Use of blood outgrowth endothelial cells for gene therapy for hemophilia A

Yi Lin, Liming Chang, Anna Solovey, John F. Healey, Pete Lollar, Robert P. Hebbel

Research output: Contribution to journalArticle

130 Citations (Scopus)

Abstract

A culture of human blood outgrowth endothelial cells (BOECs) was established from a sample of peripheral blood and was transfected using a nonviral plasmid carrying complementary DNA for modified human coagulation factor VIII (B domain deleted and replaced with green fluorescence protein). BOECs were then chemically selected, expanded, cryopreserved, and re-expanded in culture. Stably transfected BOECs were administered intravenously daily for 3 days to NOD/SCID mice at 4 cell dose levels (from 5 x 10 4 to 40 x 10 4 cells per injection). In 156 days of observation, mice showed levels of human FVIII that increased with cell dose and time. Mice in all cell dose groups achieved therapeutic levels (more than 10 ng/mL) of human FVIII, and mice in the 3 highest dose groups acquired levels that were normal (100-200 ng/mL) or even above the normal range (highest observed value, 1174 ng/mL). These levels indicate that the BOECs expanded in vivo after administration. When the mice were killed, it was found that BOEC accumulated only in bone marrow and spleen and that these cells retained endothelial phenotype and transgene expression. Cell doses used here would make scale-up to humans feasible. Thus, the use of engineered autologous BOECs, which here resulted in sustained and therapeutic levels of FVIII, may comprise an effective therapeutic strategy for use in gene therapy for hemophilia A.

Original languageEnglish (US)
Pages (from-to)457-462
Number of pages6
JournalBlood
Volume99
Issue number2
DOIs
StatePublished - Jan 15 2002
Externally publishedYes

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Gene therapy
Endothelial cells
Hemophilia A
Cell- and Tissue-Based Therapy
Genetic Therapy
Blood
Endothelial Cells
Cell culture
Inbred NOD Mouse
SCID Mice
Factor VIII
Therapeutic Uses
Transgenes
Bone Marrow Cells
Bone
Reference Values
Plasmids
Spleen
Complementary DNA
Fluorescence

ASJC Scopus subject areas

  • Hematology

Cite this

Lin, Y., Chang, L., Solovey, A., Healey, J. F., Lollar, P., & Hebbel, R. P. (2002). Use of blood outgrowth endothelial cells for gene therapy for hemophilia A. Blood, 99(2), 457-462. https://doi.org/10.1182/blood.V99.2.457

Use of blood outgrowth endothelial cells for gene therapy for hemophilia A. / Lin, Yi; Chang, Liming; Solovey, Anna; Healey, John F.; Lollar, Pete; Hebbel, Robert P.

In: Blood, Vol. 99, No. 2, 15.01.2002, p. 457-462.

Research output: Contribution to journalArticle

Lin, Y, Chang, L, Solovey, A, Healey, JF, Lollar, P & Hebbel, RP 2002, 'Use of blood outgrowth endothelial cells for gene therapy for hemophilia A', Blood, vol. 99, no. 2, pp. 457-462. https://doi.org/10.1182/blood.V99.2.457
Lin Y, Chang L, Solovey A, Healey JF, Lollar P, Hebbel RP. Use of blood outgrowth endothelial cells for gene therapy for hemophilia A. Blood. 2002 Jan 15;99(2):457-462. https://doi.org/10.1182/blood.V99.2.457
Lin, Yi ; Chang, Liming ; Solovey, Anna ; Healey, John F. ; Lollar, Pete ; Hebbel, Robert P. / Use of blood outgrowth endothelial cells for gene therapy for hemophilia A. In: Blood. 2002 ; Vol. 99, No. 2. pp. 457-462.
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