bluuebunny/crossref_metadata_2025_split huggingface dataset:
Total Therapy 2 (No Thalidomide Arm, TT2-) Is Superior to Total Therapy 1 (TT1) for Newly Diagnosed Multiple Myeloma (MM): Doubling 4-Yr Survival among Patients with Cytogenetic Abnormalities (CA) Due to Consolidation Chemotherapy (CCT) and DEX Maintenance.
Authors: Barlogie Bart , Tricot Guido , Rasmussen Erik , Anaissie Elias , van Rhee Frits , Zangari Maurizio , Hollmig Klaus , Milner Teresa , Epstein Joshua , Shaughnessy John , Crowley John 
Published: April 2020

Abstract
Background: TT2 introduced T into the frontline therapy for MM in a randomized phase III trial design (ASCO 2005). In comparison to TT1, TT2 applied: (1) more intensive chemotherapy for induction prior to and introduced CCT after melphalan 200mg/sqm-based tandem autotransplants, designed to improve survival in high-risk patients with CA; (2) DEX pulsing was added during the 1st year of interferon (IFN) maintenance therapy. We now report on the outcome of patients treated on the “no thalidomide” arm of TT2 (TT2-) in comparison to TT1, in order to evaluate the effect of dose-intensification during induction and post-transplant therapy.
Patients and Methods: 231 patients were enrolled in TT1 (median follow-up, 11yr) and 345 in TT2- (median follow-up, 3.5yr). Completion rates of 1st/2nd transplants were 195/165 (84%/71%) with TT1 and 292/235 (85%/68%) with TT2-. In TT2-, 64% started CCT and 36% received DEX consolidation (when platelets failed to recover or no benefit was documented from induction DCEP). TT2- and TT1 were compared in terms of pre-transplant-1 and final CR rates (intent-to treat), EFS/OS from treatment start, 1st transplant and from last (2nd or 1st) transplant. EFS and OS were examined in the context of baseline prognostic variables including CA.
Results: Compared to TT1, TT2- induced similar pre-Tx1 CR rates (11% vs 12%, p=0.8) and final CR rates (41.3% vs 40.7%, p=0.9). The median onset of CR was 8.9mo for TT2 vs 8.4mo for TT1 (p=0.4). 5-year EFS/OS were 45%/63% with TT2- vs 28%/57% with TT1 (p&lt;0.001/p=0.06). 4-yr post-transplant-1 EFS/OS were 48%/65% with TT2- vs 28%/56% with TT1 (p&lt;0.001/p=0.02); 4-yr post-transplant-2 EFS/OS were 46%/64% with TT2- and 31%/50% with TT1 (both p=0.01). Pre-study CA, high LDH (&gt;=190U/L), and low Hb (&lt;10g/dL) were independently, significantly associated with poorer post-transplant-2 EFS and OS (p&lt;0.05); independent of these factors, TT2- improved post-transplant-2 EFS and OS compared to TT1 (p&lt;0.001/p=0.033). In the presence of CA, TT2- with CCT vs TT2- with DEX improved 4-yr OS (measured from a 6-mo landmark post-transplant-2) from 37% (similar to 39% with TT1 [no DEX or CCT]) to 78%, which is comparable to the 65% for TT1 without CA.
Conclusion: In this historical comparison of TT1 with TT2-, the more intensive induction chemotherapy with TT2- did not improve CR. However, post-transplant CCT benefited the 1/3 of patients with CA, doubling 4-yr post-transplant OS in comparison with TT1 in this high-risk subgroup. A phase 3 randomized trial addressing the CCT concept is warranted.
Figure Figure
X-Linked Clonality Analysis and Quantitative JAK2V617F Assessment Reveals a Strong Association between Clonality and JAK2V617F in PV but Not in ET/MMM, and Identifies a Subset of JAK2V617F Negative ET and MMM Patients with Clonal Hematopoiesis.
Authors: Levine Ross L. , Belisle Claude , Wadleigh Martha , Lee Stephanie J. , Chagnon Pierre , Gilliland D. Gary , Busque Lambert 
Published: April 2020

Abstract
Polycythemia vera (PV), essential thrombocythemia (ET), and myeloid metaplasia with myelofibrosis (MMM) are categorized as a set of phenotypically related myeloproliferative disorders (MPD). Recent work has identified a somatic mutation in the JAK2 tyrosine kinase (JAK2V617F) in the majority of PV and in a subset of ET and MMM. As most, but not all, patients with MPD have clonal granulocytes, we sought to determine if there was a relationship between granulocyte clonality and JAK2V617F mutational status using a quantitative clonality assay for the human androgen receptor gene (HUMARA) and a quantitative real-time PCR assay for JAK2V617F. 168 of 190 female MPD patients were informative for the HUMARA polymorphism, and 121 patients (72%) had allele skewing consistent with clonal granulopoiesis including 80% of MMM, 75% of PV, and 67% of ET. The quantitative real-time PCR assay for JAK2V617F detected the JAK2V617F allele in 98.8% of PV, 69% of ET, and 33% of MMM, and was more sensitive than direct sequencing. A significant correlation between the degree of allele skewing and quantitative JAK2V617F allelic ratio was observed in PV (p&lt;0.0001) but not in ET (p= 0.2) or MMM (p=0.771). In addition, a subset of patients with ET and MMM with clonal granulocytes had JAK2V617F present in some, but not all, clonal granulocytes. These data suggest acquisition of the JAK2V617F mutation may be sufficient for the development of PV, but additional genetic events are necessary to cooperate with JAK2V617F in ET and MMM. Most importantly, we identified a subset of patients with ET (23%) and MMM (17%), but not PV, with clonal granulocytes who were JAK2V617F negative. A high-throughput screen for novel activating mutations in signal transduction pathways in JAK2V617F negative ET and MMM is warranted.
Complete Response (CR) with Total Therapy 2 (TT2) for Multiple Myeloma (MM): Low Frequency in Case of Prior MGUS without Adverse Consequences for Survival - Yet Critical for Disease Control in Presumed “De Novo” MM.
Authors: Fassas Athanasios , Rasmussen Erik , Shaughnessy John , Tricot Guido , Zangari Maurizio , van Rhee Frits , Anaissie Elias , Hollmig Klaus , Fox Michele , Epstein Joshua , Crowley John , Barlogie Bart 
Published: April 2020

Abstract
Background: It is now widely accepted that stringently defined CR is a prerequisite for durable EFS and OS also in MM.
Patients and Methods: The results of TT2 (4 cycles of intensive induction prior to and consolidation chemotherapy after tandem autotransplant) have recently been reported (ASCO 2005), demonstrating a significantly higher CR rate and longer EFS but not OS among those randomized to thalidomide. Baseline laboratory variables and the clinical history off 668 patients enrolled in TT2 were examined for their potential impact on CR and, in turn CR’s impact on EFS and OS. The Kaplan-Meier method was used to estimate EFS and OS. Cox regression was used to evaluate multivariate factors of CR, EFS, and OS; the prognostic value of CR and 1st and 2nd transplants were modeled as time-dependent covariates in order to adjust for the guarantee time associated with reaching these treatment markers.
Results: Of 668 patients enrolled, 45 had a documented history of prior MGUS (n=22) or smoldering MM (SMM) (n=23); 20 had a prior solitary plasmacytoma (SPC) of bone. 2-yr estimates of CR (negative immunofixation, normal bone marrow) were 47% for the 588 without history of preceding SPC/MGUS/SMM, 55% for SPC and 22% for the MGUS/SMM groups. EFS and OS at 4 yrs were 57% and 71%, independent of a preceding disorder. Multivariate analysis revealed faster onset and higher frequency of CR with only light chain MM (HR, 2.6; p&lt;.001), in the absence of MGUS/SMM (HR, 1.8; p=.049), and with thalidomide (HR, 1.7: p&lt;0.001). Cytogenetic abnormalities (CA), del13q14 and amp1q21, while adversely affecting EFS/OS, did not influence CR. On multivariate analysis for EFS and OS accounting, in addition to baseline variables, also for T, CR, 1st and 2nd transplant, and pre-MM history of MGUS/SMM, we observed that patients attaining a CR had significantly improved EFS and OS independent of key baseline variables (amp1q21, del13q14, CA, LDH, albumin) and treatment markers (T, 1st and 2nd transplants).
Conclusion: We confirm that documented MGUS/SMM pre-MM is associated with lower CR rate without impacting survival negatively, probably implying the re-establishment of the precursor condition. For the remainder, CR was critical for prolonged survival. Gene expression analysis is applied to recognize, in the absence of prior history, those patients likely having evolved from MGUS and thus contribute fundamentally to a distinction of “de novo” MM from “MGUS-evolved MM”. Those results will be presented at the meeting.
Short Telomeres Are Associated with Genetic Instability and the Occurrence of High Risk Genomic Aberrations in Chronic Lymphocytic Leukemia.
Authors: Kröber Alexander , Grabowski Pawel , Bühler Andreas , Seiler Till , Döhner Hartmut , Rosenquist Richard , Roos Göran , Stilgenbauer Stephan 
Published: April 2020

Abstract
Telomere length has been associated with the mutation status of the immunoglobulin variable heavy chain (VH) gene and the clinical course in chronic lymphocytic leukemia (CLL). In an unicentric CLL cohort of 108 patients, we have analyzed the telomere length by quantitative real time PCR, genomic aberrations by FISH with a comprehensive set of DNA probes (11q, 12q, 13q, 14q, 17p), the VH mutation status by DNA sequencing, ZAP-70 expression (clone 2F3.2, Upstate, according to Crespo et al., NEJM 2003) and CD38 expression by flow cytometry, to further study the prognostic impact and associations among these factors. A relative telomere-single-copy-gene ratio (T/S) was calculated for each sample, where low and high T/S values correspond to short and long telomere lengths, respectively. The median T/S value was 0.33 (range 0.06–1.18). There was an inverse correlation between telomere length and the following parameters: 1. VH homology (r=−0.56, p&lt;0.001), 2. CD38 expression (r=−0.44, p&lt;0.001) and 3. ZAP-70 expression (r=−0.25, p=0.01). Cases with T/S values below the median of 0.33 (short telomeres) and cases with T/S values above the median (long telomeres) had similar incidences of genomic aberrations (76 vs. 67%), 13q- (54 vs. 52%) and +12q (9 vs. 9%). In contrast, 13q- as a single aberration was significantly more frequently observed in cases with long telomeres (43 vs. 17%, p=0.006), whereas 11q- (30 vs. 9%, p=0.014), 17p- (24 vs. 0%, p&lt;0.001) and cases with two or more genomic aberrations (26 vs. 6%, p&lt;0.001) were significantly more frequent in cases with short telomeres. Compared to cases with long telomeres the treatment free survival from diagnosis (TFS) and overall survival (OS) in the group with short telomeres were significantly shorter (TFS: 29 vs. 67 months, p=0.002; OS: last observed death at 100 months, survival probability 57% vs. last observed death at 141 months, survival probability 77%, p=0.02). In conclusion, telomere length was inversely correlated with the VH mutation status, CD38 expression and ZAP-70 expression. Short telomeres were associated with genomic instability indicated by a high number of aberrations and the occurrence of 11q- and 17p- in CLL. These observations have biological and prognostic implications in CLL.
Nurselike Cells in Chronic Lymphocytic Leukemia Have Gene Expression Profiles and Functional Characteristics That Are Distinct from Those of Monocytes or Dendritic Cells.
Authors: Messmer Davorka , Endo Tomoyuki , Messmer Bradley T. , James Danelle , Zvaifler Nathan J. , Schmidt-Wolf Ingo G.H. , Tom Kipps J. 
Published: April 2020

Abstract
When CD14+ blood mononuclear cells are cultured with chronic lymphocytic leukemia (CLL) B cells they can differentiate into “nurselike” cells (NLCs), which in turn can support the survival of CLL B cells in vitro and possibly in vivo. While factors that contribute to NLC-mediated support of CLL B cell survival have been identified, it is not clear how this cell type relates to other cell types that also can differentiate from CD14+ blood cells, such as monocyte-derived dendritic cells (DCs). Prior studies have identified some phenotypic differences between NLCs, DCs, and other CD14+ blood mononuclear cells. Thus we hypothesized that these cell types may have different gene expression patterns that may relate to distinctive functional properties. To resolve this we examined the genes expressed by monocytes, NLCs, and immature DCs using Affymetrix U133 microarray analyses. Gene expression profiles were generated from the CD14+ monocyte progenitors, NLC, and DC from three different individuals. The expression profiles of DCs and NLCs differed from the CD14+ progenitors by the expression of many thousands of genes and NLC were distinguished from DCs by the expression of several hundred genes. Some of the genes expressed at higher levels in DCs relative to NLCs encode accessory molecules involved in antigen presentation. Consistent with this, we found that immature DCs were 10 times more effective than NLCs in presenting antigen to allogeneic T cells.
DCs express toll like receptors (TLR) on their cell surface that recognize pathogen components and upon exposure to TLR ligands DCs undergo a maturation process, whereby they upregulate surface molecules and gain increased T cell stimulatory capacity. The expression of TLR2, 4, and 9 was analyzed in DCs and NLCs by RT PCR. Both DCs and NLCs were found to express mRNA for TLR2 and 4, but only NLCs expressed TLR9. In concordance with this, NLCs but not DCs unregulated MHC-II after exposure to nonmethylated CpG oligodeoxinucleotides (ODN), a TLR9 agonist, whereas both cell types upregulated MHC-II after exposure to lipopolysaccharide.
Given the propensity of CD14+ cells to differentiate down a NLC pathway when co-cultured with leukemic B cells in vitro, we speculate that differentiation of CD14+ cells into NLCs may be favored in patients with CLL over differentiation into DCs. Given the relative differences in APC function of these two cell types, this may account in part for the acquired immune deficiency often observed in patients with this disease. On the other hand a stimulus like CpG ODN, might increase the ability of NLCs to activate T cells and decrease their ability to support CLL B cells survival.
Characterization of Distinct Molecular Signatures in Myeloproliferative Diseases with the JAK2V617F Mutation and Wild Type JAK2.
Authors: Ebert Benjamin L. , Levine Ross L. , Wadleigh Martha , Brunet Jean-Philippe , Pretz Jennifer L. , Buque Lambert , Lee Stephanie J. , Gilliland D. Gary , Golub Todd R. 
Published: April 2020

Abstract
The recently discovered JAK2V617F mutation provides a critical insight into the molecular pathogenesis of polycythemia vera and other myeloproliferative diseases (MPD). However, the mutations present in patients with wild type JAK2 have not been discovered, and the precise molecular consequences of JAK2 mutation have not been elucidated. We employed gene expression profiling to characterize the molecular phenotype of cells with the JAK2V617F mutation, to identify a distinct signature in cells without JAK2 mutations, and to refine a molecular taxonomy of MPDs. Using purified neutrophils from 70 patients with myeloproliferative diseases and 11 unaffected individuals, we performed gene expression profiling using oligonucleotide microarrays, sequencing of the JAK2 gene, quantitative genotyping by mass spectrometry and allele-specific quantitative PCR, and X-inactivation clonality assays. To reduce the confounding influence of normal neutrophils that are admixed with cells bearing disease-causing mutations, we examined the gene expression profiles of samples in which greater than 80% of JAK2 alleles bear the V617F mutation. PRV1, a previously identified marker of polycythemia vera, was powerfully overexpressed in neutrophils with a homozygous JAK2 mutation. In addition, cells with the JAK2 mutation had increased expression of a set of kinases, including JAK2, and decreased expression of a set of phosphatases. Cells that rely on JAK2 activation for clonal dominance may therefore derive a selective advantage from increased expression of the JAK2 gene. We next examined samples that have high clonality, and therefore relatively few normal neutrophils, but do not have a mutation in the JAK2 gene. These samples have a markedly different gene expression profile and overexpress a distinct set of kinases. The kinases overexpressed in cells with wild type JAK2 are candidates for further mutational analysis and are potential therapeutic targets. Utilizing these signatures and unsupervised analytical algorithms, the samples cluster according to their mutational status and the percentage of normal neutrophils in the sample. Our data demonstrate that the gene expression profiles of MPD samples are not uniform regardless of JAK2 genotype, implying that samples with and without JAK2 mutations have not activated the same pathway via alternative mechanisms. Moreover, we have identified a common signature in samples without JAK2 mutations that meets significance, indicating that one or a small number of mutations may play a critical role in these cells. Genotype and gene expression analyses are defining a molecular classification of myeloproliferative diseases with subtypes that have distinct therapeutic targets.
Chronic Lymphocytic Leukemia Patients with a V1-69 Gene Rearrangement Do Not Have Inferior Survival with Respect to Patients That Express Other Unmutated VH Genes.
Authors: Panovska Irina , Ivanovski Martin , Pavkovic Marica , Siljanovski Nikola , Hadzi-Pecova Liljana , Stojanovic Aleksandar , Cevreska Lidija , Efremov Dimitar G. 
Published: April 2020

Abstract
The mutational status of the immunoglobulin variable heavy chain genes (VH) is a major prognostic indicator in patients with CLL. More recently, additional prognostic categories have been identified by recognizing disease subsets which utilize unique VH genes. Examples include the V3-21 and V3-72 genes which are invariably associated with progressive and stable disease, respectively. The V1–69 gene is the most frequently rearranged VH gene in CLL and is almost always unmutated. We therefore investigated whether CLL patients that express this VH gene differ in clinical course and outcome with respect to CLL patients expressing other unmutated VH genes. The study group consisted of 106 consecutive CLLs that had been diagnosed at our institution prior to 2000. The 25 V1–69 cases (23.6%) were identified by allele-specific VH gene fingerprinting and all were found to be unmutated by nucleotide sequencing. Forty of the remaining cases were randomly selected for VH gene nucleotide sequence analysis; 19 cases expressed another unmutated VH gene, 14 cases expressed a mutated VH gene and the VH gene sequence could not be determined in 7 cases. The V1–69 cases did not differ significantly from cases with other unmutated VH genes with respect to age (median age 60 years for both subgroups), gender (male: female=21:4 for V1–69 cases and 13:6 for other VH unmutated cases, P=n.s.) and median follow-up (V1–69 cases: median 56 months, range 6 to 109; other VH unmutated cases: median 59 months, range 20 to 97, P=n.s.). The percentage of cases presenting at an advanced stage was higher in the V1–69 subgroup (36% of cases in Rai high risk category) than in the subgroup with other unmutated VH genes (5% of cases in Rai high risk category, P=0.027), whereas no significant difference was observed with respect to Ly counts and total tumor burden at diagnosis. Interestingly, 80% of the V1–69 cases compared to 42% of cases with other unmutated VH genes had received treatment immediately following diagnosis, but median time to treatment was not significantly different between the two subgroups (1 month vs. 9 months, respectively, P=0.085). To date 24 V1–69 cases (96%) and 15 cases with other unmutated VH genes (79%) have died, with median survival times of 56 and 60 months, respectively (P=0.9). For comparison, median survival of the 14 VH mutated cases from this series was 125 months (P&lt;0.0001 with respect to both subgroups with unmutated VH genes). We therefore conclude that chronic lymphocytic leukemia patients with a V1–69 gene rearrangement constitute a uniform group with unfavorable prognosis, but their survival does not differ significantly from patients with other unmutated VH genes.
Figure Figure
Possible Role of the Activation Loop Tyrosine Residue (Y842) in FLT3/ITD Signaling May Be Mediated by SHP-2.
Authors: Chen Peili , Williams Allen , Molina Henrik , Pandey Akhilesh , Small Donald 
Published: April 2020

Abstract
Internal tandem duplication (ITD) mutations in the FLT3 juxtamembrane domain lead to constitutive activation of FLT3 tyrosine kinase activity and cooperate in the oncogenic transformation of hematopoietic cells. Although numerous signaling molecules are known to be involved in FLT3/ITD signaling, the sites of interaction and mechanisms are not fully understood. In this study, FLT3/ITD protein was immunoprecipitated from TF-1/ITD cells and subjected to nanoLC-MS/MS analysis for determination of tyrosine phosphorylation sites. We found that tyrosine residue 842 (Y842) in the FLT3/ITD activation loop is constitutively phosphorylated. Mutation of this tyrosine residue to phenylalanine (FLT3/ITDY842F) reduces FLT3/ITD phosphorylation and activation of FLT3 downstream targets, including STAT5, AKT and ERK/MAPK. CEP-701, a selective FLT3 inhibitor, inhibits the proliferation of BaF3 cells expressing FLT3/ITD or FLT3/ITDY842F mutant with a similar potency. This implies CEP-701 targets FLT3/ITDY842F as effectively as FLT3/ITD. To identify the signaling pathway(s) initiated by FLT3/ITD Y842, SHP-2 and other molecules downstream of FLT3 were investigated. SHP-2 is known to be activated by FLT3 signaling and its interaction with Gab2 regulates the ERK/MAPK pathway. Previous studies also suggested that SHP-2 is possibly involved in STAT5 activation. Co-immunoprecipitation experiments revealed that SHP-2 association with the FLT3/ITDY842F mutant was considerably diminished as compared to the FLT3/ITD receptor. In addition, the interaction of SHP-2 with Gab2 adaptor protein was greatly compromised in FLT3/ITDY842F expressing BaF3 cells. Taken together, our data suggests that SHP-2 binds FLT3/ITD, either directly or indirectly, and contributes to the signaling pathways activated by mutant FLT3.
The Leukemogenic Properties of MLL Fusion Genes Do Not Require FLT3 Signaling.
Authors: Lavau Catherine , Libura Marta , Morgado Ester , Albouhair Stephanie 
Published: April 2020

Abstract
Leukemias harboring chromosomal translocations involving the Mixed Lineage Leukemia (MLL) gene express high levels of the receptor tyrosine kinase FLT3. Activation of FLT3 signaling is very frequent in human acute myeloid leukemias and its role in the transformation of myeloid cells was demonstrated experimentally. To study what role FLT3 activation played in the cellular transformation leading to MLL-rearranged leukemias we used retroviral vectors to express MLL fusion genes in flt3+/+ and flt3−/− bone marrow cells. Cohorts of mice transplanted with flt3+/+ and flt3−/− bone marrow expressing either MLL-ENL or MLL-CBP were established. All mice succumbed to AML within 60 days (MLL-ENL) or 120 days (MLL-CBP) regardless of flt3 status.
Parameters reflecting cellular proliferation (leukocytosis, spleen weights) or maturation (morphology, immunophenotype) were identical in flt3−/− versus flt3+/+ leukemic blasts. Transplantability and clonality of the leukemias were also unchanged. We showed that the FLT3 protein was expressed in flt3+/+ MLL-ENL leukemic blasts and to explore its role in cell growth we studied the sensitivity of the cells to various FLT3 inhibitors ex vivo. We found that PKC412, AG1296 and Herbymicin A had growth inhibitory activity on MLL-ENL leukemic cells in culture. However the IC50 of these drugs was much higher than those determined on cell lines expressing FLT3 mutants that constitutively activate the FLT3 signaling pathway. Furthermore, flt3−/− MLL-ENL leukemic cells were equally sensitive to the growth inhibitory activity of these drugs than flt3+/+ leukemic cells. This demonstrates that PKC412, AG1296 and Herbymicin A impair the growth of MLL cells through the inhibition of tyrosine kinases other than FLT3. Altogether our data question the value of FLT3 as a therapeutic target to fight MLL-rearranged leukemias and suggest that the efficacy of FLT3 inhibitors is in fact mediated by the interference with another pathway.