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A systematic and functional analysis of HIF-dependent splice regulator expression and pre-mRNA alternative splicing in pathologic stress-induced cardiac hypertrophy

Applicant Krek Wilhelm
Number 130505
Funding scheme Sinergia
Research institution Institut für Zellbiologie ETH Zürich
Institution of higher education ETH Zurich - ETHZ
Main discipline Cardiovascular Research
Start/End 01.01.2011 - 31.12.2013
Approved amount 1'100'000.00
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Keywords (4)

HIF; cardiac hypertrophy; splice regulators; pre-mRNA alternative splicing

Lay Summary (English)

Lead
Lay summary
Heart failure remains one of the most frequent causes of death in the industrialized world. It is the final common pathway of many forms of heart disease induced by a variety of pathological insults, including hypertension, myocardial infarction, and aortic stenosis. These stressors, if protracted or severe, lead to a progressive worsening of cardiac function and ultimately heart failure. Pathological stress-induced hypertrophy leads to cardiomyocyte growth, contractile dysfunction and heart failure. A critical and distinguishing feature of pathological hypertrophy is a shift in metabolism from fatty acid and glucose oxidation, to aerobic glycolysis and lipid accumulation. A key component mediating these changes in metabolism is hypoxia-inducible factor 1? (HIF1?). Recently, we found that alternative pre-mRNA splicing of certain genes is changed in cardiomyocytes in response to pathologic stress in an HIF1?-dependent manner. This finding adds to the growing body of evidence for key connections between alternative splicing and cell metabolism. However, no systematic and functional analysis of HIF1?-dependent splice regulator expression on splice variant usage in pathologic cardiac hypertrophy has been performed. This integrated research proposal will therefore provide novel and unique insights into the role of alternative splicing in reprogramming cardiac metabolism in response to pathologic stressors. Moreover, given the central role for HIF1? in mediating metabolic reprogramming in various disease states including cancer, our studies holds the promise to significantly alter our understanding of metabolic control in heart disease and during tumor cell evolution.
Direct link to Lay Summary Last update: 21.02.2013

Responsible applicant and co-applicants

Employees

Collaboration

Group / person Country
Types of collaboration
ETH Zurich Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Research Infrastructure

Scientific events

Active participation

Title Type of contribution Title of article or contribution Date Place Persons involved
Sensing and Signaling of Hypoxia: Interfaces with Biology and Medicine Talk given at a conference The Role of Hypoxia Inducible Factors in Cardiac and Adipose Metabolism 12.01.2014 Beckenridge/CO, United States of America Krek Wilhelm;
The reciprocal interactions of signaling pathways and non-coding RNA Talk given at a conference Advances in hypoxia signaling 15.09.2012 Ascona, Switzerland Flückiger Stefanie; Ule Jernej; Krek Wilhelm;


Associated projects

Number Title Start Funding scheme
138669 VHL tumor suppressor mechanisms 01.11.2011 Project funding
176317 Roles of ketohexokinase signaling and fructose metabolism in pancreatic tumor growth 01.11.2017 Project funding
163476 Regenerative therapy for heart disease via modulation of long noncoding RNAs 01.10.2015 Project funding
176317 Roles of ketohexokinase signaling and fructose metabolism in pancreatic tumor growth 01.11.2017 Project funding
156316 VHL tumor suppressor mechanisms 01.11.2014 Project funding
143355 Role of the Notch pathway in cardiac multipotent mesenchymal stromal cells 01.10.2012 Project funding
150837 MicroSPECT/PET/CT for preclinical molecular imaging 01.12.2013 R'EQUIP

Abstract

This Sinergia research proposal encompasses a systematic and functional analysis of the impact of HIF1a-dependent splice regulator expression on splice variant usage in pathologic cardiac hypertrophy, and it aims to define the metabolic and functional consequence of HIF1a-mediated regulation of splice variant usage in the development and maintenance of pathological hypertrophy, and in the progression to heart failure. To that end, an international, multi-institutional network of research groups has been established wherein each sub-group contributes state of the art expertise and technology towards specific components of this project.The project will be initiated through the differential expression analysis of splice regulator expression in vivo and in vitro in the context of pathological stress and as a function of HIF1a status. This phase will be characterised by the generation of the cellular and genetic material in the laboratory of Wilhelm Krek (ETH Zurich) and its expression analysis by splice-junction microarray in the group of Jernej Ule (MRC Laboratory of Molecular Biology, Cambridge, UK). Acquired data will analyzed jointly by bioinformaticians from the groups of Krek and Ule. The second phase will entail a functional in vitro, and an in vivo validation of candidate splice factors. The functional in vitro validation will be performed in the laboratory of Wilhelm Krek and will encompass a microscopy-based phenotypic screen in primary cardiomyocytes, and a secondary metabolic screen with the SeaHorse Metabolic Flux Analyzer. The in vivo validation will utilize human and mouse pathological hypertrophic material from the laboratories of Wilhelm Krek and Thierry Pedrazzini (University Lausanne), and will entail the protein and RNA expression analysis of candidate splice regulators in genetic and surgical mouse models of pathological cardiac hypertrophy, and of human diseased biopsies. Validated hypertrophy-regulated, HIF1a-sensitive candidate splice regulators from the in vitro and in vivo screen above, will be subjected to a CLIP-RNA sequencing and spice-junction microarray analysis in the laboratory of Jernej Ule, with the cellular and/or RNA material provided by the group of Wilhelm Krek. Data from this phase will demonstrate how specific hypertrophy-induced, HIF1a-sensitive splice regulators interact with the global RNA pool and provide a strong indication of critical target proteins of such interactions and their genetic and cellular consequences. Based on this data, transgenic and/or floxed mice will be acquired or generated by the groups of Thierry Pedrazzini and Wilhelm Krek. These mice will be subjected to in vivo pathological models of hypertrophy and assessed physiologically in the group of Thierry Pedrazzini. Thereafter, the cell biological, biochemical and genetic analyses of hearts derived from these mice will be performed jointly between the involved groups. In summary, we have brought together three leading research groups from Switzerland and UK, whose expertise extends from basic intracellular signalling mechanisms, to cardiac physiology and RNA technology development to address, as a team, the impact of alternative splicing on cardiac metabolism and function and its role in pathologic cardiac hypertrophy and the progression to heart failure.
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