Sickle cell anemia, an orphan disease of African Americans, is noted for its extensive morbidity and high mortality. Only one FDA approved drug is available for its pathophysiologically-based treatment. This agent, hydroxyurea, works through its ability to induce fetal hemoglobin (HbF) expression, which thwarts sickle hemoglobin polymerization. Not all patients respond to this treatment, so additional HbF inducing drugs are needed. Our goal is to create the technology for developing high throughput sickle cell anemia-specific induced pluripotent stem cells (iPS) and characterize their directly differentiated progeny. Using a novel excisable reprogramming vector we will generate 'clinical grade'human iPS cells free of any residual reprogramming transgenes. These directly differentiated sickle iPS cells will be used to produce an unlimited supply of erythroid-lineage cells to better understand HbF genetic regulation and perform pre-clinical small molecule drug screens. Specifically, we hypothesize: 1) that 'clinical grade'disease-specific iPS cells can be efficiently and reproducibly derived from peripheral blood cells and induced to normal erythroid differentiation, which includes the expected the spectrum of p-like globin gene expression;2) major HbF quantitative trait loci impact globin gene expression during embryonic, fetal, and adult erythropoiesis;3) patients with markedly elevated HbF levels serve as natural models to identify and characterize genetic variations affecting HbF production;4) high-potency inducers of HbF, either singly or in combination can be studied in iPS-derived erythroid precursor cells.
Our aims are: 1) implement an efficient and 'scalable'system for the production of sickle cell anemia-specific iPS cells and use this to recapitulate erythroid-lineage ontogeny in vitro;2) identify developmental gene expression profile differences between erythroid precursors that produce primarily HbF and those that produce primarily HbA or HbS;3) determine the effects of the known major HbF quantitative trait loci on globin gene expression in iPS cells;4) search for novel HbF genetic modifiers associated with HbF levels by examining gene expression in iPS derived erythroid cells;5) determine which novel therapeutics discovered in high throughput screens can enhance and maintain high level HBG expression in iPS-derived sickle erythroid cells. Ultimately, we hope to translate these findings into clinically efficacious treatments.

Public Health Relevance

We will make stem cells from the blood of patients with sickle cell anemia to learn how genes that make hemoglobin that is beneficial to these patients are regulated, and how these cells might be used for their treatment.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project--Cooperative Agreements (U01)
Project #
5U01HL107443-04
Application #
8691997
Study Section
Special Emphasis Panel (ZHL1)
Program Officer
Qasba, Pankaj
Project Start
2011-07-05
Project End
2016-06-30
Budget Start
2014-07-01
Budget End
2015-06-30
Support Year
4
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Boston University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
City
Boston
State
MA
Country
United States
Zip Code
02118
Park, Seonmi; Gianotti-Sommer, Andreia; Molina-Estevez, Francisco Javier et al. (2017) A Comprehensive, Ethnically Diverse Library of Sickle Cell Disease-Specific Induced Pluripotent Stem Cells. Stem Cell Reports 8:1076-1085
Kato, Gregory J; Steinberg, Martin H; Gladwin, Mark T (2017) Intravascular hemolysis and the pathophysiology of sickle cell disease. J Clin Invest 127:750-760
Habara, Alawi H; Shaikho, Elmutaz M; Steinberg, Martin H (2017) Fetal hemoglobin in sickle cell anemia: The Arab-Indian haplotype and new therapeutic agents. Am J Hematol 92:1233-1242
Morrison, Tasha A; Wilcox, Ibifiri; Luo, Hong-Yuan et al. (2017) A long noncoding RNA from the HBS1L-MYB intergenic region on chr6q23 regulates human fetal hemoglobin expression. Blood Cells Mol Dis 69:1-9
Vathipadiekal, Vinod; Farrell, John J; Wang, Shuai et al. (2016) A candidate transacting modulator of fetal hemoglobin gene expression in the Arab-Indian haplotype of sickle cell anemia. Am J Hematol 91:1118-1122
Smith, Brenden W; Stanford, Elizabeth A; Sherr, David H et al. (2016) Genome Editing of the CYP1A1 Locus in iPSCs as a Platform to Map AHR Expression throughout Human Development. Stem Cells Int 2016:2574152
Vathipadiekal, Vinod; Alsultan, Abdulrahman; Baltrusaitis, Kristin et al. (2016) Homozygosity for a haplotype in the HBG2-OR51B4 region is exclusive to Arab-Indian haplotype sickle cell anemia. Am J Hematol 91:E308-11
Steinberg, Martin H; Rodgers, Griffin P (2015) HbA2 : biology, clinical relevance and a possible target for ameliorating sickle cell disease. Br J Haematol 170:781-7
Sebastiani, P; Farrell, J J; Alsultan, A et al. (2015) BCL11A enhancer haplotypes and fetal hemoglobin in sickle cell anemia. Blood Cells Mol Dis 54:224-30
Milton, Jacqueline N; Gordeuk, Victor R; Taylor 6th, James G et al. (2014) Prediction of fetal hemoglobin in sickle cell anemia using an ensemble of genetic risk prediction models. Circ Cardiovasc Genet 7:110-5

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