Sickle cell disease (SCD) and Cooley's anemia (CA) are disorders ameliorated by continued expression of fetal hemoglobin; first shown in patients with continued expression of fetal globin due to deletional or nondeletional hereditary persistence of fetal hemoglobin (HFPH). However, others express high fetal globin in the absence of mutation, and therapeutic agents, such as hydroxyurea, induce expression of Hb F. If the rate-limiting factors involved in the regulation of fetal hemoglobin were elucidated, targeted drug discovery could succeed in developing agents with this specific effect. Our proposal focuses on three areas: 1) identify new candidate modifiers using unbiased, genome-wide microarray-based expression profiling to define genes critical to induction of Hb F in cultured human erythroid progenitors. The candidates will be assessed for identification of genes that fall into several categories, including transcription factors, signaling and chromatin remodeling molecules. 2) examine suspected regulators of Hb F expression as well as a subset of differentially-expressed transcripts comparing cord to adult erythroid cultures for functionally relevant genetic variation associated with elevated Hb F levels. Known regulators (KLFs, NFE-4 and soluble guanylate cyclase) and the most promising of the candidate transactivators identified by expression profiling will be explored to identify putative coding region and proximal promoter sequence allelic variants using both public databases and SNP discovery methods. Those with allele frequencies of 10% or more in a small group of SCD patients, will be tested in sequential cohorts of adult and pediatric patients with SCD with and without elevated Hb F. Significant QTL loci will then be examined in a group of parent-child trios and followed with a third population of SCD patients. 3) functional validation of candidate modifiers will be accomplished by knockdown technology employing morpholino antisense oligos and/or interference RNA (iRNA) using K562 cells containing a fetal globin promoter-driven GFP readout from a beta-like globin gene cluster BAC. We are confident that our strategy to identify critical regulatory molecules for Hb F regulation and our ability to quickly and reliably screen for candidate gene variants will have significant impact on the development of therapeutic intervention strategies for patients with SCD and CA.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL069256-04
Application #
6986138
Study Section
Hematology Subcommittee 2 (HEM)
Program Officer
Evans, Gregory
Project Start
2002-12-01
Project End
2007-11-30
Budget Start
2005-12-01
Budget End
2006-11-30
Support Year
4
Fiscal Year
2006
Total Cost
$369,918
Indirect Cost
Name
Thomas Jefferson University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
053284659
City
Philadelphia
State
PA
Country
United States
Zip Code
19107
Steering Committee Of The Blood And Marrow Transplant Clinical Trials Network (2016) The Blood and Marrow Transplant Clinical Trials Network: An Effective Infrastructure for Addressing Important Issues in Hematopoietic Cell Transplantation. Biol Blood Marrow Transplant 22:1747-1757
Bernardini, Laura; Alesi, Viola; Loddo, Sara et al. (2010) High-resolution SNP arrays in mental retardation diagnostics: how much do we gain? Eur J Hum Genet 18:178-85
Adachi, Kazuhiko; Ding, Min; Asakura, Toshio et al. (2009) Relationship between beta4 hydrogen bond and beta6 hydrophobic interactions during aggregate, fiber or crystal formation in oversaturated solutions of hemoglobin A and S. Arch Biochem Biophys 481:137-44
Battistella, Stefania; Damin, Francesco; Chiari, Marcella et al. (2008) Genotyping beta-globin gene mutations on copolymer-coated glass slides with the ligation detection reaction. Clin Chem 54:1657-63
Adachi, Kazuhiko; Ding, Min; Surrey, Saul (2008) Role of the beta4Thr-beta73Asp hydrogen bond in HbS polymer and domain formation from multinucleate-containing clusters. Biochemistry 47:5441-9
Adachi, Kazuhiko; Zhao, Yi; Lakka, Vinaysagar et al. (2007) Assembly of recently translated full-length and C-terminal truncated human gamma-globin chains with a pool of alpha-globin chains to form Hb F in a cell-free system. Arch Biochem Biophys 463:60-7
Fortina, Paolo; Kricka, Larry J; Graves, David J et al. (2007) Applications of nanoparticles to diagnostics and therapeutics in colorectal cancer. Trends Biotechnol 25:145-52
Schirinzi, Annalisa; Drmanac, Snezana; Dallapiccola, Bruno et al. (2006) Combinatorial sequencing-by-hybridization: analysis of the NF1 gene. Genet Test 10:8-17
Keller, M A; Addya, S; Vadigepalli, R et al. (2006) Transcriptional regulatory network analysis of developing human erythroid progenitors reveals patterns of coregulation and potential transcriptional regulators. Physiol Genomics 28:114-28
Akbar, Mohammed G K; Tamura, Yutaka; Ding, Min et al. (2006) Inhibition of hemoglobin S polymerization in vitro by a novel 15-mer EF-helix beta73 histidine-containing peptide. Biochemistry 45:8358-67

Showing the most recent 10 out of 16 publications