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VHH212 nanobody targeting the hypoxia-inducible factor 1α suppresses angiogenesis and potentiates gemcitabine therapy in pancreatic cancer in vivo
- Source :
- Cancer Biology & Medicine, Cancer Biology & Medicine, Vol 18, Iss 3, Pp 772-787 (2021)
- Publication Year :
- 2021
- Publisher :
- Compuscript, 2021.
-
Abstract
- Objective: We aimed to develop a novel anti-HIF-1α intrabody to decrease gemcitabine resistance in pancreatic cancer patients. Methods: Surface plasmon resonance and glutathione S-transferase pull-down assays were conducted to identify the binding affinity and specificity of anti-HIF-1α VHH212 [a single-domain antibody (nanobody)]. Molecular dynamics simulation was used to determine the protein-protein interactions between hypoxia-inducible factor-1α (HIF-1α) and VHH212. The real-time polymerase chain reaction (PCR) and Western blot analyses were performed to identify the expressions of HIF-1α and VEGF-A in pancreatic ductal adenocarcinoma cell lines. The efficiency of the VHH212 nanobody in inhibiting the HIF-1 signaling pathway was measured using a dual-luciferase reporter assay. Finally, a PANC-1 xenograft model was developed to evaluate the anti-tumor efficiency of combined treatment. Immunohistochemistry analysis was conducted to detect the expressions of HIF-1α and VEGF-A in tumor tissues. Results: VHH212 was stably expressed in tumor cells with low cytotoxicity, high affinity, specific subcellular localization, and neutralization of HIF-1α in the cytoplasm or nucleus. The binding affinity between VHH212 and the HIF-1α PAS-B domain was 42.7 nM. Intrabody competitive inhibition of the HIF-1α heterodimer with an aryl hydrocarbon receptor nuclear translocator was used to inhibit the HIF-1/VEGF pathway in vitro. Compared with single agent gemcitabine, co-treatment with gemcitabine and a VHH212-encoding adenovirus significantly suppressed tumor growth in the xenograft model with 80.44% tumor inhibition. Conclusions: We developed an anti-HIF-1α nanobody and showed the function of VHH212 in a preclinical murine model of PANC-1 pancreatic cancer. The combination of VHH212 and gemcitabine significantly inhibited tumor development. These results suggested that combined use of anti-HIF-1α nanobodies with first-line treatment may in the future be an effective treatment for pancreatic cancer.
- Subjects :
- Cancer Research
Aryl hydrocarbon receptor nuclear translocator
Angiogenesis
Chemosensitizer
HIF-1α
intracellular antibody
Intrabody
In vivo
Pancreatic cancer
medicine
RC254-282
biology
Chemistry
nanobody therapeutic
gemcitabine
Neoplasms. Tumors. Oncology. Including cancer and carcinogens
Cancer
medicine.disease
Gemcitabine
inhibitor
Oncology
chemosensitizer
Cancer research
biology.protein
Original Article
medicine.drug
Subjects
Details
- Language :
- English
- ISSN :
- 20953941
- Volume :
- 18
- Issue :
- 3
- Database :
- OpenAIRE
- Journal :
- Cancer Biology & Medicine
- Accession number :
- edsair.doi.dedup.....fb801dcba1eadb2b104dbfa94424c63f