bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Novel Strategy for Combining Immunotherapy with High Dose Chemotherapy and Auto-Transplantation in High Risk Multiple Myeloma.
Authors: Szmania Susann , Cottler-Fox Michele , Rosen Nancy , Viswamitra Sanjaya , Moreno Amberly , Pomtree Mindy , Barlogie Bart , Tricot Guido , van Rhee Frits 
Published: April 2020

Abstract
Although high dose chemotherapy and autologous transplantation (AT) can induce complete remission in approximately 50% of multiple myeloma (MM) patients, the majority eventually relapse. We hypothesize that a combination of AT and vaccination may sustain remissions by inducing specific cytotoxic T cells against chemotherapy resistant plasma cells. Vaccinations are typically given after AT when the immune system is very defective. We tested the novel strategy of priming anti-myeloma T-cells before AT and protecting the primed T-cells from high-dose chemotherapy by apheresis and cryopreservation of the primed cells. After high-dose chemotherapy induced cytoreduction, the primed T-cells were returned early after AT and preferentially expanded by a series of booster vaccinations (figure 1). The vaccines are monocyte derived-dendritic cells (DC) loaded with myeloma lysate (ML) and tracer antigen keyhole limpet hemocyanin (KLH), matured with trimeric CD40-ligand, administered intranodally, and followed by 5 days of low dose subcutaneous IL-2. Four patients with poor prognosis MM (characterized by abnormal metaphase cytogenetics) have been vaccinated and three are evaluable (the 4th patient is 60 days post AT2 and only 14 days post vaccine 8). ML specific immune responses were detected by delayed type hypersensitivity reactions and proliferation in 2/3 and by ELISPOT in 3/3 patients. Strong responses to KLH were observed in 3/3 patients. The pattern of response is a dramatic and rapid increase in ML specific T cell proliferation (figure 2) and IFN-g secretion after LP2 infusion plus booster vaccinations post AT, suggesting that the primed T-cells had been protected by collection and storage during AT. A high pre-existing ML specific antibody response was observed in 2/3 patients and a low antibody response pre-vaccination was clearly boosted in a third patient. The patient receiving the lowest DC dose (50 versus 115 and 139 x106) had a response to ML only detected by IFNg secretion assays, but is in near complete remission, 719 days post AT2, while disease progression was observed in the other two patients 496 and 626 days after AT. Our immune monitoring data suggest that the problem of applying vaccination in heavily treated patients may be overcome by early vaccination, collection and subsequent reinfusion of primed cells followed by repeated booster vaccination. Optimization of the vaccine regimen by increasing the frequency of vaccination or the use of highly immunogenic, well characterized tumor-associated antigens may further improve efficacy. We have an ongoing trial for high risk MM patients utilizing NY-ESO-1 and MAGE-A3 peptide vaccines based on a similar strategy to preserve immune competence.
Imatinib in the Treatment of CML Patients ≥ 65 Years Old in Late Chronic Phase: Results of a Phase II Study of the GIMEMA CML Working Party.
Authors: Bassi Simona , Castagnetti Fausto , Amabile Marilina , Giannini Barbara , Cilloni Daniela , Izzo Barbara , De Vivo Antonio , Luatti Simona , Testoni Nicoletta , Rege-Cambrin Giovanna , Iacobucci Ilaria , Gottardi Enrico , Pane Fabrizio , Salvatore Francesco , Dorigo Alessandra , Saglio Giuseppe , Martinelli Giovanni , Rosti Gianantonio , Baccarani Michele 
Published: April 2020

Abstract
Older age constitutes a poor prognostic variable in CML patients: the negative effect of age on long-term survival has been consistently observed with most effective therapeutic modalities, both drug therapies (busulfan, hydroxyurea and interferon) and allogeneic transplant. In particular, older patients treated with interferon experienced much more adverse events than younger ones. In part their poorer prognosis was probably due to poor treatment compliance and few older patients have been included in prospective studies of interferon. Actually, imatinib is the first-line treatment for CML: its efficacy is very high and it seems to be well tolerated across age groups. We performed a sub-analysis of the effects of age on response and tolerance within the phase II trial of the italian GIMEMA CML Working Party (serial n.CML/002/STI571), which included 284 late chronic phase patients, treated with imatinib (400 mg daily) after interferon failure. Following the WHO, who defines “old” a patient ≥ 65 years, we analyzed the safety and efficacy of imatinib by age group: 226/284 patients (80%) were &lt; 65 years (extr. 17–64) and 58/284 (20%) were ≥ 65 years (extr. 65–85 yrs) old. No significant differences between the two age groups were present at enrollment. Table 1 shows the responses and incidence of hematologic and non-hematologic adverse events (AEs). As expected, older patients experienced more AEs, both hematologic and non-hematologic. In part this significantly poorer tolerance probably justifies the lower response rates, both hematologic and cytogenetic. However, the overall survival was not different between the two age groups: with a median observation time of 33 months, the overall survival curves were superimposed (91%). Moreover, the rate of progression to accelerated/blastic phase was the same (10%). This study demonstrates that imatinib is greatly effective in older CML patients in late chronic phase (53% of MCR and 36% of CCR) and allows a good long-term survival. It is conceivable that treating old CML patients at the onset of the disease, as already shown by Jorge Cortes et al (Cancer2003;98:1105), could translate into response rates as the ones of younger patients.
Responses and adverse events
≥ 65 (n. 58) ≤ 65 (n226) p value Complete Hematologic response 91% 99% 0,001 Major Cytogenetic Response 53% 74% 0,003 Complete Cytogenetic Response 36% 57% 0,001 Grade III Hematologic AEs 72% 50% 0,002 Grade III+IV Hematologic AEs 86% 60% 0,0001 Grade II non Hematologic AEs 83% 68% 0,0001 Grade III + IV non Hematologic AEs 29% 10% 0,0001
High Sensitive Detection of Imatinib-Resistance Associated Mutations Based on ARMS-PCR.
Authors: Gruber Franz X.E. , Soevershaev Mikchail , Olsen Marita , Skogen Bjoern 
Published: April 2020

Abstract
Background: Point mutations in the Abl kinase domain are associated with resistance against imatinib. Strategies to overcome resistance include dose escalation, combination treatment using imatinib with conventional or other developmental agents or, in the future, imatinib may be replaced by other tyrosine kinase inhibitors which work effectively against mutated clones. Mutational profiling of the BCR-ABL kinase domain will in this scenario become an important analysis as a supplement to BCR-ABL quantitation and may provide the rational basis for therapy, once resistance is diagnosed. Our group reported recently a sensitive, single step PCR assay for quantitation of mutated clones based on the ARMS principle.
Aim: We describe an optimized, two step analysis for high sensitivity screening of mutated clones associated with resistance against imatinib targeting all P-Loop mutations, the T315I and M351T.
Methods: In a first conventional PCR-reaction a cDNA-region spanning the BCR-ABL breakpoint is amplified resulting in an isolation of the BCR-ABL kinase domain for further analysis. An aliquot is then analysed in a second PCR step, conducted on the real time PCR Taqman platform. Selectivity for the mutated clone is conferred by the amplification refractoriness of non complementary primer 3′-ends (ARMS principle). By introducing potent nucleotide-mismatches in position n-2, selectivity of the assay could be further increased. Even in the P-Loop region, which is known to be a difficult PCR template, misannealing could be reduced to an acceptable level.
Results: Assays targeting all P-Loop mutations inclusive the T315I and M351T were tested by analysis of patient samples diluted in normal cDNA and non-mutated BCR-ABL and plasmid dilutions, containing the targeted mutation in a background of wildtype plasmids. Generally a 1:1000 dilution of mutated templates could be detected (sensitivity 0.1%). For some mutations even higher sensitivity could be achieved (0.01%). The level of sensitivity is generally higher than reported for other methods described before. The first PCR step can be conducted in parallel to other PCR-based detection strategies. The second step can be run simultanously to Taqman based BCR-ABL quantitation. This makes the described assay the ideal supplement to general mutation detection approaches like D-HPLC or sequencing strategies. Compared to the single step assay we desribed before, the two step approach increases sensitivity with one or two log factors.
Conclusion: The described assay may be suitable for highly sensitive detection of mutated clones in resistant CML patients as a supplement to less sensitive general screening approaches and BCR-ABL quantitation.
Gene Expression Profiling in Quiescent Stem Cells from Normal and Chronic Myeloid Leukaemia Patients.
Authors: Graham Susan M. , Graham Gerry J. , Holyoake Tessa L. 
Published: April 2020

Abstract
Earlier studies have shown that Ph+ quiescent cells exist in chronic myeloid leukaemia (CML) (Blood (1999)94:2056) and we have previously shown that these cells are primitive in that they express the stem cell marker CD34. We have also shown that quiescent CML stem cells are insensitive to the effects of imatinib (IM Novartis Pharma) (Blood (2002) 99:319) and may present a possible source for relapse. This quiescent population therefore represents a potentially significant clinical problem and thus studies aimed at developing methods for eradicating this population are timely. In an effort to identify molecular markers of this population that may allow it to be specifically targeted during therapy, we have set out to investigate the transcriptional differences between quiescent and cycling stem cells. To this end, we have used specific stem cell enrichment and sorting protocols. Leukapheresis products from CML patients (N=5) in chronic phase at diagnosis and mobilised peripheral blood from allogeneic donors (N=3), were selected for CD34+ cells. Hoechst 33342 and Pyronin Y were used to discriminate the quiescent (G0) cells identified as Hoechstlo/Pyroninlo from the cycling cells. In combination with propidium iodide for dead cell exclusion we were able to sort 4–9x105 viable, quiescent stem cells and 4–11x106 cycling cells, which were processed for microarrays. Affymetrix gene chips (U133A) were used for the analysis and the data obtained was analysed using GeneSpring.
Number of Genes Changed in Each Comparison
3 Fold 4 Fold 5 Fold CML G0 V CML Div 37 21 10 Norm G0 V Norm Div 188 92 47 CML G0 V Norm G0 168 85 49 CML Div V Norm Div 49 27 8
Initial analysis indicates that the greatest differences in gene expression are between the normal quiescent cells (G0) and normal dividing cells (Div) and between the normal quiescent cells and CML quiescent cells. A large percentage of the genes differentially expressed between the quiescent and cycling normal cells encode regulators of the cell cycle confirming the success of the sorting strategy for quiescent and cycling cells
A selection of Genes Up-Regulated in Normal Cycling Cells Compared to G0
Gene Fold Up-regulation PCNA 3 CDC2 8 CCNB2 5 CCN1 3.5 CDC20 6 CDC25A 3.5 MCM5 3
In addition, many of the genes identified in our analysis are consistent with other published expression profiles for haemopoietic cells. Curiously, we have identified unanticipated changes in expression of cell cycle genes in the CML quiescent cells, which merit further investigation. We have also identified a number of unexpected genes as being more than 5 fold changed in the quiescent cells compared to dividing cells for both normal and CML samples. Specifically, there is a large cohort of genes preferentially expressed in quiescent normal or CML cells, which encode members of the chemokine family of proteins. Work is ongoing to establish the relevance, if any, of these genes to stem cell quiescence.
The Abl Carboxyl Terminal Region Is Required for the Induction of Acute Lymphoblastic Leukemia by V-Abl but Not for the Induction of Myeloproliferative Disease by BCR/ABL or V-Abl.
Authors: Ji Yongli , Gross Alec W. , Zhang Xiaowu , Ren Ruibao 
Published: April 2020

Abstract
The BCR/ABL oncogene, generated by the t(9;22)(q34;q11) chromosomal translocations known as the Philadelphia chromosome (Ph), is associated with virtually all human chronic myelogenous leukemia (CML), and is also associated with 20% of the adult and 5% of the pediatric cases of acute lymphoblastic leukemia (ALL). v-Abl, the oncogene of the Abelson murine leukemia virus (A-MuLV) originally found to cause pre-B cell leukemia/lymphoma when introduced into newborn mice by intraperitoneal or intravenous injection, was created by a recombination event that fused viral gag sequences to c-abl sequences, deleting the Abl SH3 domain and accumulating point mutations in the process. Using a mouse bone marrow transduction and transplantation model, we have previously shown that expression of BCR/ABL in mouse bone marrow cells rapidly (in approximately 3 weeks) induced a myeloproliferative disease (MPD) resembling human CML, while v-Abl rapidly (in approximately 4 weeks) induced B-ALL along with a minor expansion of v-Abl-infected myeloid cells. It has long been shown that the Abl carboxyl terminal region is required for the lymphoid cell transformation by v-Abl both in vivo and in vitro. We decided to investigate the role of the Abl carboxyl terminal region in BCR/ABL- and v-Abl-leukemogenesis using the mouse bone marrow transduction and transplantation model. We found that the Abl carboxyl terminal region is required for v-Abl-induced B-ALL, as expected. But, interestingly, v-Abl without C-terminus (v-Abl/p160/ΔC) induced a MPD in mice, similar to that induced by BCR/ABL, with a relative long latency (approximately 3 months in average). On the other hand, BCR/ABLΔC still induced MPD efficiently, similar as wild type BCR/ABL. These results demonstrated the Abl carboxyl terminal region is required for the induction of acute lymphoblastic leukemia by v-Abl but not for the induction of MPD by BCR/ABL or v-Abl. The results also suggested that v-Abl has a capacity to induce MPD, albeit much weaker than that of BCR/ABL and much weaker than v-Abl’s capacity to induce B-ALL. The role of the ABL carboxyl terminal region in BCR/ABL-induced B-ALL and the mechanism by which the Abl carboxyl terminal region contributes to the lineage-specific transformation are under investigation. Such research will shed lights into the mechanism of lineage-specific leukemogenesis and help to identify therapeutic targets for lymphoid malignancies.
Reduced Tissue Factor Expression in Rat Prostate Tumor Cells Limits Tumor Development in Vivo.
Authors: Song Hui , Liang Zhong , Wolter William , Suckow Mark A. , Rosen Elliot D. 
Published: April 2020

Abstract
Many tumors express procoagulant activities that contribute to the hemostatic complications associated with cancer. Tumor coagulant activities include tissue factor (TF) and cancer procoagulant (CP), a cysteine protease that activates Factor X. In current models of hemostasis, TF is critical in the initiation of coagulation following vascular injury as TF expressed on cells within the vessel wall complexes with blood-borne coagulation Factor VIIa to activate FX and FIX. In addition, TF transported to a growing thrombus by blood-borne microparticles binds FVIIa and contributes to thrombus propagation and stabilization. Recently, TF as well as other coagulation factors have been implicated in a variety of non-hemostatic processes including inflammation, angiogenesis, vascular development and cancer. To study the role of TF in tumor development we down-regulated TF expression in a prostate tumor cell line developed from the Lobund-Wistar (LW) rat. The LW rat combines histologic features of prostate cancer with the clinical features resembling clinical human disease; androgen-modulated growth, age-dependent spontaneous onset, and metastatic potential. Autochthonous tumors develop spontaneously or can be induced by treatment with MNU and testosterone. In addition, a cloned cell line from a spontaneous LW prostate tumor, PA-3, provides a transplantable tumor model. PA-3 cells were transfected with a series of plasmids expressing hairpin siRNAs designed to interfere with TF expression. The plasmids contained a U6-hairpin siRNA expression cassette, a neoR gene and EGFP. Cloned stable transformants of PA-3 cells expressing a siRNA corresponding to positions 260–278 of rat TF mRNA, PA-3[797], reliably reduced TF expression by 65% compared to control clonal PA-3 transformants, PA-3[776], expressing neoR and EGFP but no siRNA. To test the consequences of reduced TF on tumor development in vivo, 4 independently isolated PA-3[797] cell lines and 4 independently isolated PA-3[776] control lines were injected subcutaneously into LW rats (3 rats per cell line). Ten of the 12 rats injected with control cells developed detectable tumors 4 weeks post transplantation (tumor mass = 2.2g +/− 1.8g). In contrast, only 1 of the 12 rats receiving the low TF expressing PA-3[797] cells developed a small tumor (0.07 gm) suggesting TF expression is required for tumorigenesis.
No Fibrinolytic Benefit of Adjunctive Anticoagulation during Arterial Thrombolysis Induced by tPA Combined with TAFIa Inhibitor in a Rabbit Arterial Thrombosis Model.
Authors: Klement Petr , Liao Peng , Chen Frank , Weitz Jeffrey I. 
Published: April 2020

Abstract
Introduction: In a previous study, we reported that inhibition of activated TAFI (TAFIa) with potato carboxypeptidase inhibitor (PCI) enhanced tPA-induced clot lysis in a rabbit aortic thrombosis model. Building on these results, this study used the same model to determine whether the addition of anticoagulants to PCI would further enhance tPA-induced clot lysis.
Methods: After balloon injury and creation of a critical stenosis, a radiolabelled thrombus of known mass was generated in the distal aorta. Rabbits were then given tPA (0.25 mg/kg bolus followed by 0.25 mg/kg infusion over 60 min) in conjunction with PCI (30 μg/kg bolus followed by 30 μg/kg/hr) or saline (SAL, 1ml/kg bolus followed by 1 ml/kg/hr). Rabbits treated with tPA/PCI were also supplemented with hirudin (HIR, 0.2 mg/kg bolus followed by 0.2 mg/kg/hr) or heparin (H, 50 U/kg bolus followed by 50 U/kg/hr) or pentasaccharide (PENTA,1 mg/kg iv bolus) or SAL. To assess safety, standardized ear incisions were made 15 min after starting tPA and cumulative blood loss was measured over a 30 min period. This time point was chosen because both tPA and anticoagulant levels are at their peak. To assess efficacy, clot lysis was measured at 90 min by subtracting the radioactivity of the residual thrombus from that of the initial thrombus and expresing this value as a percentage of initial radioactivity.
Results: As illustrated in the table, PCI increases tPA-induced clot lysis from 71.3 to 89.2 % with a minimal effect on bleeding. The addition of anticoagulants produced no further increase in clot lysis, but all 3 of the anticoagulants tested increased blood loss.
Conclusions: These preliminary data support the concept that PCI enhances tPA-induced clot lysis to an extent where concomitant administration of anticoagulants provides no additional benefit and compromises safety.
Thrombolytic efficacy/safety profile of various treatments.
Treatment Clot lysis (%) Blood loss (μL) Values are expressed in Mean ± SEM, four rabbits per treatment group SAL 19.7 ± 2.2 15.6 ± 5.3 tPA 71.3 ± 7.1 66.8 ± 14.0 tPA + PCI 89.2 ± 3.3 104.2 ± 42.7 tPA + PCI + H 90.3 ± 5.5 134.0 ± 60.9 tPA + PCI + PENTA 90.6 ± 3.2 215.6 ± 41.5 tPA + PCI + HIR 92.0 ± 5.7 680.3 ± 536.6
Retrospective Comparison of Using or Not Using Donor Lymphocyte Transfusion in the Treatment of Hematological Relapse after Allogeneic Stem Cell Transplantation in 489 Adults with Acute Myeloid Leukemia.
Authors: Schmid Christoph , Labopin Myriam , Finke Juergen , Ehninger Gerhard , Ringden Olle , Niederwieser Dietger , Sakellari Ioanna , Urbano-Ispizua Alvaro , Ruutu Tapani , Gurman Gunham , Clarke Richard E. , Gorin Norbert , Polge Emmanuelle , Fouillard Loic , Frassoni Francesco , Kolb Hans-Jochem ,   Vanderson Rocha for the EMBT ALWP
Published: April 2020

Abstract
Relapsed AML after allogeneic SCT has a poor prognosis. So far, no standard therapy could be defined. Donor lymphocyte transfusion (DLT) has been effective in a minority, however, no data is available to identify patients who will benefit from the procedure. Neither, the outcome of patients treated with or without DLT have been compared. We retrospectively evaluated overall survival (OS) of 489 adults with de novo AML in hematological relapse after SCT, receiving DLT (n=190) or not (n=299). DLT and noDLTgroups were well balanced in terms of patient age (median:37y in both groups), donor age, cytogenetics (good:5vs7%, intermediate:83vs79%, poor:12%vs14%), WBC at diagnosis, donor type (geno-id:71vs72%, MUD:18% both, mismatched:11vs10%), status at transplantation (CR1:38vs41%, CR2:13vs15%, advanced:49vs44%), conditioning, source of stem cells, and time from transplant to relapse (5vs4.5 months). However, DLT patients had a median of 39% BM blasts, as compared to 54% for the noDLT group (p=0.03). Follow-up was 32 and 30 months. Within the DLT group, chemotherapy was additionally given in 130 cases. Nevertheless, only 33% of patients received DLT in CR or aplasia, 67% had measurable disease. AGvHD developed in 41% of patients following DLT. CR and PR were achieved in 31.1% and 4.8% of DLT patients. In a multivariate analysis, younger patient age (&lt;36 years) (HR=1.53,p=0.02) and a longer interval (&gt; 5 months) from transplant to relapse (HR=7.74,p=0.002) were associated with better OS after DLT. When comparing the outcome of patients receiving or not DLT, OS at 2 years was 10±1% for the entire cohort, 18±3% for DLT and 6±1% for noDLT (p&lt;.0001). In a multivariate analysis, use of DLT (HR=2.11,p&lt;0.0001); recipient’s age&lt;36 y (HR=1.69, p&lt;0.001); longer interval (&gt;5 months) from transplant to relapse (HR=2.40, p&lt;0.0001) and number of BM blasts (&lt;48%) at relapse (HR=1.56,p=0.002) were favorable for OS. In this retrospective analysis the results suggest that DLT may be of advantage in the treatment of AML relapse post transplant, at least in younger patients with a longer post transplant remission and relapsing with smaller amounts of blasts in BM. However, patients receiving DLT might represent a positive selection among all relapsed cases, since a considerable number from the noDLT cohort had died too early to proceed to DLT. An intetion-to-treat analysis and further prospective studies should investigate the role of DLT and other approaches, such as second reduced intensity SCT.