Varieties of interactions of anti-CD133 aptamers with cell cultures from patient glioblastoma.

in SLAS discovery : advancing life sciences R & D by Olga Antipova, Valeria Moiseenko, Fatima Dzarieva, Ekaterina Savchenko, Igor Pronin, Galina Pavlova, Alexey Kopylov

TLDR

  • The study investigates how aptamers, small molecules that can bind to specific cells, interact with cells derived from patient glioblastoma (GB). The authors use flow cytometry (FC) assessment to compare interactions of fluorescent anti-CD133 aptamers with cells, focusing on cell cultures derived from patient GB (CCPGB). The study identifies two types of interactions, target CD133-driven and off-target membrane-associated ones, that contribute to mean fluorescence intensities (MFIs). The study reveals half-saturation concentrations of 120±27 for 2'FY-RNA A15 and 180±12 for DNA Cs5 with Caco-2 cells, which is an essential step to develop aptatheranostics for GB.

Abstract

Development of aptatheranostics for glioblastoma (GB) requires investigating aptamer interactions with cells. The paper has described flow cytometry (FC) assessment of direct interactions of fluorescent anti-CD133 aptamers with cells, focusing on cell cultures derived from patient GB (CCPGB). Conventional cell lines with different levels of CD133 mRNA, Caco-2 and HCT116, were used to compare interactions with known 2'FY-RNA aptamer A15 and DNA aptamers of Ap and Cs series, labeled with FAM and Cy5. In addition, interactions of certain non-aptameric oligonucleotides were studied. In the case of antibody interactions with cells, FC signals, mean fluorescence intensities (MFIs), correlated with sizable amounts of CD133 mRNA in Caco-2 cells, and CCPGBs 107 and G01. Unexpectedly, MFI per se could not be the solid indicator of specific interactions of aptamer - CD133/cell. Instead, two types of interactions, target CD133-driven and off-target membrane-associated ones, contribute to MFI. The latter was notably observed for CCPGB Sus/fP2 with tiny CD133 mRNA amount. To prove specificity of aptamer - CD133/cell interactions, titration experiments have been performed, revealing half-saturation concentrations of 120±27 for 2'FY-RNA A15 and 180±12 for DNA Cs5 with Caco-2 cells. This knowledge is an essential step to develop aptatheranostics for GB.

Overview

  • The study investigates aptamer interactions with cells derived from patient glioblastoma (GB) using flow cytometry (FC) assessment. The authors compare interactions of fluorescent anti-CD133 aptamers with cells, focusing on cell cultures derived from patient GB (CCPGB).
  • The study aims to develop aptatheranostics for GB by investigating aptamer interactions with cells. The authors compare interactions of fluorescent anti-CD133 aptamers with cells, focusing on cell cultures derived from patient GB (CCPGB).
  • The primary objective of the study is to investigate aptamer interactions with cells derived from patient glioblastoma (GB) using flow cytometry (FC) assessment. The authors compare interactions of fluorescent anti-CD133 aptamers with cells, focusing on cell cultures derived from patient GB (CCPGB).

Comparative Analysis & Findings

  • The study compares interactions of fluorescent anti-CD133 aptamers with cells, focusing on cell cultures derived from patient GB (CCPGB).
  • The study identifies two types of interactions, target CD133-driven and off-target membrane-associated ones, that contribute to mean fluorescence intensities (MFIs).
  • The study reveals half-saturation concentrations of 120±27 for 2'FY-RNA A15 and 180±12 for DNA Cs5 with Caco-2 cells, which is an essential step to develop aptatheranostics for GB.

Implications and Future Directions

  • The study's findings provide essential knowledge for developing aptatheranostics for GB by investigating aptamer interactions with cells derived from patient GB (CCPGB).
  • The study identifies two types of interactions, target CD133-driven and off-target membrane-associated ones, that contribute to mean fluorescence intensities (MFIs).
  • The study reveals half-saturation concentrations of 120±27 for 2'FY-RNA A15 and 180±12 for DNA Cs5 with Caco-2 cells, which is an essential step to develop aptatheranostics for GB.
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