Anemia affects approximately 1.6 billion people worldwide, imposing an enormous burden on medical resources. Of the inherited anemias, the hemoglobinopathies, particularly sickle cell disease (SCD) and ?- thalassemia, stand out due to their prevalence and severity. The ability to postnatally elevate fetal hemoglobin (HbF) levels in SCD and ?-thalassemia via co-inheritance of positive genetic modifiers of HbF production or hydroxyurea (HU) treatment can significantly alleviate disease severity. However, HU is not without side effects and may even be carcinogenic. Novel therapies aimed at elevating HbF expression in adults are therefore desperately needed. Three major loci, including BCL11A, are known to modify HbF expression in humans. Together, however, they account for only ~50% of the variation in HbF levels. Hence, significant gaps in knowledge remain regarding the genetic control of HbF production. We will take advantage of two powerful mouse resources to identify novel regulators of ?-like globin switching, the mouse mutant Nan (neonatal anemia) and the newly developed high resolution Diversity Outbred (DO) mapping resource. In Nan, a single amino acid change in the second zinc finger of the erythroid Kr?ppel-like factor (EKLF) causes sequence selective disruption of binding to a subset of its target genes. The result is severe anemia accompanied by a striking failure of hemoglobin switching. Expression of embryonic ?h1 globin is upregulated 100-fold in adult Nan spleen vs. wild type via a BCL11A independent mechanism. In highly genetically diverse DO mice, expression of ?h1 globin in adults varies substantially from individual to individual. Thus, these two resources, Nan and DO mice, possess the prerequisites required to detect novel regulators of ?-like globin switching using powerful, unbiased genetic (QTL mapping) and genomic (ChIP-seq, RNA-seq) strategies.
The aim of this proposal is to identify novel genes regulating ?-like globin switching. To accomplish this aim, we will: (a) map modifiers of ?h1 expression in Nan F2 intercrosses and in DO mice;(b) perform ChIP-seq in erythroid populations to compare DNA targets differentially bound by wild type (WT) EKLF and mutant (Nan) EKLF, and RNA-seq to identify gene expression differences;and (c) analyze and integrate all data to identify, prioritize, and initiate analysis of candidate genes. ! Identifying genetic loci regulating ?h1 expression, differences in Nan- vs. WT-EKLF DNA targets, and differences in gene expression in Nan vs. WT erythroid cells will converge to identify novel regulators of ?- like globin switching, thereby providing important new therapeutic targets for the hemoglobinopathies. !

Public Health Relevance

Anemia affects approximately 1.6 billion people worldwide, imposing an enormous burden on medical resources. Notably, anemias share common etiologies in mice and man. Thus, mice are excellent model organisms in which to study mechanisms of anemia and potential therapies. Of the inherited anemias, the hemoglobinopathies, particularly sickle cell disease and ?-thalassemia, stand out due to their prevalence and severity. Elevation of fetal hemoglobin (HbF), which is normally switched off at birth, is an effective means to alleviate disease severity in these disorders. Our aim is to identiy gene variants and genetic pathways that lead to natural increases in HbF postnatally, thus identifying novel targets for the development of new therapies.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
1R01DK100692-01
Application #
8611365
Study Section
Molecular and Cellular Hematology (MCH)
Program Officer
Bishop, Terry Rogers
Project Start
2013-09-16
Project End
2014-09-15
Budget Start
2013-09-16
Budget End
2014-09-15
Support Year
1
Fiscal Year
2013
Total Cost
$262,696
Indirect Cost
$112,584
Name
Jackson Laboratory
Department
Type
DUNS #
042140483
City
Bar Harbor
State
ME
Country
United States
Zip Code
04609