TY - JOUR
T1 - Intrabody Targeting HIF-1α Mediates Transcriptional Downregulation of Target Genes Related to Solid Tumors
AU - Hu, Yaozhong
AU - Romão, Ema
AU - Vincke, Cécile
AU - Brys, Lea
AU - Elkrim, Yvon
AU - Vandevenne, Marylène
AU - Liu, Changxiao
AU - Muyldermans, Serge
PY - 2021/11/15
Y1 - 2021/11/15
N2 - 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.
AB - 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.
KW - Cell Hypoxia/genetics
KW - Down-Regulation/drug effects
KW - Escherichia coli/genetics
KW - Female
KW - Gene Expression Regulation, Neoplastic
KW - HeLa Cells
KW - Humans
KW - Hypoxia-Inducible Factor 1, alpha Subunit/antagonists & inhibitors
KW - Protein Domains/genetics
KW - Single-Domain Antibodies/genetics
KW - Transcription, Genetic/drug effects
KW - Transfection
KW - Uterine Cervical Neoplasms/genetics
UR - http://www.scopus.com/inward/record.url?scp=85118934585&partnerID=8YFLogxK
U2 - 10.3390/ijms222212335
DO - 10.3390/ijms222212335
M3 - Article
C2 - 34830219
VL - 22
JO - International Journal of Molecular Sciences
JF - International Journal of Molecular Sciences
SN - 1661-6596
IS - 22
M1 - 12335
ER -