Towards Profiling of the G-Quadruplex Targeting Drugs in the Living Human Cells Using NMR Spectroscopy

KRAFČÍK, Daniel, Eva IŠTVÁNKOVÁ, Šimon DŽATKO, Pavlína VÍŠKOVÁ, S. FOLDYNOVA-TRANTIRKOVA and Lukáš TRANTÍREK. Towards Profiling of the G-Quadruplex Targeting Drugs in the Living Human Cells Using NMR Spectroscopy. International Journal of Molecular Sciences. Basel: Multidisciplinary Digital Publishing Institute, 2021, vol. 22, No 11, p. 6042-6055. ISSN 1422-0067. Available from: https://dx.doi.org/10.3390/ijms22116042.

Basic information

• Original name: Towards Profiling of the G-Quadruplex Targeting Drugs in the Living Human Cells Using NMR Spectroscopy
• Authors: KRAFČÍK, Daniel (703 Slovakia, belonging to the institution), Eva IŠTVÁNKOVÁ (203 Czech Republic, belonging to the institution), Šimon DŽATKO (703 Slovakia, belonging to the institution), Pavlína VÍŠKOVÁ (203 Czech Republic, belonging to the institution), S. FOLDYNOVA-TRANTIRKOVA and Lukáš TRANTÍREK (203 Czech Republic, guarantor, belonging to the institution).
• Edition: International Journal of Molecular Sciences, Basel, Multidisciplinary Digital Publishing Institute, 2021, 1422-0067.

Other information

• Original language: English
• Type of outcome: Article in a journal
• Field of Study: 10608 Biochemistry and molecular biology
• Country of publisher: Switzerland
• Confidentiality degree: is not subject to a state or trade secret
• WWW: URL
• Impact factor: Impact factor: 6.208
• RIV identification code: RIV/00216224:14740/21:00120234
• Organization unit: Central European Institute of Technology
• Doi: http://dx.doi.org/10.3390/ijms22116042
• UT WoS: 000660126000001
• Keywords in English: in-cell NMR; G-quadruplex; ligand; drug; Bcl2; telomeric DNA; KRAS; BRACO19; PhenDC3; NMM
• Tags: CF NMR, rivok
• Tags: International impact, Reviewed

Abstract

Recently, the H-1-detected in-cell NMR spectroscopy has emerged as a unique tool allowing the characterization of interactions between nucleic acid-based targets and drug-like molecules in living human cells. Here, we assess the application potential of H-1 and F-19-detected in-cell NMR spectroscopy to profile drugs/ligands targeting DNA G-quadruplexes, arguably the most studied class of anti-cancer drugs targeting nucleic acids. We show that the extension of the original in-cell NMR approach is not straightforward. The severe signal broadening and overlap of H-1 in-cell NMR spectra of polymorphic G-quadruplexes and their complexes complicate their quantitative interpretation. Nevertheless, the H-1 in-cell NMR can be used to identify drugs that, despite strong interaction in vitro, lose their ability to bind G-quadruplexes in the native environment. The in-cell NMR approach is adjusted to a recently developed 3,5-bis(trifluoromethyl)phenyl probe to monitor the intracellular interaction with ligands using F-19-detected in-cell NMR. The probe allows dissecting polymorphic mixture in terms of number and relative populations of individual G-quadruplex species, including ligand-bound and unbound forms in vitro and in cellulo. Despite the probe's discussed limitations, the F-19-detected in-cell NMR appears to be a promising strategy to profile G-quadruplex-ligand interactions in the complex environment of living cells.

Links

• LM2018127, research and development project: Name: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)

Investor: Ministry of Education, Youth and Sports of the CR

• NV19-08-00450, research and development project: Name: Atomárně rozlišená NMR spektroskopie in vivo jako nástroj pro biologické testování terapeuticky významných cílů v genomové ne-kanonické DNA a jejich interakcí s léčivy ve fenotypově diverzifikovaných nádorových buňkách.

Investor: Ministry of Health of the CR