Adipose specific aptamer adipo-8 recognizes and interacts with APMAP to ameliorates fat deposition in vitro and in vivo

Life Sci. 2020 Jun 15;251:117609. doi: 10.1016/j.lfs.2020.117609. Epub 2020 Apr 6.

Abstract

Aims: To identify the target of an adipose specific aptamer adipo-8, predict the potential interaction between adipo-8 and its target, and investigate lipid-lowering effect of adipo-8 in vitro and in vivo.

Main methods: Distinct membranous protein of 3T3-L1 adipocyte pulled-down by adipo-8 was mass-spectrometry analyzed as target candidate(s), and affinity of adipo-8 to target protein-silent adipocyte was detected to validate it. Interaction between adipo-8 and target was predicted by bioinformatic analysis, further confirmed by aptamer truncation and competitive binding assay. To investigate lipid-lowering effect of adipo-8 and mechanism behind, 250 nmol/L adipo-8 or library was incubated with 3T3-L1 adipocyte or target-protein-silent adipocyte for 24 h, and 0.01 μg/g/day adipo-8 or library was administrated to high-fat-fed male mice for 21 days.

Key findings: APMAP (Adipocyte Plasma Membrane Associated Protein) was identified as adipo-8 target, and adipo-8 affinity to adipocytes was in proportional to APMAP expression. Docking model between the stem-loop structure of adipo-8 and APMAP were predicted that adipo-8 was likely to interact with APMAP at its amino-acid 275-411 sequence. Moreover, adipo-8 could ameliorate fat deposition through interaction with APMAP in vitro, and administration of adipo-8 in high-fat-diet fed mice resulted in body weight loss and blood triglyceride decrease without liver or renal dysfunction.

Significance: Adipo-8 could recognize APMAP specifically and interact with its targets to ameliorate fat deposition in vitro and in vivo. Aptamer adipo-8 has potential to act as an effective and safe targeted drug for obesity and obesity related diseases.

Keywords: APMAP; Adipogenesis; Aptamer; Obesity; Targeted therapy.

MeSH terms

  • 3T3-L1 Cells
  • Adipocytes / metabolism*
  • Adipose Tissue / metabolism*
  • Animals
  • Aptamers, Nucleotide / metabolism*
  • Diet, High-Fat
  • Male
  • Mass Spectrometry
  • Membrane Glycoproteins / metabolism*
  • Mice
  • Mice, Inbred C57BL
  • Molecular Docking Simulation

Substances

  • APMAP protein, mouse
  • Aptamers, Nucleotide
  • Membrane Glycoproteins