Block A: Membrane Biology & Biochemistry

Lipid signalling and function

10. - 14.11.2014

Gerhild van Echten-Deckert

Tel. 73 2703 E-mail: [email protected] www.limes-institut-bonn.de Programme of the week

•Monday - general discussion: cellular signalling - -1-phosphate and neurodegeneration (Morbus Alzheimer)

• Tuesday - brain specific deletion of SPL and its impact on behavivour - bioactive fatty acid derivatives: endocannabinoids and eicosanoids

• Thursday Stefanie Herresthal: : Critical Players in Alzheimer’s Disease

•Friday Gökcen Gözüm: Targggeting sppghingosine-1-ppphosphate axis in cancer Outline of objectives References:

General aspects on cell signalling Hagen et al. Subcellular origin of sphingosine-1-phosphate is essential for its toxic effect in lyase-deficient neurons. J. Biol. Chem. 284: 11346-53 (2009). Signalling via: Hagen et al. Sphingosine-1-phosphate links sphingolipid G-protein coupled receptors metabolism to neurodegeneration via a calpain-mediated mechanism. Cell Death Differ. 18: 1356-1365 (2011) Receptor tyrosine kinases httppjp://www.nature.com/cdd/journal/vaop/ncurrent/full/cdd20117a.html. NlNuclear receptors Walter, Jochen, van Echten-Deckert, Gerhild: Cross-talk of membrane and Alzheimer-related proteins. Mol. Signalling lipids: Neurodegen. 8:34-45 (2013). Fatty acids van Echten-Deckert, G., Hagen-Euteneuer, N., Karaca, I., Eicosanoids Walter, J. Sphingosine-1-phosphate: boon and bane fort he brain. Endocannabinoids Cell Physiol Biochem., 34: 148-57 (2014). Sphingolipids (S1P) van EEhtchten-Deck ert & Walt er “Sphi ngo lip ids: Cr itical pl ayers in Alzheimer’s disease” Prog Res 51, 378-93 (2012) http://dx.doi.org/10.1016/j.plipres.2012.07.001 Pathological implications: Neurodegeneration (Alzheimer) Kunkel et al. „„gTargeting the sp pghingosine-1-ppphosphate axis in cancer, inflammation and beyond“ Inflammation Nature Rev, Drug Discovery, 12:688-702 (2013). Cancer Kendall & Nicolaou „Bioactive lipid mediators in skin Inflammation and immunity“ Prog. Lipid Res., 52:141-64 (2013). Programme

•Introduction - Morbus Alzheimer: numbers and facts – Sphingolipids: history, structure, metabolism

•Sphingosine-1-phosphate (S1P) – boon or bane? - biological activity and mechanism of action – CIMES, a synthetic sphingosine analogue •S1P-lyase KO and conditional KO - molecular bases of S1P-induced neurotoxicity - S1P-lyase-deficiency and neurodegeneration - S1P-lyase-deficiency and synaptic plasticity

•Conclusion and outlook 1906: 37. Meeting of doctors for the insane of southwest Germany in Tübingen Alois Alzheimer reports about “a peculiar affection of the cerebral cortex” Auguste D.

2013: World Alzheimer Report 2013

AD most common neurodegenerative disease worldwide 36 mill. cases (2030: 77 mill. 2050: 135 mill.)

Costs worldwide: 604 Bill. USD in 2010 (1% of the global GDP) 1,117 Bill USD in 2030 Histopathological findings reported by A. Alzheimer (1906)

“Miliary foci distributed all over “Weird neurofibrillary changes, that the cortex, caused by the infiltration appeared li ke very th ick tangl es fill ing of a peculiar substance into the cortex” not only the cell body but also neuronal processes”

Images C & D are from Holtzman et al. Sci Transl. Med. 2011, 3, 1-17 Key neuropathological elements of AD

BAP-tists: senile (neuritic) plaques:extracellular aggregates of -amyloid

TAU-ists: Neurofibrillary tangles: intracellular bundles of hyperphosphorylated tau

ohcm.oxfordmedicine.com/.../graphic10017.jpeg Scheme of major proteolytic processing pathways of APP

A B

APPS- APPS-

A p3 Therapeutic approaches aimed to reduce A  prodti/duction/aggrega tion have fild!failed!   

APP APP APP APP CTF CTF

van Echten-Deckert & Walter, Prog Lipid Res 2012 Figure 3 Artistic change in an individual with Alzheimer's disease

60 years 62 years

63 years 64 years

66 years 65 years

The Lancet 2001; 357:2129-2133 Sphingolipids: History

OH OH

NH2

Johann Ludwig Wilhelm Thudichum 1884 Sphingolipids - Structure

NH2 HO

OH OH HO OH O OH HO O O OH Sphingosine AcHN OH O O O OH HOOC O O O HN CH3 HO O CH O OH 3 OH OH OH HO OH AcHN OH GM1 O

HN HO

O OH HN CH3 H C O O CH3 3 N P H3C O O Ceramide CH3 OH Ceramide

Sphingomyelin form cell type specific profiles

GlcCer Neuron GalCer Oligodendrocyte

Myelin Sheath LacCer

Axon Astrocyte

Gbose3Cer Endothelial Cells Gbose4Cer

Neuron Microglia GM3 Muscle Fiber GM2

GM1 GD3 GD1a GD1b GT1b van Echten-Deckert & Herget, BBA 2006 NH2

HO OH + CH (CH ) COSCoA 3 2 14/16 Sphingolipids: serine O palmitoyl/stearoyl-CoA

SPT (+PLP) NH Main metabolic pathways 2

HO CH3 3-ketosphinganine O

3KSR (+NADPH) De novo formation

NH2

HO CH3

OH sphinganine Degradation Sulfatide CerS1 (+stearoyl-CoA) O Recycling

HN CH3

HO GalCer CH3 dihydroceramide OH

DES O O

SMS HN CH3 HN CH 3 CH3 O

H3C O O CH3 HO CH3 N P

ceramide SMase CH3 O OH OH

CerS1 Cerase(acid)

NH2 GlcCer HO CH3

OH sphingosine

SPP SK2 LacCer O NH2

O P O CH3

O OH sphingosine-1-phosphate

Gangliosides SPL (+PLP)

phosphoethanolamine + hexa/octadecenal

van Echten-Deckert & Walter, Prog Lipid Res 2012 Biosynthesis of major brain

Glc-T Gal-T I SAT I SAT II β 1,1 β 1,4 α 2,3 α 2,8 Cer GlcCer LacCer GM3 GD3 GalNAc-T β 1,4 GlcCer GalCer

GM2 GD2 LacCer Gal-TIIT II β 1,3 Sulfatide

GM1a GD1b SAT IV α 2,3

GM3

GD1a GT1b GM2 SAT V GM1 α2,8 GD3 GD1a GD1b GT1a GQ1b GT1b

Oligo Astro Neuro a-series b-series

Glucose (Glc) Galactose (Gal) N-Acetylgalactosamine (GalNAc) Sialic Acid van Echten-Deckert & Walter, Prog Lipid Res 2012 Degradation pathways of selected glycosphingolipids affected in lysosomal storage disorders GM1 GM1- GM1--galactosidase Gangliosidosis GM2-activator, SAP-B

GM2

Tay-Sachs -hexosaminidaseA,B Sandhoff GM2-activator

GM3 sialidase Sialidosis SAP-B

Glucose (Glc) Galactose (Gal) LacCer

N-Acetylgalactosamine (GalNAc) Sialic Acid Phosphocholine GlcCer Gaucher GlcCerase SAP-C acid SMase

Cer Niemann-Pick A,B SM Farber acid Cerase SAP-C, SAP-D

Sph van Echten-Deckert & Walter, Prog Lipid Res 2012 Physiological/clinical relevance of bioactive sphingolipids Ser + FA -CoA

Sa

apppoptosis Sphingomyelin DHCer differentiation inflammation insulin resistance Ceramide C1P inflammation senescence stress response So MDR GlcCer anti-apoptosis glucose tolerance proliferation S1P migration iflinflammat ion brain function GSL angiogenesis morphogenesis wound healing tumour genesis Dual Action of S1P: Extracellular Ligand and Intracellular Second Messenger

growth factor S1P

S1P 1-5 trimeric G-proteins

Sphingosine kinase adenylate- monomeric sphingosine S1P cyclase PLC G-proteins

intracellular targets? • survival • differentiation • motility • proliferation • cytoskeleton rearrangements 2+ • Ca homeostasis • inflammation •anti-apoptosis • angiogenesis The subcellular origin of S1P is essential for its neurotoxic effect

S1P cimes Sph

PM SK1 S1PSph

(LPP?) cimesP apopttitotic SK2

SPP

ER SK2

S1PS1P apoptotic Hagen et al, JBC 2009 Summary

cimes extracellular intracellular

Z-LEHD-FMK SK2 Caspase-12 Caspase-9 Caspase-3 cimes cimesP MDL

neuronal ER-stress Ca2+ Calpain ER apoptosis S1P MDL SPP SK2 P cell cycle CDK5 Rb Sph p35 p25 reactivation

P tau

S1P

Hagen et al, Cell Death Differ, 2011 Conclusions

•Sphingosine-1-phosphate (S1P) is a neuronal death signal, when ggyenerated by SK2 and imp pgaired degradation •Calpain is an essential mediator of S1P-induced neurotoxicity •On cellular and molecular level S1P neurotoxicity parallels that of A •S1P-lyase expression is correlated with neuronal death •S1P-lyase deficiency is correlated with Alzheimer characteristics: •Hyperphhlhosphorylation of tau •Impaired APP-processing •Elevated levels of cholesteryl-ester

S1P stimulates BACE1, the rate-limiting enzyme for A production (Takasugi et al., 2011, J. Neurosci.)

Conditional knockout mouse: neuron-specific inactivation ofS1P-lyase