Targeting allosteric site of AKT by 5,7-dimethoxy-1,4-phenanthrenequinone suppresses neutrophilic inflammation

Autor: Fang Rong Chang, Chih-Ching Wu, Po-Jen Chen, Hao-Chun Hu, Tsong-Long Hwang, Chang-Yu Pan, Cheng-Yu Lin, Yann-Lii Leu, I-Ling Ko, Yung-Fong Tsai, Yang Chang Wu, Chia Lin Lee
Rok vydání: 2019
Předmět:
MAPK/ERK pathway
Models
Molecular

0301 basic medicine
Research paper
Lipopolysaccharide
Neutrophils
p38 mitogen-activated protein kinases
Anti-Inflammatory Agents
Molecular Conformation
HL-60 Cells
Inflammation
Pharmacology
Lung injury
General Biochemistry
Genetics and Molecular Biology

chemistry.chemical_compound
Structure-Activity Relationship
03 medical and health sciences
0302 clinical medicine
Acute lung injury
Cell Adhesion
medicine
Humans
Protein Interaction Domains and Motifs
Protein kinase B
business.industry
Chemistry
AKT
Neutrophil
Quinones
Degranulation
Chemotaxis
5
7-dimethoxy-1
4-phenanthrenequinone

General Medicine
Phenanthrenes
Respiratory burst
Chemotaxis
Leukocyte

030104 developmental biology
Neutrophil Infiltration
030220 oncology & carcinogenesis
Cancer research
Phosphorylation
medicine.symptom
business
Reactive Oxygen Species
Proto-Oncogene Proteins c-akt
Allosteric Site
Protein Binding
Zdroj: EBioMedicine
ISSN: 2352-3964
DOI: 10.1016/j.ebiom.2019.01.043
Popis: Background: Acute lung injury (ALI) is a severe life-threatening inflammatory disease. Neutrophil activation is a major pathogenic factor in ALI. Protein kinase B (PKB)/AKT regulates diverse cellular responses, but the significance in neutrophilic inflammation and ALI remains unknown. Methods: Human neutrophils and neutrophil-like differentiated HL-60 (dHL-60) cells were used to examine the anti-inflammatory effects of 5,7-dimethoxy-1,4-phenanthrenequinone (CLLV-1). The therapeutic potential of CLLV-1 was determined in a mouse model of lipopolysaccharide (LPS)-induced ALI. Findings: CLLV-1 inhibited respiratory burst, degranulation, adhesion, and chemotaxis in human neutrophils and dHL-60 cells. CLLV-1 inhibited the phosphorylation of AKT (Thr308 and Ser473), but not of ERK, JNK, or p38. Furthermore, CLLV-1 blocked AKT activity and covalently reacted with AKT Cys310 in vitro. The AKT309-313 peptide-CLLV-1 adducts were determined by NMR or mass spectrometry assay. The alkylation agent-conjugated AKT (reduced form) level was also inhibited by CLLV-1. Significantly, CLLV-1 ameliorated LPS-induced ALI, neutrophil infiltration, and AKT activation in mice. Interpretation: Our results identify CLLV-1 as a covalent allosteric AKT inhibitor by targeting AKT Cys310. CLLV-1 shows potent anti-inflammatory activity in human neutrophils and LPS-induced mouse ALI. Our findings provide a mechanistic framework for redox modification of AKT that may serve as a novel pharmacological target to alleviate neutrophilic inflammation. Funding Statement: This research was financial supported by the grants from the Ministry of Science Technology (MOST 106-2320-B-255-003-MY3 and MOST 104-2320-B-255-004-MY3), Ministry of Education (EMRPD1G0231 and EMRPD1H0381), and Chang Gung Memorial Hospital (CMRPF1F0011~3, CMRPF1F0061~3, CMRPF1G0241~3, and BMRP450), Taiwan. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Declaration of Interests: The authors declare that no competing interests exist. Ethics Approval Statement: The procedure of neutrophil isolation was approved by the Institutional Review Board at Chang Gung Memorial Hospital. ALI was induced by intra-tracheal spray of 2 mg/kg LPS (Escherichia coli 0111:B4) in seven to eight weeks old C57BL/6 male mice, according to the guidelines and approved by Institutional Animal Care and Use Committee of Chang Gung University, Taiwan.
Databáze: OpenAIRE
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