bluuebunny/crossref_metadata_2025 huggingface dataset:
Genetic Deletion of CCRL2 Impairs Macrophage Accumulation in Arterial Intima and Attenuates Atherosclerotic Plaque Development
Authors: Cao Yiren , Yang Fei , Tang Chaojun , Hu Shuhong , Zabel Brian Anthony , Zhu Li
Published: October 2019

Atherosclerosis is a chronic inflammatory disease of the arterial wall elicited by accumulation of LDL and leucocytes in the subendothelium at predilection sites with disturbed laminar flow. Chemokines and their receptors appear to act as critical players in atherosclerosis as they not only direct atherogenic recruitment of leucocytes but also exert cell hemostatic functions by chemokine ligand-receptor axes and their specific or combined contributions. Chemokine (C-C motif) receptor-like 2 (CCRL2) is an atypical chemokine receptor that cooperates with its ligand chemerin to play a role in cell trafficking and inflammatory response, the processes usually occur in atherosclerosis, but its role in atherosclerosis is not clear. Here we investigated the potential role of CCRL2 in atherogenesis using the classic apolipoprotein E-deficient (ApoE-/-) mouse model of atherosclerosis. Atherosclerosis-prone ApoE-/- mice were crossed with CCRL2-/- mice to obtain ApoE-/-CCRL2+/+ and ApoE-/-CCRL2-/- mice. Male mice of both genotypes were fed a standard chow diet until 8 weeks of age and then switched to a high fat diet for 16 weeks. Aortas were dissected and assessed by en face staining. Although CCRL2 deletion did not change mouse blood lipid profile and body weight, the atherosclerotic plaque area of the total aorta was reduced by 27.2% (P=0.0379) in ApoE-/-CCRL2-/- mice compared with ApoE-/-CCRL2+/+ mice with striking difference occurring in aortic arch. A reduction of lipid deposition by 32.6% (P=0.0089) was also observed in aortic root sections when CCRL2 was deleted. Further studies showed that deletion of CCRL2 reduced macrophage accumulation and polarization during the development of atherosclerosis. CCRL2 not only expresses in high levels in the plaques of ApoE-/- mice on a high fat diet but also co-localizes with macrophages and chemerin. Interestingly, using CMKLR1-/- chimeric mice, we showed that chemerin/CMKLR1/CCRL2 axis is involved in leucocyte infiltration and possibly affects plaque formation. More importantly, partial carotid artery ligation (PCL) model confirmed that disturbed blood flow-induced endothelial expression of CCRL2 modulates plaque formation. In conclusion, our results demonstrate that CCRL2 deficiency delays macrophage accumulation and the formation of atherosclerotic plaques potentially via the disruption of chemerin/CMKLR1/CCRL2 axis and the loss of CCRL2 response to the disturbed flow, which may represent a novel mechanism of atherosclerosis. This work was supported by Natural Science Foundation of China (grant 81370373 to L.Z. and 31300781 to C.T.)

Disclosures

No relevant conflicts of interest to declare.
Flow Cytometry-Based Platelet Function Testing Is Predictive of Symptom Burden in a Cohort of Bleeders
Authors: Boknäs Niklas , Ramström Sofia , Faxälv Lars , Lindahl Tomas L
Published: October 2019

Platelet function disorders (PFDs) are common in patients with mild bleeding disorders (MBDs), yet the clinical significance of laboratory findings suggestive of a PFD remain unclear due to the lack of evidence for a clear link between test results and patient phenotype. Herein, we present results from a study evaluating the potential utility of platelet function testing using whole-blood flow cytometry in a cohort of 105 patients undergoing investigation for MBD. Subjects were evaluated with a test panel comprising two different activation markers (fibrinogen binding and p-selectin exposure) and four physiologically relevant platelet agonists (ADP, PAR1-AP, PAR4-AP and CRP-XL). Abnormal test results were identified by comparison with reference ranges constructed from 24 healthy controls or the fifth percentile of the entire patient sample. We found that abnormal test results are predictive of bleeding symptom severity, and that the greatest predictive strength was achieved using a subset of the panel, comparing measurements of fibrinogen binding after activation with all four agonists with the fifth percentile of the patient sample (P = 0.00008, hazard ratio 8.7; 95 % CI 2.5-40). Our results suggest that whole-blood flow cytometry-based platelet function testing is a feasible alternative for the investigation of MBDs. We also show that platelet function testing using whole-blood flow cytometry could provide a clinically relevant quantitative assessment of platelet-related primary hemostasis.

Figure 1. Test results for each patient in comparison with reference range (A) and the fifth percentile of the entire patient cohort (B). Normal test results are colored grey, abnormal test results are colored with a continuous color gradient using the deviation from the mean divided by the standard deviation as a measure of degree of abnormality. Grey horizontal bars illustrate the number of abnormal test results for each patient. Figure 1. Test results for each patient in comparison with reference range (A) and the fifth percentile of the entire patient cohort (B). Normal test results are colored grey, abnormal test results are colored with a continuous color gradient using the deviation from the mean divided by the standard deviation as a measure of degree of abnormality. Grey horizontal bars illustrate the number of abnormal test results for each patient.

Disclosures

Lindahl: Diapensia: Equity Ownership.
Addition of Intermediate-Dose Methotrexate Is Beneficial in Terms of Both Progression-Free Survival and Overall Survival for Patients with Diffuse Large B Cell Lymphoma Receiving Either R-CHOP or CHOP
Authors: Dann Eldad J. , Goldschmidt Neta , Heffes Vered , Mashiach Tanya , Gatt Moshe E. , Horowitz Netanel A
Published: October 2019

Introduction: Patients with diffuse large B cell lymphoma (DLBCL) and high International Prognostic Index (IPI) or extra-nodal disease are at an increased risk of systemic and central nervous system (CNS) relapse. Presently, the management of DLBCL patients at the Rambam Health Care Campus (Haifa, Israel) and Hadassah Medical Center (Jerusalem, Israel) includes 6 cycles of R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone) along with 4 doses of intrathecal (IT) methotrexate (MTX), which may be followed by 2 additional cycles of an intermediate dose (3 g/m2) of intravenousMTX (ID-MTX). The current study compared progression-free survival (PFS), overall survival (OS) and CNS relapse rate in a cohort of DLBCL patients with risk factors for CNS relapse who did or did not undergo prophylactic therapy with ID-MTX.

Methods: Four hundred and eighty DLBCL patients treated at two tertiary care centers (Rambam and Hadassah) between the years 1991 and 2013 were retrospectively evaluated. The cohort included 224 females and 256 males at a median age of 58 years (range18-76). OS, PFS and CNS involvement at relapse were analyzed. The study incorporated all patients with the primary diagnosis of DLBCL and risk factors for CNS relapse, i.e., Waldeyer ring (2%), breast (1%), testicular (3%) or orbita (1%) involvement, bone marrow involvement (21%), stage IV disease (64%), LDH higher than normal (70%), ≥1 extra-nodal site (28%), IPI ≥3 (36%). Eight percent of patients had stage I-IE disease, 17% - stage II-IIE, 11% - stage III, 64% - stage IV, 29% had B symptoms. Patients with CNS involvement at diagnosis were excluded from the analysis.

Results: Four hundred and seventy three patients (99%) were treated with either CHOP or CHOP-like regimen, and 384 (80%) patients received rituximab (R). In the R-CHOP cohort, 23% of patients with IPI 0/1, 28% with IPI 2 and 42% with IPI ≥3 were given prophylaxis. One hundred and thirty patients received ID-MTX (27%), 35 patients (7%) received IT-MTX only, while no prophylaxis was administered to 315 patients (66%). The cumulative incidence of CNS relapse at 5 years was 6%, with no difference in the incidence between the patients who received prophylaxis with ID-MTX, IT-MTX or were given no prophylaxis at all. Patients with IPI ≥3 treated with ID-MTX had a decreased overall relapse rate and significantly superior OS and PFS (Table 1; Figure 1).

Conclusion: In the current study, the addition of two cycles of ID-MTX to the standard R-CHOP regimen resulted in improved 5-year PFS and OS of DLBCL patients with IPI ≥3. These data need to be further confirmed in a randomized controlled study.

Disclosures

No relevant conflicts of interest to declare.
Patterns of Microrna in Plasma Cells: From Normal Differentiation to Multiple Myeloma
Authors: Kassambara Alboukadel , Bruyer Angelique , Jourdan Michel , Robert Nicolas , Pantesco Véronique , Elemento Olivier , Klein Bernard , Moreaux Jerome
Published: October 2019

Background: MicroRNAs (miRNAs) are small noncoding RNAs that regulate gene expression either via the degradation of target mRNAs or the inhibition of protein translation. miRNAs have been involved in fine-tuning critical cellular processes and play critical roles in cell differentiation and tumor development. Furthermore, there is an increasing recognition of the role of miRNAs in multiple myeloma, a plasma cell (PC) malignancy characterized by an accumulation of malignant PCs within the bone marrow. However, little is known about miRNA expression during human PCD as well as about the full extent to which individual miRNAs regulate fundamental processes during PCD. A complete delineation of miRNA and their target expression during normal PCD is essential to understand the role of miRNAs in plasma cell malignancies.

Methods: We analyzed the expression profile of miRNAs and mRNAs during human plasma cell differentiation (PCD) to infer miRNA-target relationships, as well as in multiple myeloma tumor plasma cells. We developed a method and an R package, that uses miRNA and mRNA expression profiles across PCD cell subpopulations to infer candidate miRNA-target interactions that could be active and functional in PCD. We inferred miRNA-target relationships from sequence-based prediction, experimentally validated target interactions curated from the literature, published data from miRNA perturbation experiments and inverse expression relationships between miRNAs and their target mRNAs.

Results: Our results reveal 63 miRNAs with significant temporal changes in their expression during normal PCD. We derived a high-confidence network of 295 target relationships comprising 47 miRNAs and 147 targets. These relationships include new examples of miRNAs that are likely to coordinately regulate multiple members of critical pathways associated with PCD. Notably, we identify new miRNAs that coordinately regulate important pathways in PCD, including members of miR-30 and miR-29 families, miR-106b and miR-16, which regulates IRF4/PRDM1 axis, active DNA methylation pathway, TGF-b signaling pathway, autophagy, ZBTB4/EZH2 axis and cell cycle progression. Furthermore, our work demonstrates that 28 PCD stage-specific miRNAs are aberrantly overexpressed in multiple myeloma cells (MMCs) compared to their normal counterpart and/or are associated with high risk myeloma, suggesting that MMCs frequently acquired expression changes in miRNAs already undergoing dynamic expression modulation during normal PCD. Finally, expression of some targets of these PCD - MM miRNAs is correlated with clinical outcome of uniformly treated MM patients.

Conclusions: Altogether, our results demonstrate that miRNAs may be important in controlling PCD and malignant plasma cell biology.

Disclosures

No relevant conflicts of interest to declare.
Interference of HMGB1 Lead to Increased Vulnerability of MM Cells to Dexamethasone and Doxorubicin
Authors: Guo Xing , He Donghua , Chen Jing , Lin Xuanru , Chen Qingxiao , Yan Haimeng , Huang Xi , Li Yi , Yang Li , Zhang Enfan , Yang Yang , Wu Wenjun , He Jingsong , Huang He , Cai Zhen
Published: October 2019

Background: Multiple myeloma (MM) is the second mostly diagnosed disease among hematological malignancies after lymphoma. With the novel agents, the survival of MM patients has been improved significantly but still remains incurable because of drug resistance. Studies have found that high-mobility group box 1 protein (HMGB1) was involved in inflammation, angiogenesis, and cancer invasion progression, metastasis, and drug resistance. Our research was aimed at exploring the role of HMGB1 in MM cell proliferation and drug resistance.

Methods: First, semi-quantitative real time-polymerase chain (qRT-PCR) and western blot was used to determine the levels of HMGB1 mRNA and protein expression in MM cell lines (RPMI8226, CAG, and MM.1S) and primary MM samples. Second, MM cells were transfected with HMGB1-knockdown lentivirus and the Cell Counting Kit 8 (CCK8) assay was used to determine the proliferation of MM cells with or without chemotherapeutic drugs dexamethasone (Dex) and doxorubicin (ADM). Then cell apoptosis was detected by flow cytometry. Third, Affymetrix HTA 2.0 Array was used to compare changes in gene expression levels between HMGB1-knockdown cells and the control cells and qRT-PCR was used to verify the array results. Last, Western bolt was performed to analyze changes in signaling pathways after HMGB1 knockdown.

Results: MM cell lines and primary MM samples expressed high levels of HMGB1 mRNA and protein. Although there was no difference in MM cell proliferation between HMGB1-knockdown group and the control group (P>0.05), HMGB1-knockdown significantly enhanced inhibitory effect of chemotherapy with Dex and ADM in comparison with the wildtype HMGB1 control (P<0.05). Flow cytometry results showed that apoptosis of MM cells induced by Dex and ADM was increased when HMGB1 expression was suppressed (P<0.05). Furthermore, gene array analysis on RPMI8226 and CAG cell lines showed that anti-apoptotic genes (bcl-2, HIAP1) and MM survival related genes (DEPTOR, CXCL12) were decreased and pro-apoptotic genes (TNFRSF1B, TRAIL, CXCL10) were increased in knockdown cells compared to the controls. In addition, expression levels of genes that play important roles in signaling pathways such as mTOR, NF-κB, PI3K-AKt, and p38-MAPK were also significantly changed. The gene expression microarray results were verified by qRT-PCR and Western blot demonstrated that phosphorylation of p70S6K (substrate of mTORC1 complex) and AKT-ser473 (substrate of mTORC2 complex) were both elevated in HMGB1-knockdown MM cells compared to that in the control cells.

Conclusions: Our research showed that downregulation of HMGB1 increased sensitivity of MM cells to Dex and ADM through increasing apoptosis and regulating mTOR, NF-κB, PI3K-AKt, and p38-MAPK pathways.

Disclosures

No relevant conflicts of interest to declare.
Standard of Care of Patients with CML Treated in Community Based Oncology Group Practices in Rhineland-Palatinate (Germany)
Authors: Weide Rudolf , Rendenbach Bernhard , Grundheber Monika , Burkhard Oswald , Behringer Joachim , Maasberg Michael , Ehscheidt Peter , Strehl John Werner , Hansen Richard , Feiten Stefan
Published: October 2019

Introduction: Significant progress has been made in CML-therapy since the introduction of imatinib and other tyrosine kinase inhibitors (TKI) into clinical care. The aim of this study was to assess diagnosis, treatment and outcome of CML-patients who received their treatment in community based oncology practices in Rhineland-Palatinate and whether European LeukemiaNET-guidelines were followed.

Methods: All Ph-/BCR-ABL-positive CML-patients who were treated between 12/2001-12/2015 in 9 oncology group practices were analyzed retrospectively concerning diagnosis, treatment and outcome according to European LeukemiaNET-guidelines. Data were collected from patient files into a central data base and analyzed statistically with SPSS.

Results: 264 patients (pts) with a median age of 60 (18-90) were analyzed. 126 (48%) were female, 138 (52%) were male. At initial diagnosis bone marrow biopsy was performed in 213 pts (81%). Cytogenetics was applied in 204 pts (77%) (38% in blood, 56% in bone marrow). FISH-analysis was used in 155 pts (59%) (33% in blood, 36% in bone marrow). PCR-testing to detect a BCR-ABL1-rearrangement was applied in 200 pts (76%) (52% blood, 37% bone marrow). 258 pts (98%) were in chronic phase, 5 (2%) in accelerated phase and 1 (0.4%) in blast crisis at diagnosis. EUTOS score could be calculated in 131 pts (50%). 20% were high risk, 80% low risk. 252 pts (95%) received some form of TKI-therapy. Out of 416 TKI-therapies 308 (74%) were PCR-based monitored, 148 (36%) were monitored by cytogenetics. First line treatment was imatinib in 201 pts (80%), 51 pts (20%) received a second generation TKI. Second line treatment consisted of dasatinib in 59%, nilotinib in 32%, imatinib in 6% and bosutinib in 3%. Third line treatment was nilotinib in 56%, dasatinib in 35%, ponatinib in 6% and imatinib in 3%. 62 pts (23%) were treated within a study protocol. 13 pts (5%) received an allogeneic transplantation. Overall survival probability was 88% after 5 years and 72% after 10 years. Disease specific survival was 95% after 5 years and 86% after 10 years.

Conclusion: The overwhelming majority of CML-patients treated in oncology group practices receive standard of care as suggested by European LeukemiaNET-guidelines. Overall survival in routine care is comparable to international studies.

Disclosures

No relevant conflicts of interest to declare.
Most up-to-Date Long Term Survival Estimates for Common Hematologic Malignancies Using the Boomerang Method
Authors: Pulte Dianne , Jansen Lina , Brenner Hermann
Published: October 2019

Background: Five year survival has increased dramatically for most of the common hematologic malignancies in the early part of the 21st century, probably due to new therapeutic options. However, less is known about the effects recent changes in therapy have had on longer term survival. Here, we examine 10- and 20-year survival for patients with common hematologic malignancies including myeloma, acute lymphoblastic leukemia (ALL), acute myeloblastic leukemia (AML), chronic lymphoid leukemia (CLL), chronic myeloid leukemia (CML), non-Hodgkin lymphoma (NHL), and Hodgkin lymphoma (HL).

Methods: Data were extracted from the Surveillance, Epidemiology, and End Results (SEER9) database. Patients age 15+ with appropriate ICD-10 codes for the above malignancies were included, with exclusion of those patients diagnosed by death certificate only. The boomerang method1 was used to estimate 10 and 20-year relative survival for patients in two calendar periods, 1999-2003 and 2009-13. This recently described method has been demonstrated to more closely approximate actual observed survival for a given time period compared to other methods of estimation, including period analysis, by minimizing the contribution of earlier time periods to the estimation of survival.

Results: Ten and 20-year survival increased for each malignancy examined, with increases ranging from +4.3 percent units for 20-year survival for myeloma to +28.4 percent units for 10-year survival for CML (see Table). Ten year relative survival was greater than 50% in 2009-13 for patients with CML, CLL, NHL, and HL, at 69.9%, 59.1%, 63.4%, and 63.1%, respectively. Major decreases in survival were observed between 10- and 20-year survival estimates for all malignancies except for AML, where 10- and 20-year relative survival estimates were virtually identical in 2009-13.

Discussion: Long term survival is increasing for common hematologic malignancies, with particularly strong increases being observed for CLL, CML, and NHL. Major decreases in survival were observed between 10 and 20 years for most malignancies, including NHL and ALL, which are generally considered curable. This may be partly due to the effect of recent changes in therapy leading to underestimation of 20-year survival, even using the boomerang method, but further study of long-term outcomes in curable malignancies to determine whether there is an increased risk of specific issues later in life is indicated.

Reference:

1. Brenner H, Jansen L. Timely disclosure of progress in long-term cancer survival: the boomerang method substantially improved estimates in a comparative study. J Clin Epidemiol. 2016;70:224-32.

Disclosures

Pulte: Selexys Pharmaceuticals: Research Funding; EBSCO: Other: Review of content for Dynamed medical reference product; ApoPharma: Research Funding.
Sphingosine-1-Phosphate (S1P) Signaling through S1P Receptor 1 (S1PR1) in Macrophage Contributes to Pain in Sickle Cell Disease
Authors: Adebiyi Morayo G. , Zhang Yujin , Sun Kaiqi , Hu Xia , Zhao Shusuan , Wu Hongyu , Couturier Jacob , Lewis Dorothy L. , Walters Edgar T. , Xia Yang
Published: October 2019

Sickle cell disease (SCD) is the most common autosomal recessive hemolytic disorder affecting more than 100,000 African-Americans and millions around the world. The severe health complication of patients with SCD is chronic pain, which is the leading cause of SCD patient morbidity and hospitalization. Our proposed research builds on the results from our nonbiased high-throughput metabolomics screen, which revealed erythrocyte sphingosine-1-phosphate (S1P), a bioactive lipid, is significantly elevated in the circulation of humans and mice with SCD. We identified that erythrocyte-sphingosine kinase 1 (Sphk1), the enzyme that generates S1P, promote erythrocyte sickling and the genetic deletion of Sphk1 significantly reduce pain symptoms in SCD mice. S1P is a pleiotropic molecule that signals via five g-protein coupled receptors (GPCRs) known as S1P receptors (S1PR1-5). S1PR1 is widely expressed in many cell types, including neurons and immune cells. We treated SCD mice with FDA approved drug, FTY720, to specifically target S1PR1. We demonstrated that mice had improved mechanical and thermal pain sensitivity. Notably, using flow cytometry, we found that the percentage of macrophages infiltrated to skin and percentage of S1PR1 expression in macrophages in the skin were significantly induced in SCD mice compared to the controls. This implies that elevated S1PR1 in macrophage contributes to chronic pain in SCD mice. To test this possibility, we treated SCD mice with clodronate liposomes to deplete tissue macrophages. We identified that mice had reduced sensitivity to mechanical and thermal stimuli assessments. Altogether, we provide both genetic and pharmacological evidence that elevated plasma S1P signaling via S1PR1 likely on macrophages contributes to chronic pain in SCD. Additionally, S1PR1 is known to mediate pro-inflammatory cytokine secretion in immune cells. We identified that SCD mice have elevated levels of interleukin-6 (IL-6), a pro-inflammatory cytokine, and is likely involved in SCD pain. To study the role of IL-6 in SCD pain, we generated SCD-IL-6-/- mice and observed improved mechanical and thermal pain sensitivity in these mice. From these evidences, we hypothesize that Sphk1 mediate S1P-S1PR1 activation in macrophages and IL-6 induction to promote SCD pain. Our work addresses a knowledge gap regarding an inflammatory mediated mechanism involved in SCD pain and novel therapies to counteract pain in SCD.

Disclosures

No relevant conflicts of interest to declare.
Leukemia-Associated Macrophages Educated By Organ-Specific Leukemic Microenvironment Acquire Diverse Functions and Gene-Expression Signatures
Authors: Yang Xiao , Feng Wenli , Wang Rong , Yang Feifei , Chen Chong , Wang Lina , Zheng Guoguang
Published: October 2019

Macrophages are important components of microenvironments and participate diverse physiological and pathological processes. Though CD68+macrophages have been demonstrated a potentially clinical marker to predict adverse prognosis in lymphoma, the role of macrophages in leukemia have not been fully elucidated. Most recently, we reported leukemia-associated macrophages (LAMs), which showed leukemia-promoting effects, in Notch1-induced mouse T-ALL leukemic microenvironment. In the present study, we studied the characteristics of LAMs from BM and spleen in MLL-AF9 induced mouse AML model, and compared the gene expression diversities among LAMs from BM and SP of AML and T-ALL models.

LAMs were sorted from leukemic BM and spleen in the middle stage of AML, control macrophages were sorted from normal mouse. Then LAMs were cocultured with AML cells for 24, 48 and 72hrs. Cell count, BrdU incorporation assay and Annexin V assay were used to detect the proliferation and apoptosis of leukemia cells, respectively. Significant increase of BrdU positive and decrease of apoptotic AML cells were detected in leukemia cells cocultured with LAMs than control Mφ. Latex-beads uptaken assay and transwell assay were used to detect the phagocytosis and migration of LAMs. Significant decrease of phagocytosis and increase of migration ability were detected in LAMs than control Mφ. Furthermore, T cell coculture assay revealed that spleen LAMs suppress the CD25 and CD69 expression in both CD4+ and CD8+T cells but increase induction of Treg cells than BM LAMs. Moreover SP LAMs had a significantly higher proliferation promoting effect on leukemia cells than BM LAMs. To explore the diverse effect among LAMs from BM and spleen, we compared the transcriptional data of LAMs from BM and spleen from AML and T-ALL. The ordered list algorithm revealed the similarity score in LAM samples with same microenvironment (991.8) is higher than those with same type of leukemia (709.1), also the expression pattern analysis revealed that LAMs from leukemic BM and spleen are much more different, the phenotypic gene validated by qRT-PCR revealed that the BM LAMs are more M1-like while the spleen LAMs are more M2-like.

Our results suggested that the diverse response of LAMs to leukemia are largely determined by organ-specific leukemic microenvironment.

The work was supported by the Grants 81370634, 81570153, 81300376 from the National Natural Science Foundation of China (NSFC).

Disclosures

No relevant conflicts of interest to declare.
Peptide Vaccination Against Cytomegalovirus (CMV) Elicits Immunological and Clinical Responses after Allogeneic Stem Cell Transplantation Even from a CMV Seronegative Donor
Authors: Schmitt Michael , Schmitt Anita , Wiesneth Markus , Hückelhoven Angela , Wu Zeguang , Kuball Juergen , Wang Lei , Schauwecker Peter , Hofmann Susanne , Götz Marlies , Michels Birgit , Maccari Birgit , Wuchter Patrick , Mertens Thomas , Schnitzler Paul , Döhner Hartmut , Ho Anthony D. , Bunjes Donald , Dreger Peter , Schrezenmeier Hubert , Greiner Jochen
Published: October 2019

Introduction: Patients receiving an allogeneic stem cell graft from a cytomegalovirus (CMV) seronegative donor are prone to CMV reactivation with high risk of disease and mortality. Therefore we initiated a clinical phase I trial with a novel vaccine designed by our group: a CMV phosphoprotein 65 (CMVpp65)-derived peptide in water-in-oil emulsion (Montanide™) plus administration of granulocyte-macrophage colony stimulating factor.

Material and Methods: 10 patients after allogeneic stem cell transplantation received four vaccines s.c. at a biweekly interval. Patients were monitored for clinical course and CMVpp65 antigenemia. CMV-specific CD8+ and gamma/delta T cells were analyzed by multi-color flow cytometry. Neutralizing anti-CMV antibody assays were established and correlated to clinical parameters.

Results: Peptide vaccination was well tolerated, no drug-related serious adverse events were detected. 7 of 9 patients with CMVpp65 antigenemia cleared the CMV with enduring response > 1.5 years after 4 vaccinations and are still free from antigenemia until present. 2 patients with CMV reactivation showed persisting CMV antigenemia. One patient received prophylactic vaccination and did not develop antigenemia. An up to 6-fold increase in frequency of both CMV-specific CD8+ T cells or Vdelta2- gamma/delta T cells with a maximum of 1.32% CMV-specific CD8+ T cells ex vivo without priming and 13.7% Vdelta2- gamma/delta T cells of all CD8+ T cells was detected in five patients. Also titers of neutralizing antibodies increased in four patients up to 10-fold over the time of vaccination. Humoral and cellular immune responses correlated with clearance of the CMV load.

Conclusion: Administration of CMVpp65 peptide vaccination for patients after allogeneic stem cell transplantation at high risk for CMV reactivation was safe, well tolerated and clinically encouraging. A study in solid-organ transplant patients is ongoing.

Disclosures

Kuball: Gadeta B.V.: Membership on an entity's Board of Directors or advisory committees. Wuchter:Sanofi-Aventis: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Hexal: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees. Dreger:Gilead: Consultancy; Janssen: Consultancy; Novartis: Consultancy; Gilead: Speakers Bureau; Novartis: Speakers Bureau; Roche: Consultancy. Greiner:BMS: Research Funding.
Target Joint Status in Patients with Hemophilia a during 18 Consecutive Months of Prophylaxis with a Pegylated Full-Length Recombinant Factor VIII with Extended Half-Life
Authors: Engl Werner , Patrone Lisa , Abbuehl Brigitt E.
Published: October 2019

Introduction and Objective: Patients with severe hemophilia A repeatedly bleed into joints and subsequently develop target joints and arthropathy. BAX 855, a polyethylene glycol pegylated, full-length, recombinant factor VIII built on ADVATE, demonstrates an extended half-life, efficacy, and safety for prophylaxis and the treatment of bleeding in patients with severe hemophilia A. Target joint status was evaluated in an ad-hocanalysis of integrated efficacy data from previously treated adolescent and adult patients who participated in the pivotal (completed) and continuation (ongoing) studies.

Methods: The number of target joints, defined as a single joint with ≥3 spontaneous bleeding episodes in any consecutive 6-month period, were analyzed over 3 consecutive 6-month periods in patients who received twice weekly BAX 855 prophylaxis at 40-50 IU/kg.

Results: After approximately 6 months of twice weekly prophylaxis during the pivotal study, 101 adolescent and adult patients continued treatment in the continuation study, 51 of whom were treated with twice weekly prophylaxis for 18 consecutive months. At screening 29.4% (15/51) of these patients had no target joints, 19.6% (10) had 1 target joint, 21.6% (11) had 2, and the remaining 29.4% (15) had 3 or more. After the first 6-month treatment period, the percent of patients with no target joints increased to 66.7% (34 patients), including 19 patients in whom 1 or more target joints had resolved and 15 patients who remained target joint-free from screening. Of note, for the 15 patients with 3 or more target joints at screening, all of their target joints resolved after the first 6 months of treatment with BAX 855. This trend was maintained after the second 6-month treatment period. After the third 6-month period of twice weekly prophylaxis, the percent of patients with no target joints further increased to 82.4% (42), which included the 15 patients who remained target joint-free from screening. Nine patients had 1 or more unresolved target joint after 18 months of twice weekly prophylaxis, and in 6 of these patients, 1 to 3 other target joints had resolved. Overall, there were 89 target joints at screening which reduced to 14 after 18 months of twice weekly prophylaxis. Of those, 10 target joints changed status (eg, were present, resolved, and then re-appeared) and only 4 target joints (in 4 patients) persisted as unresolved through each of the 3 consecutive 6-month treatment periods.

Conclusions: These results demonstrate the efficacy of continuous twice weekly prophylaxis with BAX 855 for preventing and resolving the majority of target joints.

Disclosures

Engl: Shire, formerly Baxalta and Baxter: Employment, Equity Ownership. Patrone:Shire, formerly Baxalta and Baxter: Employment, Equity Ownership. Abbuehl:Shire, formerly Baxalta and Baxter: Employment, Equity Ownership.
Risk Factors for Venous Thromboembolism in Metastatic Colon Cancer Patients in the Contemporary Treatment Era: A SEER-Medicare Data Analysis
Authors: Pulluri Bhargavi , Littenberg Benjamin , Lal Inder , HOlmes Chris E. , Ades Steven
Published: October 2019

Background:The relationship between metastatic colorectal cancer (CRC) and venous thromboembolism (VTE) is not well defined in the modern treatment era. Previous population-based studies date back to a period marked by inpatient intravenous heparin therapy and inferior survival due to paucity of therapeutic anti-cancer options, and before the advent of newer therapies including oxaliplatin, irinotecan, and anti-angiogenic treatment with bevacizumab. The objectives of this retrospective study were to examine the impact of multiple putative risk factors on VTE incidence in a large representative modern cohort of older patients with metastatic CRC.

Methods:We performed a retrospective analysis of SEER-Medicare data on elderly patients with metastatic CRC diagnosed in 2004-2011. VTE and associated risk factors were analyzed using multivariate Cox proportional hazards models adjusted for sex, age at diagnosis, race, ethnicity, tumor anatomy (left/right/unknown), calendar year of diagnosis, Charlson comorbidity score, location of SEER registry and urban residence, with time-varying covariates for use of cancer therapies.

Results:Of 339,778 records, 11,086 metastatic colon cancer cases were identified. 1,338 cases had VTE with a cumulative incidence of 13% at 1 year and 19% at 3 years. The mean age was 77.9 years (range 65-106). 49.7% were women and 83.5% white. 60.5% had a Charlson comorbidity score of zero at diagnosis; 6% had scores of 6-18.

Significant predictors of VTE included female sex (Hazard Ratio (HR) 1.22; 95% Confidence Interval (CI) 1.10, 1.36; P<0.001), younger age at diagnosis (HR 1.25 per decade of age; CI 1.15, 1.37; P<0.001), urban residence (HR 1.20; CI 1.03, 1.40; P=0.02), right sided colon cancers (HR 1.19; CI, 1.06, 1.33 P=0.003), and current use of 5-fluorouracil (HR 1.33; CI 1.04, 1.70; P=0.02). The risk of VTE was significantly reduced when on bevacizumab (HR 0.77; CI 0.63, 0.93; P=.006), or irinotecan therapy (HR 0.59; CI 0.47, 0.75; P<0.001) and in those with more comorbid conditions (HR 0.97 per point of Charlson score; CI 0.935, 0.998; P=0.04).

Conclusion: The higher incidence of VTE seen in right sided colon cancers may be related to biologically aggressive tumors associated with poor survival as reported in recent studies. The lower risk of VTE in older, sicker patients was unexpected and may reflect the effect of competing mortality. In this large contemporary cohort, anti-angiogenic therapy was not associated with a higher risk of VTE; the apparent protective effects of bevacizumab and irinotecan may represent treatment assignment bias.

Disclosures

No relevant conflicts of interest to declare.
Systematic Review of Treatment for Massive and Submassive Pulmonary Embolism
Authors: Gupta Shipa , Gandara Esteban , Carrier Marc , Castellucci Lana A
Published: October 2019

Introduction: Anticoagulation is the standard treatment for pulmonary embolism (PE). In patients presenting with massive or submassive PE, the addition of systemic thrombolytics (STL) and catheter directed therapies (CDT) may also be considered. However, there is uncertainty about the benefits and risks of these treatments, and no randomized trials have compared STL to CDT.

Aim: We reviewed the risks and benefits of STL and CDT in the management of patients with massive or submassive PE.

Methods: A systematic literature search was conducted using MEDLINE, Embase, Cochrane Central Register of Controlled Trials, and bibliographies of relevant studies and reviews. Randomized trials and cohort studies were eligible. Registries and studies with fewer than 10 patients were ineligible. Pooled proportions and their associated 95% confidence intervals (CI) for mortality, major bleeding, intracranial hemorrhage (ICH), recurrent PE, and fatal recurrent PE were calculated.

Results: A total of 52 studies were included in the analyses: 19 studies evaluating STL; 18 studies using CDT; 5 studies using the AngioJet rheolytic thrombectomy system; and 10 studies using ultrasound-assisted CDT. The pooled proportion of outcomes for patients receiving STL were: 4.46% (95% CI: 3.04-6.13%) for mortality; 11.70% (95% CI: 7.78-16.90%) for major bleeding; 1.99% (95% CI: 1.06-3.20%) for ICH; 3.92% (95% CI: 2.56-5.55%) for recurrent PE; and 1.04% (95% CI: 0.42-1.94%) for fatal recurrent PE. For patients receiving CDT, the pooled proportion of outcomes were: 9.64% (95% CI: 5.93-14.13%) for mortality; 5.17% (95% CI: 2.18-9.33%) for major bleeding; 1.29% (95% CI: 0.48-2.48%) for ICH; 2.76% (95% CI: 1.36-4.63%) for recurrent PE; and 1.56% (95% CI: 0.53-3.12%) for fatal recurrent PE. For patients receiving AngioJet CDT, the pooled proportion of outcomes were: 10.18% (95% CI: 3.68-19.42%) for mortality; 9.14% (95% CI: 0.88-24.70%) for major bleeding; 2.30% (95% CI: 0.34-5.96%) for ICH; 4.84% (95% CI: 1.62-9.66%) for recurrent PE; and 4.84% (95% CI: 1.62-9.66%) for fatal recurrent PE. In patients receiving ultrasound-assisted CDT, the pooled proportion of outcomes were: 4.17% (95% CI: 1.64-7.79%) for mortality; 4.88% (95% CI: 2.48-8.05%) for major bleeding; 0.63% (95% CI: 0.10-1.59%) for ICH; 1.39% (95% CI: 0.48-2.77%) for recurrent PE; and 1.21% (95% CI: 0.35-2.56%) for fatal recurrent PE.

Conclusions: Our results suggest that all-cause mortality rates in patients with massive or submassive PE are similar between STL and ultrasound-assisted CDT and are lower than other types of CDT. In general, major bleeding rates were lower in CDT based groups than in STL. Importantly, ICH rates in CDT based treatment strategies were also lower than STL therapy. Randomized trials comparing STL and CDT would help solidify the role of these treatment strategies in patients with massive and submassive PE.

Disclosures

Carrier: BMS: Research Funding; Leo Pharma: Research Funding.
Staring inside of Idiopathic Hypereosinophilia: Identification of Clonality Using Targeted Exome Sequencing
Authors: Lee Jee-Soo , Seo Heewon , Im Kyongok , Park Si Nae , Kim Jung-Ah , Kim Seon Young , Lee Joon-hee , Kwon Sunghoon , Koh Insong , Hwang Seungwoo , Park Heung-Woo , Kang Hye-Ryun , Kim Ju Han , Kim Miyoung , Lee Dong Soon
Published: October 2019

Idiopathic hypereosinophilia (IHE) has been a disorder of uncertain etiology, characterized by persistent elevation of blood eosinophil count exceeding 1.5x103/μL without evidence of reactive and clonal causes. When end-organ damage due to eosinophilic infiltration is present, idiopathic hypereosinophilic syndrome (IHES) is diagnosed. As the name 'idiopathic' implies the uncertain etiology, the nature of eosinophilic proliferation is unclear whether benign or clonal. While T cell clonality assessment has been recommended for unexplained hypereosinophilia, this approach is not often applied to the routine practice in clinic. We hypothesized that the clonality would exist in patients classified into IHE/IHES. The bone marrow cells of 30 IHE patients were retrospectively evaluated.The T cell receptor (TCR) assay and the targeted sequencing for 88 genes related to hematologic malignancies were performed. After being analyzed, the candidate mutations were subjected to pathway analysis.Through the targeted sequencing, 140 variants in 59 genes were identified. Sixteen out of 30 patients (53.3%) harbored at least 1 candidate mutation. The most frequently affected genes were NOTCH (n=8); SCRIB and STAG2 (n=5); SH2B3 (n=4). Remarkably, we identified recurrent somatic mutations in 4 genes with 5 patients (16.7%); MPLY521X (n=2), SCRIBE795X (n=2), ASXL1G646f (n=3) and STAG2E249X (n=2). A known mutation TP53V216M was confirmed in an IHE patient. Interestingly, the TCR assay revealed that 4 of 30 patients (13.3%) had a clonal TCR rearrangement. Skin involvement was described in patients with clonal T cell population. Through the pathway analysis, the role of these genes in eosinophilia was identified. These results highly suggest that these genes are likely related to clonality of hypereosinophilia.This study revealed that somatic mutations affecting hematopoiesis and monoclonal T cell clones underlie idiopathic hypereosinophilia. It would be valuable to extend our findings on the molecular profiling of hypereosinophilia to the further study discovering the clonality of IHE.

Disclosures

No relevant conflicts of interest to declare.
Determining the Signal Source in Magnetic Resonance Imaging of Myelofibrotic Bone Marrow
Authors: Matsuura Shinobu , Patterson Shenia , Lucero Hector , Grant Aaron , Herrera Victoria L. M. , Ravid Katya
Published: October 2019

Magnetic resonance imaging (MRI) is a promising diagnostic method for evaluation of bone marrow myelofibrosis for offering non-invasive, wider-area coverage over conventional histology. Although the most striking feature in histology of myelofibrotic bone marrow is the proliferation of reticulin fibers, the spectrum of findings involves abnormalities in cell number and composition, vessel proliferation, as well as possible pre-fibrotic abnormalities in matrix such as inflammation/edema. Correlation of MRI observations with such histological findings to accurately define the pathogenic phases of myelofibrosis has been challenging. Here, we present the first MRI analysis of myelofibrosis in a mouse model (Gata-1low mice) in an attempt to correlate MRI signal observations with histological findings. T2-weighted MRI images of femur bone marrow of Gata-1low animals at different ages (12 to 46 weeks) showed high MR signal intensity, absent in age-matched wild-type controls, that is detected as early as 12 weeks of age, before onset of histologically-defined fibrosis in this mouse model. MRI acquisitions employing fat suppression, a technique that suppresses signals from lipids, together with the absence of chemical shift artifacts in the images, excluded the possibility of lipids as the source of the prominent MR signal. To determine the source of this prominent non-lipid signal in Gata-1low bone marrow, cellular composition, vascularization and vascular permeability were tested. Our findings demonstrate a multi-source nature of MRI signals in bone marrow, which evolves from young (12 weeks) to old (40 weeks) Gata-1low bone marrow consistent with the pathologic progression of myelofibrosis. Data support the importance of an integrative approach to further the development of this non-invasive modality for following the dynamics and progression of this multifactorial pathology in humans.

Disclosures

No relevant conflicts of interest to declare.
A Prognostic Scoring System for Patients with Multiple Myeloma Who Were Classified Stage II By Revised International Staging System
Authors: Jung Sung-Hoon , Kim Kihyun , Kim Jin Seok , Kim Seok Jin , Cheong June-Won , Kim Soo-Jeoong , Ahn Jae-Sook , Ahn Seo-Yeon , Yang Deok-Hwan , Kim Hyeoung-Joon , Lee Je-Jung
Published: October 2019

Background: Revised International Staging System (R-ISS) is recently proposed risk stratification algorism based on ISS, cytogenetic risk, and lactate dehydrogenase in multiple myeloma (MM). Although R-ISS had an improved prognostic power compared to ISS, there is a problem that patients with stage II increase to 1.7 times. In this study, we evaluated survival outcomes and a new prognostic tool for MM patients classified as R-ISS II.

Methods: Retrospective data from 441 patients who were initially treated with a novel agent-containing regimens were analyzed.

Results: R-ISS staging system could discriminate survival outcome more clearly compared to ISS in patients treated with novel-agents [median overall survival (OS) times of R-ISS I vs. II. Vs. III was 76.3 vs. 51.3 vs. 30.3 months, P < 0.001]. By applying R-ISS, 6.2% of patients with ISS I, and 24.3% with ISS III were reclassified into R-ISS II. Overall, 290 patients (66%) was classified into R-ISS II and had wide range of survival outcomes (0.7-114.5 months). In multivariate analysis, poor performance status (PS) (HR 1.657, 95% CI 1.159-2.370, P =0.006), del (17p13) (HR 2.227, 95% CI 1.174-4.223), P = 0.014), and thrombocytopenia (HR 1.533, 95% CI 1.033-2.276, P = 0.034) were significantly associated with OS in patients with R-ISS II. Based on this analysis, we constructed a new prognostic model consisted of poor PS (1 point), del(17p13) (2 point), and thrombocytopenia (1point). Among the 290 patients, 152 patients (52.4%) were classified as low-risk (score 0), 109 (37.5%) as intermediate risk (score 1), and 29 (10%) as high risk (scores 2 or more). The median OS times for the corresponding risk groups was 57.5 months (95% CI 40.4-74.5), 30.2 months (95% CI 20.7-39.7), and 20.9 months (95% CI 6.4-35.3) (P <0.001).

Conclusion: R-ISS is an excellent prognostic tool in MM in the novel-agent era, but there is a limitation that most of patients were classified into R-ISS II. A new prognostic model for patients with R-ISS II is needed, and prognostic scoring system based on poor PS, del(17p13), and thrombocytopenia may be useful to distinguishing survival outcomes of patients with R-ISS II.

Figure Figure.

Disclosures

No relevant conflicts of interest to declare.
Concomitant Suppression of IKZF1, IRF4 and MYC Contribute to the Anti-Tumor Activity of the BET Inhibitor, Cpi-0610, in Disease Models of Multiple Myeloma
Authors: Siu Ka Tat , Ramachandran Janani , Yee Andrew J. , Eda Homare , Santo Loredana , O'Donnell Elizabeth K. , Panaroni Cristina , Mertz Jennifer , Sims Robert J. , Cooper Michael , Raje Noopur
Published: October 2019

Inhibition of the bromodomain and extra-terminal (BET) proteins is a promising therapeutic strategy for various hematologic malignancies. Previous studies suggest that BET inhibitors constrain tumor cell proliferation and survival mainly through suppression of MYC transcription and activity. However, suppression of the transcription of additional genes also contributes to the anti-tumor activity of BET inhibitors but is less well understood.

Here we investigated the therapeutic potential of CPI-0610, a novel BET inhibitor that is currently in a phase I clinical trial in relapsed multiple myeloma (MM) (ClinicalTrials.gov Identifier: NCT02157636). CPI-0610 displays potent in vitro cytotoxicity against MM cell lines and patient-derived MM cells by inducing G1 cell cycle arrest and caspase-dependent apoptosis. Furthermore, CPI-0610-mediated BET inhibition overcomes the protective effects conferred by cytokines and bone marrow stromal cells. We also confirmed the in vivo efficacy of CPI-0610 in a MM xenograft mouse model. CPI-0610 significantly delayed tumor growth and increased the survival of MM-bearing SCID mice.

Our study found IKZF1 and IRF4 to be among the primary targets of CPI-0610, along with MYC. These findings indicate that BET inhibition not only results in a robust reduction of MYC transcription and activity but also suppresses the expression of IKZF1 and IRF4 in MM. Given that immunomodulatory drugs stabilize cereblon and facilitate Ikaros degradation in MM cells, we combined it with CPI-0610. Combination studies of CPI-0610 with lenalidomide or pomalidomide show in vitro synergism, in part due to concomitant suppression of IKZF1, IRF4, and MYC, providing a rationale for clinical testing of this drug combination in MM patients.

Disclosures

Mertz: Constellation Pharmaceuticals, Inc.: Employment, Equity Ownership. Sims:Constellation Pharmaceuticals, Inc.: Employment, Equity Ownership. Cooper:Constellation Pharmaceuticals, Inc.: Employment, Equity Ownership. Raje:Amgen: Consultancy, Membership on an entity's Board of Directors or advisory committees; Celgene: Consultancy, Membership on an entity's Board of Directors or advisory committees; Takeda: Consultancy, Membership on an entity's Board of Directors or advisory committees; Merck: Membership on an entity's Board of Directors or advisory committees; Novartis: Consultancy, Membership on an entity's Board of Directors or advisory committees; Roche: Consultancy, Membership on an entity's Board of Directors or advisory committees; BMS: Consultancy, Membership on an entity's Board of Directors or advisory committees; AstraZeneca: Research Funding; Eli Lilly: Research Funding.