bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Clinical Significance of Argonaute 2 Expression and Angiogenesis in Multiple Myeloma
Authors: Zhang Qiguo , Wu Hongyan , Ouyang Jian , Chen Bing , Dong Haibo , Zhou Rong-Fu , Zeng Hui , Zhou Min 
Published: October 2019

Abstract
Objective To assess the argonaute 2 (Ago2) expression in the bone marrow of multiple myeloma (MM) and determine its association with angiogenesis.
Methods Fifty-nine MM patients and sixteen normal controls were included. The bone marrow sections were made from plastic (methyl-methacrylate)- embedded bone marrow biopsies. The expression of Ago2 and CD34 in the bone marrow sections was detected by EnVison immunohistochemistry and the microvessel density (MVD) was then assessed. The bone marrow Ago2 expression level was also compared by western blot. The clinical significance of Ago2 expression and MVD in MM was analyzed.
Results Western blot analysis indicated that the Ago2 expression was significantly increased in the bone marrow of MM patients as compared with normal controls. As assessed by immunohistochemistry, fifty MM patients were positive for Ago2 and nine patients were negative. However, sixteen normal controls were all negative for Ago2 (χ2=42.586,P<0.001). The β2-microglobulin levels were significantly higher in the Ago2 positive group as compared with the Ago2 negative group (Z=-2.014 , P=0.042). The MVD was significantly higher in bone marrow samples of MM patients compared to the normal controls (7.89±4.88 vs. 2.16±1.32, Z=-3.283 , P<0.001). Analysis of the correlation coefficients showed that Ago2 expression was positively associated with MVD in MM patients (r=0.461, P=0.023). MVD was also significantly higher in the Ago2 positive group than in the Ago2 negative group (Z=-2.449,P=0.014).
Conclusion Ago2 expression was found to be significantly increased in the bone marrow of MM patients. Ago2 may take part in the pathogenesis of MM and mediate the angiogenesis.

Disclosures
No relevant conflicts of interest to declare.
The Impact of Genetic Abnormalities (GA) and Other Factors on Survival in Patients after 65 Years with Multiple Myeloma (MM)
Authors: Garifullin Andrei , Martynkevich Irina , Voloshin Sergei , Kuvshinov Alexei , Martynenko Ludmila , Kleina Elizaveta , Salogub Galina , Karyagina Elena , Schmidt Alexander , Abdulkadyrov Kudrat 
Published: October 2019

Abstract
Background. The increase of life expectancy in patients after 65 years with MM is the main aim of treatment. Lack of carrying out aggressive anti-multiple myeloma therapy increase influence of different factors on OS.
Aims. To compare influence GA and other different factors on overall survival in 65 and more years old patients with MM.
Methods. We retrospectively analyzed 40 patients 65 and more years (median age 71 years, range 65-86; male/female - 1:1.35). The incidences of genetic abnormalities were determined in all cases. Cytogenetic analysis was performed on bone marrow samples using standard GTG-method. Metaphase FISH analysis was performed according to the manufacturer's protocol using DNA probes: LSI 13(RB1)13q14, IGH/CCND1, IGH/FGFR3, LSI TP53 (17q13.1). Stratification of patients was carried in groups of risk according to the modified molecular classification mSMARTmod 1.0 and mSMARTmod 2.0. Patients with 2 and more chromosomal aberrations were additionally entered in high-risk group of both systems.
Results. GA in multiple myeloma after 65 years old patients were detected in 22.8% (9/40). The occurrence frequency of t(11;14) was 26.0% (6/23), del(13q) - 20.8% (5/24), t(4;14) - 4.3% (n=1/23), del(17p) - 0% (n=0/11). 33/40 (82.5%) patients entered into standard risk group, 4/40 (10%) - into intermediate risk, 3/40 (7,5%) - into high-risk. Median OS (MOS) according to mSMARTmod 1.0 in standard risk group (33/40) was 78 months, in high-risk (7/40) - 54 months. Median OS according to mSMARTmod 2.0 in standard risk group (33/40) was 78 months, in intermediate-risk (4/40) - 56 months, in high-risk (3/40) - 49 months. In patients groups without renal failure (35/40) MOS was 78 vs. 46 months with renal failure (5/40). MOS isn't reached in patients with ISS I (4/27), but MOS in patients with ISS II (13/27) and ISS III (10/27) were 50 and 54 months, respectively. MOS in patients group (10/40), who have both (bortezomib and lenolidomide) anti-myeloma agents was 110 months vs. 57 months in group (30/40) only with bortezomib-based regimen of treatment.
Conclusions. Many factors influence on OS in 65 and more years old patients with MM. However, patients, who had treatment with bortezomib and lenelidomide had the best results of OS.

Disclosures
No relevant conflicts of interest to declare.
Multiple Myeloma and Bone Marrow Microenvironment Immunohistochemical Study of the Expression of 15 Proteins Related to Myeloma Bone Disease
Authors: Flodr Patrik , Latalova Pavla , Pusciznova Petra , Pika Tomas , Bacovsky Jaroslav , Scudla Vlastimil , Minarik Jiri 
Published: October 2019

Abstract
Objective: Neoplastic milieu is an integral part of all malignant diseases including multiple myeloma and plays variable role in their development, retention/adhesivity, resistency or sensitivity to therapeutic approach, homing and also paraneoplastic manifestations. Relatively genetically stable milieu may play an important role in new specific molecular therapeutic approaches and therefore should be contextually studied with neoplastic cells as complex neoplastic tissues. The expressions of 15 proteins with close relation to the development of myeloma bone disease (MBD) were analysed in consecutive multiple myeloma specimens.
Methods: Bone marrow trephine biopsy specimens (n=57) with multiple myeloma were included in our prospective study. FFPE tissues were processed in app. 5microm sections and placed on charged slides. The indirect immunohistochemical staining was applicated after antigen retrieval and commercial primary antibodies were used for the detection of observed proteins. Standard secondary antibody and ABC method were included in visualisation. We analysed the expressions of MIP1alfa, Annexin A2, TRAP, DKK-1, RANK, RANKL, OPG, Sclerostin, Activin A, NFkappaB proteins (p50, p52, p65), p62 (sequestosome 1), MMP9 and RUNX2.
Results: Bone marrow multiple myeloma specimens showed variable positivity of MIP1alfa in 60% (cut-off point 20%), Annexin A2 in 42% (myeloma cells, cut-off point 30%) and in 74% (stromal cells, cut-off point 5%), TRAP in 28% (cut-off point 5%), DKK-1 in 23% (cut-off point 30%), RANK in 53% (cut-off point 30%), RANKL in 70%, OPG in 39% (cut-off point 5%), Sclerostin in 95% (cut-off point 90%), Activin A in 35% (cut-off point 30%), cytoplasmic positivity of p50 in 5% (cut-off point 10%), p52 in 86% (cut-off point 10%), p62 in 91% (cut-off point 10%), p65 in 89% (cut-off point 10%), positivity of MMP9 in 22% (cut-off point 30%) and positivity of RUNX2 in 56% (cut-off point 30%).
Conclusion: Our study showed variable expression of proteins related to MBD in multiple myeloma and its bone marrow microenvironment that imply biological heterogeneity, different development and stromal plasticity in this complex hemato-oncological disease. The exact and contextual knowledge of the engaged signaling pathways may suggest more specific or tailored therapeutic approaches (e.g. anti-RANKL, anti-DKK-1, anti-Sclerostin, anti-Activin A).
Supported by the grant NT 14393.

Disclosures
No relevant conflicts of interest to declare.
Distinct Clinico-Pathological Profile of Plasma Cell Leukemia Patients from North India
Authors: Sharma Praveen , Bommannan Karthik B.K , Sachdeva Man Updesh Singh , Varma Neelam , Srinivasan Radhika , Malhotra Pankaj , Varma Subhash 
Published: October 2019

Abstract
Introduction: Plasma cell leukemia (PCL) is a rare and aggressive plasma cell neoplasm where clonal plasma cells comprise ≥ 20% of peripheral blood (PB) leukocytes and/or absolute clonal PB plasma cell count is ≥ 2×109/L. Primary plasma cell leukemia (PPCL) originates de novo, whereas, secondary plasma cell leukemia (SPCL) evolves from pre-existing multiple myeloma (MM). Literature does reveal a few large case series on PCL, however, data on this rare neoplasm is very sparse from Indian sub-continent. We present clinico-pathological profile 14 cases of PCL from single tertiary care centre of north-India.
Materials and methods: Archival files were screened for period of 8 years (Jan 2007 to Dec 2014) and clinico-pathological profile, including overall survival, of patients of PCL was systematically analysed, retrospectively.
Results: Ten PPCL and four SPCL patients were traced in duration of 8 years, all belonging to north-Indian states of Punjab and Haryana. Eight PPCL and three SPCL patients had complete clinical data. Our patients had less frequent renal failure (12.5%), more frequent hepatomegaly (75%) and non-secretory type (33%) disease. Flow cytometric immunophenotyping revealed expression of CD138 (67%), CD56 (33%) and CD20 (0%), which was different from reported western data. With novel therapeutic agents, these patients fared a higher median survival of 15 months.
Conclusion: Our patients of PCL from neighbouring states of north-India have a distinct clinico-pathological profile, with better survival as compared to existing literature. The significance of our findings must be tested in a larger patient cohort and must be supported with molecular and cytogenetic investigations to unmask any significant pathogenesis.

Disclosures
No relevant conflicts of interest to declare.
SNP rs34172460 (S258A)Â in Gene CIP2A Correlates with Bortezomib Resistance in Multiple Myeloma Patients
Authors: Tang George , Li Mingjie , Sanchez Eric , Wang Cathy , Vardanyan Suzie , Lee Tiffany , Nosrati Jason , Cao Jasmin , Zahab Michael , Mehta Puja , Garzio Gigi , Chen Haiming , Berenson James 
Published: October 2019

Abstract
Introduction: The proteasome inhibitor (PI) bortezomib is a novel anticancer drug that shows activity especially for treating patients with multiple myeloma (MM). However, its clinical efficacy has been hampered by the emergence of drug-resistance phenomena, the genetic and molecular basis of which remains elusive. We propose that the resistance to bortezomib treatment may be related to a polymorphism in a gene known to be a target for bortezomib's activity which has been identified as CIP2A which is a tumor suppressor PP2A inhibitor. We have used a high resolution melting curve (HRM) approach to screen several CIP2A SNPs containing significant genetic changes that may be different between bortezomib sensitive and resistant MM patients. In this study, we evaluated SNPs in this gene in healthy subjects and MM patients who were sensitive or resistant to bortezomib-based therapy.
Methods: A panel of DNA samples was purified from MM patients (N=73) and healthy subjects (N=95). The samples were divided into three groups: bortezomib sensitive MM patients (N=61), bortezomib resistant MM patients (N=12), and healthy subjects. A single nucleotide polymorphism (SNP) of the bortezomib target genes CIP2A was selected. The PCR prime pairs for each SNP marker were designed with the software "Primer Expression 3" (Thermo Fisher Scientific). The genotype of each DNA sample was determined with the high resolution melt curve (HRM) technology on a real-time PCR instrument, StepOne Plus (Thermo Fisher Scientific).
Results: The frequency of heterozygosity and homozygosity of CIP2A, SNP rs34172460(S258A) was 0.049 (3 out of 61 cases) in the bortezomib sensitive group, while it was 0.25 (3 out of 12 cases) in the bortezomib resistant group, a 5-fold increase in the frequency of this SNP. The frequency of heterozygosity and homozygosity of this same SNP in the healthy subjects was only 0.03 (3 out of 95 samples).The minor allele frequency (MAF) in the normal population is 0.046, according to the SNP data base, compatible with what we observed in both our healthy subjects and the bortezomib sensitive MM patients.
Conclusion: This preliminary data suggests that heterozygotic and homozygotic mutants of rs34172460(S258A) may have an impact on resistance to bortezomib among patients with multiple myeloma.

Disclosures
No relevant conflicts of interest to declare.
Thymus-Derived PTH (TPTH) Is Increased after Thyroparathyroidectomy in C57BL6/Kalwrij Mice and Modulates Mouse Sensitivity to 5TGM1 Myeloma Cell Line
Authors: Zangari Maurizio , Yoo Hanna , Suva Larry , Yoon Donghoon 
Published: October 2019

Abstract
We have previously reported a significant correlation between serum PTH levels and myeloma responses to proteasome inhibition and have recently demonstrated a crucial role for the parathyroid hormone receptor 1 (PTHR1) in vitro and in vivo. In this study the effect of abrogation of parathyroid-derived PTH was tested in 5TGM1 myeloma transplanted C57BL6/KaLwRij mice. To interrogate PTH activity in vivo, thyroparathyroidectomy (TX-PTX) was performed on the C57BL6/KaLwRij mice before or after the transplantation of 5TGM1 cells. Surgical success was confirmed by the rapid loss of measureable circulating PTH. In TX-PTX mice bearing transplanted 5TGM1 tumors, serum PTH levels was noted to recover 3-4 weeks after surgery and to achieve levels consistently higher than baseline levels pre TX-PTX. These data raised the compelling possibility of an alternative PTH source. To determine the potential source of the recovered PTH levels, multiple organs were examined. Whole tissue lysates were analyzed by Western blot and revealed a faint but specific band from thymic tissue, as expected with no obvious bands detected in 13 other tissues investigated including the liver, lung, kidney, ovary and testis. In addition, murine PTH mRNA transcripts from thymus RNA were measured by Real-Time PCR, using a specifically designed primer set that spans exons 1 and 2, since intron 1 is >1kb. These primers specifically discriminate mRNA transcripts from genomic DNA and improve the assay sensitivity for the detection of PTH transcripts. Using these custom-designed primers, control thymus expressed measureable PTH mRNA transcripts that was significantly elevated in TX-PTX mice. Since the thymus is considerably bigger than the parathyroid glands, PTH was immunoprecipitated before performing Western blot analysis. Following immunoprecipitation, a distinct PTH protein band was observed from the thymus in both TX-PTX and control mice.
These data suggest that the thymus is the major source of serum PTH in TX-PTX mice and that the organ can compensate for the loss of serum PTH from the parathyroid gland. We propose that thymus-derived PTH in the TX-PTX mouse may contribute to the control of the 5TGM1 murine myeloma model.

Disclosures
Zangari: University of Arkansas for Medical Sciences: Employment; Novartis: Research Funding; Millennium: Research Funding; Onyx: Research Funding.
Minimal Residual Disease Analysis in Multiple Myeloma Patients By 6-Color Flow Cytometry: An Experience from Tertiary Care Center of North India
Authors: Sharma Praveen , Sachdeva Man Updesh Singh , Varma Neelam , Bose Parveen , Malhotra Pankaj 
Published: October 2019

Abstract
Therapeutic advances in multiple myeloma (MM) incorporating the use of high-dose melphalan, novel therapeutic immunomodulatory agents, proteasome inhibitors and supporting autologous stem-cell transplantation (ASCT) have improved response rates and overall survival. The detection of minimal residual disease (MRD) is recognized as a sensitive and rapid approach to evaluate treatment efficacy as a tool for predicting patient outcomes and guiding therapeutic decisions. MRD analysis is reflected by many different techniques, however, multiparametric flow cytometry is a sensitive, feasible and adequate method for monitoring residual disease. Studies from India related to this context are lacking. In the present study, we compare MRD levels in patients of multiple myeloma after chemotherapy/ASCT assessed by multiparametric flow cytometry, with M band status, immunofixation (IFE) and percentage of plasma cells on bone marrow aspirate.
Seventeen patients of multiple myeloma were included in the study over a duration of one year, (Male=13, Female=4) with mean age of 56.8 years (range 44-80 years). MRD was analyzed using a dual laser 6 color-flow cytometer in 9 patients of ASCT (day 100) and 8 patients on chemotherapy alone (post-induction). Pre-titrated cocktail of CD38, CD138, CD19, CD45, cytoplasmic Kappa light chain, cytoplasmic lambda light chain, CD81, CD27, CD28 CD200 and CD10 were used in 6-color combination of three tubes for MRD analysis. MRD was detectable in 5 patients, mean of 0.61% (range of 0.07 - 6.44%). M band and IFE were positive in 2 patients, each. Bone marrow plasma cells ranged from 0 to 22%. MRD levels did not show significant correlation with percentage of plasma cells in bone marrow aspirate, however it had an statistical agreement with presence or absence of serum M-band and IFE. Patients are on regular follow up for their clinical and hematological response.

Disclosures
No relevant conflicts of interest to declare.
Orthotopic Heart Transplant Facilitated Autologous Hematopoietic Stem Cell Transplantation in Light-Chain Amyloidosis
Authors: Kamble Rammurti T , Obi Gloria , Scholoff Audrey , Carrum George , Rice Lawrence , Bhimrajan Arvind , Estep Jerry 
Published: October 2019

Abstract
Dominant cardiac involvement by primary systemic amyloidosis (AL) precludes effective AL treatment and is associated with short survival. Between January 2009 and June 2015, total of 9 patients who presented with severe cardiac dysfunction as their major manifestation of AL underwent orthotopic heart transplantation (OHT). Four of these 9 patients were able to complete the planned second phase of treatment with autologous hematopoietic stem cell transplantation (ASCT) 13-16 months after OHT. Diagnosis of cardiac AL was established via endomyocardial biopsy, Congo red staining and immunohistochemistry. All patients had end stage heart failure and developed cardiogenic shock requiring intra-aortic balloon pump support (median 20 days, range 10-165) as a bridge to OHT.
The median age at AL presentation was 54 years (42-65) in 4 females and 5 males. At median follow-up of 30 months (1-54) from OHT, 6 (66 %) patients are alive (table-1). Two patients died of post-operative complications at 1 and 7 months post OHT; a 3rd patient died 36 months after OHT (23 months post ASCT) of AL progression. Four patients received ASCT at median of 13 months (13-16) after OHT. In the remaining 3 patients ASCT was not feasible due; to low DLCO; recently treated disseminated Cryptococcus; or prior ASCT.
All 4 patients with ASCT were on tacrolimus and prednisone at the time of stem cell mobilization and hematopoietic transplant; two patients were also receiving mycophenolate mofetil and valganciclovir. We collected 5.7, 6.1 and 6.2 x 106/kg CD-34+ cells in 2 days after filgrastim administration (5 ug/kg, twice, daily) and plerixafor (16 mg/kg based on day- 4 CD-34+ counts) in 3 subjects. The fourth patient initially failed to mobilize but 4.3x106/kg CD-34+ cells were subsequently obtained after stopping mycophenolate mofetil for 4 weeks. The creatinine clearance at the time of the high-dose chemotherapy given prior to ASCT was 36, 30, 41 and 43 ml/minute. All 4 patients received a renal adjusted dose of melphalan at 140mg/m2. Mycophenolate mofetil and valganciclovir were withheld during neutropenia until engraftment. No patients received post-transplant filgrastim. Patients were hospitalized for 17, 18, 18 and 20 days. Renal function remained stable during ASCT and non-hematological toxicity was limited to grade I-II apart from one grade III oral mucositis and colitis. Two patients achieved hematologic complete remission (patient2&4) while 2 patients had a partial response following ASCT. Post OHT and ASCT 3 patients are alive and well at follow-up of 54, 37 and 52 months.
The strategy of OHT followed by ASCT is therefore feasible in select patients with dominant cardiac involvement and advanced heart failure

Disclosures
No relevant conflicts of interest to declare.
Bortezomib-Induced Proinflammatory Macrophages Contribute to Multiple Myeloma Cell Aggressiveness
Authors: Beyar Katz Ofrat , Ben-Tsedek Neta , Avivi Irit , Alishekevitz Dror , Timaner Michael , Miller Valeria , Lindzen Moshit , Yarden Yosef , Shaked Yuval 
Published: October 2019

Abstract
Multiple myeloma (MM) is a chronic progressive malignancy of plasma cells. Although treatment with the novel proteasome inhibitor, bortezomib, significantly improves patient survival, some patients fail to respond due to the development of de novo resistance. Previous studies revealed that chemotherapy induces pro-tumorigenic host-mediated effects which could explain tumor re-growth and metastasis(Gingis-Velitski, Loven et al. 2011, Katz, Shaked 2014). Here we show that plasma from bortezomib-treated mice significantly increases migration, viability and proliferation of human MM cells in vitro, compared to plasma from control untreated mice. Comparable results were demonstrated with plasma obtained from patients with MM treated with bortezomib. Additionally, bortezomib induces the mobilization of pro-angiogenic bone marrow cells. Mice treated with bortezomib and subsequently intravenously injected with MM cells succumb to MM aggressiveness earlier than mice treated with the vehicle control(Figure 1). We show that pro-inflammatory macrophages contribute to MM cell aggressiveness in response to bortezomib treatment, in part by secreting interleukin-16(IL-16). Blocking IL-16 in conditioned medium obtained from bortezomib-treated macrophages generated reduced viability of MM cells in vitro. Accordingly, co-inoculation of MM cells with pro-inflammatory macrophages from bortezomib-treated mice accelerates MM disease progression. Taken together, our results suggest that, in addition to the known effective anti-tumor activity of bortezomib, this drug can induce host-driven pro-tumorigenic effects that may promote MM aggressiveness.
Figure 1. Host response to bortezomib promotes MM aggressiveness in mice. Eight week old CB.17 SCID mice were injected intravenously with 1mg/kg bortezomib or vehicle (veh). Four and 24 hours later mice were inoculated through the tail vein with 5x106 CAG-luciferase+ cells (n=6-7mice/group). Tumor growth and expansion was assessed by IVIS imaging system. Figure 1. Host response to bortezomib promotes MM aggressiveness in mice. Eight week old CB.17 SCID mice were injected intravenously with 1mg/kg bortezomib or vehicle (veh). Four and 24 hours later mice were inoculated through the tail vein with 5x106 CAG-luciferase+ cells (n=6-7mice/group). Tumor growth and expansion was assessed by IVIS imaging system.

Disclosures
No relevant conflicts of interest to declare.