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VHH212 nanobody targeting the hypoxia-inducible factor 1α suppresses angiogenesis and potentiates gemcitabine therapy in pancreatic cancer in vivo

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Cancer Biology & Medicine 2021年第3期18卷 772-787页

作者： Guangbo Kang Min Hu He Ren Jiewen Wang Xin Cheng Ruowei Li Bo Yuan Yasmine Balan Zixuan Bai He Huang Department of Biochemical Engineering School of Chemical Engineering&TechnologyTianjin UniversityTianjin 300350China Key Laboratory of Systems Bioengineering(Ministry of Education) Tianjin UniversityTianjin 300072China Department of Gastroenterology Center of Tumor Immunology and CytotherapyMedical Research Center of The Affiliated Hospital of Qingdao UniversityQingdao 266003China Department of Chemical and Biological Engineering University of OttawaOttawa KIN 6N5Canada

Objective:We aimed to develop a novel anti-HIF-1αintrabody to decrease gemcitabine resistance in pancreatic cancer ***:Surface plasmon resonance and glutathione S-transferase pull-down assays were conducted to identify the binding affinity and specificity of anti-HIF-1αVHH212[a single-domain antibody(nanobody)].Molecular dynamics simulation was used to determine the protein-protein interactions between hypoxia-inducible factor-1α(HIF-1α)and *** real-time polymerase chain reaction(PCR)and Western blot analyses were performed to identify the expressions of HIF-1αand VEGF-A in pancreatic ductal adenocarcinoma cell *** efficiency of the VHH212 nanobody in inhibiting the HIF-1 signaling pathway was measured using a dual-luciferase reporter ***,a PANC-1 xenograft model was developed to evaluate the anti-tumor efficiency of combined *** analysis was conducted to detect the expressions of HIF-1αand VEGF-A in tumor ***:VHH212 was stably expressed in tumor cells with low cytotoxicity,high affinity,specific subcellular localization,and neutralization of HIF-1αin the cytoplasm or *** binding affinity between VHH212 and the HIF-1αPAS-B domain was 42.7 n *** competitive inhibition of the HIF-1αheterodimer with an aryl hydrocarbon receptor nuclear translocator was used to inhibit the HIF-1/VEGF pathway in *** 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 ***:We developed an anti-HIF-1αnanobody and showed the function of VHH212 in a preclinical murine model of PANC-1 pancreatic *** combination of VHH212 and gemcitabine significantly inhibited tumor *** results suggested that combined use of anti-HIF-1αnanobodies with first-line treatment may in the future be an effective treatment for pancreatic cancer.

关键词： Pancreatic cancer nanobody therapeutic intracellular antibody HIF-1αinhibitor gemcitabine chemosensitizer

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