Clinical Outcome In Multiple Myeloma Research Articles

Daratumumab-Based Therapeutic Approaches and Clinical Outcomes in Multiple Myeloma and other Plasma Cell Dyscrasias: Insights from a Nationwide Real-World Chart Review Study.

Singapore leads Southeast Asia in the routine use of daratumumab for multiple myeloma and other plasma cell dyscrasias. This retrospective review analyzed 112 patients who received daratumumab between 2012 and 2020. Tolerability, and efficacy based on prior lines (PL) of therapy, cytogenetic risk group, and the presence of renal impairment were presented. Infusion-related reactions occurred in 26.8% of patients. Grades 1 and 2 hematological and non-hematological adverse events were observed in 14.3% and 33.9% of patients, respectively. After a median follow-up of 16.9 months, there was no significant difference in overall response rates (ORR) (86% versus 76.3%, p = 0.082) or depth of response (≥ complete response (CR), 35.1% versus 28.9%, p = 0.469) between myeloma patients with and without renal dysfunction. Newly diagnosed and relapsed/refractory patients had an ORR of 92% and 76.3%, and a ≥ VGPR (very good partial response) rate of 80% and 55.3%, respectively. Median progression-free survival (PFS) was better for patients with 0/1 PL compared to ≥ 2 PLs (19.8 versus 6.2 months, p < 0.001), with a deeper response (≥ CR, 38.5% versus 16.7%, p = 0.033). Forty-six and a half percentage of patients had high-risk FISH abnormalities, and those with 0/1 PL had a significantly better ORR than those with ≥ 2 PLs (83.3% vsersus 47.1%, p = 0.022), achieving an ORR similar to that of the general cohort (80.2%, p = 0.905). In conclusion, positioning daratumumab in earlier lines of therapy leads to better outcomes and may mitigate the impact of high-risk FISH abnormalities.

Comprehensive molecular profiling of multiple myeloma identifies refined copy number and expression subtypes

Multiple myeloma is a treatable, but currently incurable, hematological malignancy of plasma cells characterized by diverse and complex tumor genetics for which precision medicine approaches to treatment are lacking. The Multiple Myeloma Research Foundation’s Relating Clinical Outcomes in Multiple Myeloma to Personal Assessment of Genetic Profile study (NCT01454297) is a longitudinal, observational clinical study of newly diagnosed patients with multiple myeloma (n = 1,143) where tumor samples are characterized using whole-genome sequencing, whole-exome sequencing and RNA sequencing at diagnosis and progression, and clinical data are collected every 3 months. Analyses of the baseline cohort identified genes that are the target of recurrent gain-of-function and loss-of-function events. Consensus clustering identified 8 and 12 unique copy number and expression subtypes of myeloma, respectively, identifying high-risk genetic subtypes and elucidating many of the molecular underpinnings of these unique biological groups. Analysis of serial samples showed that 25.5% of patients transition to a high-risk expression subtype at progression. We observed robust expression of immunotherapy targets in this subtype, suggesting a potential therapeutic option.

Improving clinical outcomes in multiple myeloma: Weight maintenance strategies to reduce complications during autologous hematopoietic transplantation.

e19533 Background: Multiple myeloma (MM) is the second most prevalent hematological disease. The introduction of melphalan as high-dose therapy followed by autologous hematopoietic cell transplantation (HDT/ASCT) for younger patients has dramatically changed treatment outcomes. Monitoring daily weights has been well documented in Acute promyelocytic leukemia (APL) induction, though little has been documented in myeloma patients receiving ASCT. Monitoring daily weights can be a useful tool in maintaining patients' euvolemic status and can help to facilitate appropriate resuscitation of patients who are otherwise prone to sepsis and fluid overload. At our center over the last few years, we have standardized an approach to monitoring and maintaining the weights of MM patients at or near their pre-admission level with fluids and diuresis, as needed. We report outcomes of weight monitoring on morbidity and mortality during ASCT for MM. Methods: We conducted a retrospective chart review of MM patients receiving ASCT at our center between January 2019 and December 2022. Patients whose weights were monitored and maintained within 3 kilograms of admission weight were identified. We looked at the length of stay (LOS) and ICU transfers during this time. We reviewed the records of 205 patients induced at our center. Results: The mean age was 62 years at the time of transplant (34-78). 119 of the 205 patients identified as men (58%). Out of 205 patients, 152 patients did not have their weights maintained. Of these 152 patients, 7 were transferred to ICU. 53 patients had weights maintained, of which 1 patient was transferred to ICU (OR 0.089 (95% CI 0.018-0.6725) (p = 0.019)). The mean LOS for all patients was 12.97 days. Mean LOS was 12.41(n = 53) and 13.18(n = 152) days for patients whose weights were and were not followed, respectively (mean difference of 0.77 days) (95 % CI of -0.15 to 1.68) (p = 0.0992). 3 of the 205 patients died during admission. In these patients, weights were not maintained before ICU transfer. (OR = 0.4072 (0.0207-8.0136) (p = 0.55)). Conclusions: Fluid overload and sepsis are significant concerns in the treatment of MM. Our findings suggest that maintaining euvolemia can effectively manage patients on the floor and limit and in some cases prevent ICU transfers. Although there are limitations to our study due to its retrospective, single-center nature, the cost-effectiveness and noninvasive means of maintaining euvolemic status support its potential application to standard treatment protocols. Further prospective studies are warranted to validate these findings. [Table: see text]

Exclusive Benefits of Sustained MRD-Negative in High-Risk Multiple Myeloma

Introduction: Achievement of minimal residual disease (MRD) negativity is now recognized as a strong prognostic indicator for favorable clinical outcomes in multiple myeloma (MM) patients. Nonetheless, the optimal duration of MRD-negativity as a surrogate endpoint across different levels of sensitivity remains unclear. Additionally, it is imperative to explore whether sustained MRD-negativity holds the same prognostic value for MM patients with varying risk stratifications. Methods: In this study, 587 newly diagnosed MM patients were assessed for MRD using multiparameter flow cytometry (sensitivity of 10^-5 or 10^-6) at Zhongshan Hospital Fudan University between January 2014 and Janyary 2022. Patients were categorized as MRD-negativity not obtained (NON-MRD), obtained and sustained (SUS-MRD), or obtained and lost (LOS-MRD) based on their MRD status at given landmark point(4, 6, 12, 18, 24, and 36 months) after initiation of anti-myeloma treatment. The impact of MRD-negativity on survival outcomes was analyzed at increasing landmark times. Univariate and multivariate regression analyses were performed to examine the prognostic implications of time-dependent MRD status on overall survival in the context of different risk-stratification systems which were identified by versions of International Staging System (ISS). Results: A total of1718 MRD tests were performed on MM patients, the median number was 3. Among survival analyses at differnet landmark time, MM patients at 12 months showed greatest separation in subsequent survival between 3 groups no matter the sensitivity of 10^-5 or 10^-6 ( p=0.032, p=0.03). Pooled analysis indicated that patients who were in SUS-MRD category with a sensitivity ≥10^-5 had superior overall survival (OS) at 12 months landmark point ( p&amp;lt;0.001), with comparable survival between patients in Non-MRD and LOS-MRD category ( p=0.096, Figure 1). SUS-MRD group of MM patients at 12 months did not show better OS than the rest part patients in low-risk subgroup(ISS/R-ISS/R2-ISS=I, p=0.73, p=0.28, p=0.41,respectively). Among high-risk subgroup (ISS/R-ISS=III, R2-ISS=IV), landmark survival analyses at 12 months indicated that SUS-MRD benefited patients most, as shown the greatest OS in the SUS-MRD part ( p=0.013, p=0.011, p=0.056,respectively). Multivariate cox regression analysis showed that SUS-MRD was an independent favorable variable for patients in the ISS/R-ISS=III group ( p=0.021, HR:0.429; p=0.002, HR: 0.161). Conclusions: No matter the sensitivity of 10^-5 or 10^-6, the sustained of MRD negative lasts for 12 months is a robust marker of superior survival outcomes. MM patients who lost MRD negative has similar OS with those constant MRD positive in the long term. Benefit of Sustained MRD negative in MM limited to high risk subgroup MM patients.

The Temporal Evolution of Chromosome 1q Gain and Hyperdiploidy and Its Impact on Clinical Outcomes in Multiple Myeloma

INTRODUCTION: Multiple myeloma (MM) evolution is shaped by the acquisition and selection of distinct genomic events over time. While various temporal patterns have been identified, the precise chronological order in which key clonal genomic defining events are acquired, and their specific impact on clinical outcomes remain to be fully elucidated. METHODS: To decipher the genomic life history of MM and its impact on clinical outcomes, we interrogated the high coverage (80X) whole genome sequencing (WGS) of 378 MM patients. Overall, 319 and 59 WGS were generated from samples collected at diagnosis and at relapse, respectively. Newly diagnosed MM (NDMM) patients were treated upfront with proteosome inhibitors, lenalidomide, dexamethasone, +/- monoclonal antibodies. To estimate the relative and absolute temporal relationship among clonal events, we leveraged the molecular time approach, based on the corrected ratio between duplicated and non-duplicated single nucleotide variants (SNV). RESULTS: Overall, 300/378 (79%) patients had at least one large chromosomal gain defined as &amp;gt;1Mb with &amp;gt;50 clonal SNV eligible for molecular time analysis, for a total of 1493 events tested. Consistent with recent findings, using clock-like mutational signatures SBS1-SBS5, we estimated that the first gain in MM is typically acquired around the second or third decade of life, with a median time lag of approximately 33 years between acquisition and diagnosis. In 158/178 (89%) of hyperdiploid (HRD) patients, all trisomies of odd-numbered chromosomes were acquired simultaneously. Among patients with canonical IGH translocations and HRD, the molecular time when trisomies were acquired was usually late compared to HRD without canonical IGH translocations (p&amp;lt;0.0001). Next, we examined the temporal relationship between IGH canonical translocations and gains linked to the same IGH canonical translocation in 29 patients. We consistently observed that when IGH canonical translocations co-occurred with HRD (n=7), and/or 1q gain (n=8) in the same patient, the canonical IGH translocation was always found to be the initiating event, occurring earlier than the chromosomal gains. Interestingly, in 104/178 (58.4%) HRD patients, the earliest multi-gain event was not always restricted to odd chromosomes, but in involved other non-HRD chromosomes such as: 6p (n=43) and 1q (n=32), demonstrating that gain 1q, which is assumed to occur at progression, can also be acquired very early in disease life history. A total of 92 and 21 patients with 1q gain and 1q amp (&amp;gt;3 copies) were identified, respectively. In 14/21 (66%) with 1q amp, the first and second 1q extra copies were acquired in two distinct and independent time windows. Interestingly, patients with early 1q gain showed the same clinical outcomes as 1q amp, and inferior progression free survival outcomes compared to late 1q gain (p=0.01; Figure 1). In 18/21 1q amp the first 1q duplication was acquired early, suggesting that the adverse prognostic effect of 1q gain/amp in NDMM may be more dependent on the timing of its acquisition rather than the number of extra copies gained. To investigate late temporal patterns and differentiate selection versus acquisition of large gains at relapse after autologous stem cell transplant (ASCT), we interrogated 25/59 (42%) RRMM patients exposed to melphalan with detectable SBS-MM1. To establish the temporal order of events, we employed the melphalan mutational signature, SBS-MM1, as a unique temporal barcode (Diamond et al. Blood 2023). Overall, we observed that 21% of all loss of heterozygosity (LOH) events and 15% of all gains were acquired after ASCT (i.e., SBS-MM1 mutations were only detectable in the pre-gain/duplicated SNV). Notably, among these events, 1q gain was acquired after ASCT in 3/4 patients, indicating that the acquisition of 1q gain can occur even after administration of first-line therapy. CONCLUSIONS: In our study, we have uncovered distinct temporal patterns in the acquisition of early genomic drivers in MM. These patterns contribute to the reconstruction and better understanding of the life history of MM and may hold potential clinical and prognostic implications.

Sodium Glucose Cotransporter-2 Inhibitor Is Associated with Lower Risk of Lymphoid Malignancy: Result from Large Real-World Cohorts

Objective: Type 2 diabetes mellitus (T2DM) and cancer are among the top ten leading cause of death in the United States of America [1, 2]. It is well-known that T2DM and cancer share several common risk factors including hyperinsulinemia, obesity, chronic inflammatory status and advancing age. Recent observational studies revealed that diabetic patients are associated with modest increased risk of various blood cancers, including lymphoma and leukemia [2, 3]. Despite being a drug of choice for diabetic patients with chronic kidney disease or cardiovascular disease [4-7], contemporary data regarding the relationship sodium glucose cotransporter-2 inhibitors (SGLT-2i) and incidence of hematological malignancies remains elusive. Our study aims to assess the association between SGLT-2i and lymphoma in patients with T2DM compared to other oral antidiabetic drugs. Methods: This was a population-based cohort study based on electronic health records from 92 hospitals in the United States. Patients aged over 18 years with T2DM who were initiated treatment with SGLT-2i or DPP-4i between 2015 and 2023 were retrieved. Patients with type 1 diabetes or pre-existing malignancy were excluded. Cohorts were grouped into SGLT-2i or DPP-4i and were 1:1 propensity score matched for age, sex, ethnicity, comorbidities, and medication history. The primary outcome was newly diagnosed lymphoma and myeloma, including, Hodgkin lymphoma, B-cell lymphoma, T-cell/natural killer (NK)-cell lymphoma and plasma cell neoplasm. Hazard ratios (HRs) and 95% confidence intervals (95% Cis) were calculated using the Cox proportional hazard regression model. Kaplan-Meier analysis and log-rank tests were used to compare the outcomes across the cohorts. Results: There were a total of 1,877,388 adults with T2DM and were started on SGLT-2i or DPP-4i. The final study population after 1:1 propensity score matching included 1,399,898 patients, with 46.4% of which being females. The remaining baseline characteristics were balanced between the two group, including chronic kidney disease, heart failure and advance age. There were 5,197 incident cases of lymphomas and 1,691 incident cases of plasma cell neoplasm. SGLT-2i was associated with a significantly lower risk of developing various hematological malignancy including Hodgkin lymphoma (aHR 0.696; 95% CI 0.567-0.855), follicular lymphoma (aHR 0.798; 95% CI 0.667-0.954), non-follicular lymphoma (aHR 0.800; 95% CI 0.709-0.902), and malignant plasma cell neoplasm (aHR 0.693; 95% CI 0.624-0.768) Conclusion: The use of SGLT-2i was associated with a reduced risk of developing various lymphoma and myeloma, including Hodgkin lymphoma, non-Hodgkin lymphoma, and malignant plasma cell neoplasm, when compared to DPP-4i use. Further investigations are warranted to elucidate the mechanism and pathophysiology behind these findings. Reference 1. Cronin, K.A., et al., Annual report to the nation on the status of cancer, part 1: National cancer statistics. Cancer, 2022. 128(24): p. 4251-4284. 2. Giovannucci, E., et al., Diabetes and cancer: a consensus report. Diabetes Care, 2010. 33(7): p. 1674-85. 3. Gong, I.Y., et al., Association between diabetes and haematological malignancies: a population-based study. Diabetologia, 2021. 64(3): p. 540-551. 4. Evans, J.M., et al., Metformin and reduced risk of cancer in diabetic patients. BMJ, 2005. 330(7503): p. 1304-5. 5. Foretz, M., et al., Metformin: from mechanisms of action to therapies. Cell Metab, 2014. 20(6): p. 953-66. 6. Richardson, L.C. and L.A. Pollack, Therapy insight: Influence of type 2 diabetes on the development, treatment and outcomes of cancer. Nat Clin Pract Oncol, 2005. 2(1): p. 48-53. 7. Wu, W., et al., The association of diabetes and anti-diabetic medications with clinical outcomes in multiple myeloma. Br J Cancer, 2014. 111(3): p. 628-36.

Concomitant deletion of the short arm (del(1p13.3)) and amplification or gain (1q21) of chromosome 1 by fluorescence in situ hybridization are associated with a poor clinical outcome in multiple myeloma.

Chromosome 1 abnormalities in multiple myeloma (MM) are increasingly recognized as high risk-defining features. The authors report the prognostic value of del(1p13.3) by fluorescence in situ hybridization (FISH) at enrollment in subjects treated on total therapy clinical trials 2-6. FISH probes were generated from specific BAC DNA clones for the AHCYL1 gene locus (1p13.3) and the CKS1B locus (1q21). A total of 1133 patients were included in this analysis. Although del(1p13.3) was detected in 220 (19.4%) patients, 1q21gain or 1q21amp were observed in 300 (26.5%) and 150 (13.2%) patients, respectively. Concomitant del(1p13.3) with 1q21 gain or amp was observed in 65 (5.7%) and 29 (2.5%) patients, respectively. There was enrichment of high-risk features such as International Staging System (ISS) stage 3 disease and gene expression profiling (GEP)70 high risk (HR) in the group with del(1p13.3). Presence of del(1p13.3) confers inferior progression-free survival (PFS) and overall survival (OS). On multivariate analysis, the presence of ISS stage 3 disease, GEP70 HR, 1q21gain, and 1q21amp were independent predictors of PFS or OS. The PFS and OS of patients with combined abnormalities of del (1p13.3)/1q21gain or amp was significantly worse compared to del(1p13.3) alone and 1q21gain or 1q21 amp alone, which identifies a subset of patients with poor clinical outcomes.

Deep Transcriptome Profiling of Multiple Myeloma Using Quantitative Phenotypes.

Transcriptome studies are gaining momentum in genomic epidemiology, and the need to incorporate these data in multivariable models alongside other risk factors brings demands for new approaches. Here we describe SPECTRA, an approach to derive quantitative variables that capture the intrinsic variation in gene expression of a tissue type. We applied the SPECTRA approach to bulk RNA sequencing from malignant cells (CD138+) in patients from the Multiple Myeloma Research Foundation CoMMpass study. A set of 39 spectra variables were derived to represent multiple myeloma cells. We used these variables in predictive modeling to determine spectra-based risk scores for overall survival, progression-free survival, and time to treatment failure. Risk scores added predictive value beyond known clinical and expression risk factors and replicated in an external dataset. Spectrum variable S5, a significant predictor for all three outcomes, showed pre-ranked gene set enrichment for the unfolded protein response, a mechanism targeted by proteasome inhibitors which are a common first line agent in multiple myeloma treatment. We further used the 39 spectra variables in descriptive modeling, with significant associations found with tumor cytogenetics, race, gender, and age at diagnosis; factors known to influence multiple myeloma incidence or progression. Quantitative variables from the SPECTRA approach can predict clinical outcomes in multiple myeloma and provide a new avenue for insight into tumor differences by demographic groups. The SPECTRA approach provides a set of quantitative phenotypes that deeply profile a tissue and allows for more comprehensive modeling of gene expression with other risk factors.

MMRFVariant: Prioritizing variants in Multiple Myeloma

Background:Multiple Myeloma (MM) is a blood malignancy that occurs in the plasma cells of bone marrow. Due to the drug resistance and frequent relapses, MM remains a key area of therapeutic challenge. Analysis of variants could reveal important prognostic features in MM. A reliable interpretation on how a variant affects a protein remains yet a not trivial task. The usage of appropriate bioinformatic tools for the analysis of variants data, sourcing from the Multiple Myeloma Research Foundation (MMRF) CoMMpass (Clinical Outcomes in MM to Personal Assessment of Genetic Profile) study, a large-scale observation project focusing on MM, offers great opportunities. To the best of our knowledge, a specific tool dealing with variants data sourcing from MMRF-CoMMpass datasets does not exist yet. Material and methods:MMRFVariant is a novel R package able to identify and prioritize the disease-causing variants in MM with the aim of providing valuable insights for the MM variants interpretation. MMRFVariant deals with the datasets (IA15 release) retrieved from the MMRF-CoMMpass Study. The MM variants analysis exploits pathogenicity predictions scores and survival analysis results. Conclusion and Discussion:MMRFVariant provides five functionalities that produce graphical and tabular results: the impact-effect correlation plot, the Kaplan–Meier (KM) survival curves, the heatmap of the occurrence number and the impact table can be computed. The potentiality of the tool is highlighted in a case study in which a set of six genes is selected due to their notoriety in the MM pathogenicity: the results provided by MMRFVariant are confirmed by what is reported in other studies.

Evaluating the Impacts of CYP3A4*1B and CYP3A5*3 Variations on Pharmacokinetic Behavior and Clinical Outcomes in Multiple Myeloma Patients With Autologous Stem Cell Transplant.

There exists considerably large interpatient variability in pharmacokinetic exposure of high dose melphalan in multiple myeloma patients with hematopoietic stem-cell transplantation. In this study, we aimed to evaluate the potential impacts of CYP3A4*1B (rs2940574) and CYP3A5*3 (rs776746) variations on pharmacokinetic properties of melphalan and clinical outcomes in multiple myeloma (MM) patients. Genotypes of CYP3A4*1B (rs2940574) and CYP3A5*3 (rs776746) were determined by validated gene-specific real-time PCR (RT-PCR) assays using DNA samples from 108 MM patients; plasma concentrations of melphalan at different time points were quantified using liquid chromatography-tandem mass spectrometry (LC-MS/MS). CYP3A4*1B/*1B and CYP3A5*3/*3 carriers appeared to have a short median progression-free survival time and a higher maximum melphalan plasma concentration than non-carriers [792 vs. over 950 days, p=0.08; 9.91 (2.67, 34.03) vs. 8.66 (4.46, 17.61) mg/l, p=0.039]. CYP3A4*1B/*1B and CYP3A5*3/*3 variations might influence melphalan therapy in MM patients through yet-to-be-identified mechanisms.

Single Cell Multi-Omic Profiling of Multiple Myeloma with t(4;14) Identifies a T-Cell Population That Correlates with Clinical Outcomes

Background Multiple Myeloma (MM) is the second most common haematologic malignancy and patients with high risk MM have adverse outcomes. Translocation (4;14) occurs in 15% of MM and is associated with inferior survival. However, outcomes are heterogeneous even within this subset of patients. We set out to identify factors that correlated with progression free survival (PFS) in a cohort of t(4;14) patients by using high dimensional profiling of the malignant plasma cells (PCs) and tumor micro-environment through single cell multi-omics. Methods We analysed diagnostic bone marrow (BM) samples from 13 patients with t(4;14) MM using the ESCAPE™ platform from Singleron Biotechnologies which simultaneously measures gene and cell surface protein expression of 62 proteins in single cells. Cryopreserved BM samples were stained with antibodies and subsequently sorted on CD138 expression. The CD138 positive and negative fractions were recombined at a 1:1 ratio for analysis using the 10x Genomics 3' RNAseq kit. Resulting data were analyzed with Singleron Biotechnology's MapSuite™ single cell analytics platform. Subsequent validation of the t(4;14) findings were performed using the 5' chemistry version of ESCAPE™ with the 10x Genomics RNAseq platform. Results All patients received either bortezomib or thalidomide-based induction and had a median age of 63 years. Median PFS and overall survival (OS) were 22 and 34 months respectively. Consistent with published bulk gene expression data, MMSET was overexpressed in all PCs, while FGFR3 expression was heterogeneous. No gene or protein expression patterns within the PCs were identified that correlated with PFS or OS. In the tumor microenvironmental compartment, we identified a CD3+ population (cluster 26) that had not been previously annotated in published single cell RNA-seq data. These cells were CD45+ and CD138- at both the protein and RNA level, suggesting they are not PCs. Independent bioinformatics tools "MapCell" and "DISCO" both identified cluster 26 cells as T cells. The proportion of cluster 26 cells among non-PCs correlated negatively with PFS (P=0.04, logrank test), with samples containing a higher proportion of these cells having an inferior PFS compared to those with a lower proportion (Figure 1 panel A). We subsequently demonstrated that the T cells identified in cluster 26 could also be detected in BM samples from a separate cohort of MM patients analysed using the ESCAPE platform. These patients were treated with daratumumab in the first line or relapsed setting and included patients without t(4;14). Analysis of 15 patients from this cohort revealed that the percentage of cluster 26 cells retained its negative correlation with PFS (P=0.03, logrank test) (Figure 1 panel B).Conclusions We identified a T-cell population which may have prognostic significance in patients with t(4;14 )MM. Interestingly, this population also retains its negative correlation with PFS in a cohort of non t(4;14) patients, suggesting it may have a broader significance across cytogenetic subtypes of MM. The second cohort of patients were treated with daratumumab, hence the novel population may also be relevant in the response to anti -CD38 monoclonal antibody therapy. These data highlight the potential of single cell multi-omic analysis to identify immune micro-environmental signatures that correlate with clinical outcomes in MM. Further work is ongoing to definitively characterize these cells and explore the biological basis for their clinical impact. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Effects of AGEs, sRAGE and HMGB1 on Clinical Outcomes in Multiple Myeloma.

The receptor for advanced glycation end products (RAGE) upregulated during the onset and progression of cancer and bone-related pathologies. In this study, we aimed to investigate the role of serum advanced glycation end products (AGEs), soluble RAGE (sRAGE) and high mobility group box 1 (HMGB1), in multiple myeloma (MM). AGEs, sRAGE and HMGB1 concentrations of 54 newly diagnosed MM patients and 30 healthy volunteers were measured by ELISA. The estimations were done only once at diagnosis. The medical records of the patients were evaluated. There was no significant difference between the AGEs and sRAGE levels between the patient and control groups (p = 0.273, p = 0.313). In ROC analysis, a HMGB1 cutoff value of > 9170 pg/ml accurately discriminated MM patients (AUC = 0.672, 95% CI 0.561-0.77, p = 0.0034). AGEs level was found to be significantly higher in early-stage disease and HMGB1 in advanced disease (p = 0.022, p = 0.026). High HMGB1 levels were detected in patients whose with better first-line treatment response (p = 0.019). At 36 months, 54% of patients with low AGE were alive, compared to 79% of patients with high AGE (p = 0.055). Patients with high HMGB1 levels tended to have a longer PFS (median 43 mo [95% CI; 20.68-65.31] ) compared to patients with low HMGB1 levels (median 25 mo [95% CI; 12.39-37.6], p = 0.054). In this study, a significant elevation of serum HMGB1 level was found in MM patients. In addition, the positive effects of RAGE ligands on treatment response and prognosis were determined.

LncRNA MEG3 promotes the sensitivity of bortezomib by inhibiting autophagy in multiple myeloma

BackgroundBortezomib resistance hampers the long-term survival of multiple myeloma (MM) patients. Our previous study has proved that downregulated lncRNA MEG3 is associated with the poor clinical outcome in MM. However, the effect of MEG3 on the sensitivity of bortezomib in MM and its possible molecular mechanism remains muddled. MethodsIn this study, CCK8 and flow cytometry techniques were used to assess cell viability in MEG3 overexpressed MM cells after bortezomib treatment. The expression of autophagy-related protein LC3 and p62 were distinguished by Western blot, and the mCherry-GFP-LC3 puncta reflecting autophagy level was observed under fluorescence microscope. RNA immunoprecipitation (RIP) technology was used to detect the binding relationship of MEG3 and ATG2B to PTBP1. ResultsIncreased toxicity of bortezomib and diminished autophagy level were found in MEG3 overexpressed MM cells. Mechanistically, we discovered that RNA-binding protein PTBP1 could bind to MEG3 and ATG2B by RIP assay. Upregulation of MEG3 promoted PTBP1 expression and inhibited the expression level of ATG2B, suggesting that MEG3 recruited PTBP1 and then decayed ATG2B expression. ConclusionIn summary, our study illustrated that MEG3 increased bortezomib sensitivity by hindering autophagy through the PTBP1/ATG2B axis, providing a new therapeutic target for bortezomib-resistant MM patients.

Structural variants shape the genomic landscape and clinical outcome of multiple myeloma

Deciphering genomic architecture is key to identifying novel disease drivers and understanding the mechanisms underlying myeloma initiation and progression. In this work, using the CoMMpass dataset, we show that structural variants (SV) occur in a nonrandom fashion throughout the genome with an increased frequency in the t(4;14), RB1, or TP53 mutated cases and reduced frequency in t(11;14) cases. By mapping sites of chromosomal rearrangements to topologically associated domains and identifying significantly upregulated genes by RNAseq we identify both predicted and novel putative driver genes. These data highlight the heterogeneity of transcriptional dysregulation occurring as a consequence of both the canonical and novel structural variants. Further, it shows that the complex rearrangements chromoplexy, chromothripsis and templated insertions are common in MM with each variant having its own distinct frequency and impact on clinical outcome. Chromothripsis is associated with a significant independent negative impact on clinical outcome in newly diagnosed cases consistent with its use alongside other clinical and genetic risk factors to identify prognosis.

A Single Nucleotide Polymorphism (SNP) in the SLC22A3 Transporter Gene Is Associated With the Severity of Oral Mucositis in Multiple Myeloma Patients Receiving Autologous Stem Cell Transplant Followed by Melphalan Therapy.

It has been reported that expression of OCT3 enhanced the sensitivity to melphalan in cells, indicative of potential roles of OCT3 in melphalan transport. Herein we investigated the association of select single nucleotide polymorphisms in SLC22A3 (gene encoding OCT3) with clinical outcomes in multiple myeloma (MM) patients with hematopoietic autologous stem cell transplants followed by high-dose melphalan therapy. Melphlan concentrations in blood samples from 108 MM patients were measured using liquid chromatography-tandem mass spectrometry (LC-MS/ΜS); genotypes of rs2048327, rs1810126, and rs3088442 in these patients were determined using quatitive RT-PCR assays. Rs3088442 A variant-carriers had a significantly increased risk of severe oral mucositis in comparison with homozygous rs3088442 G-carriers with adjusted odds ratio of 4.00 (95% CI=1.25-14.7; p=0.027). Rs3088442 A carriers tended to have lower creatinine clearance (p=0.10) and higher maximum plasma concentration of melphalan (p=0.07). OCT3 might be involved in melphalan transport in MM patients.

Using MMRFBiolinks R-Package for Discovering Prognostic Markers in Multiple Myeloma.

Multiple myeloma (MM) is the second most frequent hematological malignancy in the world although the related pathogenesis remains unclear. Gene profiling studies, commonly carried out through next-generation sequencing (NGS) and Microarrays technologies, represent powerful tools for discovering prognostic markers in MM. NGS technologies have made great leaps forward both economically and technically gaining in popularity. As NGS techniques becomes simpler and cheaper, researchers choose NGS over microarrays for more of their genomic applications. However, Microarrays still provide significant benefits with respect to NGS. For instance, RNA-Seq requires more complex bioinformatic analysis with respect to Microarray as well as it lacks of standardized protocols for analysis. Therefore, a synergy between the two technologies may be well expected in the future. In order to take up this challenge, a valid tool for integrative analysis of MM data retrieved through NGS techniques is MMRFBiolinks, a new R package for integrating and analyzing datasets from the Multiple Myeloma Research Foundation (MMRF) CoMMpass (Clinical Outcomes in MM to Personal Assessment of Genetic Profile) study, available at MMRF Researcher Gateway (MMRF-RG), and at the National Cancer Institute Genomic Data Commons (NCI-GDC) Data Portal. Instead of developing a completely new package from scratch, we decided to leverage TC-GABiolinks, an R/Bioconductor package, because it provides some useful methods to access and analyze MMRF-CoMMpass data. An integrative analysis workflow based on the usage of MMRFBiolinks is illustrated.In particular, it leads towards a comparative analysis of RNA-Seq data stored at GDC Data Portal that allows to carry out a Kaplan Meier (KM ) Survival Analysis and an enrichment analysis for a Differential Gene Expression (DGE) gene set.Furthermore, it deals with MMRF-RG data for analyzing the correlation between canonical variants and treatment outcome as well as treatment class. In order to show the potential of the workflow, we present two case studies. The former deals with data of MM Bone Marrow sample types available at GDC Data Portal. The latter deals with MMRF-RG data for analyzing the correlation between canonical variants in a gene set obtained from the case study 1 and the treatment outcome as well as the treatment class.

Real-World Evidence of Epidemiology and Clinical Outcomes in Multiple Myeloma, Findings from the Registry of Hemato-Oncologic Malignancies in Colombia, Observational Study

Multiple Myeloma (MM) is a plasma cell derived clonal disorder that represents around 1% of all newly diagnosed neoplasms. Limited data regarding MM treatment in Latin America is available, and access to novel agents for a substantial portion of the population is limited by their high costs. RENEHOC is a bidirectional (retrospective and prospective) multicenter observational registry of hematological malignancies in Colombia. MM patients included up to July 2020 were analyzed on this report. 890 are reported with a median follow-up of 18 months (IQR: 7-42 months). Patients were classified by age group (≤ or > 65 years). Median age at diagnosis was 67 years (IQR: 59-75 years) and 47.1% of patients were women. 709 patients (79.6%) received Bortezomib-based schemes as part of the first line. Two hundred and fifty-two patients (28.3%) were consolidated with Autologous Stem Cell Transplantation (ASCT) in first-line. ASCT consolidation and age were the main independent factors influencing outcomes; in the non-ASCT cohort, 5-year overall survival was 48.7% (CI 41.8-55.2) compared to 80.7% (CI 73-86.4) in ASCT patients. This data depicts the reality of MM in Colombia, which likely reflects other Latin American countries, where access barriers to diagnosis and treatment are echoed in advanced stage diagnosis and a low rate of transplants. These seem to negatively impact survival despite the availability of most novel drugs approved for this disease. Thus, emphasizing the paradox that prevails in most of the region: availability without equitable access.

Clonal Hematopoiesis Prevalence Increases throughout Treatment of Newly Diagnosed Multiple Myeloma Patients

Clonal Hematopoiesis Prevalence Increases throughout Treatment of Newly Diagnosed Multiple Myeloma Patients

A Risk Scoring System for Prognosis of Multiple Myeloma

A Risk Scoring System for Prognosis of Multiple MyelomaBackground: Multiple myeloma (MM) is the second most frequently-occurring hematologic malignancy characterized by anemia, renal damage, osteolytic lesions and hypercalcemia (Kumar, et al. 2017), without specific prognostic indicators. Protein arginine methyltransferases 3 (PRMT3) is an enzyme which participates in the progression of some malignant diseases (Xiao, et al. 2017). However, the prognostic value of PRMT3 in MM remains unclear. In this study, we developed a risk scoring system based on the expression of PRMT3 to distinguish MM cohorts with different clinical characteristics.Methods: we integrated 4 datasets from The Cancer Genome Atlas (TCGA) or gene expression omnibus (GEO) and analyzed the correlation between PRMT3 expression and R-ISS stage, diseases progression, clinical characteristics or prognosis. Furthermore, we collected a cohort of newly diagnosed MM and healthy donor samples and then performed qRT-PCR to verify the expression of PRMT3. A risk scoring system was established to point out the prognostic indicator for clinical outcome of MM. The predictive power was evaluated by using Receiver Operating Characteristic (ROC) and Kaplan-Meier survival curve.Results: By extensive data analysis, we found the expression of PRMT3 was upregulated during the progression of myeloma (Figure 1 A p=0.573, 0.028, 0.02, respectively). The expression level of PRMT3 in relapsed MM patients was higher than that in newly diagnosed MM patients (Figure 1 B p=0.02, 0.016, 0.002, respectively). Meanwhile, the expression of PRMT3 was also increased in MM patients with advanced R-ISS stage (Figure 1 C p=0.001, 0.042, respectively). Moreover, the validation in a new cohort of MM samples showed the expression of PRMT3 was higher in MM patients compared to normal controls (Figure 1 D p=0.012). MM patients with high expression of PRMT3 showed prolonged Event Free Survival (EFS) and Overall Survival (OS) (Figure 2 A&B EFS: p=0.008, OS: p=0.001). Furthermore, we found the expression of PRMT3 had a positive correlation with B2M(p=0.018), HGB (p=0.001), aspirate plasma cells (p=0.002) and bone marrow biopsy plasma cells (p=0.001 Table not shown). Meanwhile, univariate and multivariate analysis showed that B2M, LDH, ALB, MRI and PRMT3 were independent adverse prognostic factors for OS in MM patients (p<0.001, p<0.001, p=0.0044, 0.0403, 0.0312, Table not shown). Finally, we established a risk scoring system which performed remarkable predicting effectiveness among MM patients. The ROC curve showed that the risk model performed well in 3-year OS (Figure 2 C AUC=0.749). A threshold score 1.26897 was recommended to distinguish the high and low risk score groups. Patients with higher risk score had a shorter OS than those with lower risk score (median 27.45 months vs. 50.13 months, p<0.001 Figure 2 D).Conclusion: Our study identified that PRMT3 was upregulated in MM patients and that increased PRMT3 was an independent adverse prognostic factor for OS in MM. The risk scoring system based on the expression of PRMT3 provided distinct insights into the prognosis of MM patients. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

OAB-033: Loss-of-function of GABARAP drives tumor resistance to bortezomib-induced immunogenic cell death in multiple myeloma

<h3>Background</h3> Resistance to immune approaches poses a major challenge to effective immunotherapy and long-term clinical outcome in multiple myeloma (MM). Here we identified loss-of-function of gamma-aminobutyric acid receptor-associated protein (GABARAP) as a tumor-intrinsic mechanism of resistance to bortezomib (BTZ)-induced immunogenic cell death (ICD), the immunogenic consequence of apoptosis resulting in specific anti-MM immunity via T-cell priming by dendritic cells (DCs). <h3>Methods</h3> We found that BTZ induces ICD in human and murine MM cell lines that is dependent on the exposure of calreticulin (CALR), which drives DCs-mediated phagocytosis of MM cells. DCs-phagocytosis is followed by a specific T cell activation, with a significant increase of CD4+ effector memory (EM), total CD8+, CD8+ EM, and CD8+ terminally differentiated EM cells. Isolated T cells after co-cultures showed the presence of MM specific CTLs that were able to efficiently induce lysis of MM cells. Our results were validated using primary cells from MM patients. Moreover, induction of a protective immune response was confirmed in vivo. Specifically, treatment of 5TGM1 tumors with BTZ induced a tumor regression in a syngeneic model; and injection of live 5TGM1WT two weeks after regression did not result in tumor development, consistent with induction of immunological memory as confirmed by ELISPOT of mouse splenocytes. We identified a specific ICD signature induced by BTZ in mice; and we found that increased expression of the human orthologs of this signature was positively correlated with OS (p=0.01) in patients enrolled in the IFM/DFCI 2009 study. Notably, these functional immunologic sequelae were abrogated after BTZ treatment of CALRKO MM cells both in vitro and in vivo, confirming the obligate role of CALR exposure in the ICD process. <h3>Results</h3> By interrogating the IFM/DFCI dataset and focusing on genes involved in ICD processes that were correlated with MM patients clinical outcome, we found that low levels of GABARAP (chr17p13.1), an autophagy regulator and putative CALR binding partner, negatively impact MM patient clinical outcome (EFS, p=0.0032); even excluding HR patients with 17p deletion (EFS, p=0.018). Interestingly, KMS11 cells carrying monoallelic deletion of GABARAP were resistant to induction of ICD by BTZ; and sensitivity was restored after overexpression of the gene. Moreover, GABARAPKO in 3 ICD-sensitive cell lines abrogated the induction of ICD by BTZ; and add-back experiments by pre-treatment with recombinant CALR or GABARAP overexpression in KO clones restored ICD. Finally, CyTOF confirmed that treatment of GABARAPKO cells with BTZ failed to activate an efficient T cell response. <h3>Conclusions</h3> our study demonstrates that loss-of-function of GABARAP, particularly in HR patients with 17p deletion, contributes to tumor immune evasion and ICD resistance. These studies provide the framework for novel combination treatments to restore anti-MM immunity and improve patient outcome in HR MM.