Correction of Sickle Cell Disease in Adult Mice by Interference with Fetal Hemoglobin Silencing

Manipulation of a transcriptional repressor promotes expression of protective fetal globin genes. Persistence of human fetal hemoglobin (HbF, α2γ2) in adults lessens the severity of sickle cell disease (SCD) and the β-thalassemias. Here, we show that the repressor BCL11A is required in vivo for silencing of γ-globin expression in adult animals, yet dispensable for red cell production. BCL11A serves as a barrier to HbF reactivation by known HbF inducing agents. In a proof-of-principle test of BCL11A as a potential therapeutic target, we demonstrate that inactivation of BCL11A in SCD transgenic mice corrects the hematologic and pathologic defects associated with SCD through high-level pancellular HbF induction. Thus, interference with HbF silencing by manipulation of a single target protein is sufficient to reverse SCD.

[1]  I. Todorov,et al.  HDAC inhibitor reduces cytokine storm and facilitates induction of chimerism that reverses lupus in anti-CD3 conditioning regimen , 2008, Proceedings of the National Academy of Sciences.

[2]  Yonggoo Kim,et al.  A New Anti-Hemoglobin F Antibody Against Synthetic Peptides for the Detection of F-Cell Precursors (F-blasts) in Bone Marrow , 2001, International journal of hematology.

[3]  Gonçalo R. Abecasis,et al.  Genome-wide association study shows BCL11A associated with persistent fetal hemoglobin and amelioration of the phenotype of β-thalassemia , 2008, Proceedings of the National Academy of Sciences.

[4]  Marius Wernig,et al.  Treatment of Sickle Cell Anemia Mouse Model with iPS Cells Generated from Autologous Skin , 2007, Science.

[5]  M Aguet,et al.  Inducible gene targeting in mice , 1995, Science.

[6]  D. Zwiers,et al.  5-Azacytidine stimulates fetal hemoglobin synthesis in anemic baboons. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[7]  R. Nagel,et al.  Second generation knockout sickle mice: the effect of HbF. , 2001, Blood.

[8]  A. Mutero,et al.  The human beta globin locus introduced by YAC transfer exhibits a specific and reproducible pattern of developmental regulation in transgenic mice. , 1997, Blood.

[9]  U. Klingmüller,et al.  A mouse model for visualization and conditional mutations in the erythroid lineage. , 2004, Blood.

[10]  A. Nienhuis,et al.  Transcriptional regulation of fetal to adult hemoglobin switching: new therapeutic opportunities. , 2011, Blood.

[11]  R. Samulski,et al.  Combination therapy utilizing shRNA knockdown and an optimized resistant transgene for rescue of diseases caused by misfolded proteins , 2011, Proceedings of the National Academy of Sciences.

[12]  Jian Xu,et al.  Transcriptional competence and the active marking of tissue-specific enhancers by defined transcription factors in embryonic and induced pluripotent stem cells. , 2009, Genes & development.

[13]  J. Hirschhorn,et al.  DNA polymorphisms at the BCL11A, HBS1L-MYB, and β-globin loci associate with fetal hemoglobin levels and pain crises in sickle cell disease , 2008, Proceedings of the National Academy of Sciences.

[14]  K. Gaensler,et al.  Methyl binding domain protein 2 mediates γ-globin gene silencing in adult human βYAC transgenic mice , 2006 .

[15]  王晓燕.  Arch Intern Med:住院增加老年人出院后骨折危险 , 2009 .

[16]  A. Kraus,et al.  Two Families Showing Interaction of Haemoglobin C or Thalassaemia with High Foetal Haemoglobin in Adults* , 1961, British medical journal.

[17]  R. Nagel,et al.  Correction of Sickle Cell Disease in Transgenic Mouse Models by Gene Therapy , 2001, Science.

[18]  Takuro Nakamura,et al.  Bcl11a is essential for normal lymphoid development , 2003, Nature Immunology.

[19]  O. Platt,et al.  Mortality in sickle cell disease. Life expectancy and risk factors for early death. , 1994, The New England journal of medicine.

[20]  S. Orkin,et al.  Transcriptional silencing of {gamma}-globin by BCL11A involves long-range interactions and cooperation with SOX6. , 2010, Genes & development.

[21]  宁北芳,et al.  疟原虫var基因转换速率变化导致抗原变异[英]／Paul H, Robert P, Christodoulou Z, et al//Proc Natl Acad Sci U S A , 2005 .

[22]  T. Townes,et al.  Knockout-transgenic mouse model of sickle cell disease. , 1997, Science.

[23]  D. Charnock-Jones,et al.  vavCre Transgenic mice: A tool for mutagenesis in hematopoietic and endothelial lineages , 2002, Genesis.

[24]  T. Kimbrough,et al.  Use of yeast artificial chromosomes (YACs) in studies of mammalian development: production of beta-globin locus YAC mice carrying human globin developmental mutants. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[25]  Stuart H. Orkin,et al.  Developmental and species-divergent globin switching are driven by BCL11A , 2009, Nature.

[26]  S. Kauppinen,et al.  Therapeutic Silencing of MicroRNA-122 in Primates with Chronic Hepatitis C Virus Infection , 2010, Science.

[27]  J. Herrick,et al.  Peculiar elongated and sickle-shaped red blood corpuscles in a case of severe anemia. 1910. , 2001, The Yale journal of biology and medicine.

[28]  J. Hirschhorn,et al.  Supporting Online Material Materials and Methods Figs. S1 to S10 Tables S1 to S7 References Human Fetal Hemoglobin Expression Is Regulated by the Developmental Stage-specific Repressor Bcl11a , 2022 .

[29]  C. Li,et al.  Model-based analysis of oligonucleotide arrays: expression index computation and outlier detection. , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[30]  P. Sperryn,et al.  Blood. , 1989, British journal of sports medicine.

[31]  E. Rubin,et al.  Transgenic knockout mice with exclusively human sickle hemoglobin and sickle cell disease. , 1997, Science.

[32]  Brad T. Sherman,et al.  DAVID: Database for Annotation, Visualization, and Integrated Discovery , 2003, Genome Biology.

[33]  T. Golub,et al.  Chemical genetic strategy identifies histone deacetylase 1 (HDAC1) and HDAC2 as therapeutic targets in sickle cell disease , 2010, Proceedings of the National Academy of Sciences.

[34]  V. Ingram,et al.  A Specific Chemical Difference Between the Globins of Normal Human and Sickle-Cell Anæmia Hæmoglobin , 1956, Nature.

[35]  Angela N Koehler,et al.  A complex task? Direct modulation of transcription factors with small molecules. , 2010, Current opinion in chemical biology.

[36]  S. Orkin,et al.  Rb intrinsically promotes erythropoiesis by coupling cell cycle exit with mitochondrial biogenesis. , 2007, Genes & development.

[37]  Raymond E. Moellering,et al.  Direct inhibition of the NOTCH transcription factor complex , 2009, Nature.

[38]  B. Pace,et al.  The HDAC inhibitors trichostatin A and suberoylanilide hydroxamic acid exhibit multiple modalities of benefit for the vascular pathobiology of sickle transgenic mice. , 2010, Blood.

[39]  T. Ley,et al.  5-azacytidine selectively increases gamma-globin synthesis in a patient with beta+ thalassemia. , 1982, The New England journal of medicine.

[40]  D. Weatherall,et al.  Hereditary persistence of fetal hemoglobin: a study of 79 affected persons in 15 Negro families in Baltimore. , 1963, Blood.

[41]  R. Rosenfeld.  Nature , 2009, Otolaryngology--head and neck surgery : official journal of American Academy of Otolaryngology-Head and Neck Surgery.

[42]  Thomas Beckers,et al.  Histone Deacetylase Inhibitors Suppress the Inducibility of Nuclear Factor-κB by Tumor Necrosis Factor-α Receptor-1 Down-regulation , 2006 .

[43]  Simon Heath,et al.  A QTL influencing F cell production maps to a gene encoding a zinc-finger protein on chromosome 2p15 , 2007, Nature Genetics.

[44]  D. Kohn,et al.  A Tale of Two SCIDs , 2011, Science Translational Medicine.

[45]  Y. Kan,et al.  Germ-line transmission and developmental regulation of a 150-kb yeast artificial chromosome containing the human beta-globin locus in transgenic mice. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[46]  J. Lieberman,et al.  Knocking down Disease with siRNAs , 2006, Cell.

[47]  S. Orkin,et al.  Therapeutic levels of fetal hemoglobin in erythroid progeny of β-thalassemic CD34+ cells after lentiviral vector-mediated gene transfer. , 2011, Blood.