PhD Scientific Days 2026

Poster Session 2.G - Pharmaceutical Sciences and Health Technologies

Developing Reporters for Monitoring Sarco/Endoplasmic Reticulum-Mitochondria Interactions

Name of the presenter

Larose Vajda, Angelique

Institute/workplace of the presenter

Semmelweis University, Department of Pharmacology and Pharmacotherapy

Authors

Angelique Larose¹
1: Semmelweis University, Department of Pharmacology and Pharmacotherapy

Text of the abstract

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 EKÖP grant