Electroporation of synthetic oligodeoxynucleotides

A novel technique for ex vivo bone marrow purging

Raymond Bergan, Frances Hakim, Gretchen N. Schwartz, Edward Kyle, Rose Cepada, Janet M. Szabo, Daniel Fowler, Ronald Gress, Len Neckers

Research output: Contribution to journalArticle

67 Citations (Scopus)

Abstract

Recent data suggest that tumor cells contaminating reinfused bone marrow may contribute to relapse in patients undergoing autologous bone marrow transplantation. Purging strategies that are able to remove these contaminating tumor cells need to be developed. This study describes how electroporation (EP) can be used to improve intracellular delivery of synthetic antisense oligodeoxynucleotides (ODNs), thereby enhancing their ability to suppress a target protein. Antisense ODNs that were introduced into cells by EP led to immediate suppression of targeted c-myc protein; this was associated with rapid cell death in the diffuse histiocytic lymphoma, U937; Burkitt's lymphoma, ST486; breast carcinoma, MCF-7; and Ewing's sarcoma, CHP-100, cell lines. Electroporation was found to have little or no detrimental effect on cells responsible for murine hematopoietic long-term reconstitution as determined from in vivo competitive repopulation studies. Using human c-myc-directed antisense ODNs as a model for the application of this approach to bone marrow purging, selective killing of human lymphoma U937 cells relative to normal human bone marrow cells was shown in cell mixing studies. In vivo studies were performed in which a survival advantage was shown for athymic mice that were inoculated with antisense-treated U937 cells as opposed to control cells. These studies suggest that EP of bone marrow may be of use in enhancing intracellular delivery of a variety of molecular/pharmaceutical agents. Taken together, these data suggest that the use of electroporation to enhance delivery of antisense ODNs is a promising new approach towards ex vivo bone marrow purging.

Original languageEnglish (US)
Pages (from-to)731-741
Number of pages11
JournalBlood
Volume88
Issue number2
StatePublished - Jul 15 1996
Externally publishedYes

Fingerprint

Bone Marrow Purging
Purging
Electroporation
Oligodeoxyribonucleotides
Bone
Cells
U937 Cells
Tumors
Bone Marrow
Proto-Oncogene Proteins c-myc
Ewing's Sarcoma
Burkitt Lymphoma
Lymphoma, Large B-Cell, Diffuse
Autologous Transplantation
Cell death
Bone Marrow Transplantation
Nude Mice
Bone Marrow Cells
Lymphoma
Neoplasms

ASJC Scopus subject areas

  • Hematology

Cite this

Bergan, R., Hakim, F., Schwartz, G. N., Kyle, E., Cepada, R., Szabo, J. M., ... Neckers, L. (1996). Electroporation of synthetic oligodeoxynucleotides: A novel technique for ex vivo bone marrow purging. Blood, 88(2), 731-741.

Electroporation of synthetic oligodeoxynucleotides : A novel technique for ex vivo bone marrow purging. / Bergan, Raymond; Hakim, Frances; Schwartz, Gretchen N.; Kyle, Edward; Cepada, Rose; Szabo, Janet M.; Fowler, Daniel; Gress, Ronald; Neckers, Len.

In: Blood, Vol. 88, No. 2, 15.07.1996, p. 731-741.

Research output: Contribution to journalArticle

Bergan, R, Hakim, F, Schwartz, GN, Kyle, E, Cepada, R, Szabo, JM, Fowler, D, Gress, R & Neckers, L 1996, 'Electroporation of synthetic oligodeoxynucleotides: A novel technique for ex vivo bone marrow purging', Blood, vol. 88, no. 2, pp. 731-741.
Bergan R, Hakim F, Schwartz GN, Kyle E, Cepada R, Szabo JM et al. Electroporation of synthetic oligodeoxynucleotides: A novel technique for ex vivo bone marrow purging. Blood. 1996 Jul 15;88(2):731-741.
Bergan, Raymond ; Hakim, Frances ; Schwartz, Gretchen N. ; Kyle, Edward ; Cepada, Rose ; Szabo, Janet M. ; Fowler, Daniel ; Gress, Ronald ; Neckers, Len. / Electroporation of synthetic oligodeoxynucleotides : A novel technique for ex vivo bone marrow purging. In: Blood. 1996 ; Vol. 88, No. 2. pp. 731-741.
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