Prevention Of Multiple Myeloma Research Articles

DNA vaccines against GPRC5D synergize with PD-1 blockade to treat multiple myeloma

Multiple myeloma (MM), a hematological malignancy of the bone marrow, remains largely incurable. The orphan G protein-coupled receptor, GPRC5D, which is uniquely expressed in plasma cells and highly expressed in MM, is a compelling candidate for immunotherapy. In this study, we investigated the efficacy of a combination of DNA vaccine encoding mouse GPRC5D and PD-1 blockade in preventing and treating MM using the 5TGM1 murine model of MM. The mouse vaccine alone was effective in preventing myeloma growth but required PD-1 antibodies to inhibit established MM tumors. We next evaluated the prophylactic and therapeutic efficacy of a nanoplasmid vector encoding human GPRC5D in several murine syngeneic tumor models. Similar results for tumor inhibition were observed, as human GPRC5D-specific T cells and antibodies were induced by DNA vaccines. Taken together, these findings underscore the potential of GPRC5D-targeted DNA vaccines as versatile platforms for the treatment and prevention of MM.

Design and Rationale of Prolonged Nightly Fasting for Multiple Myeloma Prevention (PROFAST): Protocol for a Randomized Controlled Pilot Trial.

Obesity is an established, modifiable risk factor of multiple myeloma (MM); yet, no lifestyle interventions are routinely recommended for patients with overweight or obesity with MM precursor conditions. Prolonged nightly fasting is a simple, practical dietary regimen supported by research, suggesting that the synchronization of feeding-fasting timing with sleep-wake cycles favorably affects metabolic pathways implicated in MM. We describe the design and rationale of a randomized controlled pilot trial evaluating the efficacy of a regular, prolonged nighttime fasting schedule among individuals with overweight or obesity at high risk for developing MM or a related lymphoid malignancy. We aim to investigate the effects of 4-month prolonged nightly fasting on body composition and tumor biomarkers among individuals with overweight or obesity with monoclonal gammopathy of undetermined significance (MGUS), smoldering multiple myeloma (SMM), or smoldering Waldenström macroglobulinemia (SWM). Individuals with MGUS, SMM, or SWM aged ≥18 years and a BMI of ≥25 kg/m2 are randomized to either a 14-hour nighttime fasting intervention or a healthy lifestyle education control group. Participants' baseline diet and lifestyle patterns are characterized through two 24-hour dietary recalls: questionnaires querying demographic, comorbidity, lifestyle, and quality-of-life information; and wrist actigraphy measurements for 7 days. Fasting intervention participants are supported through one-on-one telephone counseling by a health coach and automated SMS text messaging to support fasting goals. Primary end points of body composition, including visceral and subcutaneous fat (by dual-energy x-ray absorptiometry); bone marrow adiposity (by bone marrow histology); and tumor biomarkers, specifically M-proteins and serum free light-chain concentrations (by gel-based and serum free light-chain assays), are assessed at baseline and after the 4-month study period; changes therein from baseline are evaluated using a repeated measures mixed-effects model that accounts for the correlation between baseline and follow-up measures and is generally robust to missing data. Feasibilityis assessed as participant retention (percent dropout in each arm) and percentage of days participants achieved a ≥14-hour fast. The PROlonged nightly FASTing (PROFAST) study was funded in June 2022. Participant recruitment commenced in April 2023. As of July 2023, six participants consented to the study. The study is expected to be completed by April 2024, and data analysis and results are expected to be published in the first quarter of 2025. PROFAST serves as an important first step in exploring the premise that prolonged nightly fasting is a strategy to control obesity and obesity-related mechanisms of myelomagenesis. In evaluating the feasibility and impact of prolonged nightly fasting on body composition, bone marrow adipose tissue, and biomarkers of tumor burden, this pilot study may generate hypotheses regarding metabolic mechanisms underlying MM development and ultimately inform clinical and public health strategies for MM prevention. ClinicalTrials.gov NCT05565638; http://clinicaltrials.gov/ct2/show/NCT05565638. DERR1-10.2196/51368.

Understanding Patient Preferences for Intervention in Individuals with Precursor Multiple Myeloma: The Preference Study

Background: The standard of care for patients with monoclonal gammopathy of undetermined significance (MGUS) and smoldering multiple myeloma (SMM) consists of watchful waiting until progression to clinically manifest multiple myeloma. While an increasing number of clinical trials are exploring the premise that early intervention in this MGUS/SMM patient population will prevent progression to multiple myeloma, little is known about how patients might balance the risks against potential benefits of early intervention. Methods: To quantify preference for attributes of intervention in individuals with MGUS and SMM, we developed an online conjoint instrument with four attributes that we identified as being salient to clinical decision making: prevention of multiple myeloma, monthly cost, inconvenience, and risk of long-term side effects. We recruited individuals from two nationwide prospective cohorts to take the online instrument. To be eligible to participate, participants had to be age 18 or older, have a diagnosis of MGUS or SMM, and have no concurrent malignancy requiring active therapy. Logistic regression models were used to investigate the attributes that drive decisions of early treatment. In addition, we used a latent profile analysis to investigate latent classes defined by patient choices and assess patient characteristics that predict class membership. Results: Participants (n=272) were 43% male, with a mean age of 64.1 (SD: 9.7) years. Logistic regression models suggest a strong preference for choosing interventions based on a higher likelihood of multiple myeloma prevention (odds ratio [OR] per 10% change:1.86, 95% confidence interval [CI]: 1.72, 2.00) and avoidance of side effects (OR: 0.10, 95%CI: 0.08, 0.13). Latent profile analysis identified three latent profiles: the first profile defines individuals who predominantly value avoidance of side effects, the second profile characterizes individuals who value keeping costs low, and the third profile of individuals values multiple myeloma prevention above other attributes. Mean age was similar across latent profiles. A higher proportion of individuals in profile two (group prioritizing cost) did not complete college and had a household income of less than $100,000. A similar proportion of individuals with SMM prioritized avoiding side effects as they did multiple myeloma risk reduction. Conclusions: Overall, we found that low side effects and prevention of multiple myeloma are desirable intervention attributes in patients with MGUS and SMM. The prioritization of myeloma prevention vs low side effect profile was not perfectly coupled with an individual's disease burden. These findings reinforce the notion that a “one size fits all” approach to prevention and interception of multiple myeloma may not be suitable for all patients, and thus a diverse interventional portfolio may be needed to reach broad populations of patients with precursor conditions to multiple myeloma.

Thiazolidinedione Use Is Associated with a Borderline Lower Risk of Multiple Myeloma and a Significantly Lower Risk of Death in Patients with Type 2 Diabetes Mellitus in Taiwan.

Thiazolidinedione (TZD) exerts anti-proliferative effects on multiple myeloma (MM) cells. However, there has not been any human study investigating the risk of MM associated with TZD use. We used Taiwan's National Health Insurance database to identify 423,949 patients who had been newly diagnosed with diabetes mellitus between 1999 and 2005. After excluding ineligible patients, 86,999 pairs of patients with and without the use of TZD (rosiglitazone or pioglitazone) that had been matched based on propensity score were selected for a follow-up for MM until 31 December 2011. The hazard ratios for MM were estimated using Cox regression and weighted using a propensity score. After a median follow-up of 4.6 years and 4.7 years in ever users and never users of TZD, 32 and 47 cases were diagnosed with MM, respectively. A 35% lower risk (though not statistically significant) was observed among ever users (hazard ratio 0.652, 95% confidence interval: 0.416-1.023, p = 0.0625). When ever users were divided by the median (15 months) cumulative duration of TZD therapy, the hazard ratios (95% confidence interval) for the lower and upper medians were 0.706 (0.394-1.264) and 0.603 (0.346-1.051), respectively. When treated as a continuous variable, the hazard ratio for every 1-month increment of the cumulative duration was 0.980 (95% confidence interval: 0.963-0.997, p = 0.0185). In the age subgroup analysis, a significantly lower risk could be seen in the older age subgroup of ≥65 years (hazard ratio 0.550, 95% confidence interval: 0.305-0.992, p = 0.0468). Additional analyses suggested that there were no interactions between TZD and some medications and between TZD and some clinical diagnoses, and that the use of TZD as a preventive drug for MM might not be cost-effective because a number-needed-to-treat of 5800 was too large. Survival analyses suggested that ever users had a significantly lower risk of death when all patients were analyzed (hazard ratio: 0.84, 95% confidence interval: 0.81-0.87, p < 0.0001 via a log-rank test) or when patients who developed MM were analyzed (hazard ratio: 0.40, 95% confidence interval: 0.19-0.86, p = 0.0153 via a log-rank test). In Taiwanese patients with type 2 diabetes mellitus, TZD use is associated with a borderline lower risk of MM, which is more remarkable in patients aged ≥65 years. Because of the low incidence of MM, the use of TZD for the prevention of MM may not be cost-effective. Patients who have been treated with TZD may have a survival advantage. Future research is required to confirm the findings.

Obesity and myeloma: Clinical and mechanistic contributions to disease progression.

Obesity and obesogenic behaviors are positively associated with both monoclonal gammopathy of unknown significance (MGUS) and multiple myeloma (MM). As the only known modifiable risk factor, this association has emerged as a new potential target for MM prevention, but little is known about the mechanistic relationship of body weight with MM progression. Here we summarize epidemiological correlations between weight, body composition, and the various stages of myeloma disease progression and treatments, as well as the current understanding of the molecular contributions of obesity-induced changes in myeloma cell phenotype and signaling. Finally, we outline groundwork for the future characterization of the relationship between body weight patterns, the bone marrow microenvironment, and MM pathogenesis in animal models, which have the potential to impact our understanding of disease pathogenesis and inform MM prevention messages.

Single-Cell RNA-Sequencing of 245 Tumor Samples from Patients with MGUS and Smoldering Multiple Myeloma Reveals Novel Insights into Malignant Plasma Cell Biology

Introduction Patients with Monoclonal Gammopathy of Undetermined Significance (MGUS) and Smoldering Multiple Myeloma (SMM) exhibit variable risk of progression to full-blown Multiple Myeloma (MM), which cannot be fully explained by differences in tumor burden or genetic alterations. Therefore, characterizing non-genetic changes in malignant plasma cells may help to identify novel mechanisms of disease progression and improve prognostication. Our current understanding of transcriptomic alterations in patients with MGUS and SMM is based on bulk RNA-sequencing or microarray studies, which are affected by sample purity and interpatient variability. Here, we present results from the largest single-cell RNA-sequencing cohort (n=245) of tumor samples from patients with MGUS and SMM. Methods We performed single-cell RNA- and V(D)J-sequencing on 245 samples from 36 patients with MGUS, 136 patients with SMM, 37 patients with MM, and 25 healthy donors. Libraries were prepared with the Chromium Single-cell 5' and V(D)J enrichment kit by 10X Genomics and sequenced on NovaSeq at the Genomics Platform of the Broad Institute of MIT and Harvard (Cambridge, MA). CellRanger v.6.0.1. was used to extract FASTQ files and produce count matrices. Droplets with more than 15% of UMIs mapped to mitochondrial genes or fewer than 200 detected genes were removed, and doublets were identified using Scrublet, SCDS, and scDblFinder. Samples were integrated using Harmony, and plasma cells were annotated as malignant or normal based on the matched V(D)J clonotype information. Differential expression was performed at the single-cell level using Wilcoxon's rank-sum test. Results Overall, we sequenced 1,318,218 plasma cells, including 960,998 malignant and 357,220 normal plasma cells. Each patient's tumor formed a unique cluster, and tumors with similar cytogenetics clustered adjacent to each other. By comparing expression levels between malignant and normal plasma cells for each patient, we were able to identify differentially expressed genes per tumor and assess how frequently each gene is dysregulated across the cohort. This approach can provide a rationale for the prioritization of target validation and can potentially uncover clinically meaningful differences between disease stages. As hypothesized, our analysis recovered established markers of plasma cell malignancy that are useful diagnostically, such as the downregulation of CD27 (rank 1), CD99 (rank 2), and CD81 (rank 5), or therapeutically, such as the upregulation of GPRC5D (rank 9). Moreover, we discovered frequent downregulation of SH3BGRL3 (rank 3), a gene associated with cell migration, and SSR4 (rank 9), SELENOK (rank 11), OSTC (rank 15), and SPCS3 (rank 18), genes associated with protein folding and the regulation of endoplasmic reticulum (ER) stress. Interestingly, PDK1 (rank 17), an inhibitor of pyruvate dehydrogenase that is highly expressed in normal plasma cells, was frequently downregulated in malignant plasma cells, suggesting that the balance between oxidative phosphorylation and aerobic glycolysis may be different in malignant plasma cells. The most common upregulated genes were the oncogene MLLT3 (rank 1), the growth factor IGF1 (rank 2), and the proteasome regulator TJP1 (rank 3). By comparing the frequency of dysregulation between patients with SMM and MGUS, we observed that malignant plasma cells from patients with SMM were more likely to upregulate MYC, which is concordant with the known increase in MYC genetic alterations as the disease progresses. Furthermore, patients with SMM are more likely to show upregulation of CD47, which promotes immune evasion. Lastly, patients with MM were more likely to exhibit upregulation of the RNA helicase DDX21, the therapeutic target SLAMF7, the ER factor TXNDC11, and NFkB signaling factors PIM2 and IRF9, compared to patients with SMM. Discussion In conclusion, by leveraging the largest single-cell RNA-sequencing cohort of bone marrow samples from patients with MGUS and SMM, we uncovered novel insights into plasma cell malignancy, as well as differences in gene expression dysregulation between disease stages. Observations gleaned from this dataset could be used to nominate novel targets and prioritize target validation for the development of novel therapeutics for the treatment or prevention of MM.

Abstract 2962: Investigation of the relationship between obesity, weight cycling, and tumor progression in a murine myeloma model

Abstract Background: The purpose of this study was to investigate physiologic effects of body weight regulation patterns and myeloma progression in vivo. Obesity and obesogenic behaviors are positively associated with both monoclonal gammopathy of unknown significance and multiple myeloma (MM). As the only known modifiable risk factor, this association has emerged as an optimal target for MM prevention, but little is known about the mechanistic relationship of weight patterns with MM progression. In two large prospective cohorts, we have reported an association of episodes of weight cycling (WC), e.g., intentional weight loss followed by unintentional weight gain, with increased MM risk compared to weight-stable individuals (Marinac et al. JNCI Cancer Spectrum, 2019). Experimental Procedures: We used two murine MM models (C57BL/6J Vk*Myc and SCID-Beige MM.1Sluc+ xenograft); male mice were fed a control diet (CD; 10% kcal from fat), high fat diet (HFD; 60% kcal from fat), or a diet cycling regimen (DC; 4 weeks on CD then 4 weeks on HFD) for 16 weeks and thereafter injected them intravenously with tumor cells (1x106 Vk*Myc or 2x106 MM.1S). Body composition was assessed via PIXIMUS-DEXA (SCID-Beige) or NMR (C57BL/6J). Tumor burden was assessed via CD138+ FACS (Vk*Myc) or in vivo bioluminescent imaging (IVIS; MM.1S). Results: The C57BL/6J mice (n=20) on HFD consistently gained weight and sustained the gains throughout the study duration; mice on the DC regimen exhibited dramatic weight cycling responses to diet switching. We observed significant differences in body fat (%) between HFD and CD mice, as assessed by NMR (p&amp;lt;= 0.0001), while body fat fluctuated with diet in the DC mice. At the time of tumor cell injection, HFD mice had higher body weights versus CD (p&amp;lt;0.000001) and DC (p&amp;lt;0.000001) mice; however, weights of DC mice were comparable to CD despite weight fluctuation throughout the study. Three weeks after MM inoculation, we detected elevated tumor burden in DC mice vs. CD mice (n-10, p=0.0061) and a trend toward elevated tumor burden in HFD mice (p=0.145). C57BL/6J mice on the DC regimen also had greater Vk*Myc tumor burden (p=0.0413) within the bone marrow (BM), specifically. In the SCID-Beige MM.1Sluc+ model (n=10), we observed similar trends for weight gain and cycling, and greater early tumor burden in both the HFD (p&amp;lt;0.0001) and DC (p=0.0214) groups compared to CD. Tumor burden was also elevated (p=0.0004) in the heaviest mice (vs. the lightest), regardless of diet, later in the SCID-Beige study. Conclusions: Combined, these results suggest that HFD and WC may contribute to MM risk via changes in the BM microenvironment that affect MM cell homing, engraftment and early expansion. This study lays the groundwork for the future characterization of the relationship between weight patterns, the BM, and MM pathogenesis, which may affect MM prevention and treatment strategies. Citation Format: Heather Fairfield, Catherine Marinac, Mariah Farrell, Brenda Birmann, Michaela Ruth Reagan. Investigation of the relationship between obesity, weight cycling, and tumor progression in a murine myeloma model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2962.

Impact of Natural Dietary Agents on Multiple Myeloma Prevention and Treatment: Molecular Insights and Potential for Clinical Translation.

Chemoprevention is based on the use of non-toxic, pharmacologically active agents to prevent tumor progression. In this regard, natural dietary agents have been described by the most recent literature as promising tools for controlling onset and progression of malignancies. Extensive research has been so far performed to shed light on the effects of natural products on tumor growth and survival, disclosing the most relevant signal transduction pathways targeted by such compounds. Overall, anti-inflammatory, anti-oxidant and cytotoxic effects of dietary agents on tumor cells are supported either by results from epidemiological or animal studies and even by clinical trials. Multiple myeloma is a hematologic malignancy characterized by abnormal proliferation of bone marrow plasma cells and subsequent hypercalcemia, renal dysfunction, anemia, or bone disease, which remains incurable despite novel emerging therapeutic strategies. Notably, increasing evidence supports the capability of dietary natural compounds to antagonize multiple myeloma growth in preclinical models of the disease, underscoring their potential as candidate anti-cancer agents. In this review, we aim at summarizing findings on the anti-tumor activity of dietary natural products, focusing on their molecular mechanisms, which include inhibition of oncogenic signal transduction pathways and/or epigenetic modulating effects, along with their potential clinical applications against multiple myeloma and its related bone disease.

Investigation of the relationship between obesity, weight cycling, and tumor progression in a myeloma xenograft model

Obesity and obesogenic behaviors are positively associated with both monoclonal gammopathy of unknown significance and multiple myeloma (MM). As one of the only modifiable risk factors, this association has emerged as an optimal target for MM prevention, but little is known about the mechanistic relationship of weight patterns with MM disease progression. Preliminary data from epidemiologic studies suggest that repeated episodes of weight cycling (WC), resulting from intentional weight loss followed by unintentional weight gain, leads to increased risk of MM compared to weight-stable individuals.

Elucidating Under-Studied Aspects of the Link Between Obesity and Multiple Myeloma: Weight Pattern, Body Shape Trajectory, and Body Fat Distribution.

BackgroundAlthough obesity is an established modifiable risk factor for multiple myeloma (MM), several nuanced aspects of its relation to MM remain unelucidated, limiting public health and prevention messages.MethodsWe analyzed prospective data from the Nurses’ Health Study and Health Professionals Follow-Up Study to examine MM risk associated with 20-year weight patterns in adulthood, body shape trajectory from ages 5 to 60 years, and body fat distribution. For each aforementioned risk factor, we report hazard ratios (HRs) and 95% confidence intervals (CIs) for incident MM from multivariable Cox proportional-hazards models.ResultsWe documented 582 incident MM cases during 4 280 712 person-years of follow-up. Persons who exhibited extreme weight cycling, for example, those with net weight gain and one or more episodes of intentional loss of at least 20 pounds or whose cumulative intentional weight loss exceeded net weight loss with at least one episode of intentional loss of 20 pounds or more had an increased MM risk compared with individuals who maintained their weight (HR = 1.71, 95% CI = 1.05 to 2.80); the association was statistically nonsignificant after adjustment for body mass index. We identified four body shape trajectories: lean-stable, lean-increase, medium-stable, and medium-increase. MM risk was higher in the medium-increase group than in the lean-stable group (HR = 1.62, 95% CI = 1.22 to 2.14). Additionally, MM risk increased with increasing hip circumference (HR per 1-inch increase: 1.03, 95% CI = 1.01 to 1.06) but was not associated with other body fat distribution measures.ConclusionsMaintaining a lean and stable weight throughout life may provide the strongest benefit in terms of MM prevention.

MicroRNA-765 is pregulated in multiple myeloma and serves an oncogenic role by directly targeting SOX6.

Increasing evidence has revealed that microRNAs (miRNAs) are closely associated with multiple myeloma (MM) pathogenesis and progression. Therefore, an in-depth understanding of the biological functions of miRNAs in MM may be helpful for the identification of promising therapeutic techniques for patients with MM. miRNA-765 (miR-765) has been reported to be dysregulated in many types of human cancer. However, the expression pattern, specific roles and underlying mechanisms of miR-765 in MM remain largely unknown. In the present study, plasma miR-765 significantly increased in patients with MM and cell lines. The downregulation of miR-765 in MM cells attenuated proliferation and promoted apoptosis. Bioinformatics analysis predicted that SRY-Box 6 (SOX6) was a putative target of miR-765. This was experimentally verified using a luciferase reporter assay, reverse transcription-quantitative PCR and western blot analysis. Furthermore, plasma SOX6 was downregulated in patients with MM and the downregulation of SOX6 was inversely correlated with that of miR-765 expression. Furthermore, SOX6 knockdown markedly abrogated the effects of miR-765 underexpression on cell proliferation and apoptosis in MM. The current study demonstrated that miR-765 serves an oncogenic role in MM progression by directly targeting SOX6, suggesting that miR-765 may be a potential therapeutic target for MM prevention and treatment.

Public Health Approach on Multiple Myeloma Prevention in Nigeria

Public Health Approach on Multiple Myeloma Prevention in Nigeria

Multiple myeloma and physical activity: a scoping review

ObjectivesMultiple myeloma is the second most common haematological cancer. A growing body of literature is emerging that investigates the role physical activity plays in all stages of multiple myeloma (prevention...

Identification of the key genes connected with plasma cells of multiple myeloma using expression profiles.

ObjectiveTo uncover the potential regulatory mechanisms of the relevant genes that contribute to the prognosis and prevention of multiple myeloma (MM).MethodsMicroarray data (GSE13591) were downloaded, including five plasma cell samples from normal donors and 133 plasma cell samples from MM patients. Differentially expressed genes (DEGs) were identified by Student’s t-test. Functional enrichment analysis was performed for DEGs using the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. Transcription factors and tumor-associated genes were also explored by mapping genes in the TRANSFAC, the tumor suppressor gene (TSGene), and tumor-associated gene (TAG) databases. A protein–protein interaction (PPI) network and PPI subnetworks were constructed by Cytoscape software using the Search Tool for the Retrieval of Interacting Genes (STRING) database.ResultsA total of 63 DEGs (42 downregulated, 21 upregulated) were identified. Functional enrichment analysis showed that HLA-DRB1 and VCAM1 might be involved in the positive regulation of immune system processes, and HLA-DRB1 might be related to the intestinal immune network for IgA production pathway. The genes CEBPD, JUND, and ATF3 were identified as transcription factors. The top ten nodal genes in the PPI network were revealed including HLA-DRB1, VCAM1, and TFRC. In addition, genes in the PPI subnetwork, such as HLA-DRB1 and VCAM1, were enriched in the cell adhesion molecules pathway, whereas CD4 and TFRC were both enriched in the hematopoietic cell pathway.ConclusionSeveral crucial genes correlated to MM were identified, including CD4, HLA-DRB1, TFRC, and VCAM1, which might exert their roles in MM progression via immune-mediated pathways. There might be certain regulatory correlations between HLA-DRB1, CD4, and TFRC.

Dual Face of Immune Microenvironment in Myeloma

An important property of human multiple myeloma (MM) is the growth of tumor cells and their precursors in the bone marrow microenvironment. This specialized microenvironment includes a distinct component of immune cells that includes cells of innate as well as adaptive immune system. Growing body of evidence points to a central role for cross-talk between this immune microenvironment and tumor cells in regulating the growth of tumor cells in myeloma. These interactions play a dual role-with some interactions promoting tumor growth while others suppress growth of tumor cells. Cross talk between T and B cells has been implicated in normal B/plasma cell differentiation and may also play a role in pathogenesis of myeloma, explaining increased risk of plasma cell malignancies in certain settings such as Gaucher disease. The bone marrow of MM patients is often infiltrated with innate cells such as dendritic cells (DCs) and cross-talk between myeloid/plasmacytoid DCs or macrophages may promote growth and survival of MM cells. This cross-talk may also have fundamental implications for inducing genomic instability in tumor cells, such as via the induction of cytidine deaminases in tumor cells. Genomic instability is an early feature of human MM and many of the mutations found in MM tumor cells are also present in the precursor states. The immune system has the capacity to recognize these precursor lesions and even in the setting of clinical MM, the tumor bed contains T cells that can be recruited to kill tumor cells. Specific targets of tumor immunity may differ between preclinical and clinical malignancy. Immunity to certain stem cell-associated antigens are in particular associated with reduced risk of progression to clinical MM. With advanced disease, tumor infiltrating T cells encounter several immune inhibitory checkpoints which leads to suppression of function of tumor-infiltrating T cells. In addition to T cells, cells of the innate immune system such as natural killer and natural killer T cells may also be attractive targets of protective tumor immunity in MM. Immunologic approaches may also be combined with other strategies such as other immunomodulatory drugs (IMiDs) already in clinical use in MM. Recent advances in mouse modeling provide new opportunities to better understand how to harness the immune system against human MM. Such approaches have the advantage of the potential for durable control of tumor cells via immunologic memory. Such approaches are now entering the clinic as attractive targets to restore anti-tumor immunity, both in the context of therapy or prevention of MM. Disclosures Dhodapkar: Celgene: Research Funding. Off Label Use: checkpoint blockade drugs, imids.

Abstract LB-48: Cooperation of NF-κB, STAT3 and p300 on the growth and survival of plasma-cell neoplasia

Abstract Collaboration of pro-inflammatory cytokine, interleukin 6 (IL-6), with oncogenic transcription factor, MYC, has been implicated in the natural history of human B-cell and plasma-cell neoplasms such as multiple myeloma (MM) and Waldenström's macroglobulinemia (WM). To generate a mouse model of IL-6/MYC-driven B-lineage tumors, we intercrossed strain C.H2-Ld-IL6 mice, which carry a human IL6 cDNA gene under control of the major histocompatibility complex (MHC) H2-Ld promoter, with strain C.iMycEμ and C.iMycCα mice, which harbor a gene-insertion mutation that recapitulates the chromosomal T(12;15) translocation leading to deregulated expression of MYC in mouse plasmacytoma (PCT). Double-transgenic C.IL6/iMycEμ and C.IL6/iMycCα mice develop PCT with full penetrance (100% tumor incidence), short latencies (3–6 months) and remarkable phenotypic consistency. To test the hypothesis that nuclear factor-κB (NF-κB) signaling plays a role in malignant plasma-cell transformation in these mice, we performed electrophoretic mobility shift assays (EMSA) using splenic B cells from C.IL6/iMycΔEμ mice. We found that the splenocytes contained elevated levels of NF-κB and signal transducer and activator of transcription 3 (STAT3) DNA-binding activities compared to normal splenic B cells used as control. Super-shift assays demonstrated that in addition to p50, p65 and c-Rel (indicators of canonical NF-κB pathway activation), RelB was involved. This suggested the additional activation of alternative NF-kB signaling. NF-kB, pSTAT3 and co-transcription factor, p300, were shown to be physically associated in transgenic B cells. Ongoing studies are aimed at elucidating whether NF-κB/STAT3/p300 signaling underlies up-regulation of c-myc level in the course of PCT development. Using qPCR we also found that p21 – which is usually known as a tumor suppressor and direct target of NF-κB, STAT3, p53 and MYC – exhibited increased p53-independent expression in B cells obtained from C.IL6/iMycΔEμ mice, but not from C.IL6 or iMycΔEμ mice. In contrast, the expression of Aurora kinase A, a direct upstream regulator of both NF-kB and E2F1, was strongly up-regulated in B cells from C.IL6/iMycΔEμ, C.IL6 and iMycΔEμ mice. Likewise, E2F1, a direct upstream regulator of p21, and E2F3, a direct upstream regulator of both p21 and Aurora kinase A, were upregulated. Our findings suggest that p21 is up-regulated, either directly or indirectly, through Aurora kinase A and/or NF-κB/STAT3/p300 complex. Furthermore, they indicate the usefulness of the newly developed IL-6/MYC mouse tumor model for identifying potential targets for the treatment and prevention of MM and WM. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-48. doi:10.1158/1538-7445.AM2011-LB-48

Abstract B138: Disappearance of “M” component connected with cow’s milk protein-free diet

Abstract B138 The presence of “M” component, as monoclonal gammopathy of undetermined significance (MGUS) in many patients with psoriatic arthritis and in 5% healthy people older than 70 years is well known. Besides, it was stressed out that patients with psoriatic arthritis and MGUS can sometimes develop multiple myeloma. Serum from patient 83 years old, with psoriatic arthritis diagnosed at 2000 was analyzed. Determination of the total IgG, IgA and IgM immunoglobulins was done by nephelometry using commercial (Binding Site) tests. Serum protein electrophoresis was done on gel electrophoresis in barbital buffer, pH=8.6; this was followed by immunofixation using sheep antihuman antibodies (Binding Site) to light and heavy chains of immunoglobulins. Determination of serum IgA and IgG immunoreactivity to gliadin in (IU/ml) was done using commercial Binding Site ELISA test, while serum immunoreactivity to cow’s milk protein (CMP) in (AU/ml) was done by home made ELISA test, using cows milk pasteurized powder (ICN Biomedicals, Inc.) as the antigen and sheep antihuman IgA and IgG, HRP labeled antibodies (Binding Site) as secondary antibodies. Blocker was 1% bovine serum albumin. Results obtained showed the enhanced IgA and IgG immunoreactivity to CMP. The analysis of serum by electrophoresis and immunofixation reveals the presence of monoclonal IgG (λ) immunoglobulin. Upon the knowledge that the enhanced immunoreactivity especially to CMP exists the patient decided to consume the food without cow’s milk proteins. About month and half after the start of CMP free diet he has done his blood for the analysis of serum proteins again. In that time of period the patient has not taken any immunosuppressive drugs. Electrophoresis of patient sera reveals that after the CMP free diet the “M“component disappeared. These data are supported by the findings that serum anti-CMP IgG dropped from 892 AU/ml to 0 AU/ml (anti-CMP IgA also dropped from 240 AU/ml to 0 AU/ml) Besides, concentrations of total serum proteins were diminished after the CMP free diet from 73 g/l to 69 g/l, while concentration of IgG was decreased from 17.8 g/l to 11.2 g/l. Our report shows that there could be a connection between diet restricted for immunogenic protein(s) and disappearance of MGUS. Our results are along with the data from only one report dealing with a possible link between disappearance of MGUS and gluten free diet. They open the question whether the special, appropriate diet restriction regime could help in the prevention of multiple myeloma. Citation Information: Cancer Prev Res 2008;1(7 Suppl):B138.

Phenotypic Heterogeneity in Multiple Myeloma Families

To describe a series of families with familial multiple myeloma (MM). Observations were used to generate hypotheses about the role of genetic factors, the mode of inheritance of these factors, and the association of other cancers with familial MM. This observational study consisted of 39 families with multiple cases of MM or related disorders from four collaborating research centers. Each center followed its usual family study method. Probands were interviewed, and, when possible, cancers were verified by medical records and pathology review. A working pedigree was compiled on each family. Seventeen families had affected members in two or more generations, and eight families had two or more affected members in a single generation. Four families had two or more members with plasma cell dyscrasias, with or without a single case of MM. In the remaining 10 families, a single MM case occurred with a family history of other cancers. Other cancers observed in family members included hematologic malignancies and solid tumors. In families with MM in multiple generations, there was a decrease in the age at MM diagnosis in successive generations. The study of familial MM may provide insights into the pathogenesis and, ultimately, the control and prevention of MM and related disorders. Population-based epidemiologic studies are crucial, but because of the rarity of familial MM, a concerted case-finding approach may also be fruitful. Therefore, we propose an international consortium to study familial MM, and we invite all interested colleagues to participate.

Familial multiple myeloma: a family study and review of the literature.

The etiology of multiple myeloma (MM) remains obscure, although reports of familial clustering have implicated both a host susceptibility factor and environmental effects. Here we describe the medical histories of members of a family prone to MM. We developed a pedigree for an MM-prone family by using information obtained from a questionnaire. Protein immunoelectrophoresis of serum and urine from the proband and from 19 family members was performed to detect monoclonal immunoproteins. Peripheral blood obtained from the proband and from five relatives was subjected to standard cytogenetic studies to detect constitutional chromosomal abnormalities. Multifluor-fluorescence in situ hybridization (M-FISH) and standard FISH studies were performed on peripheral blood from the proband and from two other affected living relatives to determine their karyotypes and to detect clonal chromosomal abnormalities frequently seen in patients with MM. Within this family, a sibship of seven included three individuals (including the proband) with histologically verified MM and two individuals with a monoclonal gammopathy of unknown significance (MGUS), as determined by immunoelectrophoresis of serum and urine. This family also had members with acute lymphocytic leukemia, malignant melanoma, and prostate cancer. In the family members tested, we detected no constitutional chromosomal abnormality. None of the three individuals analyzed by FISH had a deletion of the retinoblastoma (Rb-1) locus, which is frequently deleted in patients with MM, and only one (the proband) had a translocation involving chromosomes 11 and 14, a clonal abnormality commonly seen in MM. The study of familial MM may provide insights into the pathogenesis and, ultimately, the control and prevention of MM and related disorders.