Intrabody targeting HIF-1α mediates transcriptional downregulation of target genes related to solid tumors

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Keywords :

HIF-1α; Nanobodies; Target genes; Transcriptional activation; HIF1A protein, human; Escherichia coli; HeLa cell line; Cell Hypoxia; Down-Regulation; Female; Gene Expression Regulation, Neoplastic; HeLa Cells; Humans; Hypoxia-Inducible Factor 1, alpha Subunit; Protein Domains; Single-Domain Antibodies; Transcription, Genetic; Transfection; Uterine Cervical Neoplasms

Abstract :

[en] Uncontrolled growth of solid tumors will result in a hallmark hypoxic condition, whereby the key transcriptional regulator of hypoxia inducible factor-1α (HIF-1α) will be stabilized to activate the transcription of target genes that are responsible for the metabolism, proliferation, and metastasis of tumor cells. Targeting and inhibiting the transcriptional activity of HIF-1 may provide an interesting strategy for cancer therapy. In the present study, an immune library and a synthetic library were constructed for the phage display selection of Nbs against recombinant PAS B domain protein (rPasB) of HIF-1α. After panning and screening, seven different nanobodies (Nbs) were selected, of which five were confirmed via immunoprecipitation to target the native HIF-1α subunit. The inhibitory effect of the selected Nbs on HIF-1 induced activation of target genes has been evaluated after intracellular expression of these Nbs in HeLa cells. The dramatic inhibition of both intrabody formats on the expression of HIF-1-related target genes has been confirmed, which indicated the inhibitory efficacy of selected Nbs on the transcriptional activity of HIF-1. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.

Disciplines :

Biochemistry, biophysics & molecular biology

Author, co-author :

Hu, Y.; Research Institute of Public Health, School of Medicine, Nankai University, Tianjin, 300071, China, Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, Brussels, 1050, Belgium

Romão, E.; Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, Brussels, 1050, Belgium

Vincke, C.; Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, Brussels, 1050, Belgium

Brys, L.; Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, Brussels, 1050, Belgium

Elkrim, Y.; Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, Brussels, 1050, Belgium

Vandevenne, Marylène ; Université de Liège - ULiège

Liu, C.; State Key Laboratory of Drug Delivery Technology and Pharmacokinetics, Tianjin Institute of Pharmaceutical Research, Tianjin, 300301, China

Muyldermans, S.; Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, Brussels, 1050, Belgium

Language :

English

Title :

Intrabody targeting HIF-1α mediates transcriptional downregulation of target genes related to solid tumors

Publication date :

2021

Journal title :

International Journal of Molecular Sciences

ISSN :

1661-6596

eISSN :

1422-0067

Publisher :

Multidisciplinary Digital Publishing Institute (MDPI), Switzerland

Volume :

Issue :

Pages :

12335

Peer reviewed :

Peer Reviewed verified by ORBi

Available on ORBi :

since 30 January 2022

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