Delivery of small interfering RNA for inhibition of endothelial cell apoptosis by hypoxia and serum deprivation

Seung-Woo Cho, Lauren Hartle, Sun Mi Son, Fan Yang, Michael Goldberg, Qiaobing Xu, Robert Langer, Daniel G. Anderson

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

RNA interference (RNAi) for anti-angiogenic or pro-apoptotic factors in endothelial cells (ECs) has great potential for the treatment of ischemic diseases by promoting angiogenesis or inhibiting apoptosis. Here, we report the utility of small interfering RNA (siRNA) in inhibiting EC apoptosis induced by tumor necrosis factor-α (TNF-α). siRNA was designed and synthesized targeting tumor necrosis factor-α receptor-1 (TNFR-1) and Src homology 2 domain-containing protein tyrosine phosphatase-1 (SHP-1). Human umbilical vein endothelial cells (HUVECs) were cultured under in vitro hypoxic and serum-deprived conditions to simulate in vivo ischemic conditions. Two days after liposomal delivery of siRNA targeting TNFR-1 and SHP-1, significant silencing of each target (TNFR-1; 76.5% and SHP-1; 97.2%) was detected. Under serum-deprived hypoxic (1% oxygen) conditions, TNF-α expression in HUVECs increased relative to normoxic (20% oxygen) and serum-containing conditions. Despite enhanced TNF-α expression, suppression of TNFR-1 or SHP-1 by siRNA delivery not only enhanced expression of angiogenic factors (KDR/Flk-1 and eNOS) and anti-apoptotic factor (Bcl-xL) but also reduced expression of a pro-apoptotic factor (Bax). Transfection of TNFR-1 or SHP-1 siRNA significantly decreased the HUVEC apoptosis while significantly enhancing HUVEC proliferation and capillary formation. The present study demonstrates that TNFR-1 and SHP-1 may be useful targets for the treatment of myocardial or hindlimb ischemia.

Original languageEnglish
Pages (from-to)158-163
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume376
Issue number1
DOIs
Publication statusPublished - 2008 Nov 7

Fingerprint

Cell Hypoxia
Tumor Necrosis Factor Receptors
Endothelial cells
Small Interfering RNA
Endothelial Cells
Human Umbilical Vein Endothelial Cells
Apoptosis
Serum
Tumor Necrosis Factor-alpha
SH2 Domain-Containing Protein Tyrosine Phosphatases
Non-Receptor Type 6 Protein Tyrosine Phosphatase
Oxygen
Protein Phosphatase 1
Protein Tyrosine Phosphatases
Angiogenesis Inducing Agents
Cell proliferation
Hindlimb
RNA Interference
Transfection
Ischemia

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Cho, Seung-Woo ; Hartle, Lauren ; Son, Sun Mi ; Yang, Fan ; Goldberg, Michael ; Xu, Qiaobing ; Langer, Robert ; Anderson, Daniel G. / Delivery of small interfering RNA for inhibition of endothelial cell apoptosis by hypoxia and serum deprivation. In: Biochemical and Biophysical Research Communications. 2008 ; Vol. 376, No. 1. pp. 158-163.
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abstract = "RNA interference (RNAi) for anti-angiogenic or pro-apoptotic factors in endothelial cells (ECs) has great potential for the treatment of ischemic diseases by promoting angiogenesis or inhibiting apoptosis. Here, we report the utility of small interfering RNA (siRNA) in inhibiting EC apoptosis induced by tumor necrosis factor-α (TNF-α). siRNA was designed and synthesized targeting tumor necrosis factor-α receptor-1 (TNFR-1) and Src homology 2 domain-containing protein tyrosine phosphatase-1 (SHP-1). Human umbilical vein endothelial cells (HUVECs) were cultured under in vitro hypoxic and serum-deprived conditions to simulate in vivo ischemic conditions. Two days after liposomal delivery of siRNA targeting TNFR-1 and SHP-1, significant silencing of each target (TNFR-1; 76.5{\%} and SHP-1; 97.2{\%}) was detected. Under serum-deprived hypoxic (1{\%} oxygen) conditions, TNF-α expression in HUVECs increased relative to normoxic (20{\%} oxygen) and serum-containing conditions. Despite enhanced TNF-α expression, suppression of TNFR-1 or SHP-1 by siRNA delivery not only enhanced expression of angiogenic factors (KDR/Flk-1 and eNOS) and anti-apoptotic factor (Bcl-xL) but also reduced expression of a pro-apoptotic factor (Bax). Transfection of TNFR-1 or SHP-1 siRNA significantly decreased the HUVEC apoptosis while significantly enhancing HUVEC proliferation and capillary formation. The present study demonstrates that TNFR-1 and SHP-1 may be useful targets for the treatment of myocardial or hindlimb ischemia.",
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Delivery of small interfering RNA for inhibition of endothelial cell apoptosis by hypoxia and serum deprivation. / Cho, Seung-Woo; Hartle, Lauren; Son, Sun Mi; Yang, Fan; Goldberg, Michael; Xu, Qiaobing; Langer, Robert; Anderson, Daniel G.

In: Biochemical and Biophysical Research Communications, Vol. 376, No. 1, 07.11.2008, p. 158-163.

Research output: Contribution to journalArticle

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