Outcomes of Medicare-age eligible NHL patients receiving RIC allogeneic transplantation: a CIBMTR analysis

The application of allogeneic hematopoietic cell transplantation (allo-HCT) in non-Hodgkin lymphoma (NHL) patients ≥65 years in the United States is limited by lack of Medicare coverage for this indication. Using the Center for International Blood and Marrow Transplant Research (CIBMTR) database, we report allo-HCT outcomes of NHL patients aged ≥65 years (older cohort; n = 446) compared with a cohort of younger NHL patients aged 55-64 years (n = 1183). We identified 1629 NHL patients undergoing a first reduced-intensity conditioning (RIC) or nonmyeloablative conditioning allo-HCT from 2008 to 2015 in the United States. Cord blood or haploidentical transplants were excluded. The median age was 68 years (range 65-77) for the older cohort vs 60 years (range 55-64) in the younger cohort. The 4-year adjusted probabilities of nonrelapse mortality (NRM), relapse/progression (R/P), progression-free survival (PFS), and overall survival (OS) of the younger and older groups were 24% vs 30% (P = .03), 41% vs 42% (P = .82), 37% vs 31% (P = .03), and 51% vs 46% (P = .07), respectively. Using multivariate analysis, compared with the younger group, the older cohort was associated with increased NRM, but there was no difference between the 2 cohorts in terms of R/P, PFS, or OS. The most common cause of death was disease relapse in both groups. In NHL patients eligible for allo-HCT, there was no difference in OS between the 2 cohorts. Age alone should not determine allo-HCT eligibility in NHL, and Medicare should expand allo-HCT coverage to older adults

Outcomes of haploidentical vs matched sibling transplantation for acute myeloid leukemia in first complete remission.

HLA-haploidentical hematopoietic cell transplantation (Haplo-HCT) using posttransplantation cyclophosphamide (PT-Cy) has improved donor availability. However, a matched sibling donor (MSD) is still considered the optimal donor. Using the Center for International Blood and Marrow Transplant Research database, we compared outcomes after Haplo-HCT vs MSD in patients with acute myeloid leukemia (AML) in first complete remission (CR1). Data from 1205 adult CR1 AML patients (2008-2015) were analyzed. A total of 336 patients underwent PT-Cy–based Haplo-HCT and 869 underwent MSD using calcineurin inhibitor–based graft-versus-host disease (GVHD) prophylaxis. The Haplo-HCT group included more reduced-intensity conditioning (65% vs 30%) and bone marrow grafts (62% vs 7%), consistent with current practice. In multivariable analysis, Haplo-HCT and MSD groups were not different with regard to overall survival (P 5 .15), leukemia-free survival (P 5 .50), nonrelapse mortality (P 5 .16), relapse (P 5 .90), or grade II-IV acute GVHD (P 5 .98). However, the Haplo-HCT group had a significantly lower rate of chronic GVHD (hazard ratio, 0.38; 95% confidence interval, 0.30-0.48; P, .001). Results of subgroup analyses by conditioning intensity and graft source suggested that the reduced incidence of chronic GVHD in Haplo-HCT is not limited to a specific graft source or conditioning intensity. Center effect and minimal residual disease–donor type interaction were not predictors of outcome. Our results indicate a lower rate of chronic GVHD after PT-Cy–based Haplo-HCT vs MSD using calcineurin inhibitor–based GVHD prophylaxis, but similar other outcomes, in patients with AML in CR1. Haplo-HCT is a viable alternative to MSD in these patients. © 2019 American Society of Hematology. All rights reserved

Insights into the role of yeast eIF2A in IRES-mediated translation.

Eukaryotic initiation factor 2A is a single polypeptide that acts to negatively regulate IRES-mediated translation during normal cellular conditions. We have found that eIF2A (encoded by YGR054w) abundance is reduced at both the mRNA and protein level during 6% ethanol stress (or 37°C heat shock) under conditions that mimic the diauxic shift in the yeast Saccharomyces cerevisiae. Furthermore, eIF2A protein is posttranslationally modified during ethanol stress. Unlike ethanol and heat shock stress, H(2)O(2) and sorbitol treatment induce the loss of eIF2A mRNA, but not protein and without protein modification. To investigate the mechanism of eIF2A function we employed immunoprecipitation-mass spectrometry and identified an interaction between eIF2A and eEF1A. The interaction between eIF2A and eEF1A increases during ethanol stress, which correlates with an increase in IRES-mediated translation from the URE2 IRES element. These data suggest that eIF2A acts as a switch to regulate IRES-mediated translation, and eEF1A may be an important mediator of translational activation during ethanol stress

Insights into the Role of Yeast eIF2A in IRES-Mediated Translation

Eukaryotic initiation factor 2A is a single polypeptide that acts to negatively regulate IRES-mediated translation during normal cellular conditions. We have found that eIF2A (encoded by YGR054w) abundance is reduced at both the mRNA and protein level during 6% ethanol stress (or 37°C heat shock) under conditions that mimic the diauxic shift in the yeast Saccharomyces cerevisiae. Furthermore, eIF2A protein is posttranslationally modified during ethanol stress. Unlike ethanol and heat shock stress, H2O2 and sorbitol treatment induce the loss of eIF2A mRNA, but not protein and without protein modification. To investigate the mechanism of eIF2A function we employed immunoprecipitation-mass spectrometry and identified an interaction between eIF2A and eEF1A. The interaction between eIF2A and eEF1A increases during ethanol stress, which correlates with an increase in IRES-mediated translation from the URE2 IRES element. These data suggest that eIF2A acts as a switch to regulate IRES-mediated translation, and eEF1A may be an important mediator of translational activation during ethanol stress.</p

Domain 3 of eEF1A is required for interaction with eIF2A.

<p><i>A,</i> The domain architecture of eEF1A is presented with amino acid demarcations at the boundaries of individual domains, and color coding of domains that is consistent throughout the figure. Bold arrows represent the location of mutations employed in <i>C</i>. Regions of eEF1A previously reported to be important for interaction with other proteins are highlighted with brackets. <i>B,</i> Domain deletions of eEF1A fused to GST were used to precipitate eIF2A-HA from eIF2A-HA yeast lysate. Western blotting for HA-tagged eIF2A was conducted on the precipitated material (upper panel). Domains of eEF1A present in the GST-fusion protein are indicated above the Western blot. GST fusion protein added to each GST-pulldown reaction was analyzed after Western blotting by coomassie staining of the membrane (lower panel). <i>C,</i> Domain 3 of eEF1A was found to be critical for the interaction between eEF1A and eIF2A. GST-fusion protein point mutants of eEF1A were used to confirm the necessity for domain 3 in the interaction between eIF2A and eEF1A using eIF2A-HA yeast. Each mutant is indicated in the figure. Western blotting followed by Coomassie staining of the membrane were conducted as described above (<i>B</i>).</p

eIF2A is post-translationally modified during 6% ethanol treatment.

<p>2D-gel electrophoresis followed by Western blotting for eIF2A from BY4684 ΔeIF2A yeast transformed with YCplac111-YP with or without the indicated stress.</p

<i>URE2</i> IRES activity increases proportional to the increase in interaction between eIF2A and eEF1A.

<p><i>URE2</i> IRES activity of the minimal IRES element (nucleotides 305–309) was evaluated as above (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0024492#pone-0024492-g007" target="_blank">Figure 7C</a>). This construct was transformed into ΔeIF2A yeast expressing HA-eIF2A from the YCplac111-YP plasmid (BY4684; ΔeIF2A+HA-eIF2A), and cultures were grown in the absence (minimal medium) or presence (6% ethanol) of 6% ethanol for one hour. Cells were harvested and β-galactosidase activity was measured. We have previously shown that there is no change in mRNA levels of this construct in an eIF2A-dependent manner <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0024492#pone.0024492-Reineke1" target="_blank">[6]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0024492#pone.0024492-Reineke2" target="_blank">[10]</a>. Error bars represent 15% of the mean, which is empirically acceptable for this type of experiment. These experiments were repeated several times.</p

Post-translationally modified eIF2A appears in 80S preinitiation complexes during 6% ethanol stress.

<p><i>A,</i> BY4684 ΔeIF2A yeast transformed with YCplac111-YP with or without 6% ethanol treatment were analyzed by polysome profiling. <i>B,</i> Subsequently, the top and 80S complex regions of the gradients (shown in <i>A</i>) were analyzed by 2D-gel electrophoresis and Western blotting with α-HA antibody to detect HA-eIF2A.</p

Proteins detected in IP-mass spectrometry experiments.

<p>HA-eIF2A protein complexes were immunoprecipitated and the protein complexes identified by mass spectrometry. Proteins detected by mass spectrometry that did not appear in the vector alone control are listed. ND under “Shown to previously interact” represents those proteins that have not been detected previously in the literature. NA under “Reference” indicates that no reference is applicable because it has not previously been detected.</p