Characterization and mechanistic studies of a novel melanoma-targeting construct containing IκBa for specific inhibition of nuclear factor-κB activity.

Hong Zhou, Yuying Liu, Lawrence H. Cheung, Sehoon Kim, Weihe Zhang, Khalid A. Mohamedali, Preetha Anand, Walter N. Hittelman, Bharat B. Aggarwal, Michael G. Rosenblum

Research output: Contribution to journalArticle

Abstract

The transcription factor nuclear factor-κB (NF-κB) is a central mediator of growth and homeostasis for both normal and neoplastic cells. IκBα is the natural intracellular inhibitor of NF-κB and can effectively complex with and thereby inhibit the biologic activity and translocation of NF-κB to the nucleus. We designed a fusion protein designated IκBα/scFvMEL composing of human IκBα and the single-chain antibody scFvMEL, targets melanoma gp240 antigen. Cells treated with IκBα/scFvMEL before irradiation showed specifically inhibition of both constitutive and radiation-induced NF-κB activity on gp240 antigen-positive A375M cells. Pretreatment of A375M cells with IκBα/scFvMEL significantly sensitized melanoma cells to ionizing radiation assessed using a clonogenic survival assay. Mechanistic studies showed that IκBα/scFvMEL, when exogenously added to A375M cells, could be coimmunoprecipitated with the p65 subunit of NF-κB. IκBα/scFvMEL inhibited in a time and/or dose-dependent manner of tumor necrosis factor α- or radiation-induced NF-κB activity in vitro. IκBα/scFvMEL was also shown to specifically inhibit the translocation of the NF-κB p65 subunit to the cell nucleus and NF-κB-mediated gene transcription. Further, initial studies showed that mice bearing well-established A375M xenografts were treated (intravenously) with IκBα/scFvMEL and showed a significant suppression of tumor growth. We also observed a decrease in levels of Bcl-2 and Bcl-XL signaling events downstream of NF-κB in the tumor model. These studies demonstrate for the first time that tumor cell-targeted delivery of IκBα may be beneficial for the treatment of melanoma when combined with standard anticancer therapies such as radiation.

Original languageEnglish (US)
Pages (from-to)766-777
Number of pages12
JournalNeoplasia (New York, N.Y.)
Volume12
Issue number10
StatePublished - Jan 1 2010

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Melanoma
Radiation
Melanoma-Specific Antigens
Single-Chain Antibodies
Neoplasms
Growth
Ionizing Radiation
Cell Nucleus
Heterografts
Homeostasis
Transcription Factors
Tumor Necrosis Factor-alpha
Antigens
Survival
Genes
Proteins

Cite this

Zhou, H., Liu, Y., Cheung, L. H., Kim, S., Zhang, W., Mohamedali, K. A., ... Rosenblum, M. G. (2010). Characterization and mechanistic studies of a novel melanoma-targeting construct containing IκBa for specific inhibition of nuclear factor-κB activity. Neoplasia (New York, N.Y.), 12(10), 766-777.

Characterization and mechanistic studies of a novel melanoma-targeting construct containing IκBa for specific inhibition of nuclear factor-κB activity. / Zhou, Hong; Liu, Yuying; Cheung, Lawrence H.; Kim, Sehoon; Zhang, Weihe; Mohamedali, Khalid A.; Anand, Preetha; Hittelman, Walter N.; Aggarwal, Bharat B.; Rosenblum, Michael G.

In: Neoplasia (New York, N.Y.), Vol. 12, No. 10, 01.01.2010, p. 766-777.

Research output: Contribution to journalArticle

Zhou, H, Liu, Y, Cheung, LH, Kim, S, Zhang, W, Mohamedali, KA, Anand, P, Hittelman, WN, Aggarwal, BB & Rosenblum, MG 2010, 'Characterization and mechanistic studies of a novel melanoma-targeting construct containing IκBa for specific inhibition of nuclear factor-κB activity.', Neoplasia (New York, N.Y.), vol. 12, no. 10, pp. 766-777.
Zhou, Hong ; Liu, Yuying ; Cheung, Lawrence H. ; Kim, Sehoon ; Zhang, Weihe ; Mohamedali, Khalid A. ; Anand, Preetha ; Hittelman, Walter N. ; Aggarwal, Bharat B. ; Rosenblum, Michael G. / Characterization and mechanistic studies of a novel melanoma-targeting construct containing IκBa for specific inhibition of nuclear factor-κB activity. In: Neoplasia (New York, N.Y.). 2010 ; Vol. 12, No. 10. pp. 766-777.
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