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Blood, 6 August 2009, Vol. 114, No. 6, pp. 1174-1185. Prepublished online as a Blood First Edition Paper on May 27, 2009; DOI 10.1182/blood-2009-01-201863. This Article Full Text Full Text (PDF) Supplemental Methods and Figures All Versions of this Article: blood-2009-01-201863v1 114/6/1174    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via CrossRef Google Scholar Articles by Perumbeti, A. Articles by Malik, P. PubMed PubMed Citation Articles by Perumbeti, A. Articles by Malik, P. Related Collections Red Cells, Iron, and Erythropoiesis Gene Therapy Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  GENE THERAPY A novel human gamma-globin gene vector for genetic correction of sickle cell anemia in a humanized sickle mouse model: critical determinants for successful correction Ajay Perumbeti1,2,*, Tomoyasu Higashimoto*,1, Fabrizia Urbinati1, Robert Franco3, Herbert J. Meiselman4, David Witte5, and Punam Malik1,2 Divisions of 1 Experimental Hematology/Cancer Biology and 2 Hematology-Oncology, Cincinnati Children's Research Foundation, Cincinnati Children's Hospital Medical Center (CCHMC), OH; 3 Division of Hematology-Oncology, University of Cincinnati College of Medicine, OH; 4 Department of Physiology, Keck School of Medicine, University of Southern California, Los Angeles; and 5 Division of Pathology, University of Cincinnati College of Medicine, OH We show that lentiviral delivery of human -globin gene under β-globin regulatory control elements in hematopoietic stem cells (HSCs) results in sufficient postnatal fetal hemoglobin (HbF) expression to correct sickle cell anemia (SCA) in the Berkeley "humanized" sickle mouse. Upon de-escalating the amount of transduced HSCs in transplant recipients, using reduced-intensity conditioning and varying gene transfer efficiency and vector copy number, we assessed critical parameters needed for correction. A systematic quantification of functional and hematologic red blood cell (RBC) indices, organ pathology, and life span was used to determine the minimal amount of HbF, F cells, HbF/F-cell, and gene-modified HSCs required for correcting the sickle phenotype. We show that long-term amelioration of disease occurred (1) when HbF exceeded 10%, F cells constituted two-thirds of the circulating RBCs, and HbF/F cell was one-third of the total hemoglobin in sickle RBCs; and (2) when approximately 20% gene-modified HSCs repopulated the marrow. Moreover, we show a novel model using reduced-intensity conditioning to determine genetically corrected HSC threshold that corrects a hematopoietic disease. These studies provide a strong preclinical model for what it would take to genetically correct SCA and are a foundation for the use of this vector in a human clinical trial. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

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