PhD Scientific Days 2025

Poster Session II. - U: Cardiovascular Medicine and Research

Developing Reporters for Monitoring Sarco/Endoplasmic Reticulum-Mitochondria Interactions

Előadó neve

Larose Vajda Angelique, MSc

Neptun code

IBXLBB

Előadó munkahelye

Semmelweis University, Department of Pharmacology and Pharmacotherapy

Előadó telefonszáma

06306910825

Előadó e-mail címe

vajda.angelique@semmelweis.hu

Az előadás címe

Developing Reporters for Monitoring Sarco/Endoplasmic Reticulum-Mitochondria Interactions

Szerző(k) neve és munkahelye

Larose Vajda Angelique MSc¹, Dr Mórotz Gábor²

1: Semmelweis University, Department of Pharmacology and Pharmacotherapy
2: Center for Pharmacology and Drug Research & Development, Semmelweis University, Budapest

Bemutatás módja

Poszter

Szekció

Poster Session II. - U: Cardiovascular Medicine and Research

Language of the presentation

English

Preferred session

Cardiovascular Medicine and Research

Összefoglaló szövege

Introduction: Myocardial infarction and heart failure are leading causes of death globally. Despite available treatments, mortality remains high. Signalling between the sarco/endoplasmic reticulum (SR/ER) and mitochondria regulate key cellular functions which are damaged in these diseases. Elucidating these signaling mechanisms is crucial for understanding disease pathogenesis and discovering new therapeutic interventions. However, precise monitoring of SR/ER-mitochondria contacts remains challenging due to requirements for highly specialized equipment, time-consuming experimental procedures, complex analysis methods, and inability to monitor living cells.
Aims: To develop a real-time monitoring system for SR/ER-mitochondria interactions in living cells by targeting the VAPB-PTPIP51 molecular tether complex, which anchors the SR/ER to mitochondria and enables organellar communication.
Methods: We utilized split luciferase complementation (NanoBit) assays to monitor VAPB-PTPIP51 interaction in real time. The ER membrane protein VAPB and outer mitochondrial membrane protein PTPIP51 were fused to luciferase fragments. We employed the weak herpes simplex virus thymidine kinase (HSV-TK) promoter to drive expression and avoid overexpression artifacts. Constructs were cloned into the Sleeping Beauty transposon system for stable integration into the genome.
Results: We successfully developed luciferase-based reporters that enable real-time monitoring of ER-mitochondria interactions in living cells. We were able to obtain expected results using the NanoBiT assay, demonstrating successful detection and quantification of VAPB-PTPIP51 interactions with robust signal-to-noise ratios and reproducible measurements.
Conclusion: Our reporters provide researchers with novel tools to monitor ER-mitochondria interactions and measure damages to organellar communication under disease-associated conditions, facilitating characterization of potential pathomechanisms in cardiac diseases.
Funding: This research is supported by the SE 250+ scholarship.

University

Semmelweis University

Supervisor

Dr Mórotz Gábor

Publication of my abstract

I do not give consent to the publication of my abstract on the website of the congress.

phd.section.field

in doctoral studies before complex exam (PhD)

Kind

Szabad

Status

elfogadva

Accepted presentation method

poszter

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

9279

Start

18:06

End

18:12