Globin vector regulatory elements are active in early hematopoietic progenitor cells

Mol Ther. 2022 Jun 1;30(6):2199-2209. doi: 10.1016/j.ymthe.2022.02.028. Epub 2022 Mar 2.


The globin genes are archetypal tissue-specific genes that are silent in most tissues but for late-stage erythroblasts upon terminal erythroid differentiation. The transcriptional activation of the β-globin gene is under the control of proximal and distal regulatory elements located on chromosome 11p15.4, including the β-globin locus control region (LCR). The incorporation of selected LCR elements in lentiviral vectors encoding β and β-like globin genes has enabled successful genetic treatment of the β-thalassemias and sickle cell disease. However, recent occurrences of benign clonal expansions in thalassemic patients and myelodysplastic syndrome in patients with sickle cell disease call attention to the non-erythroid functions of these powerful vectors. Here we demonstrate that lentivirally encoded LCR elements, in particular HS1 and HS2, can be activated in early hematopoietic cells including hematopoietic stem cells and myeloid progenitors. This activity is position-dependent and results in the transcriptional activation of a nearby reporter gene in these progenitor cell populations. We further show that flanking a globin vector with an insulator can effectively restrain this non-erythroid activity without impairing therapeutic globin expression. Globin lentiviral vectors harboring powerful LCR HS elements may thus expose to the risk of trans-activating cancer-related genes, which can be mitigated by a suitable insulator.

Keywords: ATAC-seq; beta-globin gene; erythroid expression; gene therapy; hematopoietic progenitor cells; hemoglobinopathies; lentiviral vector; locus control region; sickle cell disease; thalassemia.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, N.I.H., Extramural

MeSH terms

  • Anemia, Sickle Cell* / genetics
  • Genetic Therapy / methods
  • Genetic Vectors / genetics
  • Globins* / genetics
  • Hematopoietic Stem Cells / metabolism
  • Humans
  • beta-Globins / genetics
  • beta-Globins / metabolism


  • beta-Globins
  • Globins