From G-protein coupled receptors to heart failure and Alzheimer ’ s disease Yang Kevin Xiang Tupper Hall 2419c ykxiang@

1. From G-protein coupled receptors to heart failure and Alzheimer’s disease Yang Kevin Xiang Tupper Hall 2419c ykxiang@ucdavis.edu Department of Pharmacology UCD Medical School

2. Two diseases are explored in Xiang lab Hypertrophy and Heart Failure (No: 1 Killer in the US) Alzheimer’s Disease: a growing society burden (with aging babyboomers)

3. G-protein coupled Receptors (GPCRs) Neuropeptide Tachykinin Opioid Chemokine 2012 Nobel Prize to Brian Kobilka and Robert Lefkowitz Amine Melatonin MECA Glycoprotein Purine Adapted from Fredriksson et al MP 2003

4. Adrenergic Receptors A Heart rate, contractility, blood pressure, smooth muscle relaxation, fat and carbohydrate metabolism Memory, behavior Blood pressure, analgesia, anesthesia Blood pressure AR drugs are commonly used for asthma, hypertension, heart failure, and depression

5. Insights from molecules to mice • Single molecule studies • Receptor /G protein coupling • Posttranslational modification • Receptor dimerization • Ligand efficacy • Cellular studies • Receptor signaling transduction • Signaling crosstalk among • GPCRs and RTKs • Myocyte contraction • Cardiac stem cell growth • and differentiation • In vivo physiology • Cardiac hypertrophy & • heart failure • Memory and learning in • Alzheimer’s diseases

6. Receptor signaling crosstalk underlies cardiac hypertrophy and apoptosis in heart failure

7. Intracellular adrenergic signaling in heart Adopted from Xiao, 2003

8. Signaling cross-talks during onset (early stage) of heart failure The new idea: signaling alteration occurs before structural/morphological changes in myocardium • Prostaglandin (chronic inflammation) directly impacts cardiac contractile • Insulin (hyperinsulinemia) insults the heart directly. • IGF and adrenergic regulation of cardiac stem cell proliferation and differentiation for cardiac repairing and regeneration under ischemia/heart failure

9. Using FRET-based biosensors to study subcellular bAR signaling in diseased myocytes FSK ISO 10 mM 10 mM Plasma membrane Cytosol Sarcoplasmic reticulum Forskolin bAR bAR PDE4D5 PDE4D8 βAR PLB PLB Arrestin2 Arrestin2 AKAP AKAP PDE4D3 PDE4D5 PDE4D3 SERCA SERCA AKAP AKAP PKA PKA bAR bAR Gs Gs PDE4D8 PKA PKA RyR2 RyR2 PDE4D5 Isoproterenol AKAP AKAP AC AC PKA PKA AKAP PKA AKAP PKA Liu, PNAS, 2012

10. PGE2 and Insulin impair adrenergic signaling in cardiac myocytes PGE2 Iso EP4-R bAR PDE Insulin impairs adrenergic stimulation via enhanced Gi coupling PGE2 activates PDE4D to block cAMP diffusion from the PM to the SR under bAR stimulation Cell death Liu et al PNAS, 2012 Soto et al Circ Res, 2009 Cervantes, Circ Res, 2010

11. Many NSAIDs inhibit Cox-2, and reduce PGE2 for inflammatory responses in body NSAIDs (such as Advil) or not? Inflammation is BAD during the early stage, NSAIDs prevent cardiovascular diseases GOOD in the late stage of heart diseases, NSAIDs “kill” patients.

12. Alzheimer’s disease Adaptec from www.ahaf.org/alzdis/about/AmyloidPlaques.htm

13. The Amyloid hypothesis in Alzheimer’s disease Are there some specific cellular targets for amyloid b peptide (Ab)? Adaptec from www.ahaf.org/alzdis/about/AmyloidPlaques.htm

14. Ab interacts with CNS adrenergic system for Alzheimer’s disease NE Ab Wang et al FASEB, 2010 Wang et al JBC, 2011 Wang et all JBC, 2012, in press bAR cAMP/PKA Chronic Acute Adaptative Gene transcription AMPA Ab Neuron Apopotosis ApoE

15. Inhibition of β2AR ameliorates Aβ toxicity: may provide a new therapy for AD 80 70 ** 60 50 40 Novel object preference (%) 30 20 10 0 WT β2-KO PS1/APP β2-KO/PS1/APP A WT β2-KO PS1/APP β2-KO/PS1/APP

16. The Team University of California at Davis Sungjin Kim Qin Fu Qian Shi Dippal Parikh Rita Xu University of Illinois at Urbana Shu-Bai Liu David Cervantes Dagoberto Soto Vania De Arcangelis Ruijie Liu Dayong Wang Yongyu Wang • Funding • NIH • American Heart Association • Alzheimer’s association • Lee Cox (G.Gordavin), University of Illinois • TJ Ha (Ankur Jain), University of Illinois • Jin Zhang, John Hopkins University • Wen Chen, Temple University • Dale Abel, University of Utah Antelope Canyon, AZ, 2009

17. Insights from Molecules • Signaling complexes under physiological and pathophysiological conditions • Membrane protein oligomerization • Receptors monomers or dimers • Channels clustering, L-type Ca channels in diabetes • Post-translational modification of receptors or channels under a specific neurohormonal stimulation or disease state • The goal is to design biased drugs that selectively act on a specific pool of receptor in diseased conditions.

18. Flow Chart of SiMPull • Receptor oligomers • Receptor PTMs • Receptor binding partners • Under neurohormonal stimulation, • drugs treatment, or in diseases Jain et al Nature 2011 & Nature Protocol 2012

19. Hypertrophy and Heart Failure Adapted from Lutz et al Nature 1999

20. Schematic of SiMPull Jain et al Nature 2011

21. A New Alzheimer’s Disease Model • AD modelTg6554 X b2AR-KO • Reduced amyloid load in brain • Reduced neuron loss over time • Slow down decline of cognitive function A position on amyloid b induced b2AR signaling in neuron, and its implication in AD development, and potential therapeutic targets.

22. Studies of Single GPCR complex with SimPull • The similar signaling crosstalk may: • Affect neuronal differentiation • Enhance the memory • Reduce the blood flow for cancer cell growth • Immunoresponse A opening on study GPCR signaling cross-talk in both cardiac and neuronal cells.