Polyethyleneimine grafted with pluronic P85 enhances Ku86 antisense delivery and the ionizing radiation treatment efficacy in vivo

A. I. Belenkov, V. Y. Alakhov, A. V. Kabanov, Serguei V Vinogradov, L. C. Panasci, B. P. Monia, T. Y.K. Chow

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

In an effort to improve the efficacy of antisense delivery, we evaluated polyethyleneimine (PEI, 2 kDa) alone or grafted with nonionic amphiphilic block copolymer Pluronic® (P85) as a carrier for Ku86 antisense oligonucleotide (ASO) delivery. Ku86 is an abundant nuclear protein that plays an important role in nonhomologous DNA end joining and has implications in tumorigenesis and acquired drug resistance. Transfection of adherent and suspension cell lines with Ku86 ASOs complexed with P85-g-PEI (2 kDa) conjugates was associated with a specific decrease in Ku86 mRNA levels (EC50<75nM and EC50<250 nM, respectively, n=3). More importantly, no requirement for reduced serum conditions was necessary during transfection. In contrast, whereas Ku86 ASOs complexed with PEI (2 kDa) alone were effective in decreasing Ku86 mRNA levels in adherent cell lines (EC50<75 nM, n=3), the formulation did not produce any detectable decrease in Ku86 mRNA levels in suspension cell lines. Transfection of adherent cell lines with 500 nM Ku86 ASOs formulated with P85-g-PEI (2 kDa) was associated with a specific decrease (<10% remaining of control) in Ku86 protein expression and a two-fold increased cell death after treatment with ionizing radiation (IR). In athymic nude mice bearing subcutaneous human HT29 colon adenocarcinoma xenografts, Ku86 ASO-P85-g-PEI (2 kDa) administration (15 mg/kg, subcutaneously) with a Q1D × 7 treatment schedule, when combined with a single dose of IR (6 Gy), caused a significant inhibition of HT29 tumor growth compared with mismatch- and naked antisense-pretreated control groups (time from 200 to 1000mm3, 126.9 versus 84.18 and 87.76 days, P<0.005). A potentiation of the antitumor activity was observed in all mice treated with Ku86 ASO-P85-g-PEI (2 kDa) formulation; however, tumor growth inhibition was reversible upon treatment cessation. No morbidity/mortality or changes in histopathology were observed under this treatment regiment. Our results indicate that P85-g-PEI (2 kDa) conjugates may increase the efficacy of Ku86 ASO delivery in management of resistant malignancies, thus providing a rationale for their evaluation in cancer patients in combination with conventional anticancer therapies.

Original languageEnglish (US)
Pages (from-to)1665-1672
Number of pages8
JournalGene Therapy
Volume11
Issue number22
DOIs
StatePublished - Nov 1 2004

Fingerprint

Polyethyleneimine
Poloxamer
Antisense Oligonucleotides
Ionizing Radiation
Transfection
Cell Line
Nude Mice
Messenger RNA
Neoplasms
Suspensions
DNA End-Joining Repair
Withholding Treatment
Therapeutics
Growth
Nuclear Proteins
Drug Resistance
Heterografts
Appointments and Schedules
Colon
Carcinogenesis

Keywords

  • Antisense
  • Block copolymer
  • DNA-PK
  • Ionizing radiation
  • Ku86
  • Mouse xenografts
  • Poloxamer
  • Polyethyleneimine

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

Belenkov, A. I., Alakhov, V. Y., Kabanov, A. V., Vinogradov, S. V., Panasci, L. C., Monia, B. P., & Chow, T. Y. K. (2004). Polyethyleneimine grafted with pluronic P85 enhances Ku86 antisense delivery and the ionizing radiation treatment efficacy in vivo. Gene Therapy, 11(22), 1665-1672. https://doi.org/10.1038/sj.gt.3302355

Polyethyleneimine grafted with pluronic P85 enhances Ku86 antisense delivery and the ionizing radiation treatment efficacy in vivo. / Belenkov, A. I.; Alakhov, V. Y.; Kabanov, A. V.; Vinogradov, Serguei V; Panasci, L. C.; Monia, B. P.; Chow, T. Y.K.

In: Gene Therapy, Vol. 11, No. 22, 01.11.2004, p. 1665-1672.

Research output: Contribution to journalArticle

Belenkov, AI, Alakhov, VY, Kabanov, AV, Vinogradov, SV, Panasci, LC, Monia, BP & Chow, TYK 2004, 'Polyethyleneimine grafted with pluronic P85 enhances Ku86 antisense delivery and the ionizing radiation treatment efficacy in vivo', Gene Therapy, vol. 11, no. 22, pp. 1665-1672. https://doi.org/10.1038/sj.gt.3302355
Belenkov, A. I. ; Alakhov, V. Y. ; Kabanov, A. V. ; Vinogradov, Serguei V ; Panasci, L. C. ; Monia, B. P. ; Chow, T. Y.K. / Polyethyleneimine grafted with pluronic P85 enhances Ku86 antisense delivery and the ionizing radiation treatment efficacy in vivo. In: Gene Therapy. 2004 ; Vol. 11, No. 22. pp. 1665-1672.
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AU - Belenkov, A. I.

AU - Alakhov, V. Y.

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AU - Vinogradov, Serguei V

AU - Panasci, L. C.

AU - Monia, B. P.

AU - Chow, T. Y.K.

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