bluuebunny/crossref_metadata_2025 huggingface dataset:
Cholesterol Levels In Patients With Newly Diagnosed Chronic Lymphocytic Leukemia: A retrospective Multicenter Study
Authors: Yavasoglu Irfan , Pektas Gokhan , Yilmaz Fergün , Akgün Gülsüm , Tombak Anil , Toka Bila , Dal Sinan , Ozbas Hasan , Cetin Guven , Donmez Ayhan , Bilgir Oktay , Tiftik Naci , Ertop Sehmuz , Ayyildiz Orhan M. , Sonmez Mehmet , Kadikoylu Gurhan , Tombuloglu Murat , Bolaman Zahit
Published: October 2019

Low cholesterol levels can be detected in solid tumors and hematological malignancies such as muliple myeloma. Moreover chloesterol levels reduced in some experimental studies of patients with chronic lymphocytic leukemia (CLL). In this retrospective multicenter study, lipid levels were retrospectively evaluated in 420 (264 male and 256 female with mean age 64 ± 11 years) patients with newly diagnosed CLL, according to the International CLL study group. 71 (28 male and 43 female with mean age 55 ± 9 years) healthy subjects as control group were included to this study. Lipid parameters such as total cholesterol (TC), high density lipoprotein-cholesterol (HDL-C), very low-density lipoprotein-cholesterol (VLDL-C), and triglyceride levels were measured with enzymatic/ calorimetric method and Architect C800 instrument. Low-density lipoprotein-cholesterol (LDL-C) levels were calculated according to Friedwald formula. Lipid parameters between two groups were compared with Mann-Whitney U test. A value of p< 0.05 was accepted as statistically significant. According to Binet classification, 60% of patients were in stage A, while 25% of them were in stage C. In CLL patients, the levels of TC, HDL-C, and LDL-C were lower than those of control group (p=0.001). There was no significantly difference for triglyceride and VLDL-C levels between two groups (p>0.05) (Table-1). The levels of TC, LDL-C, and HDL-C in the patients with stage C were lower than those of both stage A and stage B (Table-2). Low cholesterol levels in patients with CLL may occur due to increased use of cholesterol by lymphocytes.Table-1Lipid parameters in control group and CLL patientsCLL (N:420)Control (N:71)P valueTC (mg/dl)175±41217±36<0.001HDL-C (mg/dl)37±1153±14<0.001LDL-C (mg/dl)108±30131±29<0.001Triglyceride (mg/dl)140±71147±68>0.05VLDL-C (mg/dl)31±1731±17>0.05Table 2Lipid parameters in the patients according to Binet classificationStage A (n:255)Stage B(n:61)Stage C(n:104)P valueTC(mg/dl)183±38179±43156±40<0.001HDL-C(mg/dl)40±1237±1232±12<0.001LDL-C (mg/dl)112±28110±3398±30<0.001Triglyceride (mg/dl)141±76136±53141±68>0.05VLDL-C(mg/dl)30±1535±3331±13>0.05

Disclosures:

Sonmez: Novartis Pharmaceuticals Corporation, Turkey: Membership on an entity’s Board of Directors or advisory committees.
The Study On The Relationship Between c-Myc and Hif-1 α In u266 Cell Line
Authors: Ma Yanping
Published: October 2019

Objective

The aim of study is to explore the relationship of C-MYC and HIF-1 α in multiple myeloma cell lines U266,and research the influence of C-MYC/HIF-1 α on VEGF, in order to find a new target for clinical treatment MM.

Methods

Transfect the siRNA c-myc plasimd,siRNA HIF-1 α plasimd and mock plasimd into U266 cell lines respectively.RT-PCR was used to detected the expression of C-MYC and HIF-1αin different group, (fetch the CT value, using 2-ΔΔCT to calculate) .The expression of protein of C-MYC.HIF-1αand VEGF was measured with Western-blot in different group,((using the ratio of purpose gene and internal reference β-actin expression as the final results).

Results

compared with the control group, the expression of C-MYC, HIF-1 α and VEGF was decreased in the siRNA c-myc group, which differences are statistically significant (p<0.05), the expression of HIF-1 α and VEGF was decreased in the siRNA c-myc group ,which differences are statistically significant (p<0.05),the expression of C-MYC have no obvious decline compared with control group.

Conclusion

The expression of HIF-1 α depends on the regulation of C-MYC gene, synthesis and secretion of VEGF depends on C-MYC/HIF-1 α, C-MYC/HIF-1 α may serve as a new target for treatment of MM.

Disclosures:

No relevant conflicts of interest to declare.
Expression Of Cancer-Testis Antigen In Multiple Myeloma
Authors: Ji Jingna , Liu Shangqin , He Li , Xu Chaoping , Hu Guiying , Zhang Yicheng , Ma Zi , Huang Jianying
Published: October 2019

In order to detect the CTAs expression in the cell lines and in patients with multiple myeloma (MM). Reverse transcriptase polymerase chain reaction (RT-PCR) detects the mRNA expression of MAGE-C1/CT7, SSX1, SSX2 and SSX4 in MM cell lines RPMI8226 and U266 and patients. Collect clinical MM patients with bone marrow specimens of 25 cases,18 cases of healthy volunteers as a control, The expression frequency in MM patients of CTA gene is parametric statistical analysis with age,gender,the amount of plasma cells,clinical stage and MM type. The CTA members we detected all express in RPMI8226 and U266 cell lines, the expression frequencies of MAGE-C1/CT7 of SSX1, SSX2 and SSX4 in 25 cases bone marrow of MM patients are as follows: 28%（7/25）、80%（20/25）、40%（10/25）、68%（17/25）. 18 cases of healthy volunteers with bone marrow does not express the gene. Four kinds of genes in the bone marrow of patients with simultaneous expression of two or more frequency 80%（20/25）, at least have an expression of the frequency 88%（22/25）. Expression of SSX1 and SSX4 in different disease stage was statistically significant(P <0.05), expression frequency was mainly for patients in the phase Ⅲhigher thanⅠand Ⅱ. Expression of MAGE-C1/CT7 and SSX2 in the period of disease was not statistically significant(P﹥0.05), Detected by the four CTA with age, gender, MM type and the volume of plasma cells was no significant difference(P﹥0.05). It was suggested MAGE-C1/CT7, SSX1, SSX2 and SSX4 gene in MM cell lines RPMI8226 and U266 and MM patients can co-express, while do not express in healthy people. In the expression frequency, SSX1 and SSX4 relate with MM clinical stage,MAGE-C1/CT7 and SSX2 do not relate with clinical stage,the 4 CTAs do not relate with gender,age,clinical type and volume of plasma cells. it provides theoretical support for the CTA vaccine in multiple myeloma immunotherapy.

Disclosures:

No relevant conflicts of interest to declare.
Bortezomib-Based Chemotherapy Regimens Can Inmprove The Complete Remission In Newly Diagnosed Multiple Myeloma Patients With Bcl-2 and Survivin Overexpression
Authors: Wen Zeng , Lifang Huang , Zhang Yicheng , Hanying Sun , Wenli Liu , Fankai Meng , Jianfeng Zhou
Published: October 2019

Compared with the traditional chemotherapeutic agents, Bortezomib-based chemotherapy regimens can increase the complete remission and improve the prognosis significantly in the newly diagnosed multiple myeloma patients. Molecular markers which can predict the chemotherapeutic efficacy and safety could help the physicians to make the right decisions and benefit the patients, and it has been one of most interesting research fields in MM. It has been reported that Survivin and Bcl-2 was involved in the adverse clinical events and therapeutic resistance, respectively. In order to investigate the relationship between the Survivin and Bcl-2 expression and the different regimens therapeutic efficacy, we retrospectively studied the proteins expression in the bone marrow biopsy specimens by inmmunohistochemistry and the efficacy of different chemotherapy regimens in the newly diagnosed MM in a single center of Tongji Hospital. Total 59 newly diagnosed MM were admitted into this study. The positive expression rate for Survivin and Bcl-2 was 35.3%（n=21）and 73.5%（n=43）, respectively. The protein expression had no relationship with the Durie-Salmon and International Staging System stratification, which suggested that Survivin and Bcl-2 were not responsible for the clinical manifestations. Bortezomib-based regimens (n=22) could effectively decrease the tumor burden and achieve response (CR+PR: 67.5% ). The non-bortezomib regimens (n=37), containing VAD(T), MP(T), TAD, were effective in the absent of Survivn and Bcl-2 expression(n=7; CR+PR 62.5% ). When Survivin and Bcl-2 were single or double positive (n=30), the newly diagnosed MM patients had no response for non-bortezomib regimens with none reaching complete remission or partial remission (p=0.0088). According to this study, we recommend the newly diagnosed MM which were Survivin and Bcl-2 single or double positive by inmmunohistochemistry received the regimens containing bortezomib for anti-myeloma therapy.

Disclosures:

No relevant conflicts of interest to declare.
Genomic Characterization Of Newly Established Two Distinct Patient-Driven Multiple Myeloma Cell Lines (SNU_MM1393_BM and SNU_MM1393_SC) From a Single Patient
Authors: Koh Youngil , Ahn Kwang-Sung , Lee Chansu , Jung Woo June , Lee Hyun Jung , Lee Dong Soon , Kim Hyo Jung , Yoon Hwi-Joong , Yoon Sung-Soo
Published: October 2019

To establish patient-derived multiple myeloma (MM) cell lines, mononuclear cells obtained from a MM patient’s bone marrow were directly injected via tail vein into a NRG/SCID mouse. Fourteen weeks after injection, tumor developed at subcutis and bone marrow (BM) in the same mouse. We separated and cultured cells from these two sites (subcutis and BM) and established two cell lines originating from a single patient (SNU_MM1393_BM and SNU_MM1393_SC). In cytogenetic analysis, karyotype of newly established two MM cell lines showed tetraploidy which is different from the karyotype of the patient (diploidy) indicating clonal evolution. In FACS analysis, the expression of CD138 and CD45 was detected in both cell lines. Response to IL-6 and soluble IL-6 receptor was different between the two cell lines. Moreover, SNU_MM1393_SC showed higher degree of resistance against proteasome inhibitor (bortezomib) compared to SNU_MM1393_BM. However, two cell lines were both sensitive to histone deacetylase inhibitor (panobinostat).

When whole exome sequencing was performed using the DNA of these two cell lines, a set of somatic mutations involving Wnt signaling and NF-kB pathway were detected in both cell lines. Additional somatic mutations of JAK1, PLCG1, IRS2, and HGS which are known to interact with JAK/STAT pathway were detected in SNU_MM1393_BM. Whereas, additional somatic mutations of EGFR, HSP90AB1, CFDR, and CALML5, which are known to interact with growth factor cell signaling pathway were detected in SNU_MM1393_SC.

These findings highlight the importance of interactions between tumor and tumor microenvironment as the myeloma progresses and will pave a way to more effective selection of targeted agents according to specific tumor characteristics obtained in the process of disease progression.

Disclosures:

No relevant conflicts of interest to declare.
Aptamer TY04 Inhibits Multiple Myeloma Cell Growth Via Cell Cycle Arrest
Authors: Liu Jing , Dai Hong-Juan , Ma Bian-Ying , Song Jian-Hui , Chen Hui-yong , Zhou Jia-Jie , Tang Shi-Jun , Yao Shan , Ye Mao , Tan Wei-Hong
Published: October 2019

Multiple myeloma (MM), also known as plasma cell myeloma, is characterized by accumulation of clonal plasma cells in the bone marrow and overproduction of monoclonal immunoglobulin (Ig) in the blood or urine. MM accounts for approximately 10% of all hematologic malignancies. Despite recent advances in the understanding and treatment of this disease, MM remains an incurable disease in the vast majority. With conventional chemotherapy, the 5-year median survival rate for MM patients is approximately 25%. Aptamers are single-stranded RNA or DNA sequences that bind to target molecules with high affinity and specificity. Compared with antibodies, aptamers have unique advantages including easy chemical synthesis and modification, low toxicity, lack of immunogenicity, and rapid tissue penetration, Based on these advantages, aptamers show great potential for therapeutic application. The aptamer TY04 is a single-stranded DNA (ssDNA) generated by a method named cell-based systematic evolution of ligands by exponential enrichment (cell-SELEX), We found TY04 strongly inhibited the growth of multiple myeloma cell lines including MM1.S, NCI-H929, KM3 and OPM2,The concentration of TY04 to inhibit 50% cell growth (IC50) on MM1.S was 3.89 μM. In contrast, TY04 had no effect on the growth of non-tumor cell lines — immortal B lymphoblastoid cell lines. Next, we used MM1.S cell line as the model to study the mechanism of TY04 anti- multiple myeloma. Flow cytometry analysis showed that TY04 with the sequence specifically bind to MM1.S cells when compared with unselected ssDNA library control. To investigate whether the target molecules of TY04 are membrane proteins on cell surface, MM1.S cells were treated with trypsin and proteinase k for 2 or 10 minutes before incubation with TY04. The result revealed that TY04 lost partly recognition ability on treated cells, indicating that the target molecules were most likely membrane proteins. Furthermore, we evaluated the cell cycle distribution of MM1.S after TY04 treatment. We found that TY04 significantly caused cell-cycle arrest in G2/M phase. The percentage of G2/M phase cells increased from 30.1±1.56 to 53.2±6.36. To identify the underlying molecular mechanism, G2/M-related proteins were assayed by flow cytometry. Following TY04 treatment, a concomitant inhibition of ERK1/2, cyclin B, CDK1 and γ-tubulin expression occurred. Meanwhile, human cell cycle PCR array was used to analyze the expression of 84 genes key to cell cycle regulation in TY04-treated MM1.S cells. Our results indicated that aptamer TY04 decreased the genes expression of CCNB1, CCNB2, BIRC5, BRCA1 and CCNH, which were involved in the progress of G2/M phase. All these results are significant in that they provide a framework for further exploring the use of TY04 as a novel anti-multiple myeloma agent.

Disclosures:

No relevant conflicts of interest to declare.
CD28 Regulates Metabolism In Multiple Myeloma For Cell Survival and Proliferation
Authors: Utley Adam T , Murray Megan , Carlson Louise M , Farren Matthew R. , Nair Jayakumar R , Lee, MD Kelvin P
Published: October 2019

Multiple myeloma is an incurable hematological malignancy of transformed plasma cells. Many cellular interactions and soluble factors have been demonstrated to play a role in myeloma pathogenesis; however, novel targets to enhance therapeutic intervention are needed. We have demonstrated that CD28 signaling in myeloma cells supports their survival during chemotherapeutic challenge in vitro and in vivo. However, the cellular mechanisms by which CD28 confers this survival advantage to myeloma cells are not completely understood.

CD28 is best characterized as the canonical T cell co-stimulatory molecule. During T cell activation, CD28 signaling induces glycolysis, a metabolic program required for T cell proliferation and functional maturation. In the absence of glycolysis, T cells utilize fatty acid oxidation for energy production through the mitochondria. However, the way in which CD28 regulates metabolism in multiple myeloma is not well understood. Here we present evidence that CD28 signaling induces glut1 expression, and that poisoning the glycolytic pathway inhibits proliferation and survival of myeloma cells.

AMPK, an energy sensitive kinase known to regulate metabolism by driving fatty acid oxidation, is normally activated when cellular energy levels are low. Interestingly, poisoning glycolysis with a glucose analogue that cannot be processed (2DG) leads to AMPK inhibition in myeloma cells. Furthermore, pharmacological activation of AMPK by AICAR, an AMP analogue, is not sufficient to rescue myeloma cell proliferation from glycolytic inhibition and in fact increases cell death (p<.01 from no treatment, p<.05 from 2DG). This evidence suggests that multiple myeloma cells are absolutely dependent upon CD28-mediated glycolysis for proliferation and survival, and that myeloma cells cannot utilize fatty acid oxidation as a subsidiary metabolic pathway for proliferation in the absence of glycolysis. This understanding will allow us to target metabolism in multiple myeloma as a novel therapeutic strategy through pharmacological targeting of the CD28 pathway. This approach can be quickly translated into the clinic, as there are FDA approved drugs which activate AMPK (Metformin) and block CD28 signaling (Abatacept).

Disclosures:

Off Label Use: Abatacept, purpose to prevent CD28-mediated cell survival in multiple myeloma Metformin, purpose to activate AMPK in driving multiple myeloma cell death.
Implementing Cybord As First-Line Therapy For Transplant-Eligible Multiple Myeloma Patients : A Single-Center Preliminary Comparaison
Authors: Duquette Dominic , Laroche Vincent
Published: October 2019

Introduction

The best induction regimen for first-line therapy in multiple myeloma patients who are eliglible for autologous stem cell transplant (ASCT) is still unknown. In 2009, we started to use CyBorD and gradually changed our first-line therapy from Vel-Dex to CyBorD based on published work from Princess Margaret Hospital in Toronto. Here, we describe the first 10 patients to have received CyBorD and compare them to an historical cohort from the same institution.

Methods

We conducted a retrospective review of patient’s charts and transplant database from 2009/10/02 to 2013/04/19. We collected information on blood labs, cytogenetics, immunofixation, response rates, adverse events and graft collection. Patients not receiving the anticipated protocol or less than 3 cycles of treatment arm were excluded from this review.

Results

All patients (100%) in both cohorts received an ASCT (CyBorD, n=10 and Vel-Dex, n=11). The 2 groups were comparable and there was 1 patient in each arm with unfavorable cytogenetics. The response rates pre ASCT were CR = 20%, >VGPR = 90% compared to CR = 0%, >VGPR = 81,8% in CyBorD and Vel-Dex patients respectively. Response rates at 100 days post-ASCT was CR = 60%, >VGPR = 100% compared to CR = 36.4%, >VGPR = 90.9% in CyBorD and Vel-Dex patients respectively. Patients in CyBorD had a median of 1 (1-2) day for graft collection and a median number of graft cells of 5,01 x 109(2,06-6,48) compared to 1 (1-2) and 4,46 x 109 (3-6,82) in the Vel-Dex group. The rate of grade 3/4 thrombocytopenia was 0%(n=0) vs 9%(n=1) in CyborD and Vel-Dex patients respectively. No case of neuropathy grade 3 or 4 was observed in the CyBorD arm.

Conclusion

CyBorD is an effective and safe induction regimen for first-line therapy in ASCT eligible multiple myeloma patients. It produces less neuropathy and has an easier schedule to administer. It is now our standard protocol for these patients but dexamethasone has been reduced to once weekly for every cycle since then.

NB: CyBorD (Cyclophosphamide 300 mg/m2 PO days 1,8,15,22; Bortezomib 1,5 mg/m2 SC days 1,8,15,22, Dexamethasone 20 mg PO bid days 1-4, 9-12, 17-20 for cycles 1-2, 20 mg PO bid once weekly for cycles 3-4.)

Disclosures:

Duquette: Janssen: Consultancy, Honoraria. Off Label Use: CyborD is a widely accepted regimen but has not been accepted by the FDA.
RARα2 Expression Confers Myeloma Stem Cell Features
Authors: Tricot Guido J. , Yang Ye , Gu Zhimin , Xu Hongwei , Huang Junwei , He Jie , Tao Yi , Zhan Fenghuang
Published: October 2019

Background

Despite the major advances in the therapy of MM, many patients ultimately relapse and die of this disease. This is most likely due to the persistence of multiple myeloma stem cells (MMSCs), which survive even after the most intensive treatments and are insensitive to non-transplant therapy. We have previously demonstrated that RARα2 expression increased in CD138 selected plasma cells of relapsed myelomas (MM) and that increased expression was linked to poor prognosis in newly diagnosed MM patients. In this study, we further explore the relationship between RARα2 and MMSCs.

Materials and Methods

RT-PCR was used to detect RARα2, induced pluripotent stem cell (iPS) gene, Wnt and Hedgehog (Hh) gene expression in MMSCs and bulk cells. Clonogenic formation assays were performed by plating RARα2 over-expressing ARP1 and OCI-MY5 MM cell lines in soft agar, treated with bortezomib, doxorubicin, etoposide, and MK-571. Side population (SP) fraction, ALDH activity and cell apoptosis in MM cells were detected by flow cytometry. Western blot were utilized to dissect RARα2-related signaling pathway. Effect of RARα2 on MM progression in vivo was also analyzed in 5TGM1 myeloma mice.

Results

Higher expression of RARα2 was identified in the multiple myeloma stem cell (MMSC) fraction. Over-expression of RARα2 in MM cell lines resulted in 1) increased drug-resistance 2) increased clonogenic potential 3) activation of both Wnt and Hedgehog (Hh) pathways 4) increased side population (SP) and aldehyde dehydrogenase1 (ALDH1) levels and 5) increased expression of iPS genes. The opposite effects were seen with RARα2 knockdown. RARα2 induced drug resistance by activating the drug efflux pump gene ABCC3 and anti-apoptotic Bcl-2 family members. Inhibition of Wnt signaling or ABCC3 function could overcome drug resistance in RARα2 over-expressing MM cells. Our In vivo study indicated that targeting Wnt and Hh pathways using CAY10404, cyclopamine or itraconazole could significantly reduce tumor burden and increase survival time in the 5TGM1 mouse model. We confirmed these findings in primary myeloma stem cells. Finally, we showed that primary myelomas with high RARα2 expression possess similar features as myeloma stem cells.

Conclusion

This study demonstrates that RARα2 plays a crucial role in MMSCs by activating Wnt and Hh pathways, regulating the transcription of iPS genes and conferring drug resistance.

Disclosures:

No relevant conflicts of interest to declare.
Immune Regulatory Properties Are a Common Feature Of Stem Cells
Authors: Di Trapani Mariano , Bassi Giulio , Ricciardi Mario , Fontana Emanuela , Bifari Francesco , Pacelli Luciano , Giacomello Luca , Pozzobon Michela , Feron Francois , De Coppi Paolo , Anversa Piero , Fumagalli Guido , Decimo Ilaria , Menard Cedric , Tarte Karin , Krampera Mauro
Published: October 2019

Allogeneic stem cell-based therapy is a promising tool for the treatment of a range of human degenerative and inflammatory diseases. Many reports highlighted the immune modulatory properties of some stem cell (SC) types, such as mesenchymal stromal cells (MSCs), but a comparative study with SCs of different origin, to assess whether immune regulation is a general SC property, is still lacking. To this aim, we applied highly standardized methods employed for MSC characterization to compare the immunological properties of bone marrow-MSCs, olfactory ecto-mesenchymal stem cells, leptomeningeal stem cells, and three different c-Kit-positive SC types, i.e. amniotic fluid SCs, cardiac SCs, and lung SCs. We found that all the analyzed human SCs share a common pattern of immunological features, in terms of expression of activation markers, modulatory activity towards immune effector cells, immunogenicity and molecular inhibitory pathways, with some SC type-related peculiarities. In addition, we found that the inhibitory behaviour is not a constitutive property of SCs, but is acquired as a consequence of immune effector cell activation, as previously described for MSCs. Thus, immune regulation is a general property of stem cells and the characterization of this phenomenon may be useful for a proper therapeutical use of SCs.

Disclosures:

No relevant conflicts of interest to declare.
Immunological Characterization Of Multipotent Mesenchymal Stromal Cells. The International Society For Cellular Therapy (ISCT) Working Proposal
Authors: Krampera Mauro , Galipeau Jacques , Shi Yufang , Tarte Karin , Sensebé Luc
Published: October 2019

The large number of experimental approaches, culture conditions, qualitative and quantitative methods, and in vitro and in vivo models employed so far to assess immune regulatory properties of multipotent mesenchymal stromal cells (MSC) has led to an excess of literature data that sometimes are poorly comparable, redundant, and even contradictory. Thus, quite paradoxically, the risk is that pre-clinical literature data may become eventually weak and scarcely useful, in both researchers’ and Regulatory Authorities’ opinion, for supporting experimentally specific MSC-based clinical trials aimed at treating autoimmune and inflammatory diseases. However, some data in this field appear more solid and reproducible and may be generally accepted to suggest reproducible immunological assays to quantify the differences in immune modulatory properties of MSCs produced according to Good Manufacturing Practice (GMP).

The MSC Committee of the International Society of Cell Therapy (ISCT) released a statement paper in 2006 that established the minimal criteria characterizing human MSC, without focusing particularly on their immunological properties. In the 7 years following the publication of this statement paper, more than 10,000 manuscripts on MSC, and many of them deal with immune regulation. To consolidate the scientific research in this field, the MSC Committee of the ISCT is publishing a working proposal paper aimed at stimulating the general discussion about the need of shared guidelines for the immunological characterization of MSCs for clinical use:

1. A standard immune plasticity assay should be implemented by using IFN-γ + TNF-α as model in vitro priming agent

2. Functional analysis of an expanded cell product may provide mechanistic insights on intra- and inter- study variance in clinical response amongst patients

3. The use of purified responders would be widely practicable and should provide more generalizable guidance on relative functional potency of MSC and as a companion to clinical trials

4. Interrogating the IDO response as part of an in vitro licensing assay should be considered central

5. Conclusions based on xenorecipient animal models on how to conduct clinical trials should be drawn with caution

6. The prospective hypothesis-driven analysis of lymphocyte populations in patients groups treated with MSC should be encouraged

7. Clinical analysis should also include the monitoring of whether injected MSCs are the target of an immune response.

Disclosures:

No relevant conflicts of interest to declare.
CD26+ (dipeptidyl peptidase IV positive) Cells In Harvest Material For Multiple Myeloma Autotransplantation
Authors: Kopinska Anna , Krawczyk-Kulis Malgorzata , Suszek-Dziaczkowska Joanna , Bieszczad Katarzyna , Jagoda Krystyna , Kyrcz-Krzemien Slawomira
Published: October 2019

Dipeptidyl peptidase IV(DDP IV) known as CD26 can play a role in the process of mobilization as well as in regeneration during hematopoietic cells transplantation. As we know from the studies conducted in vitro and in animals models DDP IV inactivates chemokine CXCL12 (stroma direct factor 1) causing interruption the connection between marrow stoma and hematopoietic cells through CXCR4 receptor.In physiological conditions on the surface of hematopoietic cells concentration of CD26 is low.The purpose of this study was to examine a correlation between the number of CD26+ and CD34+ cells collected during mobilization in multiple myeloma (MM)patients (pts) and investigate the expression of CD26+ on monoclonal cells’. The second purpose was to examine whether CD34 cells have expression of CD26 parallel.30MM pts were mobilized in our department between 2011-2013. For harvest intermediate dose of cyclophosphamid and G-CSF were administered. The number of CD26+ cells and CD34+ cells were measured using immunophenotyping method before starting the procedure in peripheral blood and in collected materialResults All pts collected sufficient for AutoHCTnumber of CD34+ cells,the medianwas 3,8 (2,05-11,1x10^6/kgb.w.).Before mobilization 49,65% of blood nucleated cells showed CD26 expression while in collected material -79,95%. The CD34+ cells in peripheral blood showed low expression of CD26 – median 4,5%. In thecollected material the expression of CD26in CD34+ cells did not increase –median 2,5%, but the number of CD34+ cells (p=0,0003)increased. On mononuclear cells (monocytes and lymphocytes) there were demonstrated also the increase ofCD26 expression after mobilization chemotherapy and G-CSF (p=0,0002).Conclusion In MM patients after mobilization chemotherapy (cyclophosphamide+G-CSF) statistically significant increase ofnumber of CD26+ cellswere observed. It will be interesting to investigate the role of CD26+ cells during autotransplantation.

Disclosures:

No relevant conflicts of interest to declare.
Recombinant Thrombomodurin For The Treatment Of Transplantation-Associated Coagulopathy After Allogeneic Stem Cell Transplantation
Authors: Ishii Kazuyoshi , Nomura Shosaku , Ito Tomoki , Katayama Yuta , Kyo Taiichi , Ota Shuichi , Seki Masanori , Chiba Shigeru , Maeda Yoshinobu , Tanimoto Mitsune , Ikezoe Takayuki , Yagi Hideo , Ishida Yoji , Fujishima Naohito , Sawada Kenichi
Published: October 2019

Background

Levels of cytokines, chemokines and soluble molecules fluctuate after allogeneic hematopoietic stem cell transplantation (HSCT). These biomarkers are possible to be a diagnostic and prognostic value for transplantation-associated coagulopathy (TAC). In the present study, we investigated the effects of recombinant thrombomodulin (rTM) on levels of the cytokines, chemokines, and soluble factors registered in the ‘SIGHT’ research. SIGHT comprises the capital letters of five complications after HSCT, namely sinusoidal obstruction syndrome (SOS same as VOD), infection, GVHD, hemophagocytic syndrome and thrombotic microangiopathy.

Methods

The subjects were 159 patients who underwent allogeneic HSCT (bone marrow = 83, peripheral blood stem cells = 31, cord blood = 45). Blood samples were collected before and after transplantation. Levels of cytokines (interleukin-6, tumor necrosis factor-α, high-mobility group box (HMGB) 1), chemokines (monocyte chemotactic protein (MCP)-1, RANTES), and soluble molecules (soluble vascular cell adhesion molecule (VCAM)-1, soluble E-selectin, plasminogen activator inhibitor-1, platelet-derived microparticles (PDMP)) were measured by enzyme-linked immunosorbent assay. The rTM was administered as a therapy for transplantation-associated coagulopathy (TAC). This protocol was completed in day 4-14 after HSCT and consisted of day doses of 380 unit/kg with every days. Control group was also used heparin or no anti-coagulation therapy.

Results

MCP-1, IL-6, and TNF-a exhibited more significant elevations on days 7–14 after HSCT. In contrast, the levels of HMGB1, sE-selectin, sVCAM-1, PAI-1 and PDMP exhibited significant changes on days 14–28. There were significant improvements in TNF-a, sE-selectin, sVCAM-1, HMGB1, PAI-1 and PDMP after rTM-treatment (n=73), but not after rTM-untreatment patients (n=86).

Conclusion

We believe one of causes for TAC is pro-inflammatory cytokine including HMGB1. For this reason, it is thought that the direct anti-inflammatory effect of rTM’s lectin domein plays an important role in therapeutic mechanism for TAC. The present findings suggest the possibility that rTM can play a therapeutic role for TAC after allogeneic HSCT.

Disclosures:

No relevant conflicts of interest to declare.
Subsequent Malignant Neoplasms After Allogeneic Hematopoietic Stem Cell Transplantation Using Reduced-Intensity Conditioning and Outpatient Conduction
Authors: Zamora-Ortiz Gabriela , Velazquez-Sanchez-de-Cima Sara , Ponce-de-Leon Sergio , Gutiérrez-Aguirre César H. , Ruiz-Delgado Guillermo J. , Gomez-Almaguer David , Ruiz-Arguelles Guillermo J.
Published: October 2019

Abstract 5502

Patients given allogeneic hematopoietic transplants (HSCT) may develop subsequent malignant neoplasms (SMN). Several variables have been identified but there are no data about the incidence of this complication in individuals given HSCT using reduced-intensity conditioning (RIC) methods. The objective of this study is to define the incidence of SMN in patients given HSCT using a RIC preparative regimen conducted on an outpatient basis. Patients given HSCT in two institutions between October 1998 and 2012 were analyzed. Overall survival was analyzed with the Kaplan-Meier procedure. Patients alive at the closure of the study or those lost to follow up were censored.  To appraise the SMN appearance, those patients dead were also regarded as censored at that moment, as well as those lost to follow up and those alive at the closing of the study. 95% confidence intervals for the survival or failure estimate were calculated with the Greenwood’s method. All the survival analyses were processed with the StataCorp 2005. Stata Statistical Software: Release 9. College Station, TX: StataCorp LP. A total of 416 allografted patients with a Karnofsky performance index of 100% were included in the study. All patients received PBSC allografts using reduce-intensity conditioning. Engraftment occurred in 350 patients (84%). The conditioning regimen was delivered as an outpatient procedure in all individuals. No patient was given radiotherapy nor antithymocyte globulin during the conditioning. Three hundred and sixty five patients (88%) were never admitted to the hospital, whereas 12% were admitted because of grade III-IV aGVHD, fever or mucositis. Median survival time was 15.7 months.  Survival at 6 months (95% CI):  66.4% (61.5- 70.8%), at 12 months:  53.3% (48.1 -58.1%), at 60 months: 30.6% (30.5-41.5%). Eight patients with a SMN were identified in the group of 416 allografted patients, the cumulative probability of SMN being 6.8 at 10 years. Since the number of expected cases in the general population is 0.62, the ratio of observed to expected cases is 3.2 (p < 0.001). This figure means that the risk of developing a malignant neoplasm in allografted individuals using our method is 3.2 times higher than that in the general population. There were three non-Hodgkin´s lymphomas (NHL), two M2 acute myelogenous leukemias (AML), one hairy cell leukemia, one tongue epidermoid carcinoma and one breast carcinoma. In conclusion, we have found a diminished incidence of SMN in this group of Mexican patients allografted with the Mexican reduced-intensity conditioning method. Possible explanations for this difference are discussed, focusing on the RIC preparative regimen.

Disclosures:

No relevant conflicts of interest to declare.
Biosimilar Compared With Originator Filgrastim For Related-Donor Allogeneic Stem Cell Mobilisation: A Prospective-Historical Control Study
Authors: Lefrère Francois , Ribeil Jean-Antoine , Turner Matthew , Hermine Olivier , Blanche Stephane , Cavazzana-Calvo Marina
Published: October 2019

Introduction

Biosimilars of filgrastim have been available for over five years In Europe, where their use now exceeds that of the originator product. However, some professional bodies have raised concerns over their use in healthy donors for allogeneic mobilisation given the absence of clinical data in this setting. Here we report the use of biosimilar filgrastim compared with a matched historical control group in allogeneic transplant.

Methods

A total of 26 healthy related-donors (parent or sibling) received biosimilar filgrastim (HX575) for allogeneic stem cell mobilisation at a single centre between 2011 and 2013. Donors and recipients were compared with a matched historical control group (n=48) who had been treated with original filgrastim (Neupogen) at the same centre between 2005 and 2011. Donors and patients in both groups were treated according to the same clinical protocol, with G-CSF 10 µg/kg/day administered to donors on days 1-5 with leukapheresis performed on day 5.

Results

Donor and recipient characteristics (age, gender, body weight) were similar in both groups. Both the biosimilar and originator groups had similar donor mean white blood cell counts at baseline (6.13 vs 6.24 x 109/l) and on day 5 (46.9 vs 45.3 x 109/l). Mean donor CD34+ cell count on day 5 was 92/µl in the biosimilar group and 88/µl in the originator group (p=0.713). Mean number of CD34+ cells per recipient body weight was 9.7 x 106 in the biosimilar group and 8.00 x 106 in the originator group (p=0.437). Occurrence and intensity of bone pain was similar in donors in both groups. The majority of recipients in both groups had acute leukemia, myeloma, lymphoma or congenital immunodeficiency syndrome and around half underwent a non-myeloablative transplant. Median time to neutrophil engraftment (>500/µl) was similar in both the biosimilar and originator groups (16.5 [range 11-44] vs 15.0 [range 9-23] days; p=0.078), as was platelet recovery (>20 g/l) (12.5 [range 8-28] vs 12.5 [range 3-38] days; p=0.990). Acute graft-versus-host disease (GVHD) occurred in 27% of patients in the biosimilar group and 38% patients in the original group, while chronic GVHD occurred in 15% and 19% of recipients, respectively.

Conclusions

Biosimilar G-CSF is as effective and well tolerated as originator G-CSF for related-donor allogeneic stem cell mobilisation. Long-term follow-up of donors is required to confirm the safety of biosimilar and originator G-CSF.

Disclosures:

Lefrère: Sandoz Biopharmaceuticals: Consultancy, Honoraria. Turner:Sandoz Biopharmaceuticals: Employment.
Acceptable Results Of Allo-HSCT From Unrelated Donors In AML Patients Who Failed To Achieve The Complete Remission
Authors: Markiewicz Miroslaw , Dzierzak-Mietla Monika , Rybicka Malwina , Gaj Magdalena , Koclega Anna , Zielinska Patrycja , Bialas Krzysztof , Kyrcz-Krzemien Slawomira
Published: October 2019

Introduction

Despite the development of a variety of new investigational therapies for patients diagnosed with acute myeloblastic leukemia who either fail to achieve remission or who relapse thereafter, identifying suitable patients for allogeneic hematopoietic stem cell transplantation (allo-HSCT) and choosing a best tolerable therapy that is most likely to succeed remains a difficult clinical problem. The rationale of this study was to evaluate results of allo-HSCT in AML patients who have failed to achieve complete remission pre-transplant.

Patients and Methods

33 pts with refractory or relapsed AML with no (18) or with partial remission (15), aged 35 (15-64) years received allo-HSCT from 10/10 (19) or 9/10 (14) HLA-matched Unrelated Donors (UD) in Hematology and BMT Center in Katowice, Poland, from Dec 2000 to Jan 2013. The myeloablative preparative regimen consisted of busulfan 4x4 mg/kg plus cyclophosphamide (Cy) 4x30 mg/kg (18 pts), TBI (12 Gy) plus Cy 3x40 mg/kg (6 pts), or treosulfan 3x14 g/m2 plus either fludarabine 5x30 mg/m2 (7 pts) or Cy 3x40 mg/kg (2 pts). Standard GVHD prophylaxis consisted of cyclosporine-A, methotrexate and pre-transplant ATG or thymoglobulin. Source of cells was peripheral blood (22 pts) or bone marrow (11 pts) with median 6.8 or 3.9 x10(6)CD34+cells/kg, respectively.

Results

All but 4 pts engrafted. The 2-year estimated overall survival rate was 47%. 17 pts died 4 (0.3-16) months following allo-HSCT due to relapse (7) infection (5), GVHD (1) GF (1) and unknown cause (3). Acute GVHD grade I, II, III and IV was observed in 15, 6, 2 and 1 pt (serious aGVHD grade III-IV in 3 pts (9%)), limited or extensive chronic GVHD was present each in 5 (20%) of 25 evaluable pts. Other complications included CMV reactivation (13), hemorrhagic cystitis (7), serious 3-4 grade mucositis (8), SOS (3) and renal failure (1). 16 pts (48.5%) are alive 45 months (4 months-10 yrs) post-transplant. No difference in survival has been observed between pts transplanted in PR or NR stages nor between pts treated with variable preparative regimens.

Conclusions

Allo-HSCT from UD with myeloablative conditioning is a feasible and effective curative therapy providing acceptable rates of long-term remission or cure in refractory or relapsed AML in patients who fail to achieve CR with conventional treatment.

Disclosures:

No relevant conflicts of interest to declare.
Allogeneic Stem Cell Transplantation For Primary Refractory Hodgkin's Disease: Ablative Vs Reduced Intensity
Authors: Ozcelik Tulay , Guvenc Serkan , Hindilerden Fehmi , Hasbal Baris , Serefhanoglu Songul , Küçükkaya Reyhan , Arat Mutlu
Published: October 2019

Minority of patients with Hodgkin’s lymphoma (HL) are primary refractory or relapse very early after first-line chemotherapy or even after high-dose chemotherapy (HDCT) and autologous HSCT. The dismal outcome of these patients was shown by many previous studies. We evaluated the role of allogeneic stem cell transplantation in this setting. We retrospectively analysed 6 consecutive patients (med age 45) with primary refractory advanced stage Hodgkin’s lymphoma transplanted between 2011-2013. The median time from initial diagnosis and from autologous HSCT to allogeneic transplantation was 44.1 mo.s and 12.4 mo.s, respectively. The donor sources were sibling in 4, unrelated in one and haploidentical in 1. The conditioning regimen was myeloablative (BU-CY) in 3 and reduced intensity (Flu-Mel±ATG) in 3 adjusted according to their age and comorbid conditions. All of them received peripheral blood stem cells. Engraftment failure was not observed. None of them were complicated with VOD or other early complications of HSCT. Day 100 transplantation related mortality was 16.7% (1 patient). The patient succumbed to carbapenemase+ klebsiella pneumonia sepsis on day +11. Median follow up of the surviving patients was 14.65 months (range, 6-26). At the last follow up 3 of them were in CR, one patient had progression after HSCT and achieved stable disease with brentuximab and DLI. One patient died from acute abdominal infection on +187 while in CR. Only one patient was complicated with both acute and chronic GVHD (unrelated HSCT). The myeloablative regimen was tolerated well and the patients achieved durable remission. The expired patients were both in RIC arm. Although the number of transplanted patients is low we can conclude that allogeneic HSCT using myeloablative regimen is safe and effective in short term. Primary refractory HL patients should be evaluated for allogeneic HSCT, as upfront or following failure of autologous HSCT and HDCT. Ablative regimen is tolerated as well as RIC, and more effective than allo RIC for disease control in the light of our current limited cohort.

Disclosures:

No relevant conflicts of interest to declare.
Autologous Stem Cell Transplantation For Acute Leukemias In Suzhou (China): Similar Outcome In CR1 Patients Compared With European Group For Blood and Marrow Transplantation (EBMT) Results: A Pair-Matched Analysis
Authors: Chen Jia , Lu Yin , Labopin Myriam , Wu De Pei , Mohty Mohamad , Gorin Norbert Claude
Published: October 2019

The first affiliated hospital of Soochow University initiated a program of stem cell transplantation for hematological malignancies in 2001 and has done until December 2011 a total of 70 autografts to consolidate acute leukemias (61 adults and 9 children).

The EBMT presently handles a registry with information on more than 450 000 transplants including over 100 000 transplants for Acute Leukemias, of which 22800 were autografts. In order to evaluate the outcome of patients transplanted in Suzhou and to compare it with results from the EBMT, we collected the necessary information on all patients transplanted in Suzhou and we did a pair matched analysis with patients reported to the EBMT registry.

The median age of the patient population autografted in Suzhou was 32 years (4-63). The median follow up was 34 months (1-136). 59 patients (47 AML, 12 ALL) were autografted in CR1 and 11 in CR2/3. The majority of patients received a non TBI conditioning (87%) and PB as a source of stem cells (87%). For those autografted in CR1, the interval from CR1 to transplant was 203 days (75-404). At 2 years, the OS, LFS, RI and NRM were 73±7%, 52± 7%, 44±8% and 5±3% for AML and 76±14%, 67± 16%, 33±17% and 0% for ALL. 48 adult patients autografted in CR1 from Suzhou were matched with 89 patients from the ALWP EBMT registry. Matching factors were age ± 3 years, Cytogenetics and the number of induction courses to reach CR1 (1 course versus more than 1). Patients from Suzhou were transplanted more recently ; the interval from diagnosis to transplant was longer (242 days vs 172 days, p<0.0001) and TBI was less frequently used (p= 0.004 ). By univariate analyses the results were: Suzhou versus EBMT: OS 76 ± 6 vs 69 ±5 % (p= 0.33), LFS 55 ± 7 vs 60 ± 5% (p= 0.44), RI 40 ± 7 vs 33 ± 5% (p= 0.27), NRM 4 ± 3 vs 7 ± 3% (p = 0.47). By multivariate analyses adjusting for interval from diagnosis to transplant, year of transplant and use of TBI, there was no significant difference for OS (HR: 0.65, 95% CI: 0.19-2.19; p=0.49), LFS (HR: 0.97, 95% CI : 0.44-2.15; p=0.95), RI ( HR: 1.02, 95% CI : 0.46-2.28; p=0.96) .

We conclude that the results from Suzhou are not statistically different from those obtained using the EBMT database. These findings as well as the observation in another study of similar outcomes following allogeneic transplants in China and within EBMT are important to consider when planning international multicenter randomized studies.

Disclosures:

No relevant conflicts of interest to declare.
Impact Of High Resolution HLA Mismatch On Outcomes Of Unrelated Peripheral Blood Stem Cell Transplantation
Authors: Park Silvia , Kim Kihyun , Jang Jun Ho , Jung Chul
Published: October 2019

Introduction

High resolution HLA disparity between donor and recipient affect transplantation outcomes of unrelated peripheral blood stem cell transplantation (PBSCT), however the importance of individual loci in a high resolution level still remains unclear. The purpose of current study was to examine the impact of high resolution HLA disparity on transplantation outcome.

Materials and methods

We retrospectively analyzed 104 pairs of donor-recipient, who received PBSCT from unrelated donors (MUDs) between January 2009 and August 2012 at the Samsung Medical Center. The result of high resolution typing of HLA-A, -B, -C and -DRB1 were reviewed. Clinical outcomes were evaluated in terms of acute/chronic GVHD, relapse and survival.

Results

All but 11 cases (89.4%) received serologically matched transplantation. Of the 104 pairs, mismatching at high resolution level was observed in 34 cases (32.7%), containing 1, 2, and 3 or more mismatches in 15 (14.4%), 14 (13.5%) and 5 (4.8%) cases. According to the HLA loci, mismatches were detected in 12 (11.5%), 10 (9.6%), 10 (9.6%) and 21 cases (20.2%) on HLA-A, -B- C, and DRB1 loci. There were no significant differences in the incidence of chronic GVHD between the 8/8 matched pairs and 7 or less/8 matched pairs. However, the incidence of grade II-IV acute GVHD was significantly higher in patients with HLA-A mismatch compared with those with full matched HLA-A (p=0.011), although it was not translated to differences in survival or relapse. Overall survival was significantly inferior in patients with mismatching on HLA-B loci (p=0.0399), and it was mainly attributable to increasing non relapse mortality (NRM) in these patients (p=0.0252).

Conclusion

The potential role of high resolution HLA disparity on the development of GVHD or transplant outcomes has not been confirmed in the present study, and further study with larger number is necessary. Of note, high-resolution mismatch on HLA-B might be associated with worse outcomes in unrelated donor PBSCT.

Disclosures:

No relevant conflicts of interest to declare.