bluuebunny/crossref_metadata_2025_split huggingface dataset:
Does AL Amyloidosis Have Unique Gene Expression Profile? Meta-Analysis Results
Authors: Kryukov Fedor , Kryukova Elena , Brozova Lucie , Pyszko Petr , Kufova Zuzana , Nemec Pavel , Kubiczkova-Besse Lenka , Jarkovsky Jiri , Hajek Roman 
Published: October 2019

Abstract
Background
Immunoglobulin light chain amyloidosis (AL amyloidosis) is a plasma cell dyscrasia characterized by deposition of amyloid fibrils in various organs and tissues, derived from monoclonal immunoglobulin light chains (LC), leading to organ dysfunction. Until now there is no systematic study of genomic expression of patients with AL amyloidosis compared to other monoclonal gammopathies.
Aim of this study was to reveal genomic expression differences between AL amyloidosis patients versus healthy donors (ND), monoclonal gammopathy of undetermined significance (MGUS), smouldering myeloma (SMM) and multiple myeloma (MM) patients.
Methods
Gene expression profiling data from GEO database (GSE6477, GSE24128) comprised of ND (n=15), MGUS (n=21), SMM (n=24), MM (n=69) and AL (n=16) patients were used. To ensure the comparability between arrays and eliminate the batch effect, quantile normalization was performed. To identify the AL patients´ gene expression profile which significantly differs in comparison with ND, MGUS, SMM or MM patients, we applied SAM algorithm for 6588 selected genes that fulfilled the condition having standard deviation above the median. Genes with fold change ≥1.5 or ≤0.5 and false discovery rate >0.001 were considered as significant. Prediction analysis of microarrays (PAM) using nearest shrunken centroids and cross validation assay was carried out to identify the smallest subsets of genes that were able to accurately predict classes.
Results
Comparison of AL patients vs. ND revealed 229 changed genes, AL vs. MGUS 126 genes, AL vs. SMM 338 genes and AL vs. MM 110 genes, respectively. Out of them, the most significantly changed 100 genes from each comparison were further analysed. Some of the genes occurred repeatedly, thus only 256 of genes from all comparisons were unique. Using PAM algorithm we found the genes that have the best predictive power for differentiation AL patients from ND, MGUS, SMM and from MM patients was as followed: RPS6, RPS14, RPS17, RPLA18A, TMEM66, EIF3L, CCDC72, GLTSCR2, NDUFS6, DUSP1, IGLJ3, IGHM, IGKV1-5IGKC///IGKV1-5///IGKV1D-8 and IGHA1///IGHA2///IGHD///IGHG1///IGHG3///IGHG4///IGHM/// IGHV3-23///IGHV4-31///IGHV4-59. Gene expression changes are presented on Figure 1.
Conclusion
Based on offered meta-analysis, 16 genes representing mostly ribosomal proteins and immunoglobulin regions allow us to detect specific aberrant clone responsible for pre-amyloid production and can serve as candidates for further detailed study.
Work was supported by grants IGA NT 12130, NT 14575 and NT 13190.
Figure 1. Gene expression for the genes which best discriminate AL patients from ND, MGUS, SMM and MM patients Figure 1. Gene expression for the genes which best discriminate AL patients from ND, MGUS, SMM and MM patients

Disclosures
Hajek: Merck: Consultancy, Honoraria; Celgene: Consultancy, Honoraria; Janssen: Honoraria.
Value Research of T-Regs Cells in Peripheral Blood of Patients with Multiple Myeloma
Authors: Golenkov Anatoly , Mitina Tatyana , Chernykh Yuliya , Lutskaya Tatyana , Mitin Alexandr , Litvina Marina 
Published: October 2019

Abstract
In this study we have assessed FOXP3 and its isoforms expression by CD4+ T cells from peripheral blood of 37 patients with various forms of multiple myeloma (MM) and 15 healthy volunteers. Newly diagnosed MM was in 17 patients; resistant/relapsed MM was registered in 13 patients and 7 patients had remission after chemotherapy. Significant role of FOXP3 expression by CD4+ T lymphocytes in pathogenesis of MM was shown. Thus, increase of FOXP3-expressing cells percent among CD4+ T cells is registered in patients with newly diagnosed MM and in resistant/relapsed patients. The main contribution to this percent increment is made by FOXP3D2 isoform expressing cells. In addition to the percent increment the number of CD4+FOXP3+ T cells is also risen in patients with newly diagnosed MM. The level of FOXP3 expression by CD4+ T cells also reflects the effectiveness of antitumor therapy, considering normalization of CD4+FOXP3+ T cells number and percent in patients with disease remission. We also discuss the possible relationship between FOXP3 mRNA post-transcriptional modification, namely alternative splicing, with stable expression of FOXP3 in T-regs cells.
The level of an expression of T-regs cells can will be applied as criterion of efficiency of antitumor therapy.

Disclosures
No relevant conflicts of interest to declare.
Leukemia-Initiating Activity in T-ALL Requires Active Hif1a and Wnt Signaling
Authors: Giambra Vincenzo , Jenkins Catherine E , Lam Sonya H , Hoofd Catherine , Belmonte Miriam , Weng Andrew P 
Published: October 2019

Abstract
Prior work has shown that NOTCH1 is a prominent oncogene in T-cell acute lymphoblastic leukemia (T-ALL) with activating NOTCH1 mutations occurring in over 50% of cases (Weng et al, Science 2004) and loss-of-function mutations in its negative regulator FBXW7 occurring in 8-15% of cases (O’Neil et al, J Exp Med 2007; Thompson et al, J Exp Med 2007). Subsequent work has shown that continued Notch signaling is required for maintenance of T-ALL leukemia stem cells (Armstrong et al, Blood 2009; Tatarek et al, Blood 2011; Giambra et al, Nat Med 2012).
Several lines of evidence have substantiated genetic interactions between the Notch and Wnt signaling pathways in various contexts, and Wnt signaling has been shown to play important roles in hematopoietic stem cell biology and also in hematopoietic cancers such as acute myelogenous leukemia (AML) and chronic myelogenous leukemia (CML) (reviewed in Luis et al, Leukemia 2012). To address what role if any Wnt signaling may play in T-ALL, we generated primary murine leukemias by viral transduction of bone marrow progenitors with activated NOTCH1, then delivered a fluorescent Wnt reporter construct (7TGP; Fuerer & Nusse, PLoS ONE 2010) by lentiviral transduction, and retransplanted the leukemias to interrogate Wnt signaling activity in vivo.
We report here that active Wnt signaling is restricted to minor subpopulations within bulk T-ALL tumors, and that these Wnt-active subsets are highly enriched for leukemia-initiating cell (LIC) activity. Moreover, using Ctnnb1loxP/loxP animals we show that inducible Cre-mediated deletion of β-catenin or enforced expression of a dominant-negative TCF construct severely compromises LIC activity. We also show that β-catenin levels are upregulated by hypoxia through Hif1a stabilization, and that deletion of Hif1a also severely compromises LIC activity. Interestingly, Wnt-active subsets are distributed diffusely throughout the marrow interstitial space suggesting that tumor infiltration induces formation of local hypoxic niches as opposed to taking up residence in pre-existing anatomic compartments with low oxygen tensions. Taken together, these results suggest a model in which hypoxic niches in vivo facilitate Hif1a-dependent accumulation of β-catenin which drives Wnt signaling and self-renewal of leukemia stem cells. Finally, we show using patient-derived xenografts that antagonism of Hif1a or Wnt signaling also compromises human LIC activity, suggesting that pharmacologic targeting of these pathways could have therapeutic application in patients with T-ALL.

Disclosures
No relevant conflicts of interest to declare.
High Serum Free Kappa Chains Are Frequently Missed By Serum Immunofixation
Authors: Sidana Surbhi , Rybicki Lisa , Reu Frederic J. , Daly Thomas 
Published: October 2019

Abstract
Background: Serum free light chains (sFLC) and immunofixation (IFE) analysis are used to detect monoclonal proteins. We noticed that some multiple myeloma (MM) patients (pts) had negative IFE results despite very high sFLC levels. This analysis was done to determine the frequency of this finding in a large cohort.
Methods: Following IRB approval, samples with simultaneous sIFE and sFLCs ordered from 1/2013 to 9/2013 were identified by querying our lab electronic database. Freelite (R) Human Kappa & Lambda Free kit (The Binding Site, Birmingham, UK) was used for sFLC and SPIFE® ImmunoFix-15 gels (Helena Laboratories, Beaumont, TX) for IFE. Clinical review was performed for a subset of patients with discrepant results between the two assays.
Results: 4404 samples from 2200 pts were identified with simultaneous sFLC and sIFE results. Overall 348 of 4404 (7.9%) samples had an abnormal sFLC ratio but a negative sIFE. Of 205 pts (457 samples) with involved serum free kappa above the expected IFE threshold (200 mg/L), 42 pts (103 samples) had negative sIFEs (20.5% pts; 22.5% samples) despite median free kappa of 556.6 mg/L (range 208.1 to 4954.4). This was much less common for involved free lambda. Only 4 of 107 pts with free lambda above 200mg/L had negative sIFEs (3.7%). Information of the nature of the plasma cell disorder was available on 40 of the 42 patients with free kappa > 200mg/L and negative IFE. In this group, 62.5% (n=25) had symptomatic MM, 12.5% (n=5) had asymptomatic MM, 5% (n=2) had light chain amyloidosis, 5% (n=2) had both amyloidosis and MM, and 15% (n=6) had MGUS or a not yet fully worked up plasma cell disorder.
Conclusions: Serum IFE, with a commonly used kit, missed about 20% of patients with free kappa above the expected IFE threshold, while free lambda was detected more reliably. Results suggest current criteria for complete remission which rely on immunofixation and bone marrow plasma cell infiltration may need to be revised for kappa light chain myeloma.

Disclosures
No relevant conflicts of interest to declare.
Reversibility of Renal Insufficiency in Patients with Newly Diagnosed Multiple Myeloma and the Role of different induction Regimens
Authors: Bao Li , Lu Jin , Huang Xiaojun 
Published: October 2019

Abstract
We retrospectively compared the impact of treatments based on thalidomide, bortezomib or tranditional regimen (VAD) on renal function recovery in a consecutive, unselected population of newly diagnosed myeloma patients presenting with renal failure. Between May 2005 and February 2014, 130 (24.3%) patients with newly diagnosed multiple myeloma and RI were treated upfront with VAD or a novel agent containing regimen and all received a similar supportive care in our single center. A complete recovery of renal function(renal complete response )was observed in 56.1% of patients treated with bortezomib, in 38.9% with thalidomide and in 28.6% with VAD (P=0.033), a significant improvement of renal function (≥renalPR (renal partial response)) was observed in 63.6% of patients treated with bortezomib, in 66.1% with thalidomide and in 42.9% with VAD (P=0.162). There has a significantly difference in the rates and quality of renal response among patients with different quality of myeloma response: renal CR rates for patients who achieved ≥VGPR, PR, MR or NR were 63.1% vs 46.8% vs 35.7% vs 25.8%, P<0.05, respectively. eGFR ≥30 ml/min and myeloma response higher than PR were independently associated with a higher probability of major renal response in the multivariate analysis. The median follow-up for all patients was 36.5 months and the median survival was 74 months. The median survival for patients of groups B,T and VAD was79 months,71 months and 39 months, respectively (P=0.007). Patients died within the first 2 months from initiation of therapy and the corresponding frequencies in groups B, T and VAD were 0%, 2.8% and 17.9%, respectively (P<0.05). In multivariate analyses on survival, myeloma response higher than PR and new agents-based therapy are identified independent prognostic factors. For newly diagnosed myeloma patients with RI, bortezomib or thalidomide based regimen should be selected promptly to achieve rapid effective myeloma rate and high rates of renal recovery.

Disclosures
No relevant conflicts of interest to declare.
The Cancer/Testis Antigens of SSX4, MAGE-A3, and CTAG2 Are Attractive Targets for Cancer Immunotherapy in Korean Patients with Multiple Myeloma
Authors: Choi Nu-Ri , Jung Sung-Hoon , Lee Hyun-Ju , Vo Manh-Cuong , Hoang My-Dung , Kim Hyeoung-Joon , Lee Je-Jung 
Published: October 2019

Abstract
Introduction: Cancer/testis antigens (CTAs) are an attractive target for cancer immunotherapy because of a tumor-restricted expression and remarkable immunogenicity. Several CTAs have been used as a source of tumor antigen in dendritic cell therapy against multiple myeloma (MM), but there was no report the CTAs in Asian patients with MM. In this study, we evaluate the expression of 10 CTAs on malignant plasma cells of bone marrow in 18 Korean patients with relapsed or refractory MM.
Materials and methods: Eighteen patients with relapsed or refractory MM were classified as four categories according to paraprotein subtypes: IgG (n=7), IgA (n=5), light chain-lambda (n=3), and light chain-kappa (n=3). The expression pattern of 10 CTAs, including NY-ESO-1, SSX2, SSX4, SSX5, MAGE-A3, MAGE-C1, MAGE-C2, BAGE2, CTAG2, and SPA7, was studied by real-time quantitative polymerase chain reaction in CD138+ cells of BM mononuclear cells (MNCs) obtained from MM patients. In addition, we compared it with expression pattern of CTAs in the MNCs from healthy normal donors and the CD138- cells of BM MNCs from MM patients as controls.
Results: In CD138+ cells of BM MNCs from the patients, five CTAs, including SSX2, SSX4, MAGE-A3, MAGEC2, and CTAG2, showed high frequency and overall 5.4 to 63.9 fold increase expression in the quantitative mRNA survey compared to MNCs from healthy donors and CD138- cells of BM MNCs from patients. Expression pattern of 5 CTAs was slightly different by paraprotein subtypes: IgA subtype - SSX4 (17.1 fold increase), MAGE-A3 (11.0 fold increase), and CTAG2 (5.9 fold increase); IgG subtype - CTAG2 (63.9 fold increase), SSX4 (40.2 fold increase), and MAGE-A3 (39.9 fold increase); lambda light chain subtype - CTAG2 (42.4 fold increase), SSX4 (29.0 fold increase), and MAGE-A3 (24.4 fold increase); kappa light chain subtype - SSX2 (6.4 fold increase), MAGE-C2 (6.2 fold increase), MAGE-A3 (5.4 fold increase), and SSX4 (5.4 fold increase).
Conclusion: This study suggests that three CTAs, such as SSX4, MAGE-A3, and CTAG2, highly expressed on malignant plasma cells are potentially promising targets for cancer immunotherapy in Korean patients with relapsed or refractory MM.

Disclosures
No relevant conflicts of interest to declare.
Colesevelam Hydrochloride for the Treatment of Lenalidomide Induced Diarrhea
Authors: Watson Melanie , Nooka Ajay K. , Gleason Charise , Valla Kelly , Kaufman Jonathan L. , Lonial Sagar 
Published: October 2019

Abstract
INTRODUCTION: Lenalidomide is an immunomodulatory drug used for both induction and maintenance in Multiple Myeloma patients. In the pooled multiple myeloma studies, 29.2% of patients reported diarrhea with lenalidomide and dexamethasone. Colesevelam hydrochloride, a drug used primarily for lipid-lowering, has shown efficacy when used for diarrhea in patients receiving lenalidomide in European studies.
METHOD: In our retrospective analysis, we have evaluated the response of twenty patients receiving lenalidomide and experiencing chronic diarrhea between October 29, 2013, and August 5, 2014. Patients were prescribed colesevelam hydrochloride, 1875mg orally twice daily. Patients were instructed to dose their lenalidomide and colesevelam separated by at least two hours. The objective is symptomatic improvement in lenalidomide induced diarrhea.
RESULTS: Twelve males and eight females on lenalidomide based therapy experiencing chronic diarrhea were prescribed colsevelam. Seventeen patients were receiving single agent lenalidomide and three patients were receiving combination therapies (elotuzumab/lenalidomide/dexamethasone:1 pt, lenalidomide/bortezomib/dexamethasone: 1 pt, and lenalidomide/cyclophosphamide/dexamethasone: 1 pt). Median age of the patients was 60 year (range: 44-77 yrs). Median prior lines of therapy were 1 (range: 1-6). Median time from initiation of lenalidomide to Colesevelam hydrochloride is 44 months (0-64 months). Eighty-five percent of patients have noted improvement and 30% experienced total resolution of symptoms. Median time from initiation of Colesevelam hydrochloride to symptomatic response is 4 weeks (range, 0-17). In one patient response was seen in less than a week.
CONCLUSION: Multiple Myeloma patients on lenalidomide therapy with chronic diarrhea noted improvement or total resolution of symptoms with the addition of colesevelam, thus allowing patients to stay on therapy, improving survival as well as quality of life.

Disclosures
Gleason: Celgene: Consultancy; Novartis: Consultancy. Lonial:Millennium: The Takeda Oncology Company: Consultancy, Research Funding; Celgene: Consultancy, Research Funding; Novartis: Consultancy, Research Funding; Bristol-Myers Squibb: Consultancy, Research Funding; Onyx Pharmaceuticals: Consultancy, Research Funding.
Cytotoxic T-Lymphocytes Recognize and Target Leukemic Stem Cells
Authors: Schneider Vanessa , Zhang Lu , Rojewski Markus , Bullinger Lars , Hofmann Susanne , Goetz Marlies , Doehner Konstanze , Doehner Hartmut , Buske Christian , Greiner Jochen 
Published: October 2019

Abstract
Leukemic stem cells (LSC) might be the source for leukemic disease self-renewal and account for disease relapse after treatment, which makes them a critical target for further therapeutic options. Treatment for acute myeloid leukemia (AML) patients became more effective, yet relapse rates are still high. Immunotherapeutic approaches might overcome the immune escape mechanisms of LSC and might be a therapeutic option. Leukemia-associated antigens (LAA) represent targets that are recognized by cytotoxic T-lymphocytes (CTL) and might be useful for specific immunotherapy.
We enriched the LSC containing fraction as CD34+CD38- cells (called LSC-EF) of 20 AML patients using FACS, analyzed expression patterns of LAA in LSC-EF and compared them to patterns in enriched HSC und bulk AML cells by microarray expression analysis and qRT-PCR. In immunoassays, we analysed the functional inhibition of LCS in colony forming assays (CFU) of AML patients by stimulated T cells.
We detected a high expression of several LAA in LSC-EF. PRAME (p = 0.0085), RHAMM (p = 0.03), WT1 (p = 0.04) and Proteinase 3 (p = 0.04) showed significant differential expression in LSC-EF compared to HSC. Survivin (p = 0.7), HAGE (p = 0.5), SSXIP2 (p = 0.4) and Aurorakinase A (p = 0.7) did not indicate a significant differential expression. BAGE does not seem to be an ideal candidate as it is significantly higher expressed on normal HSC (p = 0.0056). Further, we evaluated the expression differences of LSC-EF compared to leukemic bulk. PRAME, RHAMM and WT1 qualified as interesting potential targets as they are lower expressed on leukemic bulk than on LSC-EF. In contrast, Proteinase 3 indicates a higher expression on leukemic bulk but lower in LSC-EF. qRT-PCR confirmed our array data.
In immunoassays, T cells stimulated against LAA indicated a significant inhibition of CFUs in AML patient samples. PRAME, RHAMM and WT1 showed good immunogenic responses with CTL recognition rates in a range of 58-83%. PRAME hereby proved as most effective using two different patient samples, with a reduction of 83% colonies (p = 0.004). WT1 shows a significant reduction of 74% (p = 0.05) and RHAMM of 58% colonies (p = 0.2).
Taken together, several LAA are significantly expressed on the LSC-EF, whereas PRAME, RHAMM and WT1 seemed most promising as these LAA are differentially expressed compared to normal stem cells and stimulation of cytotoxic T cells against these LAAs inhibit LSC in CFU assays.

Disclosures
No relevant conflicts of interest to declare.
Side Population of Multiple Myeloma and Multiple Myeloma Stem Cell
Authors: Kim Kyoung Ha , Cheong Hee-Jeong , Kim Sook-Ja , Kim Se Hyung , Yoon Jina , Kim Han Jo , Kim Hyun Jung , Lee Sang-Cheol , Bae Sang Byung , Kim Chan-Kyu , Lee Namsu , Lee Kyu Taeg , Park Sung Kyu , Hong Dae Sik , Won Jong-Ho , Jun Hyun Jung , Chang Myung Hee 
Published: October 2019

Abstract
Background
Side population (SP) cells are known as enriched source of cancer initiating cells with stem cell properties. The SP cells show a distinct low-staining pattern with the Hoechst 33342 dye. Our aim was to identify SP cells in multiple myeloma (MM) cell lines as well as characteristics of SP cells. Furthermore, we investigated drugs that are effective against SP cells of MM.
Methods
SP (side population), MP (main population) cells from human multiple myeloma cell line (RPMI8226) were sorted by flow cytometry based Hoechst 33342 staining. We performed proliferative capacity in vitro, colony-forming cell assay, cell cycle analysis, drug toxicity (doxorubicin, dexamethasone, bortezomib, melphalan). Expression of stem cell markers (ABCG2, Oct4, Sox2, and Nanog) was observed by western blotting assay.
Results
Several established studies have shown a clonogenic MM cell population with CD 138-status. Therefore, we assessed whether there is correlation between CD 138 - subpopulations and SP cells. CD 138- subpopulations of MM cell lines were not consistent with SP cells. And we have found no correlation between expression of CD 19, CD20, and CD 27 and the proportion of SP cells. No difference was observed for expression of stem cell marker between the SP cells and MP cells. We examined cell cycle analysis and proliferation ability of SP and MP cells. SP cells showed a capacity for higher proliferation abilities and clonogenecity than MP cells. (Fig1,2)
We investigated target SP cells with several drugs (doxorubicin, dexamethasone, melphalan, bortezomib). Importantly, bortezomib was the most effective drug for SP cells.(Table1)
Conclusion
In our study, there was lack of evidence that SP cells was consistent with MM stem cells. No difference was observed for any other marker between SP cells and MP cells. However SP cells had higher proliferation index and clonogenicity compared with MP cells. We suggest that SP cells are rather related to resistance of drugs. Furthermore, bortezomib reduced the SP cells more effectively compared to other drugs.
Fig 1. Colony assay Fig 1. Colony assay
Fig 2. Cell growth ( X 105 ) Fig 2. Cell growth ( X 105 )

Disclosures
No relevant conflicts of interest to declare.