bluuebunny/crossref_metadata_2025 huggingface dataset:
Results of First-in-Man Phage Idiotype Vaccine Study in Patients with Multiple Myeloma.
Authors: Roehnisch Tim , Nagel Wolfgang , Boehm Thomas , Donceau Mariel , Bourquin Carole , Roehnisch Dilda A. , Rutz Beata , Straka Christian , Emmerich Bertold , Oduncu Fuat S.
Published: October 2019

Abstract 1858

Poster Board I-883

Objective:

Idiotypes (Id) expressed by tumor cells in B-cell malignancies can be regarded as tumor-specific antigens and may therefore be used as a target for vaccine immunotherapy. In this phase I/II study we have evaluated the feasibility, safety and response of phage idiotype vaccination in patients with relapsed multiple myeloma (MM) using phage displayed Id-specific proteins as immunogens and low doses of subcutaneous granulocyte-macrophage colony-stimulating factor (GM-CSF) as immunoadjuvants.

Study Design:

Patients received a total of 6 intradermal/subcutanous (i.d./s.c) immunizations of phage idiotype vaccines at day 1, 7, 14 and at week 4, 8 and 12. The vaccine dose started from 0.25 mg for the first 5 patients and was escalated to 1.25 mg (patient 6-10) and 2.5 mg (patient 11-15) if no dose-limiting toxicity was observed. Each vaccine contained 0.2 mg GM-CSF (Leukomax®) as adjuvant and 0.2 mg KLH (Immucothel®) as control antigen. They received all subcutaneous injections of Id vaccines and immunoadjuvants in an outpatient setting.

Patient Characteristics:

15 patients who had previously been treated with high-dose chemotherapy (HDT) followed by peripheral blood progenitor cell (PBPC) transplantation entered this study. All patients were in a beginning relapse after high dose therapy at the time of vaccination. Patients with fulminant relapse or patients with active infection and hematopoetic insufficiency after HDT were excluded.

Results:

The results of this first-in-man phase I/II trial for phage-based idiotype vaccination clearly demonstrated the feasibility, safety and excellent tolerability of the phage vaccination whereas local skin reactions were the most frequent side effects. Furthermore, we were able to gather precious data regarding the immunology of phage vaccines in man. We could confirm our preclinical data, that phages are good immunogens, capable of inducing a strong immune response mainly of IgG1-type and therefore enabling ADCC and CDC immune response. Altogether these data suggest, that phage idiotype vaccines has the potential to exert a beneficial effect especially in patients with minimal residual disease. Strikingly, the immune response of phages compared to the well-known KLH particles showed > 10 time higher antibody titer than KLH, which emphasize the prominent immunogenicity of the phages. Overall these data pinpoint to a promising role of phage vaccination for the immunotherapy of cancer but further studies are necessary to evaluate the full potential of phage vaccines.

Disclosures:

Roehnisch: Apalexo Biotechnologie GmbH: Research Funding. Nagel:Apalexo Biotechnologie GmbH: Research Funding. Donceau:Apalexo Biotechnologie GmbH: Research Funding. Bourquin:Apalexo Biotechnologie GmbH: Research Funding.
Oral Cyclic Melphalan and Dexamethasone in the Treatment of Patients with AL Amyloidosis; Ineligible for High-Dose Melphalan and Stem Cell Transplantation.
Authors: Sanchorawala Vaishali , Seldin David C. , Sloan John Mark , Berk John L. , Skinner Martha
Published: October 2019

Abstract 1883

Poster Board I-906

AL amyloidosis is caused by a clonal plasma cell dyscrasia and is characterized by widespread, progressive amyloid deposition leading to multisystem organ failure and death. In this disease, amyloid protein deposits are derived from monoclonal immunoglobulin light chains. Aggressive treatment of AL amyloidosis with high dose intravenous melphalan followed by autologous stem cell transplant (HDM/SCT) is effective in inducing hematologic remission and clinical improvement. However, only selected patients with AL amyloidosis are eligible for HDM/SCT due to amyloid-associated organ dysfunction. Recently, several investigators have demonstrated the efficacy of treatment with oral cyclic melphalan and dexamethasone (Mel/Dex) in inducing hematologic responses and improving survival for patients with AL amyloidosis. We report on 70 patients with AL amyloidosis who were treated with oral Mel/Dex. Oral melphalan was administered at 0.22 mg/kg/day for D1-4 and dexamethasone at 20-40 mg one day per week and repeated every month. The median age was 65 years (range, 46-84) and the median number of organ system involvement was 3 (range, 1–6). Majority of the patients (n=31) had predominant cardiac involvement. The reasons to select this regimen of Mel/Dex rather than HDM/SCT included severe cardiac involvement (n=23), age > 75 years (n=6), patient choice (n=6), severe autonomic neuropathy (n=6), poor functional status (n=5), treatment started by local physicians (n=9), complications of stem cell mobilization and collection precluding HDM (n=5) and others (n=11). Patients received a median of 4 cycles of Mel/Dex, (range 1-13). Hematologic responses were not evaluable in 22 patients due to early death, toxicity or patients failing to return for follow-up. Of 48 evaluable patients, 7 patients (15%) achieved a complete hematologic response. Median survival for these 70 patients has not yet been reached with a median follow-up of 17 months. Overall survival is 60% at 3 years. Thirteen of these patients received additional treatment for their disease. In conclusion, oral cyclic melphalan and dexamethasone can lead to hematologic complete responses in 15% of patients and to improvement in survival for patients with AL amyloidosis, who are not eligible for HDM/SCT.

Disclosures:

relevant conflicts of interest to declare.
Mechanism of Transfusion-Related Acute Lung Injury Induced by HLA Class II Antibodies.
Authors: Sachs Ulrich J , Wasel Wiebke , Reil Angelika , Bux Juergen , Bein Gregor , Santoso Sentot , Weissmann Norbert
Published: October 2019

Abstract 19

Introduction:

Transfusion-related acute lung injury (TRALI) is the leading cause of transfusion-associated fatalities in developed countries. It has been demonstrated that antibodies to human neutrophil antigens (HNA) present in a blood product can precipitate a TRALI reaction in the blood recipient. In the majority of TRALI cases, however, HLA class II antibodies rather than HNA antibodies are detected in the implicated donor. HLA class II antigens are not present on neutrophils or endothelial cells, the cellular key players in TRALI pathophysiology. Whether or not HLA class II antibodies are capable of inducing TRALI is thus under discussion. Here, we investigated a possible mechanism of HLA class II antibody-induced TRALI via monocytes. Methods and Results: Plasma from blood donors containing HLA class II antibodies (H2+plasma) is capable of activating monocytes expressing cognate antigen(s) as demonstrated by the release of IL-8, GRO-alpha, and TNF-alpha in culture supernatants. H2+plasma does not activate monocytes that do not express the cognate HLA class II antigen(s). Culture supernatants from matched pairs, but not H2+plasma itself, induce a respiratory burst reaction in neutrophils. In Boyden chamber experiments, FITC-albumin transendothelial flux is significantly increased in the presence of H2+plasma, matched monocytes, and neutrophils, but unchanged in the presence of non-matched monocytes. Diphenyleneiodonium chloride inhibits this barrier breakdown. In the ex vivo rat lung model, supernatants from matched pairs, but not from non-matched pairs, are capable of inducing TRALI in the presence, but not in the absence, of neutrophils. H2+plasma precipitates a TRALI in this model if neutrophils and matched, but not non-matched, monocytes are present. IgG-depletion of H2+plasma abolishes this effect. Conclusions: HLA class II antibodies present in donor plasma can induce TRALI in a transfusion recipient via monocyte activation. Once activated, monocytes stimulate neutrophils to produce reactive oxygen species, which lead to an increase in endothelial permeability and subsequent lung edema. This cascade depends on a match between the donor's antibodies and the recipient's HLA class II antigens and can be reproduced in a rat lung model. Our findings have important impact on current strategies that aim to prevent TRALI by donor testing and/or exclusion.

Disclosures:

No relevant conflicts of interest to declare.
Trends in Human T-Cell Leukemia Virus Type-1 (HTLV-1) Prevalence and the Incidence of Adult T-Cell Leukemia/Lymphoma (ATL) in Nagasaki, Japan: A Hospital-Based and Population-Based Study.
Authors: Iwanaga Masako , Koga Yoshito , Soda Midori , Inokuchi Naoko , Sasaki Daisuke , Hasegawa Hiroo , Yanagihara Katsunori , Yamaguchi Kazunari , Kamihira Shimeru , Yamada Yasuaki
Published: October 2019

Abstract 1920

Poster Board I-943

Introduction:

The prevalence of HTLV-1 is mostly evaluated by the age-specific seroprevalence in blood donors, and the results have been conventionally used to estimate the age-specific incidence of ATL in Japan. However, the results may be underestimated due to an age limit (16-69 yr) for donation, a healthy donor effect, and a birth cohort effect. Data concerning the birth-year specific incidence of ATL among HTLV-1 carriers other than blood donors are scarce.

Methods:

The study evaluated data of the anti-HTLV-1 antibody testing of 10,261 patients (males: 5,523, females: 4,737) who visited the Nagasaki University Hospital during 2000-2007 and data of 360 ATL cases (males: 188, females: 172) who were diagnosed in Nagasaki City (an endemic area in Japan) in a population-based Nagasaki Prefectural Cancer Registry (NPCR). To estimate birth-year specific incidence rates of ATL in population-based HTLV-1 carriers, we used the 2006 census population for Nagasaki City by applying the hospital-based seroprevalence data.

Results:

Of 10,261 patients, 1,392 (males: 653, females: 739) were HTLV-1 antibody positive. The overall HTLV-1 seroprevalence was 13.57% (95%CI: 12.90-14.23%). The seroprevalence was significantly higher in females than in males (15.60% vs. 11.82%, P<0.0001). The birth-year specific seroprevalence was 18.69% (before 1926), 17.83% (1927-1936), 15.91% (1937-1946), 13.80% (1947-1956), 9.19% (1957-1966), 4.07% (1967-1976), 2.07% (1977-1986), and 0% (after 1987) (a significantly declining trend: P <0.0001). The estimated annual number of HTLV-1 carriers by birth-year in Nagasaki city was 5257, 8093, 8151, 8083, 4434, 2180, 785, and 0, respectively. Finally, we estimated the annual incidence rate of ATLL per 100,000 HTLV-1 carriers by birth-year; 171 (before 1926), 86 (1927-1936), 41 (1937-1946), 32 (1947-1956), 11 (1957-1966), and 0 (after 1967). The crude lifetime risk of developing ATLL in HTLV-1 carriers was estimated to be 7.29% for males and 3.78% for females.

Conclusions:

The birth-year specific HTLV-1 seroprevalnces in the present study were approximately 50% higher than those previously reported in blood donors1 (for example: 6.22% in those born before 1950). Although it is possible that our results are over-estimated2, the present study suggests that there is still a large pool of elderly HTLV-1 carriers in this endemic area. Further studies are needed to investigate the mechanism of the development of ATL among HTLV-1 carriers for preventing the development. Reference: 1) Iwanaga M et al. Int J Hematol, 2009. 2) Arisawa K et al. Int J Cancer, 2000.

Disclosures:

No relevant conflicts of interest to declare.
Predictors of Early Response to Single Agent Rituximab in Patients with Indolent Lymphoma.
Authors: Cornec Divi , Tempescul Adrian , Querellou Solène , Mankai Amani , Hutin Pascal , Cymbalista Florence , Berthou Christian , Renaudineau Yves , Youinou Pierre
Published: October 2019

Abstract 1946

Poster Board I-969

STATEMENT OF PURPOSE:

Rituximab (RTX) is a chimeric antibody targeting CD20, a membranous B-cell specific protein, currently used in the treatment of B-cell malignancies and various autoimmune diseases. Despite its clear efficacy in non-Hodgkin's lymphoma, clinical and biological responses are highly variable between patients. Therefore, predictive factors are necessary to adapt therapy to each individual as early as possible.

MATERIAL AND METHODS:

We studied a set of biomarkers in a cohort of 39 patients with indolent non-Hodgkin's B-cell lymphoma (low grade follicular lymphoma or marginal zone lymphoma) and a low tumour burden, treated with single agent RTX as follows: 4 weekly 375 mg/m2 infusions, associated with short-course oral steroids. All patients benefited from FDG-PET analysis before treatment and 10 weeks later, allowing their classification into 3 early response groups: complete isotopic remission, partial response and no response. Dosage of rituximabemia, BAFF and Human Anti-Chimeric Antibodies (HACA) was performed with in-house ELISAs before each infusion and 2 months later. Polymorphisms of FcγRIIIa[158], FcγRIIa[131] and C1qA[276] genes were determined using allele-specific PCR or direct sequencing. Results were compared using Student's t-Test or two-tailed Fischer's exact test when appropriated.

RESULTS:

Referring to PET analysis, 18 patients were classified as complete isotopic remission, and 21 as partial or no response. Mean age was significantly lower in the first group (54 y+/-19 vs 66+/-13, p=0.022), as well as initial SUVmax (6.3+/-3.4 vs 10.8+/-6.8, p=0.024). Serum levels of RTX were highly variable between patients at each timepoint, but were not statistically different between the different response groups. Basal levels of BAFF were not statistically different between lymphoma patients and a control population, and did not vary after RTX-induced B cell depletion. No patient developed a HACA response during the study. Homozygous FcγRIIIa158V/V genotype was significantly associated with a complete response (p=0.037). Conversely, FcγRIIa[131] and C1qA[276] polymorphisms were not associated with the response. Genetic polymorphisms seemed to modify RTX pharmacokinetics, since homozygous FcγRIIa[131]H/H, FcγRIIIa[158]V/V and C1qA[276]G/G subjects displayed lower levels of RTX than others, without reaching statistical significance.

CONCLUSIONS:

Younger age, lower initial SUVmax and FcγRIIIa[158]V/V genotype are associated with a better early response to RTX in indolent lymphoma patients, as assessed by PET analysis. FcγR polymorphisms affect RTX kinetics probably through a modification of the efficacy of antibody dependant cell cytotoxicity. Determination of these factors could help in adapting individually therapeutic protocols.

Disclosures:

No relevant conflicts of interest to declare.
MiR-29 MicroRNAs Regulate IGF-1R Expression and Contribute Mantle Cell Lymphoma Growth and Survival.
Authors: Lin Jianhong , Zhao Jianjun , Lwin Tint , Luis Crespo , Guan Fangxia , Dessureault Sophie , Moscinski Lynn C. , Dalton William S. , Sotomayor Eduardo M. , Cheng Jin , Tao Jianguo
Published: October 2019

Abstract 1957

Poster Board I-980

Mantle cell lymphoma (MCL) is now recognized as a distinct clinicopathologic subtype of B-cell lymphomas and carries the poorest prognosis among NHL subtypes with a median survival is 3 to 4 years. Clearly better understanding of MCL oncogenesis and novel therapeutic approaches to treating MCL are needed. Mounting evidence now shows that small noncoding microRNAs (miRNAs) can play roles as oncogenes or tumor suppressor genes, suggesting their contribution to cancer development and progression. Here, we for the first time, demonstrated that that miR-29a-c is frequently deleted or down-regulated and IGF-1R is often over-expressed in primary mantle cell lymphoma cells. Expression of miR-29 inversely correlates with the expression of IGF-1R. Consistent with these results, 1) overexpression of miR-29-c down-regulated IGF-R1, 2) overexpression of miR-29c inhibited cell growth, colony formation and induced cell cycle arrest in MCL cells, 3) depletion of miR-29-c by anti-miR29-c increased the threshold for apoptosis, 4) enforced expression of miR-29c suppressed MCL growth and tumor progression in NOD/SCID mouse xenograft models, 5) miR-29 directly regulates IGF-1R expression . Furthermore cell cycle arrest induced by these miRNAs depends on the expression of IGF-1R. Our results indicate that miR-29a-c are implicated in MCL cell cycle control, survival and likely contribute to the tumorigenesis of MCL.

Disclosures:

No relevant conflicts of interest to declare.
Prevalence and Predictors of Pulmonary Arterial Hypertension in Hemoglobin E/ β-Thalassemia Patients.
Authors: Chuncharunee Suporn , Atichartakarn Vichai , Archararit Napaporn , Udomsubpayakul Umaporn , Ingsathit Atiporn , Lee Ratchanee , Tunhasiriwet Anusith , Tanomsup Supachai , Aryurachai Katcharin , Ungkanont Artit , Niparuck Pimjai , Angchaisuksiri Pantep
Published: October 2019

Abstract 2015

Poster Board I-1037

Introduction:

Pulmonary arterial hypertension (PAH) associated with thalassemia (Thal) hemoglobinopathy is now an accepted clinical entity. Most of the reports are in sickle cell disease and splenectomized β-Thal patients. Once manifested, it connotes poor prognosis. We herein present its prevalence and predictors in hemoglobin E/β-Thal (E/β-Thal) patients.

Patients and Methods:

One hundred and ten clinically stable E/β-Thal outpatients, on no medication aside from folic acid and who received no blood transfusion in the preceding 4 weeks were studied. All gave written informed consent, and study protocol was approved by the institution ethics committee on studies in humans (#0774/2548). Echocardiogram was used to estimate systolic PA pressure (SPAP). PAH was defined as an estimated SPAP ≥36 mmHg. Clinical features and laboratory data were stratified according to the presence or absence of PAH, and statistical analysis was done by STATA version 10 (Stata Corp, Texas), considering a P value <0.05 as statistically significant. Predictors of PAH were considered in univariate analysis.

Results:

There were 110 patients, 56 of whom were female and 61 were asplenic. PAH was present in 41 patients (37.3%), all of whom had normal left ventricular ejection fraction. There was no gender difference between the 2 groups (p=0.055). Selected statistically significant results are shown in the table.

Conclusions:

Prevalence of PAH in E/β-Thal patients is 37.3% without gender preponderance. Predictors are asplenia, more severe hemolysis, higher number of circulating (activated) platelets and nucleated RBCs, increased chronic low grade inflammation and increased cellular adhesion between blood and endothelial cells. These changes could facilitate development of thrombotic pulmonary arteriopathy, the underlying basis of PAH. Serum NT pro BNP assay can be utilized as a predictor or a screener of PAH in these patients.

Disclosures:

No relevant conflicts of interest to declare.
Characteristics of Transfused Platelets Do Not Affect Bleeding Outcomes in Hypoproliferative Thrombocytopenia.
Authors: Triulzi Darrell , Assmann Susan , Strauss Ronald G. , Ness Paul M. , Hess John R. , Granger Suzanne , Slichter Sherrill J.
Published: October 2019

Abstract 21

Background:

A number of studies have examined the effect of various platelet (plt) characteristics on the efficacy of plt transfusion(tx) including; source (apheresis platelets [AP] vs pooled platelets [PP]), plt ABO matching (identical [ID], minor mismatch [Min], major mismatch [Maj]), and duration of plt storage. These studies have focused on post transfusion plt counts with some evidence to support that there are differences in plt recovery based on these factors. These studies have not however examined the effect of these factors on the clinical outcome of bleeding.

Methods:

The PLADO study enrolled subjects with thrombocytopenia caused by stem cell transplantation (SCT) or chemotherapy into a blinded RCT of three different dosing strategies (lower, medium, and higher dose plt tx) to assess the effect of dose on the primary outcome of bleeding. Patients could receive either AP or PP with an attempt to keep patients on the same type plts. ABO selection and duration of storage were left to local practices. Patients were assessed daily for bleeding and, if present, assigned a WHO grade 2-4. A time to event analysis strategy was used with the outcome defined as the number of days from the first platelet transfusion to first date of bleeding. Subjects were censored on the first date they received a plt tx which differed from the first plt tx for the characteristic of interest. Analyses used Cox proportional hazard models with an adjustment for treatment group.

Results:

1351 patients were enrolled of which 1272 actually received a plt tx. 489 (38%) patients were excluded (41 given HLA selected plts, 421 with WHO Grade 2 or higher bleeding before or on the day of the first plt tx, and 27 with missing data) leaving 783 evaluable patients. The platelet source did not affect the time to first grade 2 or higher bleeding. Duration of platelet storage did not affect time to first grade 2 or higher bleeding . Because few patients received platelets stored for fewer than 3 days or more than 5 days, the analysis is shown for 3, 4 or 5 days of storage. ABO matching status did not affect time to first grade 2 or higher bleeding. A multi-predictor model using all 3 characteristics and treatment arm showed that none was significantly related to the time to grade 2 or higher bleeding.

Conclusion:

Platelet characteristics including source (AP vs PP), duration of storage for 3 to 5 days, and ABO matching, do not affect the outcome of time to bleeding in patients with SCT or chemotherapy receiving prophylactic platelet transfusions.

Disclosures:

Triulzi: Fenwal: Consultancy, Membership on an entity's Board of Directors or advisory committees. Ness:Northfield Laboratories: Consultancy, Membership on an entity's Board of Directors or advisory committees; Caridian: Consultancy, Membership on an entity's Board of Directors or advisory committees.
Influenzaviruses Are Inactivated by the Manufacturing Process of Plasma-Derived Products.
Authors: Groener Albrecht , Schäfer Wolfram
Published: October 2019

Abstract 2107

Poster Board II-84

The starting material of plasma-derived products, pooled human plasma, may contain the blood-borne viruses HIV, HCV and HBV as well as HAV and B19V. The presence of these viruses in blood has led to cases of virus transmission by blood transfusions and, in the past when these pathogens were incompletely inactivated or removed, also by plasma-derived products. There is a continuous concern for the safety of the starting material for plasma-derived products regarding emerging viruses potentially present in the donor population. These viruses may not be detected because (i) at the time of donation the donor is in the incubation period and appears healthy and (ii) no routine screening test is available to detect emerging viruses. Therefore, the manufacturing process of plasma-derived products must be able to inactivate or remove these pathogens with a high margin of safety. Currently, influenzaviruses are perceived as viruses of risk, especially the pandemic influenzavirus H1N1 and the avian influenzavirus H5N1.

We studied the inactivation of influenzaviruses by pasteurization (heat treatment at 60°C for 10 hours in aqueous solution), a virus inactivation method used in the manufacturing process of a range of plasma-derived products. The influenzavirus strains H1N1 (A/PR/8/34), H5/N2 (A/chick/Pennsylvania/1/83), H7N1 (FPV/Rostock/34) and H5N1 (A/Cygnus/Germany/06) were used in virus validation studies employing a laboratory scale of the manufacturing process. This scaled-down process was validated to mimic the production process as closely as possible, i.e. appropriate product intermediates were obtained from the production facility and spiked with virus stocks. The heat treatment was performed according to production parameters resulting in identical conditions of protein concentration, pH, temperature, time etc for different coagulation factor concentrates (VWF/FVIII; FVIII; FIX) as well as ATIII. The virus strains were propagated in a chicken cell line (CER-3, established at CSL Behring) and the virus infectivity was quantified on MDCK cells based on a cytopathogenic effect. The data demonstrate these strains of influenzaviruses are highly sensitive to heat treatment: after a heat treatment of 6 hours or less, no infectious virus could be detected. The pasteurization step demonstrated an inactivation factor for each strain of at least ≥ 4.4 log10. We plan to study additional influenzavirus strains, as well as further products, to increase the robustness of the data base showing the effective inactivation of influenzaviruses by this heat treatment method alone.

Disclosures:

Groener: CSL Behring: Employment. Schäfer:CSL Behring: Employment.
Prominin-1 Mobilisation, Collection and Immunoselection in Cancer Patients for Liver Regeneration.
Authors: Perotti Cesare , Fante Claudia Del , Viarengo Gianluca , Maestri Marcello , Salvaneschi Laura
Published: October 2019

Abstract 2141

Poster Board II-118

Background:

CD133+ (Prominin-1 positive) is a 5-transmembrane glycoprotein that identifies immature progenitor stem cells.

Immature hematopoietic stem cells retain the possibility to give origin to tissues different from hematopoietic cell lines (transdifferentiation).

Material and methods:

In this preliminary study we investigated the possibility to mobilize, collect, immunoselect and reinfuse autologous CD133+ immature stem cells in liver cancer patients. This approach was adopted to obtain, in a short time, an adequate volume increase of the disease free liver thus extending the resectability criteria of the liver, with the final goal to prolong survival.

We enrolled 4 patients with a large liver cancer and no chance of resection. The mobilization protocol consisted in: G-CSF administration (10 μg/kg/day ) for 3-5 days, peripheral blood stem cell (PBSC) monitoring starting from the 3rd day, leukapheresis (LKF) collection processing 3 blood volumes when CD133+ cells>15 μL. Patients were monitored during mobilization, collection and post collection phase for clinical status, blood pressure and bleeding. Positive CD133+ immunoselection (Miltenyi Clinimacs) was performed on LKF product after overnight storage. Quality controls on positive fraction consisted in viability and purity of CD133+ cells by cytofluorimetric analysis and clonogenic assays. Microbial tests were performed on the negative fraction.

After LKF, patients underwent right portal embolization and infusion of CD133+ cells into the opposite portal vein by a percutaneous access. Evaluation of liver regeneration was performed 30 days after stem cell infusion by spiral CT and galactose clearance. Liver resection was carried out if liver regeneration reached 30-40%.

Results:

Stem cell mobilization, LKF content and immunoselected cells are detailed in tab 1. No relevant side effects were observed. We obtained an efficient stem cell mobilization in all patients enrolled. No bacterial or fungal contamination was observed in cells infused. Results about liver regeneration and patients' follow up are detailed in table 2.

Conclusions:

Our approach to liver regeneration was feasible and safe with no relevant side effects. We observed an efficient stem cell mobilization comparable to healthy donors also in liver cancer patients. The infusion of CD133+ cells allowed a significant hepatic tissue regeneration in all patients. Controlled clinical trials are needed to confirm our preliminary results.

Disclosures:

No relevant conflicts of interest to declare.
Pentraxin-3: An Easily Measurable Soluble Factor to Improve Post-Transplant Graft Versus Host Disease Monitoring and Management.
Authors: Dander Erica , Cuccovillo Ivan , Vinci Paola , Bonanomi Sonia , Maio Lucia Di , Lucchini Giovanna , Migliavacca Maddalena , Perseghin Paolo , Rovelli Attilio , Balduzzi Adriana , Mantovani Alberto , Bottazzi Barbara , Biondi Andrea , D'Amico Giovanna
Published: October 2019

Abstract 2217

Poster Board II-194

Introduction:

Allogeneic haemopoietic-stem-cell transplantation (HSCT) is the treatment of choice for many malignant and non-malignant disorders. Despite the recent advances in post-transplantat immunosuppressive therapy, Graft-versus-Host Disease (GVHD) still represents the major life-threatening complication, developing in a substantial number of HSCT patients and resulting in poor outcome. The basis of GVHD pathophysiology are still poorly understood and its current diagnosis depends mainly on clinical manifestations and invasive biopsies. Specific biomarkers for GVHD would facilitate the early and accurate recognition of this invalidating disease as well as the monitoring of patient response to adopted anti-GVHD pharmacological treatment. With the aim to explore new reliable markers for predicting and monitoring GVHD course, we focused on pentraxin-3 (PTX-3), an acute-phase protein, that has been shown to play a crucial role in orchestrating inflammatory immune responses.

Patients and Methods:

Having obtained an informed consent, we collected plasma samples from 46 patients who received unmanipulated HSCT and from 9 healthy donors (HD) volunteers. After HSCT, 25/46 patients developed skin GVHD (18 acute GVHD and 7 chronic GVHD), while 21/46 never experienced it. Concerning GVHD patients, blood samples were collected at the day of GVHD onset/ flare, before the beginning of GVHD-specific drug therapy. PTX-3 plasma levels were monitored by ELISA assays.

Results:

Patients who did not develop GVHD after HSCT showed augmented PTX-3 plasma levels (mean=3.3 ng/ml, range=1.1-8.6 ng/ml) if compared to HD (mean=1.2 ng/ml, range=0.3-2.5 ng/ml, p<0.01). Interestingly, we observed a strong increase of PTX-3 plasma levels in patients with acute GVHD (mean=42.2 ng/ml, range=6.7-218.2 ng/ml) or with flair-ups of chronic GVHD (mean=15.8 ng/ml, range=9-44.3 ng/ml). The increase of PTX-3 levels in patients with acute and active chronic GVHD was statistically significant (p<0.01 and p<0.05 respectively) when compared to both HD and HSCT patients without GVHD.

Conclusions:

These preliminary results suggest that PTX-3 plasma levels increase very rapidly in patients experiencing active GVHD, thus candidating PTX-3 as an easily measurable soluble factor useful to corroborate clinical observations in a disease in which signs and symptoms are often protean. Further studies are needed to clarify if PTX-3 could represent a good diagnostic and/or prognostic factor rapidly indicating therapy responsiveness.

Disclosures:

No relevant conflicts of interest to declare.
Donor Statin Treatment Protects against Severe Acute Graft-Versus-Host Disease After Related Allogeneic Hematopoietic Cell Transplantation.
Authors: Rotta Marcello , Storer Barry E , Storb Rainer F , Martin Paul J , Heimfeld Shelly , Peffer Amanda , Sandmaier Brenda M , Maloney David G , Deeg H. Joachim , Appelbaum Frederick R , Mielcarek Marco
Published: October 2019

Abstract 2227

Poster Board II-204

Acute graft-versus-host disease (GVHD) contributes to morbidity and mortality after allogeneic hematopoietic cell transplantation (HCT) and more effective prevention and treatment strategies are needed. The 3-hydroxy-3-methylglutaryl coenzyme A-reductase inhibitors (statins) have been shown to possess immunomodulatory properties in vitro and in vivo. To determine whether donor or recipient statin use may affect the risk of GVHD, we retrospectively analyzed outcomes among 567 patients with hematologic malignancies who had hematopoietic cell transplantation from HLA-matched related donors at a single institution between 2001 and 2007. Compared to allografts where neither the donor nor recipient was treated with a statin at the time of transplant (n=464), statin use by the donor and not the recipient (n=75) was associated with a profoundly decreased risk of grade 3-4 acute GVHD (multivariate hazard ratio [HR], 0.28; 95= confidence interval [CI], 0.1-0.9; p=0.03) (Table). Statin use by both donor and recipient (n=12) was suggestively associated with a decreased risk of grade 3-4 acute GVHD (HR, 0.00; 95= CI, undefined; p=0.06), while statin use by the recipient and not the donor (n=16) did not confer GVHD-protection. Risks of chronic GVHD, recurrent malignancy, non-relapse mortality and overall mortality were not significantly affected by donor or recipient statin exposure. Statin-associated GVHD-protection was restricted to recipients with cyclosporine-based postgrafting immunosuppression (n=417; 74=) and was not observed among those given tacrolimus (significance of effect modification: cyclosporine vs. tacrolimus, p=0.009). These results suggest that donor statin treatment may be a promising strategy for preventing severe acute GVHD without compromising immunologic control of the underlying malignancy.

Disclosures:

No relevant conflicts of interest to declare.
Antifungal Therapy with Itraconazole Impairs Rituximab′s Anti-Lymphoma Effect by Inhibiting CD20 Recruitment to Lipid Rafts.
Authors: Bueschenfelde Christian Meyer zum , Feuerstacke Yvonne , Krainz Philipp , Herrmann Ken , Buck Andreas , Peschel Christian , Ringshausen Ingo
Published: October 2019

Abstract 2735

Poster Board II-711

Patients with hematologic malignancies have a high risk of developing invasive fungal infections. The higher risk is attributed to host defense impairment due to intensive cytotoxic chemotherapies, hematopoetic stem cell transplantation, and immunosuppressive agents. As early treatment initiation in patients with invasive fungal infections has a profound impact on mortality rates, different antifungal treatment strategies like prophylaxis, empirical, pre-emptive and targeted treatment after proven fungal infection have been developed. Azoles are the most broadly used antifungal drugs inhibiting CYP51. They play a pivotal role in the treatment and prophylaxis of systemic and dermal mycoses. Azoles inhibit the sterol 14a-demethylase activity and block sterol biosynthesis, which is lethal in unicellular organisms. However, in mammalian cells, it has been shown that they lower endogenous cholesterol production. As lipid rafts consist of sphingolipids and cholesterol and lipid rafts have been demonstrated to be crucial for rituximab induced cell death, we asked whether rituximab exhibits its full anti-lymphoma effect in the presence of azoles.

We now demonstrate that antifungal treatment with itraconazole impairs rituximab's anti-lymphoma effect both in vitro and in vivo. Using a mouse xenograft model, no tumor growth was observed in mice treated with rituximab. In contrast, in the presence of itraconazole there was tumor growth, indicating that the concentration was sufficient to antagonize rituximab′s anti-lymphoma effect. On the molecular level, CD20 raft recruitment was inhibited in the presence of itraconazole. Furthermore, the translocation of CD20 into lipid rafts has been shown to be crucial for calcium influx and apoptosis. Therefore, we tested the rituximab induced calcium flux in the absence and presence of itraconazole. Indeed, calcium flux was almost completely abolished in the presence of itraconazole. The fact that antibody treatment failure in the presence of itraconazole only affects targeted therapy against raft associated molecules i.e. CD20 and CD52 but not the non raft associated molecule CD33 further supports our proposed mechanism of action. Together, our data suggest that concomitant treatment of itraconazole and rituximab attenuates rituximab's anti-lymphoma effect. As chemotherapy by itself has high response rates in lymphoma, the loss of rituximab mediated cell death in the presence of itraconacole may be missed in our daily routine.

Disclosures:

No relevant conflicts of interest to declare.
Expression of S100A8 in Leukemic Cells Predicts Poor Survival in De Novo AML Patients.
Authors: Mossuz Pascal , Nicolas Emmanuelle , Ramus Claire , Berthier Sylvie , Arlotto Marie , Lefebvre Christine , Morel Françoise , Garin Jérome , Ifrah Norbert , Berger François , Cahn Jean-Yves
Published: October 2019

Abstract 276

Treatment strategies of acute myeloid leukaemia (AML) largely depend on their cytogenetic status. However this stratification remains insufficient for almost half of the patients, requiring subsequent molecular investigations. In our study, we used mass spectrometry-based proteomic approaches to characterize de novo-AML. Samples (blood mononuclear cells collected and frozen before any treatment) were retrospectively selected from two independent sets of newly diagnosed AML patients, included in prospective clinical trials of the GOELAMS (Groupe d'Etude Ouest-Est des Leucémies aigues). We showed that protein signature of leukemic cells defined 2 groups of patients that displayed significant variation of overall and disease free survival (Fig-1A). This proteomic classification refined cytogenetic classes. Combination of proteomic and cytogenetic allowed a new stratification highly correlated with outcome. In particular, AML with intermediate and high risk cytogenetic could be respectively subdivided according to their protein profiles in two subgroups with significantly different survival (Fig-1B). Interestingly in both type 2 and type 3 cytogenetic groups, a good proteomic profile identified subsets of patients with significantly increased probability of survival, suggesting that the weigh of a good proteomic risk might predominate over a bad predictive cytogenetic. Among proteins expressed by leukemic cells, we isolated a 10800 Da marker that retained the highest discriminative value between alive and deceased patients. The median intensity of the 10800 peak was 94.3 ± 97.7 (7.5-368.7) in living patients compared to 214.2 ± 163.7 (9.0-693.4) in dead patients (p= 0.009). Among normal cytogenetic patients, intensity levels of the marker was also significantly different between dead (217.4 ± 155.7 range 10.4-612.8) and alive patients (26.0 ± 73.1, range 6-168.1; p= 0.008) (Fig-1C). Using a thresholds of 100 (calculated by ROC curves) we were able to correctly identify 82% of patients who died (patients greater than 100) and 70% of patients who stayed alive (patients lower than 100) with a specificity of 65% and 82% respectively. These data were confirmed in a second independent set of patients. 10800 Da marker was identified by MS peptide sequencing as S100A8 (also designated MRP8 or calgranulin A), a cytosolic protein of mature granulocytes. Western blot analysis confirmed its expression mainly in AML patients with the worst prognostic but not in all AML patients, neither in some leukemic and lymphoma cell lines, arguing for a selective deregulation associated with poor prognosis. These results show that expression of S100A8 in leukemic cells at diagnosis might be considered as a predictor of low survival.

Disclosures:

No relevant conflicts of interest to declare.
The Telomerase Inhibitor RHPS4 Induces Telomere Shortening, DNA-Damage and Cell Death in AML Cells and Chemosensitises Cells to Daunorubicin.
Authors: Pallis Monica , Ojo Dotun , Richardson Jaineeta , Ronan John , Stevens Malcolm , Russell Nigel H. , Seedhouse Claire
Published: October 2019

Abstract 2760

Poster Board II-736

The quadruplex ligand RHPS4 is the lead compound in a drug discovery program at the University of Nottingham. It has been shown to bind to telomeres and inhibit telomerase, and subsequently induces growth arrest in progenitor cells from cancer cell lines whilst sparing normal haematopoietic progenitor cells. We explored its in vitro effects in AML cells, which are reported generally to have considerably shorter telomeres than normal CD34+ cells. AML cell lines were grown for 21 days in suspension culture. Primary samples were cultured for 14 days in semi-solid medium. Telomere length was measured by Southern blotting. γH2A.X was used to identify a DNA damage response, and cell viability was measured flow cytometrically with 7-amino actinomycin D. As reported in other tumour cell types, sensitivity to RHPS4 was found to be greatest in those AML cells with the shortest telomeres. In the OCI-AML3 cell line 0.3 μM RHPS4 inhibited cell growth by 50% in a 21 day clonogenic assay, accompanied by shortening of telomeres from 2.6 Kb to <1 Kb. Molm 13 cells (initial telomere length 3.2kB) also underwent telomere shortening in the presence of 0.3 μM RHPS4 (2.8Kb), whereas TF1a and U937 (both with initial telomere lengths approximately 6.5 kB) were insensitive at that concentration. After 6 days at 0.3 μM, RHPS4 was cytostatic, but at higher concentrations (1 μM) the drug was found to induce a substantial DNA damage response and loss of viability to OCI-AML3 cells. Moreover 0.3 μM RHPS4 enhanced the γH2A.X expression and cell death induced by the chemotherapy drug daunorubicin in these cells. Using 14 day clonogenic assays in primary AML samples (n=6), we found that the IC50 for RHPS4 alone was 0.7 μM. However, in the presence of 0.3 μM RHPS4, the median IC50 to daunorubicin was reduced from 19 nM to 5.5 nM. In conclusion we have determined that RHPS4 has telomere-shortening, cytostatic, cytotoxic and chemosensitising properties in AML cells.

Disclosures:

Stevens: Pharminox Ltd: director and shareholder of Pharminox Ltd which has a financial interest in RHPS4.
Clinical Experience of Decitabine in Elderly Patients with Myelodysplastic Syndromes (MDS).
Authors: Geils George F.
Published: October 2019

Abstract 2792

Poster Board II-768

Background:

Decitabine, a potent DNA hypomethylating agent, has demonstrated efficacy in MDS patients. The incidence of MDS increases with age, with limited treatment options for these patients. With advancing age, patients are generally less tolerant of treatment and suffer from more comorbidities. Important goals of therapy are to improve transfusion dependence and to delay progression of disease, with favorable toxicity. This subgroup analysis evaluated the effect of decitabine on overall response rate (ORR), duration of response, time to response, transfusion independence, and tolerability in subgroups of older patients (65-75 and >75 years of age).

Methods:

Data used in these analyses were from two previously published clinical trials (Steensma et al. J Clin Oncol, 2009 [DACO-020] and Kantarjian et al. Cancer 2006 [DACO-007]). In both trials, eligible patients were ≥18 years of age and had MDS (de novo or secondary) of any French-American-British subtype and an International Prognostic Scoring System score ≥0.5. In DACO-020, patients were treated with decitabine 20 mg/m2 IV daily for 5 consecutive days every 4 weeks. Response rate was assessed with the International Working Group (IWG 2006) criteria in DACO-020. In DACO-007, patients were treated with decitabine 15 mg/m2 IV over 3 hours every 8 hours for 3 consecutive days every 6 weeks. Response rate was assessed with the IWG 2000 criteria in DACO-007.

Results:

In DACO-020, baseline disease characteristics were generally similar between groups, except more patients in the 65-75 age group (n = 58) had RARS (19% vs. 4%) or RAEB (50% vs. 38%) and fewer had RA (12% vs. 38%) compared to patients in the >75 age group (n = 26). Further, more patients in the >75 group were transfusion dependent at baseline (77% vs. 57%).

Grade 3-4 anemia, leukopenia, neutropenia, and thrombocytopenia were less frequent (22, 5, 35, and 22%, respectively) in patients in the 65-75 age group versus patients in the >75 age group (27, 8, 46, and 35%, respectively). Remarkably, however, there were no reports of febrile neutropenia in >75 age group (26% in the 65-75 age group).

Conclusions:

In this subgroup analysis, decitabine is an effective and generally well tolerated treatment strategy, even in elderly patients with MDS. Despite the advanced age, responses were more rapid and more durable in the mature elderly than younger patients. The slightly higher hematological toxicity did not translate into increased infectious risk, with no febrile neutropenic episodes in patients over age 75 in these studies. These data suggest that the goals of therapy do not need to be stratified by age, and support future trials of decitabine in elderly and other at-risk populations with MDS.

Disclosures:

Geils: Eisai Inc. : Speakers Bureau.
A Characteristic Cytokine/Chemokine Milieu Is Present in the Bone Marrow Environment of Multiple Myeloma Patients Before and After Allogeneic Stem Cell Transplantation.
Authors: Cao Yanran , Luetkens Tim , Kobold Sebastian , Hildebrandt York , Gordic Maja , Lajmi Nesrine , Meyer Sabrina , Bartels Katrin , Zander Axel R. , Bokemeyer Carsten , Kröger Nicolaus , Atanckovic Djordje
Published: October 2019

Abstract 2795

Poster Board II-771

Background:

The interaction of Multiple Myeloma (MM) with its bone marrow (BM) microenvironment is important for the homing, survival, and proliferation of the malignant plasma cells. In this study, we aimed at answering the question which cytokines, chemokines, and growth factors are typically found in the BM of untreated MM patients as well as in MM patients after allogeneic stem cell transplantation (alloSCT) and might be involved in the pathogenesis of MM and/or Graft-versus-Host Disease (GvHD).

Design and Methods:

Applying Multiplex Bead-based Luminex technology and Quantikine ELISAs, we determined the concentrations of 34 cytokines/chemokines in the supernatants of 10 myeloma cell lines as well as in the plasma derived from BM and peripheral blood (PB) samples of 10 newly diagnosed MM patients, 20 MM patients post alloSCT, and 20 healthy donors.

Results:

Besides factors known to be secreted by myeloma cell lines (IL-1RA, IL-8, MCP-1, MIP-1α, MIP1β, MIP-3α) we observed secretion of other cytokines/chemokines such as EGF, HGF, IL2R, IL-12p40/p70, IL-22, IP-10, MIG, and RANTES. In the BM plasma samples of MM patients, we detected elevated levels of HGF, IL-2R, IL-16, EGF, IL-1RA, IP-10, MCP-1, and MIG (p<0.01 or p<0.05). Most of these factors were significantly increased in the BM compared to PB. In addition to increased levels of the factors mentioned above, the BM plasma of MM patients post alloSCT showed selectively elevated concentrations of IL-4, IL-6, IL-8, IL-12p40/p70, and eotaxin (p<0.01 or p<0.05). Eotaxin levels were particularly high in patients with chronic GvHD. Correlating the BM cytokine/chemokine levels with the clinical characteristics of MM patients, we observed that BM levels of IL-16 (r=0.829, p=0.003) and HGF (r=0.805, p=0.005) correlated positively with myeloma cell infiltration within the BM. In addition, IP-10 (r=0.770, p=0.009), HGF (r=0.645, p=0.044), and IL-6 (r=0.664, p=0.036) correlated significantly with the initial stage of disease.

Conclusions:

Our study demonstrates that myeloma cells themselves generate a significantly higher number of cytokines/chemokines than previously described. We show that characteristic cytokine/chemokine patterns exact in the BM environment of MM patients before and after alloSCT. Certain factors, such as MIP-1α, MCP-1, HGF, IL-16, IP-10, and eotaxin might not only be developed into diagnostic instruments and/or predictive biomarkers, but might also represent potential targets for future myeloma- or GvHD-specific therapies.

Disclosures:

No relevant conflicts of interest to declare.