Overall Survival In Multiple Myeloma Patients Research Articles

Radiomic nomogram for predicting high-risk cytogenetic status in multiple myeloma based on fat-suppressed T2-weighted magnetic resonance imaging

Rationale and ObjectivesRadiomics has demonstrated potential in predicting the cytogenetic status of multiple myeloma (MM). However, the role of single-sequence radiomic nomograms in predicting the high-risk cytogenetic (HRC) status of MM remains underexplored. This study aims to develop and validate radiomic nomograms based on fat-suppressed T2-weighted images (T2WI-FS) for predicting MM’s HRC status, facilitating pre-treatment decision-making and prognostic assessment. Materials and methodsA cohort of 159 MM patients was included, comprising 71 HRC and 88 non-HRC cases. Regions of interest within the most significant tumor lesions on T2WI-FS images were manually delineated, yielding 1688 features. Fourteen radiomic features were selected using 10-fold cross-validation, employing methods such as variance thresholds, Student’s t-test, redundancy analysis, and least absolute shrinkage and selection operator (LASSO). Logistic regression was utilized to develop three prediction models: a clinical model (model 1), a T2WI-FS radiomic model (model 2), and a combined clinical-radiomic model (model 3). Receiver operating characteristic (ROC) curves evaluated and compared the diagnostic performance of these models. Kaplan-Meier survival analysis and log-rank tests assessed the prognostic value of the radiomic nomograms. ResultsModels 2 and 3 demonstrated significantly greater diagnostic efficacy compared to model 1 (p < 0.05). The areas under the ROC curve for models 1, 2, and 3 were as follows: training set—0.650, 0.832, and 0.846; validation set—0.702, 0.730, and 0.757, respectively. Kaplan-Meier survival analysis indicated comparable prognostic values between the radiomic nomogram and MM cytogenetic status, with log-rank test results (p < 0.05) and concordance indices of 0.651 and 0.659, respectively; z-score test results were not statistically significant (p = 0.153). Additionally, Kaplan-Meier analysis revealed that patients in the non-HRC group, low-RS group, and aged ≤ 60 years exhibited the longest overall survival, while those in the HRC group, high-RS group, and aged > 60 years demonstrated the shortest overall survival (p = 0.004, Log-rank test). ConclusionsRadiomic nomograms are capable of predicting the HRC status in MM. The cytogenetic status, radiomics model Rad score, and age collectively influence the overall survival of MM patients. These factors potentially contribute to pre-treatment clinical decision-making and prognostic assessment.

Circulating plasma cells as a predictive biomarker in Multiple myeloma: an updated systematic review and meta-analysis

Background Circulating plasma cells (CPCs) are defined by the presence of peripheral blood clonal plasma cells, which would contribute to the progression and dissemination of multiple myeloma (MM). An increasing number of studies have demonstrated the predictive potential of CPCs in the past few years. Therefore, there is a growing need for an updated meta-analysis to identify the specific relationship between CPCs and the prognosis of MM based on the current research status. Methods The PubMed, Embase, and Cochrane Library databases were screened to determine eligible studies from inception to November 5, 2023. Publications that reported the prognostic value of CPCs in MM patients were included. Hazard ratios (HRs) with 95% confidence intervals (CIs) of overall survival (OS) and progression-free survival (PFS) were extracted to pool the results. Subgroup analyses were performed based on region, sample size, cut-off value, detection time, initial treatment, and data type. The association between CPCs level and clinicopathological characteristics, including the International Staging System (ISS), Revised-ISS (R-ISS), and cytogenetic abnormalities were also evaluated. Statistical analyses were conducted using STATA 17.0 software. Results Twenty-two studies with a total of 5637 myeloma patients were enrolled in the current meta-analysis. The results indicated that myeloma patients with elevated CPCs were expected to have a poor OS (HR = 2.19, 95% CI: 1.81–2.66, p < 0.001) and PFS (HR = 2.45, 95% CI: 1.93–3.12, p < 0.001). Subgroup analyses did not alter the prognostic role of CPCs, regardless of region, sample size, cut-off value, detection time, initial treatment, or data type. Moreover, the increased CPCs were significantly related to advanced tumour stage (ISS III vs. ISS I–II: pooled OR = 2.89, 95% CI: 2.41–3.46, p < 0.001; R-ISS III vs. R-ISS I–II: pooled OR = 3.65, 95% CI: 2.43–5.50, p < 0.001) and high-risk cytogenetics (high-risk vs. standard-risk: OR = 2.22, 95% CI: 1.60–3.08, p < 0.001). Conclusion Our meta-analysis confirmed that the increased number of CPCs had a negative impact on the PFS and OS of MM patients. Therefore, CPCs could be a promising prognostic biomarker that helps with risk stratification and disease monitoring.

Abstract 3345: PP1 phosphatase complex at the hub of the integrated stress response: A potential therapeutic target in multiple myeloma

Abstract Protein phosphatase-1 (PP1) is a class of eukaryotic enzyme that catalyzes more than half of the phosphoserine/threonine dephosphorylation reactions, and plays essential roles in cell division, glycogen metabolism and protein synthesis. The substrate specificity of PP1 holoenzyme complex is mediated by PP1-interacting proteins and its catalytic subunits. Of note, PPP1R15B/CReP and PPP1R15A/GADD34 are regulatory subunits of PP1 complex that regulates global protein synthesis and mRNA translation through eIF2α dephosphorylation. Notably, PPP1R15B is constitutively expressed, whereas the expression of PPP1R15A is stress-inducible. Previous studies showed that multiple myeloma (MM), a hematological malignancy arising from immunoglobulin-secreting plasma cells, is characterized by continual ER stress and high dependency on the unfolded protein response (UPR). The integrated stress response (ISR) activated by eIF2α phosphorylation is the fastest reaction pathway of the UPR; however, its functional role and therapeutic potential in MM remains largely unknown. In this study, we performed gene expression analysis and super-enhancer (SE) profiling in MM cell lines, primary patient samples, normal CD138+ plasma cells and memory B cells. we found that PPP1R15B is highly expressed in MM cells as compared to healthy controls, but did not observe significant upregulation of PPP1R15A in MM cells. In addition, higher expression of PPP1R15B predicted poor overall survival of MM patients, suggesting its clinical relevance in MM pathogenesis. Depletion of PPP1R15B significantly reduced cell viability, proliferation and induced G2/M phase cell cycle arrest. Gene Ontology analysis of downregulated genes in response to PPP1R15B knockdown indicates significant association with the UPR pathway. Immunoblot analysis showed activation of ER stress pro-apoptotic executors PUMA, NOXA, and cleavage of caspase-12 in PPP1R15B knockdown cells. Furthermore, we examined the therapeutic effects of eIF2α dephosphorylation inhibitors, Salubrinal and Raphin1, on MM cells. Raphin1 is a selective inhibitor of PPP1R15B that binds strongly to the PPP1R15B-PP1c complex, whereas Salubrinal inhibits both PPP1R15A-PP1c and PPP1R15B-PP1c complex. Our data showed that both Salubrinal and Raphin1 inhibited MM cell proliferation in a dose-dependent manner, while sparing normal plasma cells. MM cell lines with elevated PPP1R15B expression level were more sensitive to Salubrinal or Raphin1 than those with lower PPP1R15B level. Similar changes in the expression patterns of ISR-related genes were observed in response to eIF2α dephosphorylation inhibitors. Taken together, our study suggests that targeting PPP1R15B-PP1c complex or maintaining eIF2α hyperphosphorylation may serve as a new therapeutic approach in MM, which warrants further investigation. Citation Format: Sinan Xiong, Jianbiao Zhou, Tze King Tan, Sabrina Hui-Min Toh, Tae-Hoon Chung, Takaomi Sanda, Wee Joo Chng. PP1 phosphatase complex at the hub of the integrated stress response: A potential therapeutic target in multiple myeloma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3345.

PHF19 before and post induction treatment possess favorable potency of reflecting treatment response to protease inhibitors, event-free survival, and overall survival in multiple myeloma patients

ABSTRACT Objective Plant homeodomain finger protein 19 (PHF19) regulates hematopoietic stem cell differentiation and promotes multiple myeloma (MM) progression. This study intended to explore the potency of PHF19 at baseline and post induction treatment in estimating treatment response to protease inhibitors and survival in MM patients. Methods This retrospective study screened 69 MM patients who received protease inhibitors with bone marrow (BM) samples available at both baseline and post induction treatment. Twenty healthy BM donors were included as healthy controls (HCs). PHF19 in plasma cells from BM was quantified by reverse transcription-quantitative polymerase chain reaction. Results PHF19 at baseline and post induction treatment in MM patients were increased than in HCs. In MM patients, PHF19 was declined post induction treatment. Elevated PHF19 at baseline and post induction treatment were correlated with renal impairment, beta-2-microglobulin ≥5.5 mg/L, t (4; 14), higher international staging system (ISS) stage, and higher revised ISS (R-ISS) stage. Concerning treatment response, PHF19 at baseline and post induction treatment were negatively associated with complete response and overall response rate. Notably, abnormal PHF19 (above 95% quantile value of PHF19 in HCs) at baseline and post induction treatment were linked with shortened event-free survival (EFS) and overall survival (OS). After adjustment, abnormal PHF19 post induction treatment was independently related to shortened EFS (hazard ratio = 2.474) and OS (hazard ratio = 3.124). Conclusion PHF19 is aberrantly high and declines post induction therapy, which simultaneously reflects unfavorable treatment response to protease inhibitors as well as shorter EFS and OS in MM patients.

Update on the outcome of M-protein screening program of multiple myeloma in China: A 7-year cohort study.

To improve the early detection rate of multiple myeloma (MM), the M-protein screening system has been performed in the hospital population at Zhongshan Hospital Fudan University since 2014, with electrophoretic-based monoclonal immunoglobulin (M-protein) screening integrated into the blood biochemistry panel. This study updated 7-year follow-up findings of MM patients diagnosed by screening-driven and symptom-driven approaches. The retrospective study compared the characteristics and outcomes of patients diagnosed through two patterns by reviewing the plasma cell disease database from January 2014 to October 2021. The screening-driven group included patients diagnosed through the screening system during workups of unrelated medical conditions or routine checkups. In contrast, patients who visited or were referred to the hematological department due to myeloma-related end-organ damage were categorized into the symptom-driven group. There were 3,110,218 serum protein electrophoresis (SPEP) tests performed during 7 years, with 1.95% (60,609) patients yielding positive SPEP results. Of 911 confirmed MM cases (excluding concurrent amyloidosis), 366 were assigned to the screening-driven group, while 545 were to the symptom-driven group. Compared to the symptom-driven group, the screening group had more IgG subtypes, earlier International Stage System stages, fewer disease-related symptoms, lower ECOG scores, less extramedullary disease, a lower percentage of bone marrow plasma cells, and a lower level of lactate dehydrogenase. Frontline response results of two groups were similar. Patients detected through screening had a significantly improved median progression-free survival (PFS) than the symptom-driven group (62.2 vs. 24.9 months, p < 0.001, HR: 2.12, 95% CIs: 1.69-2.65), with median follow-ups of 32.6 and 27.4 months. Furthermore, the median overall survival (OS) was significantly longer in patients of the screening group (not reached vs. 62.3 months, p < 0.001, HR: 2.49, 95% CIs: 1.81-3.41). After being adjusted for well-acknowledged myeloma prognostic factors, the screening-driven diagnostic pattern remained an independent prognostic factor indicating improved PFS and OS in MM patients. Routine M-protein screening for MM in the hospital population results in an earlier diagnosis and better patient outcomes.

Muscle and fat composition in patients with newly diagnosed multiple myeloma

Measures of muscle and adipose tissue mass have been associated with outcomes in several malignancies, but studies in multiple myeloma (MM) are inconsistent. The aim of this study was to evaluate the association between muscle and fat areas and radiodensity, and overall survival (OS) in patients with newly diagnosed MM. We included 341 patients diagnosed with MM from 2010–2019 who had an 18F-fluorodeoxyglucose positron emission tomography/computed tomography at diagnosis. A cross-sectional image at the third lumbar vertebrae was segmented into muscle and fat components. Median follow up was 5.7 years. There was no association between sarcopenia and baseline disease characteristics or OS. Low muscle radiodensity was associated with higher disease stage, anemia, and renal failure. OS was 5.6 vs. 9.0 years in patients with muscle radiodensity in the lower vs. middle/upper tertiles, respectively (P = 0.02). High subcutaneous adipose tissue (SAT) radiodensity was associated with higher stage, anemia, thrombocytopenia, hypercalcemia, renal failure, and high LDH. OS was 5.4 years vs. not reached in patients with SAT radiodensity in the upper vs. middle/lower tertiles, respectively (P = 0.001). In conclusion, sarcopenia was not associated with OS in MM patients. High SAT radiodensity and low muscle radiodensity were associated with advanced disease stage and adverse laboratory characteristics.

PSMC2 Overexpression Enhances Proteasome Activity, Mitigates ER Stress, and Correleates with Reduced Overall Survival in Multiple Myeloma

Background: Multiple myeloma (MM) remains a highly treatable but generally incurable plasma cell dyscrasia. These plasma cells synthesize massive amounts of paraprotein relying heavily on the ubiquitin proteasome system to maintain homeostasis and are exquisitely sensitive to proteasome inhibition to avoid terminal ER stress. We postulated that increased expression of clinically relevant proteasome subunits would correlate with reduced overall survival in MM patients treated with proteasome inhibitors. Objective: To correlate proteasome gene expression levels to clinicals outcomes in both the phase III APEX trial and the MMRF COMMPASS Trial. To model the effects of these hits on MMCL UPS function. Results: We identified that 19S regulatory ATPases PSMC2 and PSMC6 correlated with poorer OS, outperforming the proteasome inhibitor (PI) target, PSMB5, in this analysis. We focused on modeling the more frequently upregulated PSMC2. We created PSMC2 overexpressing multiple myeloma cell lines, noting PSMC2 is readily incorporated in the higher order proteasome complexes. PSMC2 overexpressing cells exhibit higher levels of 20S proteasome activity (1.6 fold increase in ARH77 and 3 fold increase in U266 cell line compared to control, p&amp;lt; 0.01 in each case) and are more resistant to PI challenge (Bortezomib LD 50 of 30 nM v 15 nM in U266 overexpressed and control cells respectively, Bortezomib LD 50 of 18 nM v 10 nM in ARH77 overexpressed v control cells respectively). PIs are known to increase ER stress, and PSMC2 overexpressing cells are better able to mitigate PI mediated ER stress with PSMC2 overexpressing cells exhibiting less induction of ER stress sensors p-IRE1α and ATF4 compared to PSMC2 knockdown cells on bortezomib or tunicamycin challenge as quantitated by western blotting and utilizing an fluorescent ER stress sensor that models the conversion of XBP1u to XBP1s by pIRE1α. Mechanistically, PSMC subunits play a key role in the recognition and degradation of misfolded ER proteins, and increased 19S subunits may allow more effective ER associated protein degradation (ERAD) Conclusions: Taken together, our results suggest PSMC2 overexpression correlates with reduced OS in MM patients treated with bortezomib and that PSMC2 may represent an actionable therapeutic target to prevent or overcome PI resistance. PSMC2 overexpression increases proteasome activity in MM cells, mitigates ER stress on PI challenge and promotes PI resistance. mitigates ER stress upon PI challenge, and portending poorer clinical prognosis and more drug resistance, suggesting that 19S targeted agents may help overcome drug resistance in MM. Figure 1 A. PSMC2 overexpression increases chymotrypsin-like proteasome activity at baseline in U266 and ARH77 cells with a similar effect of residual proteasome activity after challenge with bortezomib or carfilzomib. B. ARH77 and U266 FLAG and vector cells were challenged with bortezomib for 18 hrs and then assayed by annexin PI staining. LD50 increased from 10 nm to 15 nm in ARH77 cells and from 12 to 25 in U266 Cells.

Identification and Validation of High-Risk Genes Contributing to Poor Prognosis in Multiple Myeloma Patients with chr1q Gain/Amplification

The gain/amplification of 1q21 (1q gain/amp) is a common cytogenetic abnormality (CA) in multiple myeloma (MM) and implies poor outcome. Despite the dysregulation of numerous genes located in chromosome 1q due to gain/amplification, the pivotal genes influencing the prognosis of multiple myeloma remain undefined. Gene expression matrices and clinical data of multiple myeloma (MM) patients were acquired from publicly available databases, including the Gene Expression Omnibus (GEO) and CoMMpass. A comparative analysis was performed between MM patients with and without 1q gain/amplification to identify genes that exhibited differential expression due to this genetic alteration. The Least Absolute Shrinkage and Selection Operator (LASSO) technique and multivariate Cox regression analysis were employed to select potential candidate genes strongly associated with an unfavorable prognosis in MM patients. The overall survival of MM patients was assessed using the Kaplan-Meier method, and survival curves were compared using the log-rank test. In the training set comprising 413 MM patients from the GSE4581 database, 93 individuals (22.5%) displayed 1q gain/amplification. Differential gene expression analysis identified 541 upregulated genes in patients with 1q gain/amp, with 187 of these genes located on chromosome 1q. Subsequently, the 187 upregulated genes on chromosome 1q underwent LASSO and multivariate Cox regression analysis. As a result, ten genes (ANP32E, ARPC5, CDC42BPA, CDKN2C, CHD1L, COAS, ISG20L2, NUF2, TAGLN2, UBE2T) were identified as significant contributors to poor overall survival in MM patients. Additional analysis utilizing single-cell RNA sequencing (scRNAseq) was conducted on plasma cells from 12 MM patients and ten healthy donors from the GSE117156 dataset. The findings unveiled elevated expression levels of CDKN2C, ISG20L2, NUF2 and UBE2T in malignant plasma cells compared to those from healthy donors. The prognostic significance of the four high-risk genes (CDKN2C, ISG20L2, NUF2 and UBE2T) was further confirmed in the CoMMpass database, which comprises 858 MM patients. Although patients with 1q gain/amp showed more inferior survival ( P=0.012), without the four high-risk genes overexpression in patients with 1q gain/amp showed the similar survival with patients without 1q abnormal( P=0.74). Furthermore, in the CoMMpass database, we observed a notable pattern wherein patients with higher expression levels of the four high-risk genes (N=27) were more prevalent in the late stage, with 51% of them classified at stage III. Conversely, only 27% of patients with low expression levels of the four high-risk genes were at stage III. Notably, no significant differences were observed in terms of age, sex, or race between the two groups. In summary, our findings demonstrate that overexpression of CDKN2C, ISG20L2, NUF2 and UBE2T contribute to the poor outcome in multiple myeloma patients with 1q gain/ amplification.

Exploring Autologous Hematopoietic Stem Cell Transplantation (HSCT) Utilization and HSCT Refusal Trends in Multiple Myeloma Patients: A Comprehensive Examination of NCDB Data from 2004 to 2020

Background: Autologous hematopoietic stem cell transplantation (HSCT) remains standard of care in multiple myeloma (MM) in eligible patients and is proven to improve survival. However, a significant number of patients can decline HSCT as part of their treatment. Understanding the patterns and factors behind HSCT refusal is crucial for optimizing patient care and tailoring effective interventions. Methods: A total of 43,653 patients MM patients recommended for HSCT from 2004 to 2020 were identified via the NCDB, after excluding cases with incomplete values. KM curves compared overall survival (OS) between HSCT and non-HSCT patients, with both groups receiving induction chemotherapy. Factors influencing HSCT refusal were determined using multivariate logistic regression with backward elimination. SAS version 9.4 was used to analyze the data. Results: Among 43,653 patients recommended for HSCT only 1.95% (850 individuals) refused HSCT and 98.05% received HSCT. The patient characteristics are presented in Table 1. As noted in Figure 1, the median OS of MM patients who opted for induction chemotherapy followed by HSCT, had a much higher OS of 124 months, compared to the MM patients who declined HSCT and had chemotherapy alone with a median OS of 95 months. On multivariate logistic regression analysis patients aged 60-69 and those aged 70 and above demonstrated higher odds of refusing HSCT compared to those below 50 years, with OR of 1.50 (p = 0.0120) and 3.26 (p &amp;lt; 0.0001), respectively. Male patients exhibited decreased odds of HSCT refusal compared to females, with an OR of 0.84 (p = 0.0329). Black patients showed a higher likelihood of HSCT refusal than White patients (OR = 1.38, p = 0.0022). Hispanic patients were less likely to refuse HSCT compared to non-Hispanic patients (OR = 0.65, p = 0.0296). Patients without insurance, those covered by Medicaid/other government insurance and Medicare, exhibited increased odds of HSCT refusal compared to private insurance with ORs of 2.23 (p = 0.0066), 1.38 (p = 0.0366) and 1.34 (p = 0.0048), respectively. Moreover, patients with a Charlson-Deyo score of 2 or 3 had a higher likelihood of HSCT refusal compared to those with a score of 0 (OR = 1.48, p = 0.0024). Patients treated at non-academic facilities were more likely to refuse HSCT than those treated at academic facilities (OR = 2.67, p &amp;lt; 0.0001). Furthermore, patients with a median household income less than $38,000 had increased odds of HSCT refusal compared to those earning $63,000 or more (OR = 1.62, p = 0.0002). Patients with incomes in the ranges of $38,000-$47,999 and $48,000-$62,999 also demonstrated higher odds of refusal with ORs of 1.39 (p = 0.0043) and 1.34 (p = 0.0050), respectively. The year of diagnosis exhibited a significant association with the likelihood of HSCT refusal: patients diagnosed in 2004-2007, 2008-2011 and 2012-2015 had lower odds of refusing HSCT compared to those diagnosed in the reference period (2016-2020), with ORs of 0.16 (p &amp;lt; 0.0001), 0.56 (p &amp;lt; 0.0001) and 0.75 (p = 0.0024), respectively. Geographic location significantly influenced HSCT refusal, with the South Atlantic region (SA) as the reference category. Lower odds of refusal were observed in West South Central (OR 0.29, p &amp;lt; 0.0001), Mountain (OR 0.53, p = 0.001), East South Central (OR 0.63, p = 0.025), and East North Central (OR 0.72, p = 0.01) regions. Conclusion: Our analysis of large real-world database shows that the HSCT significantly improves survival in MM and should be recommended for eligible patients. Furthermore, the study shows that the HSCT refusal rate in real-world is low at ~2% but still represents a missed opportunity to provide standard of care with room for improvement. Older patients (age &amp;gt;60), females, Blacks, non-Hispanics, patients with non-private health insurance, higher comorbidities, non-academic facilities, and lower income (&amp;lt;$63,000/yr), were found to be associated with increased odds of declining HSCT. There was also geographic variation across the USA with higher HSCT refusal in the South Atlantic region compared to other regions. In addition, the increasing trend in HSCT refusal from 2004 to 2020 is likely due to advances in MM therapeutics due to the availability of novel agents and anti-CD38-immunotherapy. Our study suggests significant racial, economic, and geographic variation regarding the utilization of HSCT across the US which should be further studied in prospective studies.

High serum miR-223-3p expression level predicts complete response and prolonged overall survival in multiple myeloma patients undergoing autologous hematopoietic stem cell transplantation.

AHSCT is the treatment of choice for newly diagnosed patients with transplant-eligible multiple myeloma (MM). However, considerable variability in response to autologous hematopoietic stem cell transplantation (AHSCT) results in only 50% of patients achieving complete response (CR) after AHSCT, which is directly associated with improved progression-free and overall survival (OS). In this study, we aimed to investigate the potential predictive role of selected serum miRNAs in MM patients who underwent AHSCT. Serum expression level of 6 miRNAs: miR-221-3p, miR-15b-5p, miR-223-3p, miR-320c, miR-361-3p, and miR-150-5p was evaluated in 51 patients who underwent AHSCT. Blood samples were collected at two time points: before conditioning chemotherapy (T1) and fourteen days after transplant (+14) (T2). All selected miRNAs significantly changed their expression level across the procedure- two were up-regulated after AHSCT: hsa-miR-320c (FC 1.42, p<0.0001) and hsa-miR-361-3p (FC 1.35, p=0.0168); four were down-regulated: hsa-miR-15b-5p (FC 0.53, p<0.0001), hsa-miR-221-3p (FC 0.78, p=0.0004), hsa-miR-223-3p (FC 0.74, p=0.0015) and hsa-miR-150-5p (FC 0.75, p=0.0080). Notably, before AHSCT, hsa-miR-223-3p was down-regulated in International Staging System (ISS) III patients (FC=0.76, p=0.0155), and hsa-miR-320c was up-regulated (FC=1.27, p=0.0470). These differences became non-significant after AHSCT. Eight (15.69%) patients achieved CR before AHSCT and 17 patients (33.33%) at +100 days after AHSCT. In multivariate logistic regression analysis, achievement of CR after induction and hsa-miR-223-3p at T1 were independent predictors of CR after AHSCT. In multivariate Cox regression analysis, hsa-miR-223-3p at T1 expression level was associated with prolonged OS (HR 0.06, 95%CI: 0.00 - 0.99, p=0.0488). Serum expression of has-miR-223-3p is a predictor of CR and prolonged OS in MM patients undergoing AHSCT.

GPER1 Activation Exerts Anti-Tumor Activity in Multiple Myeloma.

G protein-coupled estrogen receptor 1 (GPER1) activation is emerging as a promising therapeutic strategy against several cancer types. While GPER targeting has been widely studied in the context of solid tumors, its effect on hematological malignancies remains to be fully understood. Here, we show that GPER1 mRNA is down-regulated in plasma cells from overt multiple myeloma (MM) and plasma cell leukemia patients as compared to normal donors or pre-malignant conditions (monoclonal gammopathy of undetermined significance and smoldering MM); moreover, lower GPER1 expression associates with worse overall survival of MM patients. Using the clinically applicable GPER1-selective agonist G-1, we demonstrate that the pharmacological activation of GPER1 triggered in vitro anti-MM activity through apoptosis induction, also overcoming the protective effects exerted by bone marrow stromal cells. Noteworthy, G-1 treatment reduced in vivo MM growth in two distinct xenograft models, even bearing bortezomib-resistant MM cells. Mechanistically, G-1 upregulated the miR-29b oncosuppressive network, blunting an established miR-29b-Sp1 feedback loop operative in MM cells. Overall, this study highlights the druggability of GPER1 in MM, providing the first preclinical framework for further development of GPER1 agonists to treat this malignancy.

Identification and validation of a cellular senescence-related lncRNA signature for prognostic prediction in patients with multiple myeloma

ABSTRACT Multiple myeloma (MM) is the second most common hematologic malignancy, which primarily occurs in the elderly. Cellular senescence is considered to be closely associated with the occurrence and progression of malignant tumors including MM, and lncRNA can mediate the process of cellular senescence by regulating key signaling pathways such as p53/p21 and p16/RB. However, the role of cellular senescence related lncRNAs (CSRLs) in MM development has never been reported. Herein, we identified 11 CSRLs (AC004918.5, AC103858.1, AC245100.4, ACBD3-AS1, AL441992.2, ATP2A1-AS1, CCDC18-AS1, LINC00996, TMEM161B-AS1, RP11-706O15.1, and SMURF2P1) to build the CSRLs risk model, which was confirmed to be highly associated with overall survival (OS) of MM patients. We further demonstrated the strong prognostic value of the risk model in MM patients receiving different regimens, especially for those with three-drug combination of bortezomib, lenalidomide, and dexamethasone (VRd) as first-line therapy. Not only that, our risk model also excels in predicting the OS of MM patients at 1, 2, and 3 years. In order to verify the function of these CSRLs in MM, we selected the lncRNA ATP2A1-AS1 which presented the largest expression difference between high-risk groups and low-risk groups for subsequent analysis and validation. Finally, we found that down-regulation of ATP2A1-AS1 can promote cellular senescence in MM cell lines. In conclusion, the CSRLs risk model established in present study provides a novel and more accurate method for predicting MM patients’ prognosis and identifies a new target for MM therapeutic intervention.

A Machine Learning Model to Predict Survival and Therapeutic Responses in Multiple Myeloma.

Multiple myeloma (MM) is a highly heterogeneous hematologic tumor. Ubiquitin proteasome pathways (UPP) play a vital role in its initiation and development. We used cox regression analysis and least absolute shrinkage and selector operation (LASSO) to select ubiquitin proteasome pathway associated genes (UPPGs) correlated with the overall survival (OS) of MM patients in a Gene Expression Omnibus (GEO) dataset, and we formed this into ubiquitin proteasome pathway risk score (UPPRS). The association between clinical outcomes and responses triggered by proteasome inhibitors (PIs) and UPPRS were evaluated. MMRF CoMMpass was used for validation. We applied machine learning algorithms to MM clinical and UPPRS in the whole cohort to make a prognostic nomogram. Single-cell data and vitro experiments were performed to unravel the mechanism and functions of UPPRS. UPPRS consisting of 9 genes showed a strong ability to predict OS in MM patients. Additionally, UPPRS can be used to sort out the patients who would gain more benefits from PIs. A machine learning model incorporating UPPRS and International Staging System (ISS) improved survival prediction in both datasets compared to the revisions of ISS. At the single-cell level, high-risk UPPRS myeloma cells exhibited increased cell adhesion. Targeted UPPGs effectively inhibited myeloma cells in vitro. The UPP genes risk score is a helpful tool for risk stratification in MM patients, particularly those treated with PIs.

Omics Analysis of Ferroptosis and Establishment of Prognostic Model for multiple myeloma Patients

To explore the role of ferroptosis-related genes in multiple myeloma(MM) through TCGA database and FerrDb, and build a prognostic model of ferroptosis-related genes for MM patients. Using the TCGA database containing clinical information and gene expression profile data of 764 patients with MM and the FerrDb database including ferroptosis-related genes, the differentially expressed ferroptosis-related genes were screened by wilcox.test function. The prognostic model of ferroptosis-related genes was established by Lasso regression, and the Kaplan-Meier survival curve was drawn. Then COX regression analysis was used to screen independent prognostic factors. Finally, the differential genes between high-risk and low-risk patients were screened, and enrichment analysis was used to explore the mechanism of the relationship between ferroptosis and prognosis in MM. 36 differential genes related to ferroptosis were screened out from bone marrow samples of 764 MM patients and 4 normal people, including 12 up-regulated genes and 24 down-regulated genes. Six prognosis-related genes (GCLM, GLS2, SLC7A11, AIFM2, ACO1, G6PD) were screened out by Lasso regression and the prognostic model with ferroptosis-related genes of MM was established. Kaplan-Meier survival curve analysis showed that the survival rate between high risk group and low risk group was significantly different(P<0.01). Univariate COX regression analysis showed that age, sex, ISS stage and risk score were significantly correlated with overall survival of MM patients(P<0.05), while multivariate COX regression analysis showed that age, ISS stage and risk score were independent prognostic indicators for MM patients (P<0.05). GO and KEGG enrichment analysis showed that the ferroptosis-related genes was mainly related to neutrophil degranulation and migration, cytokine activity and regulation, cell component, antigen processing and presentation, complement and coagulation cascades, haematopoietic cell lineage and so on, which may affect the prognosis of patients. Ferroptosis-related genes change significantly during the pathogenesis of MM. The prognostic model of ferroptosis-related genes can be used to predict the survival of MM patients, but the mechanism of the potential function of ferroptosis-related genes needs to be confirmed by further clinical studies.

Analysis of Long-Term Prognostic Factors Based on Nutritional Status in Patients with Multiple Myeloma

To explore the prognostic factors of patients with multiple myeloma (MM) based on nutritional status. The Controlling Nutritional Status (CONUT) score and clinical parameters at diagnosis of 203 newly diagnosed MM patients hospitalized in the department of hematology, Wuxi People's Hospital from January 1, 2007 to June 30, 2019 were analyzed retrospectively. The best cut-off value was determined by ROC curve, and the patients were divided into high CONUT group (>6.5 points) and low CONUT group (≤6.5 points); through COX regression multivariate analysis of overall survival (OS) time, CONUT, ISS stage, LDH and treatment response were selected for multiparameter prognostic stratification. The OS of MM patients in high CONUT group was shorter. The low-risk group (≤2 points) of the multiparameter risk stratification had longer OS time and progression-free survival (PFS) time compared with the high-risk group (>2 points), and it was also effective for different age or karyotype subgroups, new drug groups containing bortezomib and transplant-ineligible subgroup. The risk stratification of MM patients based on CONUT, ISS stage, LDH and treatment response is worthy of clinical application.

The Correlation of miR-671-5p with Diagnosis and Prognosis in Multiple Myeloma.

This study aimed to investigate the value of miR-671-5p in multiple myeloma (MM) in diagnostics and prognosis and developed a potential biomarker to improve the prognosis of MM. Plasma cells were isolated from bone marrow samples of 80 MM patients, in which miR-671-5p levels were determined. The correlation between miR-671-5p expression with serum creatinine, β-2-microglobulin, lactate dehydrogenase, bone lesions, International Staging System staging, chromosomal abnormalities, and albumin was analyzed. The association between miR-671-5p expression with progression-free survival and overall survival in MM patients was determined. miR-671-5p expression was reduced and predicted an increased risk of MM. miR-671-5p expression was negatively correlated with serum creatinine, β-2-microglobulin, lactate dehydrogenase, bone lesions, International Staging System staging, and chromosomal abnormalities, and positively correlated with albumin. miR-671-5p expression was augmented in complete response patients and overall response rate patients, and differentiated CR and ORR patients from Non-CR and Non-ORR patients. Furthermore, miR-671-5p low expression was associated with unfavorable progression-free survival and overall survival in MM patients. In a word, miR-671-5p is associated with worsening clinical properties, increased ISS staging, unfavorable chromosomal abnormalities, and poor prognosis in MM patients.

Expression of SIL-2R in Patients with Multiple Myeloma and Its Clinical Significance

To investigate the expression and clinical significance of soluble interleukin-2 receptor(sIL-2R) in patients with multiple myeloma(MM). 54 newly diagnosed MM patients in the Second Affiliated Hospital of Fujian Medical University from February 2020 to December 2021 were selected as the observation group, and 60 healthy people in our hospital in the same period were selected as the control group. The expression levels of sIL-2R in the serum of the two groups were detected by enzyme-linked immunosorbent assay. The differences of sIL-2R expression level among different clinical parameter groups in MM patients were compared. The clinical parameters include:gender, age, ISS stage, hemoglobin, albumin, serum creatinine, lactate dehydrogenase and β2-microglobulin, blood calcium, bone marrow plasma cell ratio and treatment response. The relationship between sIL-2R expression level and progression-free survival(PFS) and overall survival(OS) in MM patients were analyzed. The expression of serum SIL-2R in MM patients was significantly higher than that in healthy control group (P<0.05). The expression of sIL-2R in MM patients who did not achieve complete remission(CR) was significantly higher than those of CR patients (P=0.037). There was no significant difference in the expression of serum sIL-2R between the groups of different sex, age, ISS stage, hemoglobin concentration, albumin content, serum creatinine level, lactate dehydrogenase level, the content of β2-microglobulin, the concentration of blood calcium, and the proportion of bone marrow plasma cells(P>0.05). The PFS of sIL-2R high expression group(15 months) was shorter than that of sIL-2R low expression group (22 months), which was significant difference (P=0.041). But there was no significant difference in OS between sIL-2R high expression group and sIL-2R low expression group (P=0.124). Univariate analysis results showed that the high expression of serum sIL-2R was associated with poor PFS in MM patients. Multivariate analysis results showed that the high expression of serum sIL-2R was still an independent adverse prognostic factor for PFS in MM patients, However, the expression of serum sIL-2R was not statistically significant in evaluating OS in MM patients by univariate and multivariate analysis. The expression of serum sIL-2R in MM patients was significantly higher than that in healthy people. Serum sIL-2R is an independent prognostic factor of PFS in MM patients.

Update on the Outcome of M-Protein Screening Program of Multiple Myeloma in China: A 7-Year Cohort Study

Background: To improve the early detection rate of multiple myeloma(MM), the M-protein screening system has been performed in hospital population at Zhongshan Hospital Fudan University since 2014, with electrophoretic-based screening integrated into routine blood tests. This study updated the 7-year follow-up findings of MM patients diagnosed by screening-driven and end-organ symptom-driven pattern. Methods: The retrospective study compared the characteristics and results of two alternative diagnosis patterns by reviewing the plasma cell disease database of MM patients diagnosed between January 2014 and October 2021 at hospital. Screening-driven patients were classified as having an incidental finding of M-protein during workup of unrelated medical conditions or routine body check-up, while symptom-driven patients exhibited myeloma-related end-organ damage's symptoms. Results: Over 300 million serum protein electrophoresis tests were performed during 7 years, with 1-2 percent yielding positive results, prompting patients to seek further examination. Of 911 confirmed MM cases (concurrent amyloidosis excluded), 366 were assigned to the screening-driven group while 545 were to the symptom-driven group. In comparison to the symptom-driven group, the screening group had more IgG phenotypes (P=0.001), earlier ISS stages(P<0.001), fewer disease-related symptoms (hypercalcemia, renal insufficiency, anemia and bone lesion), lower ECOG scores (P<0.001), less extramedullary disease (P<0.001), a lower rate of bone marrow plasma cells(P<0.001), and lower level of lactate dehydrogenase(LDH)(P=0.018) at baseline. Regarding first-line therapy, it was found that while both groups had comparable exposure to proteasome inhibitor and immunomodulatory drugs as well as rates of the best efficacy, the screening group was more likely to have >2-drug regimens and ≧ 6 induction cycles than the other group. Patients detected through screening had a significantly longer median progression-free survival (PFS) time than the symptom-driven group (62.2 VS 24.9 months) with median follow-ups of 32.9 and 27.8 months, respectively. Furthermore, the screening group did not achieve the median overall survival (OS) time, which was 62.3 months in the symptom group. In multivariate cox analysis, the screening-driven diagnostic pattern remains an independent prognostic factor indicating favorable PFS and OS in MM patients after adjusting for well-acknowledge myeloma prognostic factors. Conclusion: Routine M-protein screening for MM in hospital patients results in an earlier diagnosis and better patient outcomes in China. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Survival of Patients with Multiple Myeloma Diagnosed with Second Primary Malignancies: An ASCO Cancerlinq Analysis

Background: While there is significant literature regarding the incidence of secondary primary malignancies (SPMs) in patients with multiple myeloma (MM), there is a relative paucity of data regarding prognosis and mortality in this population. This study aimed to utilize real-world data from the ASCO CancerLinq MM registry to determine outcomes of MM patients with SPMs. Methods: CancerLinQ, backed by the American Society of Clinical Oncology (ASCO) and launched in 2014, is a cancer registry comprised of over 100 oncology practices and hospitals in the United States. For this retrospective analysis, we included MM patients who were diagnosed with MM between 2009 and 2021. ICD-9 and ICD-10 codes were utilized for SPM diagnoses of interest. Overall survival from the date of initial multiple myeloma diagnosis was the primary endpoint, and was compared between MM patients with versus without later diagnosis of a SPM. Analyses were conducted with STATA Version 17.0 (College Station, TX); critical alpha level was set at .05. Kaplan-Meier curves were generated to demonstrate overall survival (OS). Median OS and hazard ratios (HR) were calculated for all patients later diagnosed during the reporting period with a SPM versus without, and by specific SPMs versus all patients without that SPM. Results: There were 34,234 patients meeting inclusion criteria, of whom 10.62% developed any SPM (n=3,634). Results showed longer median OS in all patients subsequently diagnosed with an SPM (130 months vs. 103 months, HR: 0.741, 95% CI 0.690-0.796, p<.001). Analyzed by specific SPM, median OS was prolonged in patients later diagnosed with non-melanoma cutaneous cancers compared to those without (141 months vs. 105 months, HR 0.472 p<.001; 95% CI 0.398-0.559), breast cancer (median OS not reached vs. 105 months, HR 0.584, 95% CI 0.475-0.717; p<.001), prostate cancer (median OS not reached vs 105 months, HR 0.599, 95% CI 0.483-0.743, p<.001), non-Hodgkin lymphoma (median OS not reached vs 105 months, HR 0.702, 95% CI 0.594-0.831, p<.001),diffuse large B cell lymphoma (median OS not reached vs. 106 months, HR .712, 95% CI .516-.984, p=.04), and melanoma (median OS not reached vs 105 months, HR 0.743, 95% CI 0.611-0.904, p=.003). Survival was not significantly affected in patients diagnosed with colorectal cancer, acute lymphoblastic leukemia, or chronic lymphocytic leukemia. Survival was worse in patients diagnosed with acute myeloid leukemia (67 months vs 106 months, HR 1.522, 95% CI 1.267-1.828, p<.001) and lung cancer (76 months vs 106 months, HR 1.374, 95% CI 1.157-1.632, p<.001). Discussion: This is the first study to utilize a real-world dataset to characterize OS in MM patients subsequently diagnosed with an SPM. Counterintuitively, patients diagnosed with any SPM were more likely to have lived longer than patients not diagnosed with an SPM. This observation could be multifactorial and could reflect patients with more indolent disease biology or those who received better treatments, resulting in higher longevity and immortal time bias. Further study and results analyzing the timing of SPM diagnosis (post MM) and its subsequent impact on OS are forthcoming and will be presented. The finding that patients diagnosed with AML lived shorter likely is indicative of dramatically worse outcomes of secondary AML in the MM population. The shorter survival of patients diagnosed with lung cancer may also represent causal signal. The finding that both AML and lung cancer as SPMs resulted in shorter survival is consistent with what would be expected in the general population, and serves as a good internal control. In summary, this study is the first to shed light on prognosis of MM patients diagnosed with various SPMs. Disclosure: The findings and opinions contained herein are those of the authors and do not represent the views/opinions of the United States Government, the United States Air Force, United States Navy, Walter Reed National Military Medical Center, or the Department of Defense. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

EZH2 Targeting Induces CD38 Upregulation and Response to Anti-CD38 Antibodies in Multiple Myeloma

Multiple myeloma (MM) is the second most common hematological malignancy characterized by the accumulation of tumor plasma cells within the bone marrow. MM is characterized by high molecular and clinical heterogeneity. During the last 10 years new therapeutic classes including targeted immunotherapies significantly improved the overall survival of MM patients. However, drug resistance and relapse remain major challenges in MM. In MM, marked reduction of CD38 expression via exocytosis, endocytosis, degradation of the antigen-antibody complex and trogocytosis has been associated with resistance to CD38 targeted immunotherapies. Our group previously described a link and a negative correlation between CD38 and EZH2 (Enhancer of Zeste homolog 2) expression during normal B to plasma cell differentiation in association with a transcriptional control of CD38 expression involving the polycomb PRC2 complex (Herviou et al. Leukemia. 2019). Here we identified a significant negative correlation between CD38 expression and EZH2 expression in 2 independent cohorts of MM patients (p<0.05; Montpellier cohort n=198; Compass cohort n=631). Thus, we hypothesized that PRC2 targeting using EZH2 inhibitor could result in CD38 re-expression to overcome daratumumab resistance. We first characterized the membrane CD38 expression by calculating the Antibody Binding Capacity in our unique panel of 40 Human Myeloma Cell lines (HMCL) representative of MM heterogeneity. We observed a heterogeneous expression of the membrane CD38 expression on MM cell lines with cell lines demonstrating no membrane expression and cell lines demonstrating high membrane expression. Next, we selected 3 HMCLs with low (JJN3), intermediate (XG2), and high (XG7) CD38 expression, and treated these cell lines with sublethal concentration of EZH2 inhibitor (EPZ-6438) for 9 days. CD38 membrane expression was characterized at Day 3, Day 6 and Day 9. Interestingly, EZP-6438 treatment induced upregulation of cell surface CD38 expression on all the HMCLs tested with the highest effect measured at day 9. Similar re-expression pattern was observed by western blot at these timing in the JJN3 CD38low cell line. Remarkably, ChIPseq analyses underlined a significant association of H3K27me3 mark at CD38 promoter in CD38neg and CD38low cell lines compared to CD38highcells. In addition, we investigated the efficiency of CD38 monoclonal antibodies (Daratumumab and Isatuximab) lysis through ADCC (Antibody dependent cellular cytotoxicity) assay after 9 days of EPZ-6438 pretreatment. Using JJN3 CD38low HMCL, EPZ-6438 pretreatment significantly improved the ADCC lysis induced by low doses of Daratumumab and Isatuximab treatment, in presence of purified Natural Killer (NK) from healthy donor, together with CD38 re-expression (p<0.05; n=3). Finally, we searched to validate these results using primary MM cells from patients at relapse after Daratumumab treatment. EPZ-6438 pretreatment resulted in a significant CD38 membrane expression upregulation with a 2 fold difference compared to the control condition together with improved ADCC toxicity of anti-CD38 antibodies in vitro. Altogether we demonstrated that treatment of MM cells with EZH2 inhibitor leads to significant upregulation of membrane CD38 expression in cell lines and primary MM cells from patients. CD38 re-expression was linked to an improvement of Daratumumab and Isatuximab ADCC efficiency. Thus, EZH2 targeting may be of therapeutic interest to overcome resistance to anti-CD38 targeted immunotherapies in Multiple Myeloma.