Genetic ablation of Acp1 (Lmptp) in mice prevents heart failure
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1 Genetic ablation of Acp1 (Lmptp) in mice prevents heart failure Coralie Poizat, Ph.D. Director, Cardiovascular Research Program KFSHRC-Riyadh Saudi Heart Failure Working Group Jeddah, 5 December 2015
2 Cardiovascular Diseases: a burden for our society Leading cause of mortality in the Eastern and Western world Numbers from WHO 17.3 million people died of CVDs in 2008 Estimated 23.3 million deaths in 2030 Saudi Arabia: CVDs account for 25% of all deaths 80% of the Saudi population is under age 40: incidence of deaths from CVDs will rise Economic cost estimated at SAR 80 billion in 2030
3 Genetic mechanisms regulating cardiac hypertrophy & heart failure Dilated Hypertrophic
4 Major signaling pathways in cardiac hypertrophy Shp2 (PTPN22) Deletion causes DCM PTP1B Deletion protects against HF PTEN Inactivation protects against I/R LMPTP? Bianca et al., 2010
5 Low Molecular Weight Protein Tyrosine Phosphatase (LMPTP) The smallest PTPs ~18KDa Ubiquitous, cytosolic Encoded by a single gene ACP1 3 major isoforms Gene UTR Protein mrna LMPTP A= IF1 LMPTP B= IF2 LMPTP C
6 In vivo function of Lmptp a key regulator of metabolic homeostasis Regulates insulin signaling by dephosphorylating Tyr on IR ACP1 gene highly polymorphic and polymorphism affects enzymatic activity in human Low LMPTP activity associated with protection against metabolic syndrome in human Obese mice treated with anti-lmptp anti-sense oligonucleotides have improved insulin sensitivity and reduced hyperglycemia Role of LMPTP in the heart?
7 Lmptp expression during cardiac development and after pressure overload (in mice) Western blot Sham WT Western blot WT Lmptp GAPDH Lmptp Tubulin P1 P 7 D30 High resolution echocardiography
8 Increased Lmptp expression in human heart failure Western blot Trichrome staining Controls Failing (DCM) LMPTP GAPDH Control LMPTP/GAPDH CT *** 1 2 DCM Failing
9 Deletion of Lmptp in mice +/+ +/- -/- Lmptp GAPDH
10 Acp1 -/- mice develop normally and show no sign of pathology at baseline Growth curve 35 Weight (g) Acp1 +/+ Acp1 -/- Acp1 +/+ Acp1 -/ * Acp1 +/+ Acp1 -/ Age (weeks)
11 Pressure-overload-hypertrophy () Ligation New York-Presbyterian Echocardiography Week Sacrifice Tissue harvesting Biochemical analyses
12 Preserved cardiac dimension and function in Acp1 -/- mice after Ventricular septum thickness (mm) End- systolic volume (ml) Septum thickness * ** *** weeks post-op End syst Vol weeks post-op Posterior wall thickness (mm) Ejection fraction (%) * PWT ** ** * weeks post-op EF *** *** weeks post-op +/+ sham -/- sham -/- +/+
13 Reduced cardiac hypertrophy in Acp1 -/- hearts +/+ sham +/+ WGA staining Acp1 +/+ Acp1 -/- sham 200 µm 200 µm -/- sham -/- 200 µm 200 µm
14 Reduced fibrosis and heart failure markers in Acp1 -/- hearts Trichrome staining Acp1 +/+ Acp1 -/- sham 100 µm 100 µm 100 µm 100 µm
15 Gene expression profiling in Acp1 -/- hearts -/- Heat Map Gene numbers Up-regulated Down-regulated +/+ -/- sh +/+ sh Hypertrophy 1 Stenosis2 Dilation3 Fibrosis4 Output5 -log(p value)
16 Deleterious signaling pathways are minimally up-regulated in Acp1 -/- hearts Short-term (10 min & 24 hours) +/+ -/- 10min 24h 10min 24h Sham Sham Sham Sham Gαq/11 CaMKIIδ p-camkiiδ PLCβ3 p- PLCβ3
17 Protective signaling pathways are upregulated in Acp1 -/- hearts Canonical pathways IR phosphorylation 10min 24h +/+ -/- +/+ -/- -log(p value) IP: IRβ Sham Sham Sham Sham Insulin receptor 1 WB: IRβ PKA 2 NF-kB 3 Ephrin receptor 4 WB: 4G10 IP: IgG WB: IR IGF-15 WB: 4G10
18 Summary Lmptp deficiency protects against pressureoverload hypertrophy and heart failure Protective mechanism involves minimal activation of PLC/CaMKII signaling and activation of the IR, PKA and NFkB pathways
19 Future work Mechanism: Generate conditional deletion of Acp1 in cardiomyocytes, fibroblasts and vessels Screen for small molecule inhibitors of LMPTP to be tested in mice as therapy to treat heart failure
20 Acknowledgments Cardiovascular Research Program KFSHRC Fallou Wade Kamar Al-Haffar Salma Awad Mahmoud Muhammad Kunhi Israa Zahid Qussay Marashli Karim Belhaj KFSHRC Dr Falah Al-Mohanna Dr Abdullah Assiri Dilek Colak Ahmad Abusaleem Khalid AlKhatib King Saud University Dr Waleed Al-Habeeb International Haruhiro Toko Pearl Quijada Mark Sussman Stephanie Stanford Nunzio Bottini
21 Genetic mechanisms regulating cardiac hypertrophy & heart failure Dilation Myocardial infarction Dilated cardiomyopathy RV LV Pathological Chronic hypertension Aortic valve stenosis Developmental Physiological Chronic exercise Pregnancy Extensive fibrosis, cell death, advanced cardiac dysfunction Fibrosis ± cardiac dysfunction No cardiac dysfunction Heineke and Molkentin, 2006
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