PhD Scientific Days 2025

Poster Session II. - B: Molecular Medicine

Selection of Cadherin-11 Specific Aptamers

Előadó neve

Ms. Tar Alexandra

Neptun code

CODKM6

Előadó munkahelye

Department of Molecular Biology, Institute of Biochemistry and Molecular Biology, Semmelweis University

Előadó telefonszáma

+36209426869

Előadó e-mail címe

tar.alexandra@semmelweis.hu

Az előadás címe

Selection of Cadherin-11 Specific Aptamers

Szerző(k) neve és munkahelye

Alexandra Tar¹, Jessica Lange², Brigitta Kállai¹, Krisztina Percze¹, Thomas Karonitsch², Felix Kartnig², Hans Kiener², Daniel Aletaha², Ákos Harkai¹, Anna Tory¹, Leonhard Heinz², Peter Mandl², Tamás Mészáros¹

1: Department of Molecular Biology, Institute of Biochemistry and Molecular Biology, Semmelweis University
2: Division of Rheumatology, Department of Internal Medicine III, Medical University of Vienna

Bemutatás módja

Poszter

Szekció

Poster Session II. - B: Molecular Medicine

Language of the presentation

Hungarian

Preferred session

Molecular Medicine

Összefoglaló szövege

Introduction
Rheumatoid arthritis (RA) is an autoimmune disease characterised by structural joint damage and functional impairment. Fibroblast-like synoviocytes (FLSs) contribute to the healthy synovial membrane; however, under pathological conditions they become activated and play a crucial role in the development of synovitis, the chronic inflammation of the synovial membrane. FLSs express cadherin-11 (CDH11), a vital protein in cell-cell adhesion and a potential therapeutic target in RA.
Aims
Our aim is to generate CDH11-selective, modified nucleotide holding aptamers by toggle-SELEX and functionally characterise the enriched oligonucleotide candidates.
Method
CDH11-selective aptamer candidates were produced using toggle-SELEX. Subsequently, the enriched library was inserted into a cloning vector and examined by colony PCR, lab-on-a-chip nucleic acid electrophoresis, and Sanger sequencing. In addition, next-generation sequencing (NGS) was used to analyse the oligonucleotide pool after each selection step, including the initial library. The NGS data was analysed using AptaSUITE in our laboratory and by our collaborating, generative AI applying company (PentaBind). Wet-lab screening of the sequences was performed using AlphaScreen interaction analysis. MicroScale Thermophoresis (MST) was applied to further characterise the interaction of aptamer candidates with CDH11 and CDH13.
Results
The sequencing data analysis revealed the enrichment of specific oligonucleotides and various sequence-structure motifs. Generative AI analysis identified additional aptamer candidates using uniqueness rank score. AlphaScreen demonstrated that most studied candidates could interact with the in vitro translated CDH11. MST results showed that four candidates exhibited low micromolar binding affinity to CDH11. Besides, one candidate displayed clearly higher affinity to CDH11 than to CDH13, suggesting specificity. This candidate also performed well in artificial synovial fluid.
Conclusion
The top-performing aptamer candidate exhibited specific binding to CDH11 with low micromolar affinity and preserved its efficacy in artificial synovial fluid. Currently, we are modifying the nucleic acid sequence of this aptamer to further enhance its performance.
Funding
OTKA-ANN-139564
TKP2021-EGA-24
tar.alexandra@semmelweis.hu
Semmelweis University, Molecular Medicine Division
Tamás Mészáros

University

Semmelweis University

Supervisor

Tamás Mészáros

Publication of my abstract

I give consent to the publication of my abstract on the website of the congress.

phd.section.field

in doctoral studies after complex exam (PhD)

Kind

Szabad

Status

elfogadva

Accepted presentation method

poszter

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

7431

Start

18:36

End

18:42