRNA expression levels. Culture of U266, RPMI8226 and CD138(+) cells with exogenous IL-6 in vitro induced high HO-1 expression levels and allowed them to resist lenalidomide. It is hypothesized that this was probably attributable to IL-6-mediated activation of the Janus kinase 2-signal transducer and activator of transcription 3 pathway. In contrast, without IL-6 coculture, enhanced HO-1 expression in U266, RPMI8226 and bone marrow CD138(+) cells from MM patients significantly inreased IL-6 mRNA expression levels and facilitated autocrine IL-6 production. The findings were associated with high HO-1 expression-enhanced p38 mitogen-activated protein kinase phosphorylation. Reduced HO-1 expression sensitized MM cells to lenalidomide. Therefore, we postulated that IL-6 in the bone marrow microenvironment of MM patients stimulated high HO-1 expression in MM cells and their resistance to lenalidomide. High HO-1 expression also stimulated autocrine IL-6 production, and exacerbated drug resistance and disease. This study supports the use of HO-1 as a possible marker for both MM prognosis and drug resistance, and as a potential therapeutic target.

High Expression of AF1q Is an Adverse Prognostic Factor in Multiple Myeloma Patients Treated with Autologous Stem Cell Transplantation

Introduction: Multiple myeloma is a heterogeneous hematological malignancy. Investigation on the poor prognostic factors contributes the development of the effective stratified treatment.AF1q is an oncogene which expressed in leukemia cells, located in 1q21. The gene is well known as a one of the partner of 11q23. Recently, the association between the over expression of AF1q and the tumor progression has been reported by several investigators.We studied the expression of AF1q in patients with multiple myeloma and analyzed the prognostic value.Method: Newly diagnosed multiple myeloma patients in National Center for Global Health and Medicine hospital from January 2001 to March 2013 were studied. Patients were treated with vincristine-adriamycin-dexamethason or bortezomib-dexamethason induction therapy followed by autologous stem cell transplantation using high dose melphalan.The expression of AF1q was evaluated using the immunostain of bone marrow clot samples at the diagnosis of multiple myeloma. The expression of AF1q was graded from "-" to "+++". The clinical response, progression free survival, and overall survival were analyzed.Results: Clinical data and bone marrow clot samples of 61 patients were analyzed. Mean age was 55.5 years old, and 52.5% was male. The grades of AF1q expression were 2 of "-", 18 of "+", 17 of "++", and 24 of "+++". We defined the cases with "-" and "+" as low expression, "++" and "+++" as high expression. Very good partial response or better was obtained after completion of autologous stem cell transplantation in 45% of patients with low AF1q expression and 48.8% of high expression, there was no statistically significant difference.Survival analysis using Log Rank method showed that high expression of AF1q was associated with significantly shorter progression free survival (p=0.004), but not overall survival.Conclusion: In this study, we found that the high expression of AF1q was an adverse prognostic factor in multiple myeloma. [Display omitted] DisclosuresHagiwara:Celgene Corporation: Membership on an entity's Board of Directors or advisory committees.

De Novo Nethyltransferases, DNMT3a and DNMT3b Are Underexpressed in Multiple Myeloma

Background and aims: Presently, there is growing evidence that along with the important role of genetic abnormalities, epigenetic aberrations are relevant factors in multiple myeloma (MM). As was recently found, genome-wide analysis of DNA methylation reveals epigenetic alterations in plasma cells from patients with MM and individuals with monoclonal gammopathy of undetermined significance (MGUS). MGUS is characterized by predominant hypomethylation. Transformation into MM is accompanied by progressive hypermethylation with maximum methylation seen in relapsed disease. DNA methyltransferases (DNMTs) catalyze DNA methylation through transfer of methyl group to cytosine of the CpG dinucleotides, resulting in 5-methylcytostine. DNMT1 maintains patterns of methylated cytosine residues in human genome. DNMT3A and DNMT3B are de novo DNA methyltransferases, whose role is to maintain new methylation pattern that forms due to formation of the cancer.Methods: 30 bone-marrow aspirates from individuals with MGUS or MM patients before the treatment initiation were used. The cDNA was synthesized using 100 ng of total RNA in a 20 µl reaction volume (Roche, Diagnostics, Basel, Switzerland). Quantification of DNMT1, DNMT3a and DNMT3b levels by TaqMan® probes (Life Technologies, Grand Island, NY) with Xceed qPCR Master Mix (IAB, BioTech-Europe, Czech Republic) was performed. For normalization, the GAPDH was used.Results: Although MM is characterized by widespread alterations in DNA methylation, we observed that DNMT3a and DNMT3b de novo methyltransferases were underexpressed in both, MGUS individuals and MM patients when compared to DNMT1 expression level (Figure 1). The transcribed genes have increased levels of 5-hydroxymethylcytosine, then the DNMTs activities might compensate for active hydroxymethylation - demethylation.Conclusions: Our results confirm that the expression of de novo DNA methyltransferases is deregulated in MM cell lines. The presented analysis is first of its kind that was performed on human myeloma cell lines, especially with the focus on the residual expression of Dnmt3a.With support of the grant NT14393. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Pre-Transplant Beta2-Microglobulin Is an Independent Biomarker of Acute Graft-Versus-Host-Disease after Allogeneic Hematopoietic Cell Transplant in a Single Institutional Study

Introduction: Refinement of strategies capable to stratify the risk of acute Graft-versus-Host Disease (aGVHD) before HCT could facilitate critical clinical decisions in in the course of HCT, and improve studies about preventive and therapeutic interventions. The HCT comorbidity index (HCT-CI) is a relatively simple index that, accordign to recent data, is also capable to stratify the risk of aGVHD. More recently, it has been shown that the addition of laboratory biomarkers such as ferritin, albumin, C-reactive protein and platelet count, measured in the pre-transplant period, can further refine the accuracy of this index. β2-microglobulin (β2-m) is a component of the MHC class I complex that lies beside the α3 chain of this molecule and lacks a transmembrane region. The best characterized function of β2-m is the stabilization of the tertiary structure of the MHC class I. However, the protein also seems to be involved in iron metabolism and proliferation of cancer cells. β2-m is widely used as a prognostic factor in multiple myeloma. Interestingly, it also represents a biomarker of frailty in the elderly (Annweiler et al, 2011). In our Institution, β2-m is systematically measured before HCT in all patients since 2010. Based on the role of this protein in immune regulation, and on its association with frailty and other important comorbidities, we hypothesized that pre-transplant β2-m levels could be a biomarker of aGVHD.Methods: This is a retrospective single-center observational diagnostic accuracy study. All consecutive patients (pts) submitted to allogeneic HCT in our Institution (n=103) between January 2010 and February 2014 were included in the analysis. Included pts underwent one single HCT using all types of conditioning, and unrelated or related HLA-compatible donors. Baseline levels of β2-m were systematically collected approximately 30 days before HSCT (along with several other laboratory tests evaluated in the pre-HCT visit), in the outpatient setting. β2-m was measured in the same clinical laboratory and by the same methodology (immunonephelometry) during the study period. Clinical and laboratory data were obtained from the medical records. Univariate and multivariate Cox regression were applied for searching independent variables influencing the risk for aGVHD. The Kaplan-Meier method and log-rank test were used for the calculation of overall survival (OS). aGVHD and TRM incidence were calculated considering early death and relapse as a competitive event (for acute GvHD), and relapse (for TRM), using the Gray's test to compare the curves.Results: β2-m levels were available for 97 pts with a median age of 44 years (16-68). High and low-dose conditioning were used in 59 and 38 pts respectively. Overall survival at 12 and 60 months were 60% (95%CI: 50-70%) and 44% (95%CI: 30-58%), respectively. TRM at 12 months was 20% (95%CI: 12-28%). Twenty-six (34%) pts presented aGVHD at a median of 62 days (20-140), of which 16 presented grade II aGVHD and 10 pts grade III-IV aGVHD. The cumulative incidence of aGVHD was 29% (95%CI: 19-29%) in 100 days. Higher levels of β2-m were observed in pts who developed aGVHD when compared to patients without this complication (p = 0.008). β2-m was also higher in older pts, in patients with more advanced disease, and submitted to low dose conditioning. In the multivariate Cox regression model for aGVHD, β2-m remained significant in different models incorporating variables such as age, conditioning dose, early x advanced disease, HCT-CI, diagnosis of lymphoproliferative diseases and ferritin, and adjusted for age and for the presence of lymphoproliferative disease (table 1). No association was observed between β2-m with TRM and OS.Conclusion: the strong association between pre-transplant β2-m and the occurrence of aGVHD suggests that this protein could represent an additional biomarker of aGVHD. Larger multicenter studies are required to validate our results, and define whether the incorporation of β2-m levels into aGVHD risk stratification models could add prognostic value, and to understand the biological mechanisms underlying this association.Table 1Multivariate analysis of the risk of aGVHDVariableHRp-valueCI 95%Age-Adjustedβ2-m3.720.031.12-12.3Lymphoproliferative disease-adjustedβ2-m4.440.031.12-17.5No adjustmentβ2-m4.970.011.40-17.6HCT-CI ≥22.790.041.05-7.42 DisclosuresNo relevant conflicts of interest to declare.

Potential role of FLT3-ligand in the angiogenic process of multiple myeloma

The aim of the study was to evaluate serum levels of FLT3-ligand (FLT3-L), a soluble molecule in bone marrow (BM), participating actively in hematopoiesis, in relation with angiogenic factors in multiple myeloma (MM) patients. We measured, in 70 patients with active MM and in 38 of them who responded to conventional therapy, serum levels of FLT3-L, along with known angiogenic factors, such as VEGF, endoglin, TNF-alpha and HGF (with ELISA) and BM microvascular density (MVD), estimating the immunohistochemical expression of CD31. All pre-treatment values were higher in active MM patients compared to controls (p<0.001 for all cases), in parallel with both International Staging System and Durie–Salmon stages (p<0.001 for all cases). Moreover, levels of FLT3-L correlated positively with all soluble angiogenic factors, as well with MVD (p<0.0001 for all cases). Post-treatment values of FLT3-L decreased significantly in responders to therapy (p<0.001). The underlying relation of MM angiogenesis with FLT3-L may result from the fact that BM microvasculature is a major source of FLT3-L, both in BM niche and probably in peripheral blood. Our results suggest that serum levels of FLT3-L may be used as angiogenic marker in MM patients.

High expression of AF1q is an adverse prognostic factor in multiple myeloma patients treated with autologous stem cell transplantation

Abstract Introduction: Multiple myeloma is a heterogeneous hematological malignancy. Investigation on the poor prognostic factors contributes the development of the effective stratified treatment. AF1q is an oncogene which expressed in leukemia cells, located in 1q21. The gene is well known as a one of the partner of 11q23. Recently, the association between the over expression of AF1q and the tumor progression has been reported by several investigators. We studied the expression of AF1q in patients with multiple myeloma and analyzed the prognostic value. Method: Newly diagnosed multiple myeloma patients in National Center for Global Health and Medicine hospital from January 2001 to March 2013 were studied. Patients were treated with vincristine-adriamycin-dexamethason or bortezomib-dexamethason induction therapy followed by autologous stem cell transplantation using high dose melphalan. The expression of AF1q was evaluated using the immunostain of bone marrow clot samples at the diagnosis of multiple myeloma. The expression of AF1q was graded from - to +++. The clinical response, progression free survival, and overall survival were analyzed. Results: Clinical data and bone marrow clot samples of 61 patients were analyzed. Mean age was 55.5 years old, and 52.5% was male. The grades of AF1q expression were 2 of -, 18 of +, 17 of ++, and 24 of +++. We defined the cases with - and + as low expression, ++ and +++ as high expression. Very good partial response or better was obtained after completion of autologous stem cell transplantation in 45% of patients with low AF1q expression and 48.8% of high expression, there was no statistically significant difference. Survival analysis using Log Rank method showed that high expression of AF1q was associated with significantly shorter progression free survival (p=0.004), but not overall survival. Conclusion: In this study, we found that the high expression of AF1q was an adverse prognostic factor in multiple myeloma. Download : Download high-res image (56KB) Download : Download full-size image Figure 1 . Disclosures Hagiwara: Celgene Corporation: Membership on an entity's Board of Directors or advisory committees.

Cytogenetic Abnormalities of Undetermined Clinical Sinigicance Correlate with Poor Prognosic Factors in Multiple Myeloma

Abstract Introduction: The prognosis of multiple myeloma (MM) depends on the underlying cytogenetic subtype. Recurrent cytogenetic changes including hyperdiploidy of odd-numbered chromosomes, -13/13q- and balanced translocations involving IGH have already been incorporated into risk stratification strategies and are used in standard clinical practice. Nevertheless, MM often exhibits a heterogenous and complex karyotype, and many other chromosomal abnormalities of uncertain clinical significance are observed. We performed a comprehensive analysis for the relationship between cytogenetic abnormalities of undetermined prognostic significance and known prognostic factors in MM. Materials and Methods: Conventional G-banding was performed in 333 newly diagnosed MM. Karyotypes were described using the most recent version of the International System for Human Cytogenetic Nomenclature. Translocations with breakpoints at p10, p11, q10, or q11 were all regarded as whole arm (centromere to centromere) translocations. Poor prognostic indicators as follows: hemoglobin (Hb) level 12 mg/dL, IgG level > 7 g/dL or IgA level > 5 g/dL, b2-microglobulin level > 5.5 mg/L, and albumin level 2 mg/dL, IgA type, lambda type, elevated free light chain level > 100 mg/dL, abnormal kappa/lambda ratio of > 4.0 or Results: Among the 333 MM patients, 212 showed a normal or good/intermediate karyotype (195 with normal karyotype; 17 with abnormal karyotype associated with good or intermediate prognosis); these served as the control group in this study. The remaining 121 had other cytogenetic abnormalities (with or without hyperdiploidy, or translocations associated with good or intermediate prognosis). Chromosomes 1p (12.0%), 1q (18.9%), 6q (7.2%), 8q (6.9%), 11q (5.4%), 12p (5.1%), 14q (9.3%) and 19q (5.1%).showed frequent abnormalities. Common breakpoints were p10–12, p13, p22 in 1p, q10–12, q21, q25, q32 in 1q; q21, q25 in 6q; q10, q22, q24 in 8q; q10, q13 in 11q; p10–12 in 12p; q24, q32in 14q and q10, q13.3 in 19q ( Figure 1 ). Certain chromosomal abnormalities were associated with known prognostic factors (p-value 5 g/dL or IgA level of> 3 g/dL; 14q - kappa or lambda type chains > 100 mg/dL; 1p, 1q, 6q, and 19q - Hb 2.0 mg/dL; 1p, 1q, 6q, or 19q - b2-microglobulin > 5.5 mg/L; 1p, 8q, 11q, and 14q - calcium > 12 mg/dL; 1p, 3p, 12p, or 14q – elevated LDH level. Conclusion: We analyzed the distribution and clinical significance of the less commonly occurring cytogenetic abnormalities for the first time. Abnormalities in chromosomes 1p, 1q, 3p, 6q, 8q, 11q, 12q, 14q, and 19q were associated with clinical factors associated with a poor prognosis. Not all chromosomes showing frequent aberrations correlated with prognostic factors, indicating that it is not the presence of a cytogenetic abnormality in itself, but the particular chromosome(s) involved in the abnormality that is the key factor that determines the initial clinical characteristics of MM. Download : Download high-res image (123KB) Download : Download full-size image Figure 1 . Diagrammatic representation of the chromosomal breakpoints involved in structural abnormalities in multiple myeloma. Triangles represent breakpoints in interstitial or telomeric regions and diamonds represent breakpoints in centromeric or pericentromeric (p10-p11 or q10-q11) regions. Disclosures No relevant conflicts of interest to declare.

High Serum Lactate Dehydrogenase Level Used as a Prognostic Factor in Multiple Myeloma

High Serum Lactate Dehydrogenase Level Used as a Prognostic Factor in Multiple Myeloma

Serum Free Light Chain Assessment Early After Stem Cell Transplantation as a Prognostic Factor in Multiple Myeloma.

Multiple myeloma is an incurable cancer commonly treated with stem cell transplantation (SCT). Response is traditionally evaluated 100 days after SCT, both to allow for hematopoietic reconstitution and due to immunoglobulins' long half-lives. Free light chains (FLC) have significantly shorter half-lives and may provide evidence of response or treatment failure earlier after SCT. We retrospectively studied 83 consecutive patients with multiple myeloma who underwent SCT and found 69 who had FLC measured 30 or 60 days after SCT. Using conventional FLC response criteria, we considered a patient to be at high risk for early relapse if he or she failed to experience a partial response by day 30 or 60. After a median overall follow-up of only 335 days, these high-risk patients had significantly shorter progression-free survival (median, 98 vs. 335 days, P = .001) and overall survival (366 days vs. median not reached, P = .016). Early FLC assessment either 1 or 2 months after SCT using standard FLC response criteria was able to identify a subset of patients at high risk of early relapse, and these patients may benefit from earlier interventions.

Is this the time to introduce minimal residual disease in multiple myeloma clinical practice?

Increasing therapeutic options and prolonged survival in multiple myeloma have raised interest in the concept of depth of response and its importance to predict patients' outcomes. Although the efficacy of current treatment approaches has greatly improved in the past decade, the definition of complete response (CR) remains unaltered and continues to use conventional serological and morphologic techniques. That notwithstanding, there is growing interest in minimal residual disease (MRD) monitoring, which has emerged in recent years as one of the most relevant prognostic factors in multiple myeloma. MRD can be assessed both inside (e.g., immunophenotypic and molecular techniques) and outside the bone marrow (e.g., PET/CT). Here, we focus on flow- and molecular-based assays by which different cooperative groups have demonstrated the efficacy of MRD assessment to predict outcomes even among patients in CR, and irrespectively of disease risk. Although further standardization is still required, the time has come to implement MRD monitoring in prospective clinical trials as a sensitive tool to evaluate treatment efficacy and for risk-adapted treatment, particularly in the consolidation and maintenance settings. Here, we present a comprehensive and critical review on the methodologic aspects, specific characteristics, and clinical significance of MRD monitoring by flow cytometry, PCR, and next-generation sequencing.

The flow cytometry-defined light chain cytoplasmic immunoglobulin index and an associated 12-gene expression signature are independent prognostic factors in multiple myeloma.

As part of Total Therapy (TT) 3b, baseline marrow aspirates were subjected to two-color flow cytometry of nuclear DNA content and cytoplasmic immunoglobulin (DNA/CIG) as well as plasma cell gene expression profiling (GEP). DNA/CIG-derived parameters, GEP and standard clinical variables were examined for their effects on overall survival (OS) and progression-free survival (PFS). Among DNA/CIG parameters, the percentage of the light chain-restricted (LCR) cells and their cytoplasmic immunoglobulin index (CIg) were linked to poor outcome. In the absence of GEP data, low CIg <2.8, albumin <3.5 g/dl and age ⩾65 years were significantly associated with inferior OS and PFS. When GEP information was included, low CIg survived the model along with GEP70-defined high risk and low albumin. Low CIg was linked to beta-2-microglobulin >5.5 mg/l, a percentage of LCR cells exceeding 50%, C-reactive protein ⩾8 mg/l and GEP-derived high centrosome index. Further analysis revealed an association of low CIg with 12 gene probes implicated in cell cycle regulation, differentiation and drug transportation from which a risk score was developed in TT3b that held prognostic significance also in TT3a, TT2 and HOVON trials, thus validating its general applicability. Low CIg is a powerful new prognostic variable and has identified potentially drug-able targets.

Serum β2-microglobulin is frequently elevated in type 1 Gaucher patients

β2-Microglobulin is the major prognostic factor in multiple myeloma, a known comorbidity of Gaucher disease. We evaluated herein serum β2-microglobulin levels of 31 type 1 Gaucher patients; for 8/31 patients, pre- and post-treatment comparisons were made. Thirteen patients (on treatment=6) had high levels of β2-microglobulin, and showed higher chitotriosidase activity and Severity Score Index, and lower concentration of platelets, than patients with normal levels. Levels of β2-microglobulin correlated with chitotriosidase activity (ρ=0.65; p<0.01), platelets (ρ=−0.42; p=0.02) and α1- (ρ=0.43; p=0.02) and α2-protein bands (ρ=−0.40; p=0.03). Regarding pre- and post-treatment values, median β2-microglobulin levels decreased after treatment (pre-=2931ng/mL; post-=1970ng/mL; p<0.01). Our data suggest that levels of serum β2-microglobulin are frequently elevated in type 1 Gaucher patients, correlate with severity of the disease and decrease after treatment.

Cytogenetic Abnormalities of Undetermined Clinical Significance Correlate with Poor Prognosic Factors in Multiple Myeloma

Introduction: The prognosis of multiple myeloma (MM) depends on the underlying cytogenetic subtype. Recurrent cytogenetic changes including hyperdiploidy of odd-numbered chromosomes, -13/13q- and balanced translocations involving IGH have already been incorporated into risk stratification strategies and are used in standard clinical practice. Nevertheless, MM often exhibits a heterogenous and complex karyotype, and many other chromosomal abnormalities of uncertain clinical significance are observed. We performed a comprehensive analysis for the relationship between cytogenetic abnormalities of undetermined prognostic significance and known prognostic factors in MM.Materials and Methods: Conventional G-banding was performed in 333 newly diagnosed MM. Karyotypes were described using the most recent version of the International System for Human Cytogenetic Nomenclature. Translocations with breakpoints at p10, p11, q10, or q11 were all regarded as whole arm (centromere to centromere) translocations. Poor prognostic indicators as follows: hemoglobin (Hb) level < 10 g/dL, calcium level > 12 mg/dL, IgG level > 7 g/dL or IgA level > 5 g/dL, b2-microglobulin level > 5.5 mg/L, and albumin level < 3.5 g/dL. Creatinine level > 2 mg/dL, IgA type, lambda type, elevated free light chain level > 100 mg/dL, abnormal kappa/lambda ratio of > 4.0 or < 0.5, and elevated LDH level were also included.Results: Among the 333 MM patients, 212 showed a normal or good/intermediate karyotype (195 with normal karyotype; 17 with abnormal karyotype associated with good or intermediate prognosis); these served as the control group in this study. The remaining 121 had other cytogenetic abnormalities (with or without hyperdiploidy, or translocations associated with good or intermediate prognosis). Chromosomes 1p (12.0%), 1q (18.9%), 6q (7.2%), 8q (6.9%), 11q (5.4%), 12p (5.1%), 14q (9.3%) and 19q (5.1%).showed frequent abnormalities. Common breakpoints were p10–12, p13, p22 in 1p, q10–12, q21, q25, q32 in 1q; q21, q25 in 6q; q10, q22, q24 in 8q; q10, q13 in 11q; p10–12 in 12p; q24, q32in 14q and q10, q13.3 in 19q (Figure 1). Certain chromosomal abnormalities were associated with known prognostic factors (p-value < 0.05): 8q - IgA type; 1q - IgG level of > 5 g/dL or IgA level of> 3 g/dL; 14q - kappa or lambda type chains > 100 mg/dL; 1p, 1q, 6q, and 19q - Hb < 10.0 g/dL; 1p and 12q - albumin < 3.5 g/dL; 14q and 19q –creatinine > 2.0 mg/dL; 1p, 1q, 6q, or 19q - b2-microglobulin > 5.5 mg/L; 1p, 8q, 11q, and 14q - calcium > 12 mg/dL; 1p, 3p, 12p, or 14q – elevated LDH level.Conclusion: We analyzed the distribution and clinical significance of the less commonly occurring cytogenetic abnormalities for the first time. Abnormalities in chromosomes 1p, 1q, 3p, 6q, 8q, 11q, 12q, 14q, and 19q were associated with clinical factors associated with a poor prognosis. Not all chromosomes showing frequent aberrations correlated with prognostic factors, indicating that it is not the presence of a cytogenetic abnormality in itself, but the particular chromosome(s) involved in the abnormality that is the key factor that determines the initial clinical characteristics of MM. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

CD200 May be a Potential Target for Therapy in Standard Risk Childhood ALL

Multiagent therapies for children with acute lymphoblastic leukaemia (ALL) have resulted in impressive remission rates of around 90%. Unfortunately, relapse rates remain high and most children who relapse will not survive. Innovative antibody and chimeric antigen receptor (CAR) modified T-cell therapies have shown promising results in ALL. However, this is a heterogeneous disease and several groups have shown ALL evolves in a branching fashion. Moreover, some leukaemia stem cell (LSC) populations do not express antigens that are the targets of developing therapies such as CD19, CD20, and CD22. Consequently, these cells may be largely unaffected and could provide a reservoir of resistant cells, from which relapse may develop. The diversity of reported LSC populations may be a reason for treatment failure and improvements in therapy will require targeting of all cells that have the potential to initiate and maintain this disease. In an attempt to identify more appropriate therapeutic targets we conducted an extensive microarray screen of several LSC subpopulations and bulk leukaemia cells from a group of paediatric patients with favourable cytogenetic abnormalities. This screen identified a number of genes that were up-regulated in all LSC subpopulations compared to levels in normal haemopoietic cells. The up-regulated genes encode for cell surface antigens, including the type-1 membrane protein, CD200 and signalling pathways. Expression of CD200 was 12-fold higher in bulk ALL cells compared to normal BM cells (p=0.001). CD200 is a key immunosuppressive molecule that has been implicated as a poor prognostic factor in multiple myeloma and acute myeloid leukaemia. To validate the microarray results, we assessed expression of CD200 in 20 BCP-ALL cases (11 pre B, 9 c-ALL) by flow cytometry and in functional assays. Fifteen of these cases were classified as MRD low risk and had favourable cytogenetic abnormalities and 5 were classified as high risk by MRD and cytogenetic abnormalities. ALL cells were stained with antibodies against CD34, CD19 and CD200 and cells were sorted on the basis of expression/lack of expression of CD34/CD200. The functional ability of the sorted populations was assessed in NSG mice and serial transplantation experiments were performed to assess self-renewal capacity. Overall, the flow cytometric analyses confirmed the microarray data in that expression of CD200 was high in this patient cohort (60±6.6%) compared to normal BM (0.1%±0.04, p ≤ 0.001), regardless of cytogenetic abnormality or MRD risk status. In standard risk cases, BM engraftment was achieved with the CD34+/CD200+ (2-66% human leukaemia) and CD34-/CD200+ (10-26%) subpopulations only, using inocula of 4x104-107 cells. No engraftment was attained using similar numbers of CD200- cells, regardless of expression of CD34 or CD19 in this group. Interestingly, when cells from the 5 patients defined as high risk were inoculated, engraftment was achieved using all four sorted subpopulations. The levels of leukaemia engraftment in these cases ranged from 2-95% using 1x103-5x106 cells and levels were similar across all subpopulations in individual patient samples. Results from serial transplantation studies to date demonstrate the sorted subpopulations were capable of self-renewal. As with the primary transplants, all four subpopulations from high risk samples engrafted while this ability was restricted to CD200+ subpopulations in standard risk cases. FISH analyses have confirmed all the engrafting cells had an abnormal karyotype. These data provide further evidence that surface antigen immunophenotype is not a good predictor of function in paediatric ALL, particularly in high risk cases. However, in standard risk cases it may be possible to eliminate LSC by targeting CD200. Treating human chronic lymphocytic leukaemia cells with anti-CD200 antibodies has been shown to prevent engraftment in NSG mice. Our data suggests that investigation of the effects of anti-CD200 in standard risk childhood ALL are warranted. DisclosuresNo relevant conflicts of interest to declare.

Abstract 3852: Obesity is associated with clinical characteristics in African American multiple myeloma patients

Abstract African-American ethnicity, male sex, older age and obesity are accepted risk factors for multiple myeloma (MM). Obesity early in life is a risk factor for many cancers, including MM; most studies have focused on populations of European origin. African-Americans have a higher prevalence of obesity than other populations, and may have a distinct genetic contribution to this condition. We established a multi-center study to investigate possible explanations for the excess risk of MM among African-Americans. The aim of the present case-case analysis was to determine whether body mass index (BMI) was associated with clinical characteristics at presentation in African-American MM patients. 1,065 patients diagnosed with active MM from January 1, 2009 through September 30, 2013 were recruited through nine outpatient centers at academic medical centers and 3 SEER population-based cancer registries. Information on weight and height at 20 years of age and at 5 years prior to diagnosis was obtained from questionnaires. Clinical information collected from the medical records and interviews included age at diagnosis, stage, percent plasmacytosis on bone marrow biopsy, B2 microglobulin level, immunoglobulin class and presence of lytic bone lesions. Molecular profiling abstracted from pathology reports included chromosomal gain/deletions (1P-, 1Q+, +3, +5, +7, +9, +11, 12P-, Del(13), +15, Del(17P), Del(17P13)), translocations (4;14), (11;14), and (14;16 ). Analysis of Variance (ANOVA) was used to test the null hypothesis that age at diagnosis was similar across BMI categories. A T-test was used to test the null hypothesis that the mean age at diagnosis in underweight/normal patients was not different than that for overweight and obese patients. The Mantel-Haenszel Chi square test for trend was used to test the null hypothesis that the proportion of cytogenetic and molecular abnormalities was similar across BMI categories. The study population consists of 306 African American male and 396 African American female MM patients. In both, obesity at 20 years of age was associated with younger age at diagnosis, compared to a normal/underweight (p=0.0015). Among males only, obesity at 5 years prior to diagnosis was also associated with younger age at diagnosis (p=0.03). A trend between increasing BMI and molecular characteristics associated with a poor prognosis was suggested. We observed a strong association between obesity at age 20 and younger age at diagnosis among males and females, and in males only at 5 years prior to diagnosis. Obesity is one of the few known potentially modifiable risk factors for MM. Younger age at diagnosis reflects an earlier accumulation of either or both genetic and environmental risk factors. We also observed a link between obesity at each of the two time points and specific molecular markers of poor prognosis. Additional studies are needed to determine biological significance of the results. Citation Format: Amie E. Hwang, Sikander Ailwadhi, Carol Ann Huff, Leon Bernal-Mizrachi, Christopher A. Haiman, Edward Peters, Seema Singhal, Karen Pawlish, Cathryn Bock, Todd Zimmerman, David J. Van Den Berg, David V. Conti, Brenda B. Birmann, Jayesh Mehta, John J. Graff, Daniel O. Stram, Niquelle Brown, Yang Yu, Moosa Azadian, Laurence Kolonel, Brian E. Henderson, Ann Mohrbacher, Graham Colditz, Brian-C Chiu, Michael Tomasson, Jeffrey Zonder, Robert Z. Orlowski, Sagar Lonial, Wendy Cozen. Obesity is associated with clinical characteristics in African American multiple myeloma patients. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3852. doi:10.1158/1538-7445.AM2014-3852