bluuebunny/crossref_metadata_2025_split huggingface dataset:
18F-FDG-PET/CT and Osteolytic Bone Disease in Multiple Myeloma
Authors: Østergaard Brian , Mortensen Julie R. , Nielsen Anne L , Asmussen Jon T. , Gerke Oke , Plesner Torben , Høilund-Carlsen Poul Flemming , Abildgaard Niels 
Published: November 2019

Abstract
Introduction: FDG-PET/CT is a promising methodology for staging, prognostication, and response evaluation in multiple myeloma (MM). The number of focal FDG lesions and the intensity of FDG uptake (standard uptake value, SUV) at diagnosis are informative of disease aggressiveness. Osteolytic bone disease is a hallmark of MM. Hypothetically; increased focal metabolic activity will precede, induce and correlate to osteolytic lesions. We aimed to elucidate the association between focal FDG positive lesions and osteolytic bone disease in MM patients.
Methods: Twenty-two newly diagnosed MM patients, 16 males and 6 females aged 53-81years, were prospectively enrolled and studied with a standardized baseline FDG-PET/CT at diagnosis. A nuclear medicine and a radiology specialist evaluated the PET and CT parts separately and independently. Focal FDG positive lesions were assessed by dedicated software (ROVER™, ABX, Germany) to yield volume and SUV measures. All osteolytic lesions identified on CT were noted by size and localization.
Results: FDG-PET and CT together identified a total of 390 lesions. We found more osteolytic lesions on CT (335) than FDG-positive lesions (169); the concordance between the modalities was 30%. 34% (114/335) of lytic lesions were FDG-positive, whereas ⅔ (114/169) of focal FDG-PET positive lesions were lytic according to CT. We found a significant correlation between higher FDG uptake measured as SUVpeak and osteolytic lesions >10 mm, and focal lesions with high FDG uptake had more pronounced osteolysis than focal lesions with less FDG uptake. Still sparse follow-up data did not allow analysis of the predictive value of focal PET positivity and the risk of development of future osteolysis. More compulsive data will be presented at the meeting.
Conclusion: FDG-PET/CT offers dual information including metabolic activity (FDG-avidity) and degree of lytic bone disease (low dose whole body CT) in MM. About ⅔ (221/335) of osteolytic lesions were FDG-negative, whereas ⅔ (114/169) of FDG-positive lesions had a lytic component.

Disclosures
Plesner: Janssen: Membership on an entity's Board of Directors or advisory committees, Research Funding.
Interleukin-6 Drives Multiple Myeloma Progression By up-Regulating of CD147/Emmprin Expression
Authors: Hu Jinsong , Lei Li , Wang Yanmeng , Wang Ke , Hu Xiaoyan , Wang Aiying , Vanderkerken Karin 
Published: November 2019

Abstract
Multiple myeloma (MM) is a B-cell malignancy characterized by the clonal proliferation of plasma cells in the bone marrow (BM). CD147, also known as extracellular matrix metalloproteinase inducer (EMMPRIN), is a type I transmembrane glycoprotein that belongs to the immunoglobulin superfamily. CD147 is expressed in a variety of tissues, and is involved in a number of physiological and pathological processes. With regard to MM, it was recently documented that the aberrantly elevated expression of CD147 has been tightly correlated with MM cell colonization and proliferation. However, it is still unclear what mechanism is involved in the dysregulation of CD147 expression in MM. In this study, we found that CD147 can be up-regulated by interleukin-6 (IL-6), which is derived from either autocrine or paracrine sources and plays an essential role in the malignant progression of MM. When serum-starved MM cell lines RPMI8226, MM1.S and NCIH929 were stimulated with 20ng/mL of IL-6 for 24 hours, we found that the expression of CD147 on MM cell membrane was significantly up-regulated, as determined by flow cytometric analysis. Moreover, when using the IL-6 autocrine MM cell line U266, CD147 level was found to be continuously increased in serum-starved MM cells in a time-dependent manner. Interestingly, the increase of CD147 in U266 cells was found to be abrogated when using neutralizing antibody to block the secreted IL-6. Next, we investigated the molecular mechanisms involved in the IL-6-mediated upregulation of CD147 in MM cells. We confirmed that IL-6 can activate the JAK/STAT3 signaling pathway, inducing the phosphorylation of STAT3 at Tyrosine 705 in RPMI8226 and MM1.S cells. Further studies using ChIP analysis demonstrated that there is a STAT3 binding site at the position −659 to −650 in the promoter of the CD147 gene. In addition, when inhibiting of STAT3 phosphorylation with a specific inhibitor S3I-201, the CD147 expression in MM cells was accordingly decreased. Finally, we found that siRNA mediated knock-down of CD147 in RPMI8226 and MM1.S cells can decrease the proliferation and migration of MM cells induced by IL-6 and Cyclophilin A (CypA). In conclusion, our data provide evidence that IL-6 can up-regulate CD147 expression in a STAT3-dependent manner, and offer a compelling rationale for exploring this axis as a therapeutic target for MM.

Disclosures
No relevant conflicts of interest to declare.
Molecular and Biological Characteristics of Tumour Substrate in Patients with Extramedullary Multiple Myeloma
Authors: Firsova Maya V. , Mendeleeva Larisa P. , Kovrigina Alla M. , Deineko Natalia L. , Solovev Maxim V. , Nareyko Maria V. , Kuzmina Larisa A. , Savchenko Valery G. 
Published: November 2019

Abstract
Introduction. At the time of the diagnosis of MM the incidence of extramedullary disease (EMD) varies from 7 to 18%, and in case of à relapse, it is observed in more than 20% of patients. Clinical progression of MM with EMD is characterised by highly aggressive tumour, worse prognosis and often by resistance to therapy, low overall survival rate.
Aim of the study. To evaluate and to compare molecular and biological characteristics of tumour cells in bone marrow and extramedullary plasmacytoma in primary MM patients.
Materials and methods. The study enrolled 28 patients with primary MM (18 males, 10 females) 23-59 years old (median value: 52 years old). The disease was diagnosed in accordance with the IMWG criteria (2014). All patients underwent bone marrow trephine biopsy. 13 patients had EMD including 10 patients with soft-tissue plasmacytomas accompanied by cortical layer involvement and the tumour mass spreading from bone. 3 patients had extramedullary foci in stomach, brain, soft submandibular tissues. In all cases a tumour biopsy was taken which confirmed the presence of plasma cell infiltration. Paraffin block slices from trephine biopsy material and tumour biopsy material were used to perform an immunohistochemistry analysis with an antibody panel to CD56, Ki67, CXCR4, CD 166.
Results. The immunohistochemistry analysis revealed more frequent expression of CD56 (80% vs. 38.5%, p=0.05) and high Ki-67 expression (53.3% vs. 0%, p = 0.002) in bone marrow of MM patients without EMD in comparison with those with plasmacytomas. At the same time patients without EMD demonstrated CXCR4 expression by myeloma cells in bone marrow, that was significantly higher compared with MM patients with plasmacytomas (100% vs. 46.2%, p = 0.001) (Table 1).
Immunohistochemical parameters of plasma cells in bone marrow and plasmacytoma of MM patients with EMD were compared. A higher level of CD56, Ki-67, CXCR4 expression in plasma cells from tumour biopsy was observed, although the differences were not statistically significant. The CD166 expression level in bone marrow and plasmacytoma was similar.
Conclusion. Immunohistochemistry analysis of the tumour substrate revealed that MM patients with and without EMD demonstrate different molecular and biological characteristics of bone marrow. Lack of CXCR4 expression in bone marrow may be the evidence of poor prognosis for the MM patients with the higher risk of EMD occurrence.

Disclosures
No relevant conflicts of interest to declare.
The Atypical Gtpase Rhou Lies Downstream IL6/STAT3 and Regulates Myeloma Plasma Cells Adhesion/Motility
Authors: Nunes Canovas Sara , Carrino Marilena , Manni Sabrina , Quotti Tubi Laura , Manzoni Martina , Barilà Gregorio , Trentin Livio , Zambello Renato , Neri Antonino , Semenzato Gianpietro , Piazza Francesco 
Published: November 2019

Abstract
RhoU is an atypical G protein of the Rho GTPase family endowed with a very high intrinsic guanine nucleotide exchange activity ensuring a prevalent GTP-loaded conformation. It is present at very low levels in most cells, however some reports have shown that the Wnt-1 and STAT3 pathways might regulate its expression which resulted highly increased in malignant tumors. Since the IL6R/STAT3 signaling is of critical in Multiple Myeloma (MM) growth, homing and adhesion to marrow stroma and RhoU GTPase may mediate the induction of filopodia formation and stress fiber dissolution, we aimed at studying the role of this protein in MM malignancy.
mRNA expression studies unraveled that MM patients have heterogeneous levels of RhoU ranging from -0.7 fold change to 18.5 fold change when normalized over controls. By analyzing available gene expression data sets, we found that RhoU expression changes were associated with distinctive expression of 557 genes. We also found a correlation between RhoU levels and genetic abnormalities that associate with a poor prognosis in MM patients, namely the t(4;14), del(13) and 1q gain. RhoU was positively modulated by IL-6 and by soluble factors from bone marrow stromal cells' culture. Its expression was down-modulated after STAT3 inhibition with the chemical inhibitor Stattic, even in the presence of IL-6. The treatment with Stattic also decreased MM cells' migration capability. Remarkably, RhoU knock down by siRNA caused changes in MM cells' phenotype and deranged adhesion/migration functional properties. Lastly we have looked into changes in RhoU expression after the treatment of MM cells with Lenalidomide, known to act on MM microenvironment and alter MM cells' adhesion to protective bone marrow niches. Surprisingly, treatment of MM cell lines or from patients with Lenalidomide led to an upregulation of RhoU expression in the cells before onset of apoptosis after 72 hours.
Our data point to a functional role in cell adhesion, migration and shape for RhoU GTPase in a subset of MM, in particular in cases with genetic abnormalities correlating with a poor prognosis. RhoU is downstream from STAT3/IL-6 and likely from growth factor delivered by bone marrow stromal cells. The upregulation of RhoU expression after treatment with Lenalidomide was associated to a stall of the cellular cytoskeleton and subsequent apoptosis. Thus RhoU might be important in MM pathogenesis and could become a suitable target to disrupt the MM plasma cell interaction with protective bone marrow niches.

Disclosures
No relevant conflicts of interest to declare.
Rigosertib, a Pan RAS Inhibitor, Decreases Mouse and Human Myeloma Cell Growth in Preclinical Models
Authors: Zangari Maurizio , Shin Ikjae , Branca Antonio , Yoo Hanna , Fallo Sophia , Davies Faith E , Hoffman Benjamin , Fruchtman Steven M. , Yoon Donghoon 
Published: November 2019

Abstract
Introduction
Multiple myeloma (MM) is a B cell cancer mainly characterized by proliferation of malignant plasma cells in the bone marrow, presence of a monoclonal serum immunoglobulin, and occurrence of osteolytic lesions. RAS genes encode a family of small GTPase proteins ubiquitously expressed in all cell lineages and organs that transmit signals for cell growth, differentiation, and survival. Mutations in RAS genes often lead to constitutively activated RAS signaling, resulting in cancer. Various RAS mutations have been described in myeloma patients. RAS mutations are rarely found in pre-malignant stages as MGUS (monoclonal gammopathy of undetermined significance). These findings suggest that RAS mutations may play an important role in malignant transformation of MM.
Rigosertib (ON 01910.Na) is an inhibitor of RAS-related signaling pathways associated with cancer cell division, growth, and survival. Rigosertib binds directly to the RAS-binding domain in RAS effector proteins resulting in disruption of RAS/effector interactions and inhibition of multiple RAS-driven signaling pathways. Clinical trials with rigosertib have been conducted in MDS and solid tumors, but the effect of rigosertib in MM has not yet been fully characterized.
Methods
Various mouse/human MM cell lines (5TGM1, KMS28, RPMI8226, JJN3, OPM2) were tested for their growth with treatment of rigosertib at different concentrations (1, 10, 50, 100, 500 nM) for 4 days.
For a mouse MM model study, 0.5e6 Luciferase infected 5TGM1 cells were injected via tail vein into C57BL6/KaLwRij mouse. Rigosertib (100 mg/kg QD) was administrated via intraperitoneal injection from 14 days post-transplantation. The MM growths were weekly monitored by in vivo IVIS imaging after luciferin (Luciferase substrate) injection.
Results
All cells (5TGM1, KMS28, RPMI8226, JJN3, OPM2) tested showed growth inhibition with rigosertib in a dose-dependent manner. MM progression assessed by in vivo imaging was found to be significantly suppressed by rigosertib treatment as shown in Figure. Furthermore, a survival analysis demonstrated a positive trend towards increased life expectancy with rigosertib treatment (Median survival; control (35 days) vs treated (44.5 days)).
Conclusion
Our results demonstrate that rigosertib suppresses MM growth in both human/mouse cell lines and a mouse MM model and suggest additional exploration of rigosertib alone or in combination with other MM treatments.
Figure 1 Figure 1.

Disclosures
Davies: Takeda: Consultancy, Honoraria; Celgene: Consultancy, Honoraria; Janssen: Consultancy, Honoraria. Hoffman:Onconova: Employment. Fruchtman:Onconova: Employment.
Repurposing Leflunomide for Multiple Myeloma Treatment
Authors: Buettner Ralf , Morales Corey , Perret Natalie , Palmer Joycelynne , Krishnan Amrita , Rosenzweig Michael , Rosen Steven T. 
Published: November 2019

Abstract
Background: Despite recent advances in treatment that have improved the prognosis for patients with multiple myeloma (MM), the disease remains incurable. There is a need for MM treatments with new mechanisms of action. Leflunomide, a commercially available oral immunosuppressive agent that has been FDA-approved since 1998 for the treatment of rheumatoid arthritis (RA) was evaluated as a potential MM therapy. The primary mechanism of action is de novo inhibition of pyrimidine synthesis by targeting dihydroorotate dehydrogenase (DHODH), and thus achieving an anti-proliferative effect in B- and T-lymphocytes. A secondary mechanism of action is inhibition of cytokine and growth factor receptor-associated tyrosine kinase activity. Methods and Results: Pre-clinical studies of teriflunomide, the active metabolite of leflunomide, showed that it inhibited cell growth and induced apoptosis in MM cell lines (MM.1S, MM.1R, U266, H929, RPMI-8226) and primary MM patients' (CD138+) plasma cells at clinically achievable concentrations (50-200 uM) in a time- and dose-dependent manner. We also found that teriflunomide induces cell-cycle arrest in both, glucocorticoid-sensitive (MM.1S) and resistant (MM.1R) MM cell lines at <200 uM. In addition, teriflunomide and dexamethasone synergized in the in vitro growth inhibition of MM cell line MM.1S. To identify MM-associated mRNAs and miRNAs whose expression levels are frequently altered upon teriflunomide exposure, MM cell lines (RPMI-8226, U266, MM.1S, NCI-H929) and CD138-enriched primary plasma cells from two MM patient samples were treated with 200 µM teriflunomide or DMSO control for 24 h before extraction and purification of mRNA and microRNA. mRNA-seq and miRNA-seq analysis from teriflunomide-treated MM samples revealed that similar changes were present between patient samples and cell lines. A total of 382 genes were found to be differentially expressed (225 upregulated, 157 downregulated). Upregulated genes included those that participate in defense response and negative regulation of cell growth. Genes involved in mitosis, rRNA biogenesis/processing, and immune response were generally downregulated. Analysis of microRNA-seq data from these samples revealed five differentially expressed, mostly newly discovered miRNAs that have unknown function.
Conclusions: Leflunomide and its analogues demonstrated anti MM effects in vitro as well as synergy with dexamethasone. Based on our promising pre-clinical results we have initiated a single-agent phase I/II clinical trial in patients with relapsed/refractory MM.

Disclosures
No relevant conflicts of interest to declare.
CD38 Deregulation As Strategy to Make Multiple Myeloma Cells More Sensitive to NAD+ Depleting Agents
Authors: Cea Michele , Cagnetta Antonia , Soncini Debora , Minetto Paola , Bergamaschi Micaela , Rivoli Giulia , Canepa Letizia , Gobbi Marco , Lemoli Roberto M 
Published: November 2019

Abstract
Background: The upregulated NAD+ biosynthesis, which is needed to face increased proliferation and metabolic processes, represents an important feature distinguishing cancer cells from their normal counterparts. As a result, the NAD+ biosynthetic apparatus emerges as highly promising therapeutic target for tumors, as suggested by the use of nicotinamide phosphoribosyltransferase (NAMPT) inhibitors in a number of malignancies, including Multiple Myeloma (MM) and leukemia (Cea et al. Blood 2012; Cagnetta et al. Blood 2013; Cagnetta et al. CCR 2015). The ecto-enzyme CD38 is emerging as a novel therapeutic target for patients with hematological malignancies, including MM, with several monoclonal antibodies already tested in clinical trials with promising results. Importantly, CD38 by regulating intracellular NAD+ stores acts as member of the intricate network supporting metabolic reprogramming associated with cancer. Based on these assumptions, here we explored CD38 activity as innovative strategy to enhance the anti-tumor activity of NAMPT inhibition in Multiple Myeloma cells.
Methods: A panel of different MM cell lines and primary cells, both sensitive and resistant to conventional and novel anti-MM therapies, was used in the study. The effects of Nampt inhibition was evaluated in presence of CD38-gene editing (loss/gain of approaches) by using CTG assay and Annexin-V/propidium iodide staining. Next, the anti-MM effects of chemicals affecting CD38 activity were also evaluated in combination with low doses of Nampt inhibitors. Mechanistic studies were performed with Western-blotting, lentivirus-mediated shRNAs and enzymatic assays.
Results and Conclusions: By using different approaches, we found that CD38 deregulation makes MM cells more vulnerable to NAD+ depleting agents. Ongoing mechanistic studies suggest the central role played by energetic metabolism in the observed synergism. Overall our data provide the mechanistic preclinical rationale to enhance anti-MM activity of Nampt targeting agents, in order to both overcome drug resistance and improve patients outcome.

Disclosures
Gobbi: Mundipharma: Consultancy, Research Funding; Takeda: Consultancy; Roche: Honoraria; Celgene: Consultancy; Gilead: Honoraria; Novartis: Consultancy, Research Funding; Janssen: Consultancy, Honoraria.
Safety and Eficacy of Low Dose- Bortezomib Plus Melphalan/Prednisone (velcadito protocol) in Elderly Patintes Not Eligible to Transplant in Newly Dignosed Multiple Myeloma. Restrospective Study of 26 Patients
Authors: Gusmão Breno Moreno , Blumenschein Danielle Isadora , Persona Ernesto Perez , Guinea De Castro Jose Maria , Lopes Iracema Esteves , Hamerschlak Nelson 
Published: November 2019

Abstract
Introduction: Protocols based on bortezomib have become the standard therapy in the elderly and not eligible for transplantation. The VISTA study showed OR 71%, CR 30%, regardless of adverse factors such as age, renal failure or cytogenetic changes, but the patient had a high rate neuropathy (44%). The Spanish group and the Italian group modified the dose of bortezomib achieving rates of CR 22% and 20%, reducing neurotoxicity 8% and 5% respectively.
We present the experience of 26 newly diagnosed patients treated with low-dose bortezomib associdados the melphalan / prednisone (Velcadito protocol).
Objective: The aim of this study is to assess the effectiveness and safety of low-dose bortezomib in elderly patients not eligible to transplant. We evaluated according to the criteria IMWG.
Results: We evaluated 26 patients treated at the Hospital Universitario de Alava (Spain) and Hospital Israelita Albert Einstein (Brazil) among 2004 to 2016. The patients received 1 cycle with bortezomib 1.3mg/m2 (days 1,4,8,11) associated with melphalan 9 mg/m2 (1- 4 days) and prednisone 60mg/m2 (1- 4 days) 28 days cycle. From the 2nd cycle the patient received low doses of bortezomib (1 mg / m2, days 1 and 4) no change in other drugs and treatment regimen.
The mean age was 78 years. 50% men/ 50% women. 2 patients had poor cytogenetics Pronóstico, 1 intermediate and 23 standard. According to the IMWG: ISS1 = 7 (26%) ISS 2 = 10 (38.5%), ISS3 = 6 (23%). 38,4% (n=10) of patientes had neuropathy but only 11,5%(n=3) grade 3-4. 61,5% (n=16) presented as neutropenia toxicity, 19,2% (n=5) with grade 3 (neutrophils <1000). 15,3% (n=4) had thrombocytopenia with grade greater than 3. Response rate was 100%, and if we consider at least PR was 92% (CR 38.5% VGPR 31%). The global survival had median of 80 months (95% confidence interval 57,8 - 102,1%) and the survival free of progression had median of 37 months (95% confidence interval 15 - 58,9%) with follow-up of 91 months.
Conclusion: The use of velcadito protocol is safe, very effective and seems to be a good option for elderly patients with newly diagnose MM. A pharmacoeconomic analysis is recommended to assess the impact of the cost of treatment. Being a prospective clinical trial where an analysis of this impact of all patients treated with subcutaneous bortezomib beyond the characteristics of this study.

Disclosures
No relevant conflicts of interest to declare.