ABSTRACT
Diamond-Blackfan Anemia (DBA) is a bone marrow failure disorder characterized by low red blood cell count. Mutations in ribosomal protein genes have been identified in approximately half of all DBA cases. Corticosteriod therapy and bone marrow transplantation are common treatment options for patients; however, significant risks and complications are associated with these treatment options. Therefore, novel therapeutic approaches are needed for treating DBA. Sotatercept (ACE-011, and its murine ortholog RAP-011) acts as an activin receptor type IIA ligand trap, increasing hemoglobin and hematocrit in pharmacologic models, in healthy volunteers, and in patients with ß-thalassemia, by expanding late-stage erythroblasts through a mechanism distinct from erythropoietin. Here, we evaluated the effects of RAP-011 in zebrafish models of RPL11 ribosome deficiency. Treatment with RAP-011 dramatically restored hemoglobin levels caused by ribosome stress. In zebrafish embryos, RAP-011 likely stimulates erythropoietic activity by sequestering lefty1 from erythroid cells. These findings identify lefty1 as a signaling component in the development of erythroid cells and rationalize the use of sotatercept in DBA patients.
Subject(s)
Anemia, Diamond-Blackfan/drug therapy , Erythropoiesis/drug effects , Left-Right Determination Factors/antagonists & inhibitors , Recombinant Fusion Proteins/therapeutic use , Zebrafish Proteins/antagonists & inhibitors , Activin Receptors, Type II/antagonists & inhibitors , Activin Receptors, Type II/blood , Anemia, Diamond-Blackfan/blood , Anemia, Diamond-Blackfan/genetics , Animals , Disease Models, Animal , Erythropoiesis/genetics , Gene Knockdown Techniques , Genes, p53 , Humans , Left-Right Determination Factors/blood , Left-Right Determination Factors/genetics , Ligands , Ribosomal Proteins/blood , Ribosomal Proteins/deficiency , Ribosomal Proteins/genetics , Signal Transduction/drug effects , Zebrafish , Zebrafish Proteins/blood , Zebrafish Proteins/genetics , beta-Thalassemia/blood , beta-Thalassemia/drug therapyABSTRACT
As a NODAL pathway inhibitor, EBAF plays a critical role during mammalian cardiac development. As recent tests that have been conducted on gene-targeted mice indicate, its expression is frequently altered where cardiac defects are present. We aimed to explore the EBAF expression pattern and molecular mechanism of EBAF gene for VSD genesis. In this report, we show that the average expression of EBAF in the disease tissues of VSD patients was lower than the expression in normal fetuses without VSD. Further study showed that the expression pattern of EBAF was potentially involved in cardiomyocyte apoptosis by Annexin-V and RT-PCR assays. We also found that abnormal activation of NODAL-PITX2C pathway was associated with down-regulation of EBAF. By luciferase reporter assays, we find that EBAF expression is mediated by transcriptional factors smad2 and cited2. In addition, ChIP assays showed that histone acetyltransferase p300 is involved in the activation of EBAF through inducing hyperacetylation of histone H4 at the EBAF promoter. Co-immunoprecipitation also indicates that the expression of EBAF is regulated by a transcriptional complex including p300, smad2, and cited2. This study revealed a novel regulator mechanism of EBAF, which may be a potential molecular target for halting the onset of VSDs. They also indicate that smad2, cited2, and p300 may play important roles in modulating the confirmation of ventricular septal defects.
