bluuebunny/crossref_metadata_2025 huggingface dataset:
Clinical Results of Gemox-R in Mantle CELL Lymphoma: The Role of Oxaliplatin
Authors: García-Recio Marta , Gutierrez Antonio , Obrador-Hervia Antonia , García Mañó Lucia , Bento Leyre , Besalduch Juan , Rodriguez Jose , Sampol Antonia , Villalonga Priam , Fernandez Silvia
Published: October 2019

Introduction: Mantle cell lymphoma (MCL) is mostly incurable. The current standard therapy achieves a high rate of complete remission (CR), but the pattern of continuous relapses still marks this disease as a challenge. We previously reported the efficacy of GemOx-R, a combination regimen of gemcitabine, oxaliplatin and rituximab, in patients with refractory and relapsing MCL. Our aim is to confirm our previous results in a larger retrospective series and evaluate the efficacy of each component of GemOx-R in a panel of MCL cell lines and in patient-derived primary cells.

Methods: Between 2003 and 2015, 30 patients with MCL were included in a retrospective study of treatment with GemOx-R from the University Hospital Son Espases: 10 cases frontline and 20 in the salvage setting. Frontline cohort was consolidated with radioimmunotherapy and received maintenance therapy with rituximab. The translational study was performed in established cell lines as well as primary MCL lines from patients by cell viability, cell cycle, apoptosis and western blot analysis. Drug synergy was determined by the isobologram and combination index methods.

Results: This is a high risk series of patients: median age 70 years, 87% stage IV and 86% intermediate or high risk MIPI. Overall response rate and CRR was 80% and 60% in the frontline cohort as well as 85 % and 60% for salvage patients, respectively. Median progression-free survival was 28 months in the entire series: 66 and 22 months, respectively, for the two cohorts. Median overall survival was 34 months in the entire series: not reached and 20 months, respectively, for the two cohorts. Grade 3 and 4 toxicity was as follows: neutropenia (63%), anemia (34%) and thrombocytopenia (30%) as well as 24% of grade 1 and 2 neurotoxicity. Cell viability and apoptosis analysis showed that oxaliplatin is the most effective drug in this regimen in contrast to the poor responses induced by gemcitabine and rituximab. Oxaliplatin had a profound effect on cellular viability, consistent with the induction of caspase activityand the downregulation of pro-survival proteins. We further present synergistic efficacy of oxaliplatin combined with cytarabine in MCL cells.

Conclusions: (1) GemOx-R shows excellent results in MCL both in the frontline and salvage settings considering the high risk patients included. (2) Oxaliplatin is the most effective drug in GemOx-R; (3) oxaliplatin has a robust in vitro activity comparable to that of cytarabine, and the combination of both oxaliplatin and cytarabine shows a significant synergism; (4) taken together, our findings suggest that oxaliplatin alone or combined with cytarabine could constitute a new or alternative backbone for promising new regimens in MCL.

Disclosures

No relevant conflicts of interest to declare.
Phase II Study of Ofatumumab in Relapsed Nodular Lymphocyte-Predominant Hodgkin Lymphoma: A Report from the German Hodgkin Study Group
Authors: Eichenauer Dennis A. , Goergen Helen , Pluetschow Annette , Behringer Karolin , Kreissl Stefanie , Bröckelmann Paul , Fuchs Michael , Böll Boris , von Tresckow Bastian , Borchmann Peter , Engert Andreas
Published: October 2019

Background:

Nodular lymphocyte-predominant Hodgkin lymphoma (NLPHL) accounts for approximately 5% of all Hodgkin lymphoma (HL) cases. One hallmark of NLPHL is the consistent expression of CD20 on the malignant lymphocyte predominant (LP) cells. To shed more light on the role of anti-CD20 antibody treatment in relapsed NLPHL, we conducted a phase II study evaluating the fully humanized anti-CD20 antibody ofatumumab in 28 patients. Treatment consisted of 8 weekly doses (week 1: 300 mg, week 2-8: 1000 mg) of the antibody.

Results:

The median age of study patients was 45 years (range: 22-68) and the majority were male (64%). A median of 1 line of therapy (range: 1-5) had been applied prior to study treatment and 7/28 patients (25%) already had rituximab-containing treatment. At the final restaging 3 months after the end of treatment, response was documented in 27/28 patients (96%; 95%-CI: 84%-100%). After a median follow-up of 26 months, 1-year and 2-year progression-free survival (PFS) estimates were 93% and 80%, respectively. No patient died. Transformation into aggressive non-Hodgkin lymphoma (NHL) occurred in 2/28 patients (7.1%). No grade III/IV toxic events were observed.

Conclusion:

In summary, the anti-CD20 antibody ofatumumab represents a highly active and well tolerated treatment option in relapsed NLPHL. Longer follow-up is required for final conclusions.

Disclosures

Off Label Use: Ofatumumab in lymphocyte-predominant Hodgkin lymphoma. von Tresckow:Takeda: Consultancy; Celgene: Other: honoraria for preparation of scientific educational events; Novartis: Consultancy, Other: Travel and accomodation, Research Funding; Amgen: Other: honoraria for preparation of scientific educational events. Borchmann:Millennium: Research Funding. Engert:Takeda: Consultancy, Research Funding.
Selective Inhibitors of Nuclear Export (SINE) Compounds Suppress Both HIV Replication and AIDS Related Lymphoma
Authors: Daelemans Dirk , Boons Eline , Vanstreels Els , Jacquemyn Maarten , Nogueira Tatiane C , Neggers Jasper E , Vercruysse Thomas , van den Oord Joost , Tamir Sharon , Kauffman Michael , Shacham Sharon , Landesman Yosef , Snoeck Robert , Pannecouque Christophe , Andrei Graciela
Published: October 2019

Infection with human immunodeficiency virus (HIV) compromises the immune system leaving infected individuals vulnerable to other pathologies including cancer. HIV-infected patients have an increased risk of cancer development as compared to the general population. Primary effusion lymphoma (PEL), a high-grade non-Hodgkin lymphoma with very poor prognosis, typically affects individuals infected with HIV in advanced stages of acquired immune deficiency syndrome (AIDS). While the use of anti-HIV drugs is associated with better prognosis, treating cancer in HIV-infected patients is often challenging due to potential drug interactions between anti-cancer agents and antiretroviral drugs. We demonstrate that inhibition of the exportin-1 (CRM1/XPO1) mediated nuclear protein transport by SINE compounds results in effective suppression of HIV replication, which was quantified by measuring virus-associated core antigen (p24) by ELISA. In addition by using MTT, assays, Annexin staining, FACS and immunoblot analysis we demonstrated that XPO1 inhibition induced cell cycle arrest and apoptosis in PEL cell lines (BC-1, BCBL-1, and JSC-1) at similar doses. SINE blocked the nuclear export of the late viral partially and unspliced RNA species encoding for the viral structural proteins. In BC-1 PEL cells inhibition of XPO1 resulted in the nuclear accumulation of p53 tumor suppressor protein as well as inhibition of NF-κB activity. In vivo, oral administration twice a week for 4 weeks, 20 mg/kg of the clinical SINE compound selinexor (currently in phase I/II clinical trials) led to a significant suppression of the BC-1 PEL growth in vivo. These findings provide a strong basis for inhibiting XPO1 as a novel strategy for the combined treatment of HIV and PEL or other AIDS-related cancers.

Disclosures

Tamir: Karyopharm: Employment. Kauffman:Karyopharm: Employment, Equity Ownership. Shacham:Karyopharm: Employment, Equity Ownership. Landesman:Karyopharm: Employment.
Histone Deacetylase Inhibitor SAHA Mediates Epigenetic Silencing of KIT D816V Mutated Systemic Mastocytosis Primary Mast Cells and Selective Apoptosis of Mutated Mast Cells
Authors: Abdulkadir Hani , Grootens Jennine , Kjellander Matilda , Hellstrom Lindberg Eva , Nilsson Gunnar , Ungerstedt Johanna
Published: October 2019

Systemic mastocytosis (SM) is a myeloproliferative disease for which there is currently no specific therapy. Over 90% of the patients carry the D816V point mutation that renders the KIT receptor constitutively active. In the current study, we assessed the sensitivity of mast cell line HMC1.2 and primary SM patient mast cells to histone deacetylase inhibitors, and found that SAHA is most efficient. SAHA induced a rapid downregulation of KIT mRNA, with a subsequent reduction in total KIT protein as well as cell surface KIT. This was followed by major mast cell apoptosis. Primary SM patient mast cells cultured ex vivo were even more sensitive to SAHA than HMC1.2 cells, whereas healthy subject mast cells were unaffected. There was a correlation between cell death and SM disease severity, where cell death was more pronounced in the case of aggressive disease, with almost 100% cell death among mast cells from the mast cell leukemia patient. Using ChIP qPCR, we found that the level of active chromatin mark H3K18ac/totalH3 decreased significantly in the KIT region, due to an increase in H3 density. This epigenetic silencing was specific to the KIT region and not seen in control genes upstream and downstream of KIT. Primary analysis of ChIP-seq data for histone marks H3K4me3 and H3K27me3, demonstrates a downregulation of transcription factors involved in activation of KIT receptor, such as MAPK, for the SAHA treated samples. This indicates an indirect epigenetic silencing of KIT. Our results therefore demonstrate that SAHA epigenetically silences KIT, and work is ongoing to elucidate the exact mechanisms of KIT regulation. Altogether, SAHA maybe a specific treatment for SM.

Disclosures

No relevant conflicts of interest to declare.
Kras Is Critical for CD8 T Cell Antiviral Function
Authors: Chen Yuhong , Zheng Yongwei , You Xiaona , Xin Gang , Yu Mei , Fu Guoping , Zhou Fen , Zhu Wen , Cui Weiguo , Zhang Jing , Wen Renren , Wang Demin
Published: October 2019

Small GTPases regulate multiple signaling pathways and individual Ras member can have distinct biological function. To overcome embryonic lethality of Kras-deficient mice, we generated and examined mice with hematopoietic- and T cell-specific deletion of Kras. In VavCreKrasfl/fl mice with hematopoietic deletion of Kras, thymic T-cell development was normal based on the presence of normal populations of total, CD4- CD8-, CD4+ CD8+, CD4+ and CD8+ thymocytes. The populations of splenic CD4+ and CD8+ T cells were also comparable between VavCreKrasfl/fl relative to control mice. In addition, no consistent defects in the 3 H-thymidine incorporation rate of Kras-deficient splenic CD4+ or CD8+ T cells in response to anti-CD3 or anti-CD3 plus IL-2 was detected. Nonetheless, we studied the effect of Kras deficiency on CD8 T-cell immune response to acute infection of the Armstrong strain of lymphocytic choriomeningitis virus (LCMV). Sub-lethally irradiated Rag1-deficient mice transplanted with bone marrow (BM) cells from VavCreKrasfl/fl or control mice were subjected to LCMV infection. Infection-induced expansion of CD8 T cells and generation of LCMV epitope gp33-specific CD8 T cells were markedly reduced in the recipients that received the BM from VavCreKrasfl/fl relative to control mice. Following in vitro stimulation with the LCMV epitope gp33, the induction of IFNg-expressing CD8 T cells from LCMV-infected recipients that received the BM from VavCreKrasfl/fl mice was dramatically reduced. Further, BM chimeric mice with CD8 T cell-specific deficiency generated by transplantation of lethally irradiated CD8 T cell-depleted CD45.1 congenic mice with a mixture of BM cells from VavCreKrasfl/fl mice and BM cells from CD8 T cell-deficient mice exhibited an impaired CD8 T-cell immune response to LCMV infection. Lastly, we examined the role of Kras in TCR signaling. The level of total TCR-activated Ras (Ras-GTP) was markedly reduced in Kras-deficient relative to control CD8 T cells. Importantly, TCR-induced ERK1/2 activation was impaired in Kras-deficient relative to control CD8 T cells. Consistently, TCR-induced activation of Raf-1 and MEK1/2 was markedly reduced in mutant CD8 T cells. However, TCR-induced JNK and p38 activation as well as Ca2+ flux were normal in Kras-deficient CD8 T cells. Of note, TCR-induced activation of Ca2+ flux, JNK and p38 as well as ERK1/2, MEK1/2 and Raf1 was normal in Kras-deficient relative to control CD4 cells. Taken together, these data demonstrate that Kras is dispensable for T cell development or TCR-induced proliferation of CD4 or CD8 T cells in vitro, but regulates TCR-induced activation of the Raf-1/MEK/ERK pathway in CD8 but not CD4 T cells and intrinsically controls CD8 T-cell immune response to viral infection.

Disclosures

No relevant conflicts of interest to declare.
SFRP4-Mediated Inhibition of Canonical Wnt-Signaling in DLBCL
Authors: Koch Raphael , Chapuy Bjoern
Published: October 2019

Introduction: Wnt-signaling enhances proliferation in a variety of malignancies including diffuse large B-cell lymphoma (DLBCL) and exosomes, small secreted microvesicles, are known to carry active wnt-molecules to mediate wnt-signaling. We found consistent suppression of SFRP4, an inhibitor of canonical wnt-signaling in DLBCL, elucidated it's mechanism of action and addressed it´s impact on regulating proliferation of diffuse large B-cell lymphomas. We conducted a screen of chemical wnt-inhibitors and assessed wnt-addiction in DLBCL cell lines.

Material & Methods: The Cancer Cell Line Encyclopedia (CCLE) was used to assess the transcript abundace of SFRP4 in 18 DLBCL cell lines. SFRP4-expression was confirmed using western blot and immunofluorescence in representative DLBCL cell lines. Purified exosomes of DLBCL cell lines, recombinant human SFRP4 and SFRP4-shRNA were used to address the biological mechanism of SFRP4 action. 5-Azacytidine was used to restore SFRP4-expression. Furthermore shRNA mediated knockdown of beta-catenin and pharmacological inhibition of canonical wnt-signaling using XAV-939, IWP-2, C59 and UM206 were used to address wnt-addiction in DLBCL cell lines.

Results: We found SFRP4 to be significantly suppressed in a variety of malignancies, including DLBCL, with promotor methylation as a known mechanism of suppression. SFRP4 was able to suppress exosomal mediated canonical wnt-signaling. SFRP4 inhibited proliferation in DLBCL cell lines and enhanced cytotoxicity of doxorubicin. Treatment with 5-Azacytidine restored expression of SFRP4, induced decline of beta-catenin and suppressed cell proliferation. Knock down of beta-catenin and pharmacological perturbation of canonical wnt-signaling identified wnt-addiction in beta-catenin expressing cell lines in the absence of a MYC translocation or amplification.

Conclusions: SFRP4 suppresses proliferation of DLBCL cell lines by antagonizing exosomal mediated canonical wnt-signaling. Pharmacological inhibition of canonical wnt-signaling significantly decreased proliferation of DLBCL cell lines in the absence of genomic alterations involving MYC.

Disclosures

No relevant conflicts of interest to declare.
Pcbp1 and Ncoa4 Regulate the Flux of Iron through Ferritin in Developing Erythroid Cells
Authors: Ryu Moon-Suhn , Wohlschlegel James , Philpott Caroline C.
Published: October 2019

In general human cell models, Poly(rC)-binding proteins (Pcbp) mediate the delivery and integration of iron into the iron storage protein. Conversely, nuclear receptor coactivator 4 (Ncoa4) initiates the liberation of ferritin iron by localizing the protein to autophagosomes. The erythron is the major consumer of body iron. Whether these regulators of ferritin iron are functional in developing red cells remains unclear, and thus was investigated. Both Pcbp1 and Ncoa4 were readily detectable in immunoprecipitates of erythroid ferritin. The peak of Pcbp1-binding to ferritin preceded the stage of maximal Ncoa4-ferritin interaction, which coincided with the induction of β-globin, during erythroid maturation. Both Pcbp1 and Ncoa4 deficiency resulted in lower iron incorporation into heme and hemoglobin by differentiation, without compromised Alas2 gene expression. In Pcbp1-deficent cells, less iron loading into ferritin accompanied the erythropoietic defects. Ncoa4-lacking cells exhibited a massive accumulation of iron in ferritin. No apparent interaction between Pcbp2 and ferritin was detectable in erythroid cells. However, Pcbp2 depletion affected ferritin mineralization in a manner opposite to Pcbp1 deficiency. This was associated with an increase in Pcbp1-ferritin complex formation detected by coimmunoprecipitation. Supplemental iron led to decreases in the amount of both Pcbp1 and Ncoa4 interacting with ferritin. The iron-responsiveness of Ncoa4's ferritin-binding activity was primarily due to post-transcriptional changes in protein abundance. The down-regulation of Ncoa4 protein by high iron required intact lysosome activity, and the expression of an E3 ubiquitin protein ligase, Herc2. Collectively, the present studies provide evidence of the metallochaperone activity of Pcbp1 for ferritin mineralization, and the involvement of Ncoa4 in the mobilization process of stored iron in erythroid cells. Furthermore, the impaired hemoglobinization by insufficiency of these genes identifies ferritin as an essential source of iron for heme production during terminal erythroid differentiation.

Disclosures

No relevant conflicts of interest to declare.
Fam210b Is Required for Optimal Cellular and Mitochondrial Iron Uptake during Erythroid Differentiation
Authors: Yien Yvette Y , Chen Caiyong , Shi Jiahai , Li Liangtao , Bauer Daniel E. , Huston Nicholas , Kingsley Paul D. , Ablain Julien , Zon Leonard I , Palis James , Orkin Stuart H. , Kaplan Jerry , Phillips John D , Lodish Harvey F. , Paw Barry
Published: October 2019

Red cells synthesize large quantities of heme during terminal differentiation. Central to erythropoiesis is the transport and trafficking of iron within the cell. Despite the importance of iron transport during erythroid heme synthesis, the molecules involved in intracellular trafficking of iron are largely unknown. In a screen for genes that are up-regulated during erythroid terminal differentiation, we identified FAM210B, a predicted multi-pass transmembrane mitochondrial protein as an essential component of mitochondrial iron transport during erythroid differentiation.

In zebrafish and mice, Fam210b mRNA is enriched in differentiating erythroid cells and liver (fetal and adult), which are tissues that require large amounts of iron for heme synthesis. Here, we report that FAM210B facilitates mitochondrial iron import during erythroid differentiation and is essential for hemoglobin synthesis. Zebrafish are anemic when fam210b is silenced using anti-sense morpholinos (Fig. A). CRISPR knockout of Fam210b caused a heme synthesis defect in differentiating Friend murine erythroleukemia (MEL) cells. PPIX levels in Fam210b deficient cells are normal, demonstrating that Fam210b does not participate in synthesis of the heme tetrapyrrole ring. Consistent with this result, supplementation of Fam210b deficient MEL cells with either aminolevulinic acid, the first committed substrate of the heme synthesis pathway or a chemical analog of protoporphyrin IX failed to chemically complement the heme synthesis defect. While Fam210b was not required for basal housekeeping heme synthesis, Fam210b deficientcells showed defective total cellular and mitochondrial iron uptake during erythroid differentiation (Fig. B). As a result, Fam210b deficient cells had defective hemoglobinization. Supplementation of Fam210b-/- MEL cells with non-transferrin iron chelates restored erythroid differentiation and hemoglobin synthesis; whereas, similar chemical complementation could not be achieved in the Tmem14c-/- cells, which have a primary defect in tetrapyrrole transport. (Fig. C). Our findings reveal that FAM210B is required for optimal mitochondrial iron import during erythroid differentiation for hemoglobin synthesis. It may therefore function as a genetic modifier for mitochondriopathies, anemias or porphyrias.

Figure 1. Figure 1.

Disclosures

Bauer: Biogen: Research Funding; Editas Medicine: Consultancy. Orkin:Editas Inc.: Consultancy.
A Germline Mutation in ERBB3 Predisposes to Inherited Erythroid Myelodysplasia/Erythroleukemia
Authors: Braunstein Evan M. , Li Ruijuan , Sobreira Nara , Gocke Christopher , Brodsky Robert A. , Valle David , Cheng Linzhao
Published: October 2019

Background: Myelodisplastic syndrome (MDS) and acute myeloid leukemia (AML) are usually sporadic diseases. However, rare familial cases have helped to identify human disease genes and provided insight into hematopoiesis and leukemogenesis. Further, the ability to identify familial MDS/AML disease genes has improved due to advances in next generation sequencing technology. In addition to RUNX1, GATA2, CEBPA, germline mutations in both ETV6 and DDX41 were recently shown to predispose to myeloid hematologic malignancies.

Methods: We investigated a large family with an inherited MDS/AML marked by erythroid hyperplasia in all affected individuals. A family pedigree was consistent with an autosomal dominant mode of inheritance with high penetrance. Whole exome sequencing was performed on germline DNA of the proband and multiple unaffected family members in order to identify the inherited genetic mutation predisposing to disease. Candidate single nucleotide variants (SNVs) were obtained after excluding all known SNVs with a variant allele frequency greater than 0.01, then prioritized and screened using DNA from a second affected individual.

Results: An Ala1337Thr missense variant in the ERBB3 gene, a member of the EGFR tyrosine kinase receptor family, was identified as the potential pathologic germline mutation in this family. Further sequencing identified the presence of this variant in two additional affected individuals and its absence in eight unaffected family members, indicating that it co-segregates with disease. Functional validation of this ERBB3 variant was performed using an erythroleukemia cell line. Overexpression of the ERBB3 A1337T mutant in TF1 cells conveyed a block in erythroid differentiation compared to cells expressing unmutated ERBB3, measured by the lack of upregulation of Glycophorin A expression in the presence of erythropoietin. This activity was dependent upon the presence of Neuregulin-1β, the ERBB3 receptor ligand, and expression of the co-receptor ERBB2. In addition, ERBB3 mutated cells displayed a 1.3-fold growth advantage over unmutated ERBB3 in the presence of the Neuregulin-1β.

Conclusions: This work has identified a novel mutation that predisposes to MDS/AML and adds ERBB3 to the growing list of disease predisposition genes in hematologic malignancies. It will allow for the early diagnosis of individuals with inherited erythroid MDS/AML prior to the onset of disease. In addition, it allows for the characterization of a gene with a likely crucial role in erythropoiesis as well as hematologic disease.

Figure 1. Figure 1.

Disclosures

Brodsky: Alexion Pharmaceuticals: Membership on an entity's Board of Directors or advisory committees.
CLL Disease Severity Is Enhanced in Tcl1 Mice Deficient for Pro-Apoptotic Regulator Noxa
Authors: Slinger Erik , Wensveen Felix , Kater Arnon P. , Eldering Eric
Published: October 2019

Despite the development of new treatment strategies, chronic lymphocytic leukemia (CLL) remains an incurable disease. Apoptosis dysregulation caused by environmental signaling, predominantly within lymph nodes, plays a key role in CLL maintenance and treatment resistance. In earlier work, we demonstrated that expression of the pro-apoptotic Bcl-2 family memberNoxa is increased in circulating CLL cells compared to normal B-cells (1). Moreover, Noxa levels are decreased in CLL cells that reside in the lymph node while its key anti-apoptotic binding partner, Mcl-1, shows a reverse trend (2). This suggests that the decreased Noxa/MCL-1 ratio within the microenvironment may contribute to enhanced CLL survival. Here we aimed at elucidating a functional role for Noxa in CLL in the context of a murine model by crossing Noxa deficient mice (Noxa-KO) with Eμ.Tcl1-Tg mice (TCL1) mice.

We first established that also in Tcl1 mice the Noxa/Mcl-1 ratio was increased PB vs. LN cells. The phenotype and distribution of leukemic cells was similar in Tcl1 and Noxa-KO/tcl1 mice. Interestingly, the accumulation of CLL cells in the peripheral blood was accelerated in Noxa-KO/TCL1 mice. This was accompanied by a reduction of apoptotic cells in the spleen of Noxa-KO/TCL1 mice. In accordance with these findings, survival of Noxa-KO/Tcl1 mice was decreased compared to the Tcl1 mice (342 days vs. 396 days, p<0.001, Figure 1). Our data suggest a role for Noxa in CLL as a tumor suppressor, and provides a rationale for the use of compounds which may alter the Noxa/Mcl1 balance in patients.

1. W. J. Mackus et al., Chronic lymphocytic leukemia cells display p53-dependent drug-induced Puma upregulation. Leukemia19, 427-434 (2005).

2. L. A. Smit et al., Differential Noxa/Mcl-1 balance in peripheral versus lymph node chronic lymphocytic leukemia cells correlates with survival capacity. Blood109, 1660-1668 (2007).

Figure 1. Survival analysis of Noxa-KO/TCL1 mice and TCL1 mice. The Kaplan-Meier curves were analyzed by a Log-Rank test. Figure 1. Survival analysis of Noxa-KO/TCL1 mice and TCL1 mice. The Kaplan-Meier curves were analyzed by a Log-Rank test.

Disclosures

No relevant conflicts of interest to declare.
Prevalence of Oligoclonal Bands in Multiple Myeloma Patients Who Achieved Better Results Than Very Good Partial Response after Treatment with Standard or High Doses Chemotherapy-Final Analysis
Authors: Vieira Luiza soares , Crusoe Edvan de queiroz , Almeida Manuella de S. Sampaio , Sousa Lais , Perez ana Lucia , Cury Priscilla , Rossato Marina , Higashi Fabiana , Dias ana Luiza , Castro anna Carolina , Chiattone Carlos , Hungria Vania T M
Published: October 2019

Introduction - Oligoclonal bands (OB) are monoclonal proteins distinct from those originally identified in the multiple myeloma (MM) diagnosis. Some authors consider that appearance of these bands confers a better prognosis and may be linked to immune reconstitution. There is no data of the exact prevalence of OB emergence in patients with very good partial response (VGPR) or better after different treatment schedules. Objectives - To determine the prevalence of OB in MM patients treated with or without high-dose chemotherapy that obtained at least VGPR and its prognostic value. Methods- This is a retrospective and prospective cohort study. Data were collected from records of patients that achieved at least VGPR to identify the OB emergence. Subsequently, new sample collections from the positive patients were made in order to monitor the progress and duration of the maintenance of these bands. Results-Median follow-up was 42m and 101 patients were included. Median age was 58y (29-87) and 55% were male. IgG was the most frequent component (60%). Durie-Salmon IIIA/B was identified in 92% of the population; ISS was 33% in stage I, 30% in stage II, and 31% in stage III. The prevalence of OB identified by SPE and IF was 50.5% (51 cases), with a higher prevalence in those who underwent transplantation and those who achieved complete response (p=0.00139 and p=0.0368, respectively). Progression free survival (PFS) was longer in the OB group (45.4m x 34.7m p = 0.0075). Conclusion - The OB prevalence in this population was 50.5% and oligoclonality resulted in a longer PFS.

Figure 1. Figure 1.

Disclosures

No relevant conflicts of interest to declare.
Allogeneic Myeloma GVAX with Lenalidomide Enhances Progression Free Survival through the Generation of Tumor Specific Immunity in Patients in Near Complete Remission
Authors: Borrello Ivan M. , Noonan Kimberly , Huff Carol Ann , Ferguson Anna , Sidorski Amy , Rudraraju Lakshmi , Cimbro Raffaello , Marchionni Luigi
Published: October 2019

Background: The depth of response to therapy in myeloma (MM) correlates with improved progression free survival. We have previously shown that lenalidomide (Len) can augment vaccine efficacy to the pneumococcal conjugate vaccine (PCV), PrevnarÒ. We extend those observations to examine whether vaccinating patients on Len in a near complete remission (nCR) (negative M-spike, IFE positive) could further deepen the clinical response and generate measurable myeloma specific immunity.

Design: Patients on a Len-containing regimen (VRD, Rd, BiRD or R) that achieved and maintained a nCR for 4-6 months were eligible for the study. Patients continued only on single agent Len and received 4 GVAX vaccinations consisting of two allogeneic MM lines: H929, U266 admixed with K562 transduced to express GM-CSF as well as PCV. Patients received 3 monthly vaccines and a boost at 6 months. Immune monitoring was performed on BM samples obtained at baseline, 3 months and 1 year

Results: To date 32 patients have been screened. 17 patients initially in a nCR were ineligible for vaccination: 3 (18%) had disease progression, 7 (42%) entered into an IFE negative CR, and 7 (42%) maintained a nCRduring the observation period but opted not to enroll. 15 patients have been enrolled and completed their vaccinations. Patient characteristics are shown in the Table. Of note, none possessed high-risk features by ISS or FISH. Median follow-up for the study is 34.0 months. Median progression free survival (PFS) of the cohort of vaccinated patients has not been reached whereas the PFS in the observation arm that remained on the multidrug Len-containing therapy was 17.9 months (p<0.001). Vaccination in the setting of a rising M-spike was less likely to induce a durable remission with a median PFS of 14.3 months (p<0.003). Laboratory analysis showed that the patients achieving a CR had greater expression of PD-1 on CD4 and CD8 cells at baseline in the BM. Furthermore, durable responses were associated with the development and persistence of MM-specific immunity.

Conclusions: Vaccination in combination with Len in patients with minimal residual disease generates potent MM-specific immunity and appears to significantly extend the PFS. Vaccination in a nCR with a poly-antigenic approach such as GVAX in combination with Len-induced immunomodulation shows promising early clinical activity that warrants further investigation as an approach to inducing and maintaining durable clinical remissions.

Table 1. Vaccination(n=15) Observation(n=16) Age 69 (55-81) 66 (40-83) FISH (high risk) 0% 0% ISS Stage III 2 (13%) 3 (19%) IFE negative 0 (0%) 7 (42%) Prior Therapies 1.8 (1-4) 1.8 (1-3)

Disclosures

Borrello: Celgene: Research Funding. Noonan:Celgene: Speakers Bureau.
Influence of Matched Sibling Donor's Availability and Stem Cell Transplantation on Treatment Outcome for Acute Lymphoblastic Leukemia; Study on 294 Patients
Authors: Alfraih Feras , Chebbo Wahiba , Kim Dennis Dong Hwan , Aldawsari Ghuzayel , Alsharif Fahad Z. , Alzahrani Hazza A , Mohamed Said Y , Rasheed Walid , Hanbali Amr , Ahmed Syed Osman , Shaheen Marwan , Chaudhri Naeem A. , Al Mohareb Fahad , Aljurf Mahmoud
Published: October 2019

Introduction

Predicting treatment outcome of acute leukemia has been an important issue. Many factors have been elucidated. We evaluated the impact of donor's availability on treatment outcome including mortality in patients with acute lymphoblastic leukemia (ALL).

Methods

A total of 294 patients with ALL were evaluated after receiving chemotherapy between year 2001 and July 2014 at our center. Patients were assessed for the need of hematopoietic stem cell transplants (HSCT), availability of HLA sibling match donor and the impact on overall outcome. Indications for transplantation were defined as (1) WBC->100,000 for T-ALL, >30 for B-ALL with, (2) cytogenetic abnormalities of t(9:22), (4:11) or (1:19), (3) relapse or primary refractory disease or (4) MRD positivity. Patients were divided into 3 categories, group A with an indication for HSCT and available donor (HSCT+/D+), group B with HSCT indication but no available donor (HSCT+/D-) and group C with no indication for HSCT regardless of donor status (HSCT-).

Results

The median age was 20 (14-63 years). 95 (32%) were female while 198 (68%) were male. 276 (86%) patients were newly diagnosed while 18 (14%) were relapsed. Immuno-phenotype was B for 191 (65%), T for 91 (31%) versus mixed lineage for 12 (4%). 33 (11%) patients were positive for Philadelphia chromosome. Median WBC at diagnosis was 23.2X109/L CNS involvement was positive in 26 (8%) patients.

With a median follow-up of 60 months for survivors (range 2 to 116.5 months), 148 (50%) patients showed HSCT+/D+, HSCT+/D- in 79 (27%) and HSCT- in 67 (23%) patients.

the 5-year OS for HSCT+/D-, HSCT+/D+ and HSCT- were 29%, 57% and 55% (p value <0.001), (figure A) while DFS was 25%, 44% and 44% (p value =0.004), respectively. No statistical significance difference was noted on NRM between the groups. On multivariate analysis for OS, donor unavailability was confirmed as an independent risk factor (p<0.001, HR 2.21).

Conclusion

Our study suggested that unavailability of donor is an independent risk factor on OS. The use of alternative donor and building national /regional unrelated donor registry is highly warranted even in countries with high probability of finding matched sibling donor.

Figure 1. Figure 1.

Disclosures

Kim: Bristol-Myers Squibb: Consultancy, Research Funding; Novartis Pharmaceuticals: Consultancy, Research Funding.
Zoledronic Acid Inhibits Cell Growth of Multiple Myeloma Cells and Shows Synergistic Antimyeloma Effects with Bortezomib Via CD38 /cADPR /Ca2+ /Ras /Pakt /NF-Kb /Pim-2 Pathway
Authors: Fu Rong , Liu Zhaoyun , Liu Hui , Song Jia , Wang Guojin , Wang Yihao , Li Lijuan , Shao Zonghong
Published: October 2019

The Third-generation biphosphonate Zoledronic acid (ZOL) can exhibit direct antitumour activity which is widely used in multiple myeloma (MM) patients with bone events. However,it remains unknown whether Zol alone or combined with Bortezomib(B) has any inhibitory effect on myeloma cells. In this study, cell line RPMI-8226 were treated with ZOL alone at various concentrations and combined with Bortezomib to observe the effect and probable molecular mechanisms.Cell proliferation,cell apoptosis,cell phenotype(CD38+ and CD138+),RT-PCR, measurement of intracellular [Ca2+] concentration and Western blot analyses were performed.We found that ZOL alone strongly inhibited proliferation of multiple myeloma cells in vitro, and induced their apoptosis.The same results were obtained in the myeloma cell group treated with ZOL combined with Bortezomib after 24h instead of 48h.On the mechanism study, we found that Zol alone or combined with Bortezomib in 24h suppressed the mean fluorescence intensity(MFI) of CD38 and the mRNA of cADPR,NF-Kb,pim-2 ,and the signal pathway protein Ras/pAkt/NF-Kb/pim-2 and the intracellular [Ca2+]i concentration.However, it did not show the same results in 48h. In conclusion, Zol inhibited cell growth of multiple myeloma cells probably via suppressing CD38/cADPR/Ca2+/Ras/pAkt/NF-Kb/pim-2 pathway.Combination of Zol with Bortezomib within 24 hours maybe more benefitial to alleviate tumour load in MM patients.

Disclosures

No relevant conflicts of interest to declare.
In the Novel Agents Era, Is the International Staging System Still Has the Prognostic Value in Patients with Renal Impairment?
Authors: Jung Sung-Hoon , Kim Hee Kyung , Kim Seok Jin , Lee Seung-Shin , Ahn Jae-Sook , Kim Yeo-Kyeoung , Yang Deok-Hwan , Kim Hyeoung-Joon , Lee Je-Jung , Kim Kihyun
Published: October 2019

The International Staging System (ISS) is a robust prognostic tool for patients with symptomatic multiple myeloma (MM). In this study, we evaluated the prognostic value of ISS in patients with renal impairment who were treated with novel agents. We retrospectively analyzed 456 MM patients who were treated upfront with novel-agent containing regimen. The degree of renal failure was classified according to the National Foundation Kidney Disease Outcome Quality Initiative chronic kidney disease (CKD) classification. Renal impairment (RI) was defined as an eGFR< 60 mL/min/1.73m2 (CKD3-5). Of the all patients, 207 (45.4%) patients had stage 3-5 CKD at initial diagnosis. The β2-microglobulin level were significantly negatively correlated with GFR (r = - 0.536, P < 0.001), and median β2-microglobulin level of patients with CKD 3-5 was 7,600 mg/L (range, 668-80,000 mg/L). Sixty-seven percent of patients with CKD3-5 had ISS-3, and 89.8% of patients with CKD 4-5 had ISS-3. Patients with CKD 3-5 had poor overall survival outcomes compared to CKD 1-2 in univariate and not in multivariate analysis. ISS was powerful prognostic tool in all patients (P=0.003), but ISS had no prognostic impact in patients with CKD 3-5 (P = 0.897). Furthermore, patients with ISS 3 showed favorable overall survival compared to patients with ISS 1 or 2 in CKD 3. By multivariate analysis, cytogenetic high risk (HR 2.232, 95% CI 1.319-3.777, P =0.003) and hypercalcemia ≥11.5 g/dl (HR 2.036, 95% CI 1.170-3.544, P=0.012) were significantly associated with overall survival. In conclusion, β2-microglobulin level increase in patients with RI, and most of patients with RI had ISS-3. In addition, the ISS had no prognostic value in patients with RI in the novel agent era. New prognostic tool including cytogenetic risk will be needed for patients with RI.

Disclosures

No relevant conflicts of interest to declare.
High Level of Soluble Interleukin-2 Receptor in Serum Predicts Treatment Resistance and Poor Progression Free Survival in Multiple Myeloma
Authors: Wang Liang , Xia Zhongjun
Published: October 2019

Background: The IL-2/IL-2 receptor (IL-2R) system plays a central role in maintaining normal T cell immunity, and its disturbance is associated with several hematologic disorders. Studies have found in several types of lymphoma that abnormal amounts of soluble IL-2R (sIL-2R) may result in imbalance of the IL-2/IL-2R system and hence of the T cell immunoregulation. Whether the level of sIL-2R in blood could predict treatment response and survival outcomes or not needs to be investigated in multiple myeloma (MM) patients.

Methods: The level of sIL-2R in serum was measured using enzyme-linked immunosorbent assay (ELISA) in 81 patients with newly diagnosed MM. 26 patients (32.1%) were treated with bortezomib-based regimens and 55 patients (67.9%) received old drugs-based regimens.

Results: The mean concentration of sIL-2R for myeloma patients was 8.51ng/ml, significantly higher than that of healthy controls (0.56ng/ml, p<0.0001). The best cutoff value for sIL-2R in predicting high risk for disease progression is 6.049ng/ml with an area under curve (AUC) of 0.665 (p = 0.013). 36 patients (44.4%) were classified as higher sIL-2R level group (>6.049ng/ml), and 45 patients (55.6%) as lower group (=<6.049ng/ml). The overall response rate (ORR) was 60.0% in lower sIL-2R level group, higher than that in higher level group (41.7%, p=0.156). The 3-year progression free survival (PFS) and overall survival (OS) rate was 16% and 64%, respectively. In a multivariate survival analysis including ECOG performance status score, treatment response, and sIL-2R level, it was found that all these three parameters were significantly independent prognostic factors for PFS (p=0.032, 0.016, and 0.043, respectively), but none factors maintained their value in predicting OS. Subgroup analysis revealed that high level of sIL-2R correlated with significantly inferior PFS in patients treated with bortezomib-based regimens (p=0.004).

Conclusions: Serum sIL-2R level is a valuable biomarker for predicting treatment response and an independent prognostic factor for PFS, indicating novel drugs targeting the imbalance of IL-2/IL-2R system may be a promising strategy in MM.

Disclosures

No relevant conflicts of interest to declare.
Notch Is a Novel Critical Signaling Pathway Regulating Responses of T Cell and Antigen Presenting Cells in Multiple Murine aGVHD Models
Authors: Luo Xiaodan , Qin Pengfei , Wang Chunyan , Huang Zhenqian , Tan Huo
Published: October 2019

Introduction: Acute graft-versus-host disease (aGVHD) is a potentially life-threatening complication mediated by both host-derived antigen presenting cells (APCs) and donor T cells after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Despite prophylaxis and treatments, aGVHD stell occurs in many allo-HSCT patients. The role of Notch1 signal inhibition becomes more and more important in aGVHD study. This study is to investigate the role of Notch1 inhibition by γ-secretase inhibitor DAPT in murine aGVHD model.

Methods: We established a C57BL/6 BALB/c murine aGVHD model. γ-secretase inhibitor-DAPT is used to inhibit Notch1 signal in vivo and in vitro before transplantation. The degree of clinical and histopathologic GVHD is assessed by aGVHD scores and body weight. The functions of host-derived APCs and donor T cells are analyzed by flow cytometry, ELISA and PCR.

Results: All mice survived at least 14 days after transplantation and all of them developed aGVHD (n=20). The expression of Hes-1, as one of the target genes of Notch1 signal pathway, decreased significantly after DAPT inhibition. Body weight of mice in control groups decreased significantly compared to mice with Notch1 inhibition by DAPT after transplantation. Notch1 inhibited recipients produced markedly decreased amounts of the pro-inflammatory cytokines IFN-γ. The expressions of CD4 and Foxp3 increased while CD11c, CD80 and CD86 decreased after Notch1 inhibition.

Conclusions: These results indicate that Notch is a novel critical signaling pathway regulating responses of T cell and antigen presenting cells in multiple murine aGVHD models. Notch signaling inhibition appears to limit the harmful effects of aGVHD.

Disclosures

No relevant conflicts of interest to declare.
Clinical Characteristic and Prognosis of Post-Transplant Lymphoproliferative Disorder in Pediatric Heart Transplant Recipients in Japan
Authors: Miyamura Takako , Hashii Yoshiko , Fukushima Norihide , Kogaki Shigetoyo , Yoshida Hisao , Takahashi Kunihiko , Narita Jun , Ozono Keiichi
Published: October 2019

Purpose: Post-transplant lymphoproliferative disorder (PTLD) caused by Epstein-Barr virus (EBV) has recently has been recognized as a serious complication of various organ transplantations. In our institute, we have followed-up 34 pediatric patients after heart transplantation. We retrospectively investigated clinical features of PTLD in heart transplant recipients.

Methods: The evaluation period was from July 2004 to June 2015. Six patients with PTLD (4 boys, 2 girls) were diagnosed by blood examination, lymph node histology and EBV viral load in peripheral blood and invasive organ.

Results: Median age at onset of PTLD was 4.5 years old (range, 3.2-16.2 years). Median time from transplantation to onset was 18 months (range, 7-45 months). Bone marrow and central nervous system invasion were not detected in all cases. Four patients were EBV-seronegative recipients for EBV-seropositive donors. We performed the lymphoma-based chemotherapy combined with the anti-CD20 monoclonal antibody rituximab and achieved remission. As a result, five patients remained alive in remission, and one patient died of therapy-related toxicity. Although one patient relapsed after chemotherapy, she achieved second remission after multimodal chemotherapy combined with rituximab. Abdominal lymph node involvement was detected in four cases, and one patient experienced severe abdominal symptoms including intestinal perforation.

Conclusion: EBV serostatus and the period from transplantation to onset of PTLD appear to represent important prognostic factors. Early detection followed by pre-emptive reduction of immunosuppressive agents is essential to the management of PTLD, but may increase the risk of fatal rejection. We administered chemotherapy combined with rituximab, and this treatment strategy might be effective. However, severe fatal toxicity developed in one patient, and severe abdominal symptoms due to tumor lysis were noted in many cases. These findings suggested the importance of carefully selecting the induction treatment and perform various supportive therapies. We should establish appropriate treatment strategies according to prognostic factors because of the distinctive characteristics of PTLD by accumulating more cases.

Disclosures

No relevant conflicts of interest to declare.
Frequency of Invasive Fungal Infections in Patients Undergoing Allogeneic Stem Cell Transplantation Conditioned with Reduced BUCY2 and Antifungal Prophylaxis with Low Dose Amphotericin B
Authors: Armillas Canseco Flor Maria , Rivera Franco Monica M , Leon Rodriguez Eucario , Terrazas Marin Ricardo Antonio
Published: October 2019

Introduction: Invasive fungal infection (IFI) is a major cause of morbidity and mortality in patients with allogeneic stem cell transplantation (allo-SCT). One well-known risk factor for fungal infection includes bowel mucosal damage due to conditioning chemotherapy regimens. The use of reduced-intensity conditioning may favorably impact the epidemiology of IFI after allo-SCT. Data for IFI in this population are scarce. On the other hand, despite the low incidence of IFI with the use of the new antifungal drugs, the costs remain to be high and sometimes unaffordable for the patients.

Objective: To analyze the frequency of invasive fungal infections in patients who underwent stem cell transplantation conditioned with reduced BUCY2, at INCMNSZ, from November 1998 to December 2014.

Material and methods: A retrospective analysis was performed in 58 patients receiving reduced BUCY2 as part of their SCT conditioning regimen. Most of the patients received antifungal prophylaxis with low dose of amphotericin B (˂20 mg/day) during the neutropenia following transplant.

Results: Fifty eight patients undergoing allo-SCT with conditioning regimen reduced BUCY2, from November 1998 to December 2014, were included. Patients (male, 57%) had a median age of 39 years (range 17-67). The median follow-up was 90 months. The patients had a following range of underlying diseases: myelodysplastic syndrome (n=14, 24.1%), chronic myeloid leukemia (CML, n=14, 24.1%), acute myeloid leukemia (AML, n=12, 21%), acute lymphoblastic leukemia (LLA, n=10, 17%), lymphomas (n=3, 5.2%), myelofibrosis (n=2, 3.4%), or others (n=3, 5.2%). All patients were conditioned with 12mg/kg of busulfan and 80mg/kg of cyclophosphamide. 22% of patients presented mucositis grade III-IV. 85% (50/59) of patients received fungal prophylaxis with low dose amphotericin B. Four patients (6.8%) presented IFI during the first 100 days post-transplant, and one (1.7%) presented late IFI. The mortality secondary to IFI was 5%. Transplant related mortality (TRM) was 17%.

Conclusion: From the beginnings of our transplant program we have had a low incidence of IFI and low TRM, with the prophylactic use of low dose amphotericin B and the modified conditioning regimen reduced BUCY2, compared to the reported literature. The use of reduced BUCY2 and low dose amphotericin B can be cost-effective in medical centers in developing countries.

Disclosures

No relevant conflicts of interest to declare.