Erythroid lineage chromatin accessibility maps facilitate identification and validation of NFIX as a fetal hemoglobin repressor.

Human genetics has validated de-repression of fetal gamma globin (HBG) in adult erythroblasts as a powerful therapeutic paradigm in diseases involving defective adult beta globin (HBB)1. To identify factors involved in the switch from HBG to HBB expression, we performed Assay for Transposase Accessible Chromatin with high-throughput sequencing (ATAC-seq)2 on sorted erythroid lineage cells derived from bone marrow (BM) or cord blood (CB), representing adult and fetal states, respectively. BM to CB cell ATAC-seq profile comparisons revealed genome-wide enrichment of NFI DNA binding motifs and increased NFIX promoter chromatin accessibility, suggesting that NFIX may repress HBG. NFIX knockdown in BM cells increased HBG mRNA and fetal hemoglobin (HbF) protein levels, coincident with increased chromatin accessibility and decreased DNA methylation at the HBG promoter. Conversely, overexpression of NFIX in CB cells reduced HbF levels. Identification and validation of NFIX as a new target for HbF activation has implications in the development of therapeutics for hemoglobinopathies.

Reward motivation more consistently modulates memory for younger compared to older adults in a directed forgetting task.

Remembering and forgetting are both important processes of a healthy memory system, but both processes can show age-related decline. Reward anticipation is effective at improving remembering in both younger and older adults, but little is known about the effects of incentives on forgetting. In four online experiments, we examined whether reward motivation modulates intentional remembering and forgetting in younger and older adults, and systematically varied the presentation of reward cues during encoding to test whether the temporal dynamics of reward anticipation are important for directed forgetting performance. Both age groups showed directed forgetting effects such that participants remembered more items they were instructed to remember than instructed to forget, but across experiments, we found no evidence that reward incentives improved forgetting in either age group. Younger adults consistently exhibited reward-modulated memory across experiments and varying the timing of the reward cue had little impact on performance. Older adults displayed inconsistent effects of reward on memory, only when reward anticipation was elicited closer to the middle of the experimental trial did it enhance memory in this task. Overall, the findings from the current set of experiments indicate that reward anticipation improved memory, but not forgetting, and most consistently for younger adults, compared to older adults. Further, older adults' cognitive performance may be more sensitive to the placement and timing of reward anticipation in the experimental trial perhaps due to the time course of reward anticipation and interactions with the hippocampus that may show age-related change. (PsycInfo Database Record (c) 2023 APA, all rights reserved).

Influence of Reward Motivation on Directed Forgetting in Younger and Older Adults.

An important feature of the memory system is the ability to forget, but aging is associated with declines in the ability to intentionally forget potentially due to declines in cognitive control. Despite cognitive deficits, older adults are sensitive to affective manipulations, such as reward motivation, and reward anticipation can improve older adults' memory performance. The goal of the current studies was to examine the effect of reward motivation on directed remembering and forgetting. Participants were healthy CloudResearch/Turk Prime workers aged 18-35 and 60-85. In Experiment 1, we conducted a typical item-method directed forgetting task using neutral words presented one at a time followed by a to-be-remembered (TBR) or to-be-forgotten (TBF) cue. A recognition memory test followed that included all words from the encoding task, as well as new words. We replicated prior findings of better memory for TBR compared to TBF items, but not typical age-related differences in recognition of TBF items. In Experiments 2-4, we repeated this paradigm except that in the second block of trials, each word was presented with a high ($0.75) or low ($0.01) reward cue indicating the value that could be earned if the item was successfully Remembered or Forgotten (depending on cue). During recognition, correct responses to target items (both TBR and TBF) resulted in the associated reward, but incorrect "old" responses resulted in a loss of $0.50. In three experiments, high rewards led to better memory for younger and older adults compared to low rewards, regardless of the directed cue to remember or forget the word. In Experiments 3 and 4, older adults showed typical deficits in directed forgetting, but this was across reward conditions. For older adults, there was no evidence that including reward motivation improved cognitive control abilities as high value reward anticipation did not improve directed forgetting. Instead, in line with hypotheses, high compared to low value reward anticipation leads to engagement of processes that result in better memory regardless of the TBR or TBF cue, and reward anticipation bolsters memory in a relatively automatic, rather than strategic, fashion that overrides one's ability to cognitively control encoding processes.

Small-Molecule PAPD5 Inhibitors Restore Telomerase Activity in Patient Stem Cells.

Genetic lesions that reduce telomerase activity inhibit stem cell replication and cause a range of incurable diseases, including dyskeratosis congenita (DC) and pulmonary fibrosis (PF). Modalities to restore telomerase in stem cells throughout the body remain unclear. Here, we describe small-molecule PAPD5 inhibitors that demonstrate telomere restoration in vitro, in stem cell models, and in vivo. PAPD5 is a non-canonical polymerase that oligoadenylates and destabilizes telomerase RNA component (TERC). We identified BCH001, a specific PAPD5 inhibitor that restored telomerase activity and telomere length in DC patient induced pluripotent stem cells. When human blood stem cells engineered to carry DC-causing PARN mutations were xenotransplanted into immunodeficient mice, oral treatment with a repurposed PAPD5 inhibitor, the dihydroquinolizinone RG7834, rescued TERC 3' end maturation and telomere length. These findings pave the way for developing systemic telomere therapeutics to counteract stem cell exhaustion in DC, PF, and possibly other aging-related diseases.

Posttranscriptional manipulation of TERC reverses molecular hallmarks of telomere disease.

The telomerase RNA component (TERC) is a critical determinant of cellular self-renewal. Poly(A)-specific ribonuclease (PARN) is required for posttranscriptional maturation of TERC. PARN mutations lead to incomplete 3' end processing and increased destruction of nascent TERC RNA transcripts, resulting in telomerase deficiency and telomere diseases. Here, we determined that overexpression of TERC increased telomere length in PARN-deficient cells and hypothesized that decreasing posttranscriptional 3' oligo-adenylation of TERC would counteract the deleterious effects of PARN mutations. Inhibition of the noncanonical poly(A) polymerase PAP-associated domain-containing 5 (PAPD5) increased TERC levels in PARN-mutant patient cells. PAPD5 inhibition was also associated with increases in TERC stability, telomerase activity, and telomere elongation. Our results demonstrate that manipulating posttranscriptional regulatory pathways may be a potential strategy to reverse the molecular hallmarks of telomere disease.

Poly(A)-specific ribonuclease (PARN) mediates 3'-end maturation of the telomerase RNA component.

Mutations in the PARN gene (encoding poly(A)-specific ribonuclease) cause telomere diseases including familial idiopathic pulmonary fibrosis (IPF) and dyskeratosis congenita, but how PARN deficiency impairs telomere maintenance is unclear. Here, using somatic cells and induced pluripotent stem cells (iPSCs) from patients with dyskeratosis congenita with PARN mutations, we show that PARN is required for the 3'-end maturation of the telomerase RNA component (TERC). Patient-derived cells as well as immortalized cells in which PARN is disrupted show decreased levels of TERC. Deep sequencing of TERC RNA 3' termini shows that PARN is required for removal of post-transcriptionally acquired oligo(A) tails that target nuclear RNAs for degradation. Diminished TERC levels and the increased proportion of oligo(A) forms of TERC are normalized by restoring PARN, which is limiting for TERC maturation in cells. Our results demonstrate a new role for PARN in the biogenesis of TERC and provide a mechanism linking PARN mutations to telomere diseases.