Cardiovascular Trait (CVT) Consortium Patricia Munroe

Primary traits of interest Hypertension Heart Failure Cardiac arrhythmias (ECG)

Aims of the CVT consortium  To characterise a series of KO mouse models derived from genes selected from human GWAS studies and functional experimentation.  Specific objectives:  To establish whether the KO mouse has the CV phenotype we predicted based on gene selection.  To establish additional phenotypic deviations from normal, including target-organ damage to heart, brain and kidney.  To characterise the functional consequences and pathways affected by absence of the KO gene.

Consortium phenotype expertise and interests BP/hypertension/CAD ECG measures/arrhythmias LVD, LVH and Heart Failure New members Alistair Poole (UK) Tim Warner (UK) Cristiana Ruhrberg (UK) Ines Pineda-Torra (UK) Ross Breckenridge (UK) CVT consortium members are from UK, NL, FR and USA

CV screening by IMPC CV screening Standard screens Heart weight, electrolytes, cardiac histopathology, eye fundus (retinal vessels) Additional screens requested (under consideration) Blood pressure (e.g., tail cuff) ECG (ECGenie) ECHO/US? Additional blood clinical chemistry - serum creatinine and natriuretic peptides (BNP and/or ANP- when test validated, NT-proBNP) Additional urine collection and chemistry (preferably 24hours at metabolic cage, alternatively urine collection during anesthesia/sacrifice) - urinary creatinine and urinary albumin (and/or total protein) Additional organ collection at necropsy - Heart (dissection into atria and ventricles, and left verses right heart weight), kidney, brain, aorta, liver Kidney and lung weight/body weight or tibia length ratio Kidney histopathology Challenges / Niche screens - High saline water or high-salt diet (if indicated by trait)

Flow chart of the experiments

Telemetry SBP/DBP/day night averages ECG measures: HR, HR variability, duration, arrhythmias ECHO Measures of ventricular function, structure and dimensions EP (sinus node recovery time, atrial and ventricular arrhythmia inducibility etc ECG (needle electrodes in anaesthesised animals, P-wave, PR, QRS, QTc) ‘ in vitro’ cellular EP (single cell EP, measures of K+, Ca+ and Na+ currents) Consortium secondary screens

‘in vitro’ vascular function Organ baths will be used and aorta and resistance arteries will be studied using pharmacological compounds Langendorrf perfusion Assessment of basic heart physiology using experimental perturbations, eg. ischemia- reperfusion, or EP studies Metabolic cages Collection of urine, allowing measures of output and electrolytes. Histology and molecular techniques Inflammatory responses Eg., assessment of leucocyte recruitment paradigm under baseline and stimulated conditions Consortium secondary screens

Acute pharmacological Including acute provocations/challenges eg, Ang II Chronic pharmacological/induced hypertension Chronic pharmacological interventions including injections. Induced HT models including high salt intake, AngII etc Exercise Exercise induced CV changes, using running wheel Myocardial infarction e.g., ligation of left coronary artery. Pressure and volume overload e.g., pressure overload to induce cardiac hypertrophy. Volume overload to induce cardiac dilatation or failure Models and challenges

Current activities Developing packages of work for year 1 –How best to distribute CV phenotyping between centres? –Creation of working groups per gene of interest –Obtaining additional functional data on genes of interest for inclusion in grants –Funding options for technical staff and CV phenotyping New member interests –Development of HT platelet phenotyping protocol (A. Poole), and a platelet aggregation assay (T. Warner) –Inclusion of CAD genes from GWAS (N. Samani) –Lipids and metabolomics (M. Ala-Korpela)

Additional slides