bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Phase I Study of Bortezomib in Combination with ICE (BICE) in Patients with Relapsed/Refractory Classical Hodgkin Lymphoma
Authors: Fanale Michelle A. , Fayad Luis E. , Pro Barbara , Samaniego Felipe , Zachariah Gracy , Nunez Cesar A. , Ji Yuan , Younes Anas 
Published: April 2020

Abstract
Background: The proteasome inhibitor bortezomib induces cell cycle arrest and apoptosis of Hodgkin lymphoma (HL) cells. We have demonstrated that bortezomib’s single agent efficacy was limited in heavily pretreated HL patients. However, preclinical work has shown that bortezomib may synergize with chemotherapy. Thus, we evaluated the combination of bortezomib plus ICE chemotherapy in patients with relapsed/refractory HL.
Methods: Eligibility:
Relapsed/refractory classical HL, Only 1 prior chemotherapy regimen, Prior treatment with anthracycline-containing regimen, No prior ASCT.
Regimen: ICE (Moskowitz, C et al 2001). Bortezomib on D1 and D4 at dose levels of 0 = 1 mg/m2, 1 = 1.3 mg/m2, 2 = 1.5 mg/m2. Neulasta given on D5. Cycles were repeated on D14 if platelet count is ≥ 100,000/mm3 and ANC is ≥ 1,000/mm3. After 3 cycles CTs and PET/CT were performed to evaluate response. Hematologic DLT was defined as grade 4 thrombocytopenia or neutropenia lasting greater than 2 weeks.
Results: 13 patients enrolled (6 primary refractory and 7 relapsed HL). Median age 32 (range 21–39). 3 patients were treated at dose levels 0 and 1 and 7 patients were treated at dose level 2. 12 patients were evaluable for toxicity and response. The ORR was 75% with 25% CR (1 primary refractory, 2 relapsed) and 50% PR (1 primary refractory, 5 relapsed). PET/CT scans showed negativity in all patients who responded to treatment. 8/9 patients who responded to treatment underwent ASCT (1 patient declined ASCT). All patients who underwent ASCT had successful pheresis and engraftment. At median follow-up of 12 months all 8 patients treated with ASCT are free of relapse. Treatment was well tolerated with no DLTs noted. Reversible grade 4 neutropenia and thrombocytopenia occurred respectively in 33% and 50% of the patients. Median day for retreatment for cycle 2 was D20 and for cycle 3 was D21. There have been no toxicities of peripheral neuropathy, febrile neutropenia, or infection.
Conclusions: Our data suggests favorable tolerability and efficacy of bortezomib plus ICE in patients with relapsed/refractory Hodgkin lymphoma. We are planning to confirm these promising results in a randomized phase II trial.
Phase I Trial of Bortezomib in Combination with Rituximab-HyperCVAD/Methotrexate and Cytarabine for Untreated Mantle Cell Lymphoma
Authors: Romaguera Jorge , Fayad Luis , McLaughlin Peter , Pro Barbara , Rodriguez Maria Alma , Wang Michael , Weaver Pamela , Hartig Kimberly , Kwak Larry , Feldman Tatyana , Smith Judy , Ford Peggy , Goldberg Stuart , Pecora Andrew , Goy Andre 
Published: April 2020

Abstract
Background: Though mantle cell lymphoma (MCL) outcome has improved over the last two decades - especially with dose-intense or high-dose therapy - MCL patients still relapse particularly in &gt;65yo. Bortezomib has shown single agent activity of 33% in relapsed MCL and has been safely combined with conventional as well as intensified doxorubicin-containing regimens, but not with high doses of the methotrexate/cytarabine combination, where there is concern about lung toxicity.
Materials and methods: MCL pts from ages 18 through 79, with adequate organ function unless due to lymphoma, and without HIV-1 infection or any significant medical/mental condition were treated under IRB-approved study using R-HCVAD alternating with R-M/A (JCO October 1, 2005) with doxorubicin given by bolus. Bortezomib was added to R-HCVAD as a fixed dose of 1.3 mg/m2 IV given at the end of the first infusion of cyclophosphamide on day 2 and after doxorubicin bolus on day 5. Bortezomib was added to R-M/A after rituximab on day 1 and on day 6, in increasing doses of 0.7, 1, and 1.3 mg/m2 in cohorts of 3 patients. As an additional precaution, the first cohort of patients received a −1 dose level of the cytarabine adjusted by age. Dose limiting toxicity was defined initially as a drop of 10% from baseline in diffusing lung capacity (DLCO) and later increased to 25% (grade 1 toxicity according to National Cancer Institute criteria) after the first cycle of R-MA with bortezomib.
Results: Since April 2007, sixteen patients have been entered on the trial, ages 40 to 70 years (median 58). Currently 2/3 patients have been entered in the last cohort. There has been no DLT. After 59 cycles, toxicities according to NCI criteria were as follows:
BR-HCVAD Phase I Toxicity in 59 cycles
Variable Grade1 (%) Grade2 (%) Grade3 (%) Grade4 (%) *First R-M/A cycle of each patient. 15/16 patients evaluable. Thrombocytopenia 7 (12) 3 (5) 8 (14) 26 (44) Neutropenia 3 (5) 5 (8) 4 (7) 28 (47) Anemia 11 (19) 23 (39) 11 (19) 2 (3) Neutropenic fever 0 0 5 (8) 1 (2) Dyspnea 0 0 0 0 DLCO* 2 (3) 0 0 0 Neurological 5 (8) Sensory 1 (2) 0 0
As expected, responses are excellent with 11 complete responders and 4 partial responders out of 15 evaluable patients, of whom 3 are still on treatment.
Conclusion: Bortezomib can be safely combined with R-M/A. Accrual is ongoing and more patients and dose levels will be reported on at the meeting.
Factor V as Anticoagulant Cofactor for Activated Protein C in Factor Va Inactivation
Authors: Cramer Thomas J , Griffin John H. , Gale Andrew J. 
Published: April 2020

Abstract
Factor V (FV) is a cofactor that promotes inactivation of activated factor VIII (FVIIIa) by the activated protein C and protein S complex (APC/protein S). Cleavage in FV at Arg506 is required for proteolytic inactivation of FVa, but also for the anticoagulant function of FV as cofactor for APC in the inactivation of FVIIIa. This is demonstrated by the well known FVLeiden mutant with Arg506 mutated to glutamine (Q506), causing APC resistance due to both impaired sensitivity of Q506FVa to APC and reduced cofactor activity of Q506FV for APC inactivation of FVIIIa. However, FVIIIa loses activity rapidly due to dissociation of the A2 domain, and this may be the primary mechanism of FVIIIa inactivation. Thus, we question whether the APC-mediated inactivation of FVIIIa is relevant to the FVLeiden thrombophilic phenotype. Rather, we hypothesized that FV can function as an anticoagulant cofactor for the APC/protein S complex in the inactivation of activated FV (FVa). To test this hypothesis, we designed a coagulation assay initiated by tissue factor that was sensitive to FV but was insensitive to FVIII. FV was titrated into FV deficient plasma and clotting times were measured in absence and presence of APC to determine an APC sensitivity ratio (APCsr). An increase in the APCsr was observed as the level of FV was increased, suggesting an anticoagulant function of FV. Similar titrations were done with Q506FV, showing no increase in clotting time when APC was present and an APCsr of 1.0 in the presence of Q506 FV. Control experiments confirmed that this clotting assay was insensitive to the presence or absence of FVIII; thus, these assays were reflecting FVa inactivation. The potential anticoagulant effect of FV as cofactor for APC in FVa inactivation was further investigated by monitoring proteolysis of purified FVa by APC over time using SDS PAGE. Recombinant purified FVa was labeled with a fluorescent dye, and then subjected to proteolysis by APC/protein S in the absence or presence of FV in a time course. The resulting FVa fragments seen on SDS gels reflected the known cleavages at Arg306 and Arg506, and the FVa cleavage products were quantified by digital fluorescent scanning of the gel. FV stimulated a small but statistically insignificant increase in the rate of FVa cleavage by APC/protein S. Thus, in our experimental conditions, we found a significant anticoagulant effect of FV in clotting assays that were sensitive to FV but not sensitive to FVIII whereas in purified reaction mixtures there was not a significant enhancement by FV of APC proteolysis of FVa. These data contrasting FV’s apparent APC-cofactor activities between plasma and purified reaction mixtures lead us to speculate that other factors or mechanisms present in plasma also contribute to the anticoagulant function of APC in a FV dependent manner.
Relapse of Children with Aplastic Anemia after Immunosuppressive Therapy
Authors: Kamio Takuya , Ito Etsuro , Ohara Akira , Kosaka Yoshiyuki , Tsuchida Masahiro , Mugishima Hideo , Yabe Hiromasa , Morimoto Akira , Ohga Shouichi , Yagasaki Hiroshi , Bessho Fumio , Nakahata Tatsutoshi , Kojima Seiji 
Published: April 2020

Abstract
Although the therapeutic outcome for acquired aplastic anemia (AA) has improved markedly with the introduction of immunosuppressive therapy (IST) with antithymocyte globulin (ATG) and cyclosporine A (CyA), a significant portion of patients treated with IST subsequently relapse and require second-line therapy. However, detailed analysis of the relapse cases has not been reported. In this study, we assessed the relapse rate, response to second-line treatment after relapse and prognosis. From November 1992 to July 2007, 473 newly diagnosed AA children (213 female, 260 male) entered two consecutive prospective studies (AA-92 and AA-97). The median age was 8 years, ranging from 1 to 18 years. Patients with very severe (n = 210), severe (n = 149) and non-severe disease (n = 114) received initial IST consisting of ATG and/or CyA. At six months after the initial IST, 280 patients (59.2%) achieved CR (n = 94) or PR (n=186). Relapse is defined by conversion to no response (NR) from partial or complete response (PR/CR). Among the 280 patients who responded to IST, 49 (17.5%) relapsed. The cumulative incidence of relapse was 19.6% at 10 years and the median time to relapse was 18 months, ranging 7 to 138 months. Among the 49 patients who relapsed, 22 received a second round of IST with ATG and CyA. Fourteen patients (63.6%) responded to the second round. However, three relapsed, and two developed paroxysmal nocturnal hemoglobinuria. Eight patients received CyA after relapse. Four patients responded to CyA, but two relapsed subsequently. Hematopoietic stem cell transplantation (HSCT) was attempted in 26 patients who relapsed after initial responses. Before HSCT, nine received a second course of IST with ATG and CyA and two received CyA. Fifteen patients progressed directly to HSCT. Bone marrow transplantation (BMT) from an HLA-matched sibling (n =7), HLA-matched family member (n = 1), or HLA-mismatched family member (n = 2) was performed in 10 patients, and all are presently alive. BMT from an unrelated donor was attempted in 13 patients, and four died of complications related to BMT. Cord blood transplantation from an unrelated donor was attempted in two patients and two patients are alive. Of the 26 patients who received HSCT, 21 are alive and well, 22 to 137 months (median 72 months) following transplantation. There were seven deaths out of total 49 who initially relapsed. The causes of death were HSCT-related complications (n = 5), MDS/acute myelogenous leukemia (n = 1), bacteremia (n = 1). Eight of 9 patients who received HSCT following a second round of IST are alive and well. The present study suggests that a second round of IST should be offered to the patients who relapse, and HSCT should be considered for the patients who fail to respond to the second round of IST.
Comprehensive Ultra High Resolution Analysis of Copy Number and Allele Status Including Novel Microdeletions Spanning NF1 in Acute Myelogenous Leukemia
Authors: Malek Sami , Ouillette Peter , Wright Whitney , Dressel Amanda , Karp Judy , Bockenstedt Paula , Al-Zoubi Ammar , Talpaz Moshe , Kujawski Lisa , Shedden Kerby , Shakhan Sajid , Li Cheng , Erba Harry , Parkin Brian 
Published: April 2020

Abstract
Karyotypic abnormalities are of dominant importance in AML risk prognostication and therapy selection. A comprehensive description of subchromosomal genomic copy number changes and allele status together with gene mutation analysis and identification of chromosomal translocations is needed to fully harvest the prognostic and biological power of genomic changes in AML.
We have analyzed DNA from 96 AML-derived pure blast populations (purified using column-based multi parameter negative selection followed by multi-gated FACS sorting) compared with paired buccal DNA using the Affymetrix 6.0 SNP platform. To support data analysis and display, we have developed the software tools PLUT and LOH tool version 2 and have refined dChipSNP. Data were supplemented with blast karyotypes and mutation status of Flt3, NPM1, p53, N-ras and K-ras.
Results: AML cases carried between zero and thirty-four subchromosomal losses and gains. Approximately 23% of all cases had ≥3 subchromosomal lesions and approximately 50% of all cases had no such lesions. Of the 22 cases with complex and hypercomplex genomic changes by SNP profiling, ~50% had mutations in p53 exons 5–9 and ~50% were p53 wild-type by sequence analysis. Monoallelic deletion of p53 as part of various deletions at 17p was found in 7 of 96 (7%) cases. Importantly, 7/96=7% additional cases demonstrated UPD at 17p which spanned the p53 locus. Of all 14/96=15% of cases with LOH at 17p (with and without copy loss) 9/14=64% carried p53 mutations and 11/14=79% had complex karyotypes. Of the AML cases with NPM1 mutations (19 of 96 cases or 20%), 75% carried no detectable subchromosomal lesions, while 25% carried between 1 and 4 such lesions.
Previously unidentified microdeletions resulting in monoallelic NF1 loss were identified on 17p in a total of 10 of 96 (10%) cases, thus suggesting activation of the Ras pathway independent of Ras mutations (all affected cases had wild-type N-ras and K-ras) in a substantial subset of AML cases. Mutational analysis of all coding exons of NF1 on the retained allele in the affected cases is ongoing.
Additional recurrent microdeletions were identified on chromosomes 3p and 3q as well as 12p, the latter encompassing the genes ETV6 and p27, as previously reported.
Finally, analysis of the prognostically and therapeutically important deletions 5q and 7q identified large deletions without recurrent microdeletions. Regions of minimal loss on 5q have been delineated.
The T(4;11) Fusion Protein MLL/AF4 Regulates TERT Expression
Authors: Gessner Andreas , Thomas Maria , Castro Patricia Garrido , Heidenreich Olaf , Greil Johann 
Published: April 2020

Abstract
About 50% of all infants suffering from acute lymphoblastic leukaemia (ALL) show the translocation t(4;11)(q21;q23) which creates the fusion genes MLL/AF4 and AF4/MLL. This reciprocal translocation identifies a therapy resistant form of leukaemia with a poor prognosis. In order to gain a better insight into the molecular mechanisms of t(4;11) leukaemias we used the two ALL cell lines SEM and RS4;11, both harbouring the t(4;11) translocation, however with different fusion sites. Specific small interfering RNAs (siRNAs) against the two fusion site variants of the MLL/AF4 fusion transcript were designed and transfected into the respective cells via electroporation, using very mild conditions. Serial electroporations at two-day intervals resulted in sustained depletion of the MLL/AF4 transcript up to 70–80%, and depending on the experimental setup, cells were analysed after two or three electroporations. Knock-down of MLL/AF4 resulted in strong inhibition of proliferation and clonogenicity in the two t(4;11)-positive cell lines SEM and RS4;11, along with induction of apoptosis. MLL/AF4 depletion resulted in a 65% decrease in telomerase activity in both SEM and RS4;11 cells, with telomerase reverse transcriptase (TERT) expression being reduced twofold on both transcript and protein levels. Notably, TERT reduction was even stronger (90% depletion) when apoptosis caused by MLL/AF4 knock-down was suppressed with the caspase inhibitor zVAD. In contrast, levels of the RNA component of the telomerase, TERC, were not affected by MLL/AF4 knock-down. Additionally, MLL/AF4 knock-down was associated with reduced expression of several members of the HOXA gene cluster, HOXA6 (65% reduction), HOXA7 (85% reduction), HOXA9 (60% reduction) and HOXA10 (75% reduction). Interestingly, siRNA-mediated knock-down of the MLL/AF4 target gene HOXA7 also induced apoptosis and resulted in a 70% decrease of TERT levels in two t(4;11) positive cell lines without affecting MLL/AF4. Chromatin immunoprecipitation assays revealed HOXA7 binding to the promoter of TERT. Therefore, MLL/AF4 regulates TERT expression, at least in part, via HOXA7. These data suggest that t(4;11) positive cells with substantially lower TERT expression undergo apoptosis, and that TERT may play an antiapoptotic role in t(4;11) positive ALL. Furthermore, these studies identify TERT as a putative new therapeutic target in this therapy-resistant infant leukaemia.
The in Vivo Response of Patients Suffering from Severe Congenital Neutropenia (CN) to G-CSF Is Due to “Emergency” Granulopoiesis Induced by C/EBPβ
Authors: Dan Lan , Thakur Basant Kumar , Skokowa Julia , Welte Karl 
Published: April 2020

Abstract
C/EBP transcription factors are crucial for the regulation of granulopoiesis in vitro and in vivo. C/EBPa is considered to be the master regulator of “steady state” granulopoiesis via upregulation of several myeloid genes (e.g. ELA2, CSFR3, etc.). The absence of C/ EBPa results in a complete loss of neutrophils. We were able to show that in patients with severe congenital neutropenia (CN) harbouring HAX-1 or ELA2 mutations C/EBPa is severely abrogated secondary to defective expression of LEF-1 (Skokowa J, et al. Nat Med.12, 1191–7 (2006)). Therefore, we were interested, whether other transcription factors are capable of substituting C/EBPa, since these patients respond to G-CSF with slight increase in neutrophils from less than 200/ul to above 1500/ul depending on the dose of G-CSF. C/EBPβ has recently been shown to be required for cytokine induced “emergency” granulopoiesis (Hirai H, et al. Nat Immunol.7, 732-9 (2006)). Therefore, we investigated the expression pattern of C/EBPβ during G-CSF treatment of CN patients. Indeed, C/EBPβ mRNA was upregulated 2.8-fold in CD33+ myeloid cells from CN patients by G-CSF treatment, as compared to healthy individuals. It was associated with upregulation of G-CSFR mRNA and –protein expression as well as ligand binding to G-CSFR in myeloid cells, and elevated levels of biologically active G-CSF in serum from CN patients. To confirm C/EBPβ-dependent activation of G-CSFR and G-CSF gene expression, we performed reporter gene assays in CD34+ bone marrow hematopoietic progenitor cells from two CN patients co-transfected with C/EBPβ and reporter constructs containing upstream regulatory regions of G-CSF or G-CSFR genes, −1470bp and −670bp, respectively. Indeed, C/EBPβ activated G-CSFR promoter 3.2-fold and G-CSF promoter 5.4-fold. These data demonstrate that in CN patients, G-CSF induces C/EBPa independent granulopoiesis and that C/EBPβ is required for the response to G-CSF treatment in these patients. C/EBPβ leads in response to G-CSF to induction of differentiation of neutrophil precursors to mature neutrophils in vivo. Our hypothesis is that in CN the steady state granulopoiesis is abrogated whereas the emergency granulopoiesis still leads to sufficient numbers of neutrophils.
Loss of Id4 Accelerates CLL Progression in TCL1 Mice
Authors: Chen Shih-Shih , Johnson Amy J. , Claus Rainer , Sablitzky Fred , Plass Christoph , Byrd John C. 
Published: April 2020

Abstract
The inhibitor of DNA binding protein 4 (ID4) is a member of dominant negative basic helix loop helix (bHLH) transcription factor family that has a HLH domain but lacking DNA-binding domain. Methylation of ID4 in B-cell lineage malignancies including CLL and germinal center non-Hodgkin lymphomas (GC-NHLs) has been described. To date, the impact of Id4 loss in CLL has not been demonstrated. We utilized the TCL1 transgenic mouse model of human CLL to study the importance of loss of ID4 in disease progression. We demonstrated that ID4 was silenced by methylation only at the time mice had symptomatic leukemia. We therefore sought to investigate if earlier loss of Id4 would accelerate CLL progression. We generated mice with haploid loss of Id4 in TCL1 transgenic background by breeding homozygous TCL1 transgenic mice and mice with heterozygous Id4 mutant gene. By comparing to the control littermates with heterozygous TCL1 oncogene, mice with the loss of Id4 occur to have significantly accelerated CLL disease progression with shorter overall survival (12 versus 16 months, p-value&lt;0.0001). The pathological analysis on Id4 mutant mice demonstrated that the CLL symptoms such as the elevated white blood cell counts, enlarged spleen and lymphoid tissues. We then performed the microarray studies on 1 month old TCL1 mice with control or mutant Id4 to identify ID4-dependent transcriptional changes in primary B-cells. Among the identified targets, genes involved in cell proliferation and anti-apoptosis were then verified. Supportively, B-cells from Id4 mutant TCL1 mice have significantly (p-value&lt;0.001) diminished apoptosis in response to dexamethasone treatment. Additionally, both significant in vitro (p-value&lt;0.001) and in vivo (p-value=0.025) increased proliferation of Id4 mutant TCL1 B-cells after the stimulation by CpG685NO168 was demonstrated. These data provide support that Id4 acts as tumor suppressor in transformed B-lymphocytes and is silenced through the process of methylation. The Effort to identify binding partners of Id4 in CLL cells and targeting its re-expression is warranted.