julilili42/papers huggingface dataset:
Role of Consolidation Therapy in the Treatment of Patients up to 60 Years with High Risk AML.
2005-11-16

Abstract In the prospective multicenter AML 01/99 trial, 484 patients with acute myeloid leukemia up to 60 years (385 with de novo and 99 with secondary AML) were treated with risk-adapted therapy. Patients with CBF-leukemias [t(8;21) or inv(16)] or normal karyotype and good response (GR) to induction I (≤5% blasts in day 15 BM) were considered standard risk (SR), all other patients as high risk (HR). Patients with t(15;17) were excluded. Induction course I consisted of standard dose araC, idarubicine and etoposide (IVA-I). Patients with GR to IVA-I continued with IVA-II on day 21 as double induction. In patients with bad response (&amp;gt;5% bone marrow blasts on day 15), the second cycle consisted of FlAG/Ida. Double induction was followed by an early consolidation with intermediate dose araC. As late consolidation, HR patients with an HLA-matched sibling were scheduled for a matched related donor (MRD) transplantation whereas patients without a family donor should undergo autologous peripheral blood stem cell transplantation (autoPBSCT). PBSC were mobilized after early consolidation. A pilot study within the 01/99 trial showed superior results for matched unrelated donor (MUD) transplantation compared to patients who had undergone autoPBSCT. Therefore, since 2003 all HR patients without a sibling donor were scheduled for MUD transplantation. 234 patients were classified as HR, of whom 137 (58%) achieved complete remission. 31 (13%) of the HR patients died during induction. After 70 months, overall survival (OS) was significantly worse for HR patients (28%) than for SR patients (51%). For HR patients, there was no difference in OS and relapse-free survival (RFS) between those patients classified as HR because of bad response to IVA-1 (n = 76), unfavourable karyotype (n = 83) or both characteristics (n = 75). Moreover, RFS was neither different between HR patients with de novo or secondary AML nor between HR patients with normal karyotype, complex karyotype or other high risk cytogenetics. Regarding the different regimens for late consolidation, the “as treated” analysis of the HR patients revealed that OS and RFS was significantly better after an MRD transplantation (n=34, OS 68%, RFS 65%) than after autoPBSCT (n=26, OS 23%, RFS 7%). OS and RFS for the 29 patients who received MUD transplantation was also significantly better than after autoPBSCT with 56% and 52% respectively. In conclusion, HR patients as defined in our study do not benefit from an autologous PBSCT even after intensive double induction and early consolidation. Therefore, an allogeneic transplantation from a related or unrelated donor should be performed.
Inhibition of Hypusination of Eukaryotic Initiation Factor 5a (eIF-5A) as a Novel a Synergistic Treatment Strategy in Imatinib-Treated BCR-ABL Positive Leukemias Identified by a Global Proteomics Approach.
2005-11-16

Abstract Imatinib (IM) is highly effective in the treatment of CML and remains the gold standard non-transplant therapeutic option. However, resistance to IM develops frequently, particularly in late stage disease. The aim of this study was to discover novel cellular Bcr-Abl targets leading to the identification of potential novel treatment options that would synergise with IM. Protein expression analysis of K562 leukemic cells after IM treatment was performed using 2-dimensional gel electrophoresis, matrix-assisted laser desorption/ionization - time of flight mass spectrometry (MALDI-TOF) and nano electrospray ionization tandem mass spectrometry (ESI-MS/MS). Differential protein expression of selected proteins was confirmed by Western blot analysis. Among others, we detected the IM-dependent down-regulation of the eukaryotic translation initiation factor 5A (eIF-5A), which is essential for cell proliferation. EIF-5a represents the only known eukaryotic protein activated by a unique, 2-step post-translational modification called hypusination, a process that can be specifically inhibited by hypusination inhibitors (HI), e.g. ciclopirox and GC7. We demonstrated that selective HI elicit a strong synergistic effect with IM on both cellular cytotoxicity (using MTT assay) and apoptosis (using PI staining) of Bcr-Abl positive leukemic cell lines and even more strikingly on primary CD34+ cells from CML patients at diagnosis. The effect of HI on CD34+ cells from healthy donors and CML patients was compared by short-term expansion in serum free medium using high resolution cell cycle analysis and carboxy-fluorescein succinimidyl ester (CFSE) labelling to track cell division. We observed a substantial anti-proliferative effect of the drug combination on Bcr-Abl positive as compared to Bcr-Abl negative cells. However, we were unable to demonstrate a significant cytotoxic effect of the combination on the quiescent Ph+ stem cell population. Despite this, HI exerted strong antiprolifertive effects not only on Ba/Fp210 but also on resistant Ba/F3p210 mutants T315I, E255V and M351T. In conclusion, IM induced significant changes of global cellular protein expression in K562 cells, including down-regulation of eIF-5A. Inhibition of eIF-5A hypusination represents a promising target for combined therapy in Bcr-Abl positive leukemias. Ongoing studies are investigating whether novel, more specific, HI in combination with IM (and/or 2nd generation Bcr-Abl inihibitors e.g AMN107, BMS-354825) will be able to exert these effects on the quiescent Bcr-Abl+ stem cell compartment, and if these novel combinations are effective in IM-resistant cells.
Effective Ex Vivo-Generation and Quantification of Leukemia-Derived DC Has To Precede a Specific T-Cell Stimulation for Adoptive Immunotherapy in AML and MDS.
2005-11-16

Abstract The presentation of leukemic antigens can be improved in AML and MDS by in vitro conversion of leukemic cells in leukemia-derived DC (DCleu), thereby forming a platform for the generation of leukemia-specific cytotoxic lymphocytes (CTL). In preliminary analyses we could already define optimal serum-free culture conditions, generate and characterize DC from 55 MDS- and 99 AML-cases and characterize their T-cell activating function (Kufner S. 2005 I-III). DC/DCleu were quantified according to their surface DC /blast -marker profiles. However in 6/30% of cases with AML/MDS less than 10% of DC could be generated and only 50–60% of leukemic cells were convertible to DCleu. Therefore we tested 5 alternative methods to improve the harvest of DC/DCleu in 99 AML and 55 MDS-patients (1) ‘MCM-mimic’ (Lee, 2003), 2) ‘Ca-Ionophore’ (Houtenbos, 2003), 3) ‘Picibanil’ (Sato, 2003), 4) ‘Cytokines’ (Westers, 2003) and 5) ‘Poly I:C’ (Rouas, 2004)). 1. Although the percentual harvest of DC/DCleu was comparable in all of the methods (28–40% DC, with 45–65% of those DC being DCleu and about 47% of blasts being convertible to DCleu) not every method was successful in individual patients. 2. However we could demonstrate that DC can be generated from every sample with at least one of the following methods: MCM-mimic, Ca-Ionophore and Picibanil. 3. Highest DC/DCleu yield was seen in monocytoid differentiated FAB-types and was independent of the cytogenetic risk. 4. Many ‘DC-surface markers’ can be aready expressed on naive blasts in AML/MDS-patients before there conversion to DCleu. Moreover the ‘DC-marker expression’ is variable in individual patients and culture methods. In summary our data show 1. that the generation of DC/DCleu is possible independent from the karyotype in every patient under serum-free conditions with at least one of the 3 methods MCM-mimic, Ca-Ionophore and Picibanil. 2. The overall percentual harvest of DCleu from MNC-fractions after culture in the 5 methods compared is low. Possibly a combination of the optimal method with optimal ‘DC- and blast markers’ in every single case could improve the detectability and quantification of DC/DCleu and residual blasts. This contributes to improve the generability and quality of DCleu to stimulate and expand anti-leukemia-directed T-cells for the immunotherapy of AML and MDS.
Long Term Remission in Follicular Lymphoma (FL) and Mantle Cell Lymphoma (MCL) with Rituximab Consolidation after High-Dose Chemotherapy and Autologous Stem Cell Transplantation: Extended Follow-Up of a Multicenter Phase II Study.
2005-11-16

Abstract We investigated the efficacy of rituximab given after high-dose therapy (HDT) and autologous peripheral blood stem cell transplantation (ASCT) in order to prevent relapse by clearance of minimal residual disease in patients with advanced FL and MCL. Eligibility included newly diagnosed stage III/IV t(14;18)+ FL or newly diagnosed or relapsed patients with stage III/IV t(11;14)+ MCL. 35% of patients had elevated LDH levels, 23% had B-symptoms. The age-adjusted IPI risk category was low-intermediate in 65% and high-intermediate in 35% of the patients. Initial cytoreduction consisted of VACOP-B (6 weeks) or CHOP chemotherapy, followed by G-CSF supported VIP-(E) or DexaBEAM therapy. PBSC were harvested, and positive selection of CD34+ cells was performed. HDT consisted of TBI (6x2 Gy) and cyclophosphamide (120 mg/kg), followed by CD34+ PBSCT (7.16x106 CD34+/kg; range 2.4–22.7). A median of 62 days (range 38–108) after transplantation, rituximab was given at a dose of 375 mg/m2 weekly for 4 weeks. No additional treatment was given thereafter. Between October 1997 and July 2001, 31 patients were included. Median age at transplantation was 49 (31–60) years. One patient died early after HDT due to neutropenic septicemia before rituximab administration. Therefore, only 30 patients were evaluable for safety and efficacy of rituximab after HDT and ASCT (FL n=20, MCL n=10). Twenty seven were evaluable for MRD. In the intent-to-treat (ITT) analysis, 7 treatment failures were observed: two FL patients relapsed at 25 and 33 months post transplant, four MCL patients relapsed at 24, 38, 47 and 48 months post transplant, and one MCL patient died unexpectedly in complete remission (CR) at 19 months post transplant. One of the MCL patients died at 37 months post transplant due to progressive disease. Currently, 28 of 30 patients are alive. With an extended median follow up of 61 months (18.7 – 87.5 months) after ASCT, the progression-free survival (PFS) for the ITT population is 76.7%, the median PFS has not yet been reached. In the FL population, the PFS at 5 years is 90% (95% CI: 65.6 – 97.4) whereas in the MCL population, the PFS is 46.2% (95% CI: 14.7 – 73.3) (log rank p=0.0168). The overall survival (OS) at 5 years for the ITT population is 93.3% (95% CI; 75.6 – 98.3) with 100% OS in the FL patients and 79.4% (95% CI: 39.6 – 94.4) (log rank p=0.0400) in MCL patients. In summary, at an extended follow-up of 5 years, we show that one 4-week course of Rituximab consolidation after HDT and ASCT induces prolonged remissions in both FL and MCL. Futher improvement in MCL therapy is warranted.
Targeting MLL-AF4 with Short Interfering RNAs (siRNAs) Induces Apoptosis Related Genes APAF-1 and SEPT4 in t(4;11)-Positive Human Leukemic Cells.
2005-11-16

Abstract The chromosomal translocation t(4;11) marks infant acute lymphoblastic leukemia associated with a particularly dismal prognosis. We have recently shown that transfection of leukemic cells with MLL-AF4 siRNAs reduces leukemia-associated morbidity and mortality in xenotransplanted SCID mice suggesting that MLL-AF4 depletion negatively affects leukemia-initiating cells. In cell culture, transient suppression of MLL-AF4 expression impairs the proliferation of both t(4;11) cell lines SEM and RS4;11 more than tenfold. Clonogenicity of SEM cells was reduced fivefold and of RS4;11 cells more than twofold. One major reason for the antiproliferative consequences is the induction of apoptosis associated with caspase-3 activation and diminished BCL-XL expression. Caspase-3 was induced 3-fold and BCL-xL showed a 60% reduction after siRNA treatment compared to Mock-transfected controls in SEM cells. Analysis of intrinsic signaling pathways of apoptosis showed that MLL-AF4 depletion induces the expression of the proapoptotic genes APAF-1 and SEPT4 (ARTS). In both cell lines APAF-1 was induced more than twofold and SEPT4 more than fourfold. Our findings demonstrate that MLL-AF4 negatively interferes with an intrinsic apoptotic response and is, thus, important for the survival of these highly aggressive and chemoresistant leukemic cells. SiRNA-mediated MLL-AF4 inhibition may sensitize these cells for apoptosis-inducing agents. Therefore, complementing existing treatment strategies with a targeted inhibition of MLL-AF4 expression may lead to an improved and more specific treatment of this therapy-resistant leukemia.
Prospective Randomized Comparison of High Dose AraC and Autologous Peripheral Blood Stem Cell Transplantation as Late Consolidation for Patients ≤60 Years with Standard Risk AML.
2005-11-16

Abstract 484 patients aged ≤60 years with de novo (n=385) or secondary (n=99) acute myeloid leukemia (AML) were treated with risk-adapted therapy. Patients with t(8;21), inv(16) or normal karyotype and good response (GR) to induction I (≤5% blasts in day 15 BM) were considered standard risk (SR), all others as high risk (HR). Induction I consisted of standard dose araC, idarubicine and etoposide (IVA-I). Patients with GR to IVA-I continued with IVA-II on day 21 as double induction. Patients with bad response (&amp;gt;5% blasts in day 15 BM) received FlAG/Ida as second cycle. Double induction was followed by early consolidation with intermediate dose araC. As late consolidation, SR patients with normal karyotype and an HLA-matched sibling received matched related donor (MRD) transplantation. The remaining SR patients were randomized between high dose araC (HD-araC; 12 x 3g/m2)/daunorubicine or an autologous peripheral blood stem cell transplantation (autoPBCSCT) with PBSC mobilized after early consolidation. 90% of SR patients (n=250) achieved complete remission (CR) in contrast to 58% of HR patients (n=234) (overall CR rate 75%). 4% of the SR patients died during induction. At 70 months, overall survival (OS) was significantly better for SR (51%) than for HR patients (28%). Within the SR group, relapse free survival at 50 months was not different between patients with normal karyotype (n=165), inv(16) (n=30) or t(8;21) (n=30). 55 SR patients were randomized to receive HD-araC and 59 to autoPBSCT. Distribution of the main characteristics were well balanced (age, performance status, cytogenetics, FLT3 mutation status, de novo or secondary AML). At 63 months, OS was 62% for patients treated with autoPBSCT and 59% for those receiving HD-araC (p=0.91, intent-to-treat). Median duration of neutropenia (&amp;lt;500/μl) was 8 days for autoPBSCT and 20 days for HD-araC (p&amp;lt;0.05). This corresponded to a significantly higher rate of septicemia (20% vs. 10%) and pneumonia (13% vs. 3%) after HD-araC. The duration of thrombocytopenia was 22 days after HD-araC and 11 days after autoPBSCT (p&amp;lt;0.01). 14 patients died in the HD-araC group (4 in CR and 10 from relapse) and 14 patients died after autoPBSCT (1 in CR and 13 from relapse). In patients with normal karyotype, OS at 67 months for the SR patients receiving MRD transplantation (n = 37) was 61% with no significant difference to HD-araC or autoPBSCT. In conclusion, OS is similar in SR AML patients after autoPBSCT, HD-araC or MRD transplantation. In SR patients without an HLA-identical sibling donor, autoPBSCT instead of HD-araC is recommended for late consolidation due to the reduced treatment related toxicity.
Intradermal RNA-Vaccination of Patients with Metastatic Renal Cell Carcinoma.
2005-11-16

Abstract Background: The use of in vitro transcribed RNA isolated from tumor cells or coding for defined tumor associated antigens was shown to be a very powerfull method to generate antigen specific T cells upon transfection into dendritic cells (DC). More recently it was demonstrated in a mouse model that application of RNA intra dermaly can elicit both, CD8 and CD4 mediated immune responses. Furthermore, in Her-2-neu transgenic mice vaccinations with naked RNA were able to elicit a specific immune response in vivo and delay the development of breast cancer in treated animals. Based on these results we analyzed the clinical and immunological responses in patients with metastatic renal cell carcinoma (RCC) vaccinated with mRNA encoding for tumor associated antigens (TAA) using two different treatment arms. Methods: In vitro transcribed RNA was generated using plasmids coding for the tumor antigens MUC1, CEA, Her-2/neu, telomerase, survivin and MAGE-1. RNA coding for HbsAg and Influenza matrix protein were included as controls. Vaccinations were performed intradermally on day 1,14, 28 and 42 in a first group of patients and on day 0–3, 7–10, 28 and 42 in a second group (intensified arm). Vaccinations were repeated afterwards monthly until tumor progression. One day after each RNA injection patients additionally received 1 injection of GM-CSF (250 μg) sc. The enhancement of T cell precursor was monitored using IFN-g ELISPOT and tetramer staining.. Results: 25 patients were included in this study. In 3 patients regression of metastases was induced and 6 patients had a stabilization of the disease. Specific CD8 and CD4 T cell responses in vivo were detected in the first analyzed patients. The treatment was well tolerated with no severe side effects. In most cases erythema and induration were observed after injections of GM-CSF. One patient developed an allergic exanthema after GM-CSF application. Conclusions: This study demonstrates that intradermal RNA-vaccination can be effective in the treatment of metastatic RCC and induces clinical and immunological responses.
Severe Impairment of Dendritic Cell Allostimulatory Activity by Sendai Virus Vectors Is Overcome by Matrix Protein Gene Deletion.
2005-11-16

Abstract Delivery of antigens to dendritic cells (DCs) plays a pivotal role in the induction of efficient immune responses ranging from immunity to tolerance. The observation that certain viral pathogens are able to infect DCs has led to a concept where applications of recombinant viruses are employed for antigen delivery with the potential benefit of inducing potent antigen specific T-cell responses directed against multiple epitopes. As a prerequisite for such an application, the infection of DCs by recombinant viruses should not interfere with their stimulatory capacity. In this context we could show, that an emerging neagtive-strand RNA viral vector system based on Sendai virus (SeV) is able to efficiently infect monocyte derived human DCs (moDCs). However, after infection with SeV wildtype the responses of DCs to bacterial lipopolysaccharide (LPS) as a powerful mediator of DC maturation characterized by upregulation of CD83, MHC and costimulatory molecules, enhanced secretion of cytokines and the T cell stimulatory capacity were severly impaired. This was not due to a higher production of IL-10 or inhibition of the nuclear localized expression of Rel-B and c-rel, members of the NF-kB transcription factor family, but was rather the result of an increased apoptosis induction of DC. Interestingly, using various recombinant SeV vectors being devoid of single viral genes we were able to identify the SeV matrix (M) protein as a key component in moDC functional impairment after viral infection. Consequently, usage of M-deficient SeV vectors preserved the allostimulatory activity in infected moDCs despite an efficient expression of all other virally encoded genes, thereby identifying M-deficient vectors as a highly potent tool for the genetic manipulation of DCs.
Genetic Polymorphisms in Chemokine Receptor 5, Interleukin 10 and Monocyte Chemoattractant Protein 1 Contribute to Cytomegalovirus Reactivation and Disease after Allogeneic Stem Cell Transplantation.
2005-11-16

Abstract Human Cytomegalovirus (HCMV) infection is still one of the most challenging complications after allogeneic stem cell transplantation (alloSCT). Here, we demonstrate that defined single nucleotide polymorphisms (SNPs) are associated with the susceptibility to HCMV infection after alloSCT. Genotyping of 83 patients (with HCMV reactivation) and 71 controls (no HCMV reactivation) was performed. All donors and SCT recipients were Caucasians. Patients suffered from chronic myeloproliferative disorders (n = 16) and AML /ALL /SAA /NHL /MDS (n = 117). For 21 patients, underlying disease could not be determined retrospectively. Genotyping was performed by PyroseqencingTM on the PSQTM HS96A System (Biotage AB, Uppsala Sweden) or on ABI 7900 ABI PRISM® Sequence Detection System using TaqMan® Assay-By-DesignTM and Assay-On-Demand TM (Applied Biosystems). By Armitage’s-Trend-Test, no significant association for 78 /87 markers and the occurrence of HCMV reactivation or disease (p &amp;gt; 0.05) was found. However, 9 markers showed a significant association with HCMV reactivation and /or disease. These markers are located in CCR5, MCP 1and IL10. If comparing individuals with HCMV-DNAemia versus no reactivation regardless of disease, 5 markers showed a significant association (in MCP 1and IL10). Two markers of MCP 1were significantly associated with HCMV reactivation, but none of the markers analyzed in CCR5 and IL10. Thus, we presume that HCMV reactivation is associated with polymorphisms in MCP1, whereas disease might be related to polymorphisms in CCR5 and IL10. Analyzing the clinical parameters, we found an association between CD34 selection and HCMV reactivation (p = 0.0378) or HCMV disease (p = 0.001). Furthermore, severe acute GvHD was significantly related to HCMV disease (p = 0.0015), chronic GvHD was significantly associated to HCMV disease only if reactivation versus disease was compared (p = 0.034). Treatment with corticosteroids led to HCMV reactivation (p = 0.036). In conclusion, screening of alloSCT patients for the presence of defined polymorphisms may help to predict the individual risk of HCMV reactivation and disease, and thus justifying additional antiviral prophylaxis.