Completed Grants and Sponsored Research

This research is designed to test the hypothesis that NHEJ-mediated targeted indel production itself would be a favorable therapeutic genome editing strategy for sickle cell disease by disruption of endogenous β-globin gene cluster sequences that repress the expression of fetal hemoglobin.

The goal of this research is to study various features of the early blood cells that are collected during the plerixafor trial. We will test whether the plerixafor-mobilized cells are suitable for gene therapy and gene editing.

The goal of this project is to explore genetic determinants of human red blood cell traits that modulate erythroid enhancer elements. The aims are: to compare deletion of trait-associated enhancers to non-trait associated enhancers to investigate the genetic architecture of erythroid traits; and to disrupt trait-associated enhancers to identify variants and genes underlying erythroid traits.

The goals of this research are to utilize a high-resolution, high-throughput genome editing approach to identify genetically implicated noncoding sequences (in addition to the GWAS-marked BCL11A enhancer) associated with HbF level and to define trans-acting factors that physically and functionally interact with critical noncoding sequences at BCL11A and HBS1L-MYB.

This research aims to investigate the effects of disrupting the BCL 11 A erythroid enhancer in HSCs and ensure that the modified cells can produce all types of blood cells while also leading to elevated levels of HbF. In addition, we will investigate whether targeting a more specific, enriched HSC population will allow for increased rates of editing and a more effective therapy. The work proposed here will provide important advancements in creating a gene therapy treatment for the 13-hemoglobinopathies as well as improving the use of genome editing technologies in hematopoietic stem cells.

The goal of this proposal is to identify mechanisms that regulate HbF level and could serve as rational targets for efforts to re-induce HbF for the β-hemoglobin disorders.

In this proposal, we will identify minimal sequences at the β-globin gene cluster necessary for BCL11A to repress γ-globin in β-thalassemia erythroid cells; and compare methodologies for safely and effectively performing genome editing of these repressive sequences in primary human β-thalassemia HSCs.

The goals of this research are to discover molecular mechanisms underpinning hemoglobin switching. Specific aims include: to investigate mechanisms of BCL11A transcriptional regulation promoting the fetal-to-adult developmental transition; and to functionally evaluate regulatory elements at the BCL11A locus.

The goal of this project is to explore whether a specific domain(s) of NuRD (Nucleosome Remodeling Deacetylase) subunits is necessary for γ-globin silencing and serves as a potential target for HbF reactivation therapy. The long-term goal of this project is to develop a compound(s) that induces HbF by targeting the epigenetic complex regulating globin switching.

Molecular identification of Psickle in reticulocytes generated from HUDEP-2 cells gene-edited to express HbS. This project is designed to enable production of a human cellular model of HbS expression using gene editing and then initial characterization of Psickle activity in HbA and HbS expressing erythroid cells.

Specific Aim 1. Utilize a custom pooled CRISPR guide RNA library to perform high-resolution mutagenesis to determine minimal non-coding sequences at the B-globin gene cluster necessary for HbF repression. Specific Aim 2. Determine efficiency and specificity of genome editing human HSOCs by measuring rate of on-and off-target editing and impact on HbF level and henatopolesis.