Directing in Vitro Selection towards G-quadruplex-forming Aptamers to Inhibit HMGB1 Pathological Activity.
Angew Chem Int Ed Engl
; 63(16): e202319828, 2024 04 15.
Article
em En
| MEDLINE
| ID: mdl-38358301
ABSTRACT
In the search for novel, effective inhibitors of High-Mobility Group Box1 (HMGB1)-a protein involved in various inflammatory and autoimmune diseases as well as in cancer-we herein discovered a set of anti-HMGB1 G-quadruplex(G4)-forming aptamers by using an in vitro selection procedure applied to a doped library of guanine-rich oligonucleotides. The selected DNA sequences were then studied in a pseudo-physiological buffer mimicking the extracellular medium, where HMGB1 exerts its pathological activity, using spectroscopic, electrophoretic, and chromatographic techniques. All the oligonucleotides proved to fold into monomeric G4s and in some cases also dimeric species, stable at physiological temperature. Remarkably, the protein preferentially recognized the sequences forming dimeric parallel G4 structures, as evidenced by a properly designed chemiluminescent binding assay which also highlighted a good selectivity of these aptamers for HMGB1. Moreover, all aptamers showed anti-HMGB1 activity, inhibiting protein-induced cell migration. The acquired data allowed identifying L12 as the best anti-HMGB1 aptamer, featured by high thermal and enzymatic stability, no toxicity at least up to 5â
µM concentration on healthy cells, along with potent anti-HMGB1 activity (IC50 ca. 28â
nM) and good binding affinity for the protein, thus indicating it as a very promising lead candidate for in vivo studies.
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Base de dados:
MEDLINE
Assunto principal:
Proteína HMGB1
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Aptâmeros de Nucleotídeos
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Quadruplex G
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article