ELŐADÁS KIVONAT CLASSROOM LECTURE HANDOUT. financed by the program

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nemzetközi versenyképességének erősítésére program keretében finanszírozott ELŐADÁS KIVONAT CLASSROOM LECTURE HANDOUT financed by the program

Hungarian universities Dátum / Date: 2017. FEBRUÁR 8.

1 TÁMOP C-13/1/KONV projekt Az élettudományi-klinikai felsőoktatás gyakorlatorientált és hallgatóbarát korszerűsítése a vidéki képzőhelyek nemzetközi versenyképességének erősítésére program keretében finanszírozott ELŐADÁS KIVONAT CLASSROOM LECTURE HANDOUT financed by the program Practice-oriented, student-friendly modernization of the biomedical education for strengthening the international competitiveness of the rural Hungarian universities Dátum / Date: FEBRUÁR 8. / FEBRUARY 8, 2017 Helyszín / Place: MTA SZBK Biofizika Tanácsterem / Lecture room of the Institute of Biophysics, BRC SZEGED, TEMESVÁRI KRT. 62. Az előadás címe / Title of the presentation: LIPIDS IN HEALTH AND DISEASE Előadó / Speaker: GÁBOR BALOGH Biological Research Centre Address: H-6726 Szeged, Temesvári krt. 62. Mail: H-6701 Szeged, POB

Lipids in health and disease Gábor Balogh Laboratory of Molecular Stress Biology Lipidomics Research Group Institute of Biochemistry BRC HAS 2017 balogh.gabor@brc.mta.

as endogenous receptor agonists Lipids in inflammation Lipidomics: How to quantify several hundreds of lipid species in a couple of minutes? Stress lipidomics Neurolipidomics & imaging MS 1.

& intelligent knife Lipid biomarkers of diabetes Membrane lipid therapy New directions: towards single cell lipidomics Modern view of biological membranes The complex world of lipids Lipid metabolism

2 Lipids in health and disease Gábor Balogh Laboratory of Molecular Stress Biology Lipidomics Research Group Institute of Biochemistry BRC HAS 2017 Modern view of biological membranes The complex world of lipids Lipid metabolism The origin of lipid diversity The function of microdomains Lipids in Calcium regulation Lipids in apoptosis Lipids as endogenous receptor agonists Lipids in inflammation Lipidomics: How to quantify several hundreds of lipid species in a couple of minutes? Stress lipidomics Neurolipidomics & imaging MS 1. Lipid biomarkers of cancer: imaging MS 2. & intelligent knife Lipid biomarkers of diabetes Membrane lipid therapy New directions: towards single cell lipidomics Modern view of biological membranes The complex world of lipids Lipid metabolism The origin of lipid diversity The function of microdomains Lipids in Calcium regulation Lipids in apoptosis Lipids as endogenous receptor agonists Lipids in inflammation Lipidomics: How to quantify several hundreds of lipid species in a couple of minutes? Stress lipidomics Neurolipidomics & imaging MS 1. Lipid biomarkers of cancer: imaging MS 2. & intelligent knife Lipid biomarkers of diabetes Membrane lipid therapy New directions: towards single cell lipidomics 1

3 Classical textbook view Subcellular distribution of lipids Sites of lipid synthesis Redrawn after van Meer et al.,

Complexity, leaflet asymmetry www.rbclab.

4 Complexity, leaflet asymmetry Lateral segregation Seeger/Heimburg: Monte-Carlo simulation Modern view of biological membranes The complex world of lipids Lipid metabolism The origin of lipid diversity The function of microdomains Lipids in Calcium regulation Lipids in apoptosis Lipids as endogenous receptor agonists Lipids in inflammation Lipidomics: How to quantify several hundreds of lipid species in a couple of minutes? Stress lipidomics Neurolipidomics & imaging MS 1. Lipid biomarkers of cancer: imaging MS 2. & intelligent knife Lipid biomarkers of diabetes Membrane lipid therapy New directions: towards single cell lipidomics 3

5 Lipids OH H O O H/Radyl H/Radyl The complex world of lipids Fatty acids trans cis saturated unsaturated 4

6 Fatty acid nomenclature ω α Δ9 desaturation ω3 desaturation ω6 desaturation ω3 desaturation essential fatty acids (mammals) Lipid backbones H 3 C OH CH 3 CH 3 HO OH OH R NH 2 OH CH 3 CH 3 Glycerol Sphingoid base Sterane Phospholipids - headgroups CL anionic 5

7 6

8 Inverted hexagonal (H II ) and micellar hexagonal (H I ) phases Tresset PMC Biophysics :3 micelle Bristol model makers Lipid diversity Structure The complexity of lipidome 7

Lipid diversity Structure and function Fully substituted Storage lipids O O O FA1 FA2 FA3 TG R Energy storage and lipid transport Membrane lipids O O P FA1 FA2 Choline OH NH FA P Choline Membrane

lipids Lipid metabolism The origin of lipid diversity The function of microdomains Lipids in Calcium regulation Lipids in apoptosis Lipids as endogenous receptor agonists Lipids in inflammation

9 Lipid diversity Structure and function Fully substituted Storage lipids O O O FA1 FA2 FA3 TG R Energy storage and lipid transport Membrane lipids O O P FA1 FA2 Choline OH NH FA P Choline Membrane structure and function PC SM Partially substituted Signal lipids O O FA1 FA2 R OH NH FA Cell signalling and metabolic homeostasis OH DG OH Cer Modern view of biological membranes The complex world of lipids Lipid metabolism The origin of lipid diversity The function of microdomains Lipids in Calcium regulation Lipids in apoptosis Lipids as endogenous receptor agonists Lipids in inflammation Lipidomics: How to quantify several hundreds of lipid species in a couple of minutes? Stress lipidomics Neurolipidomics & imaging MS 1. Lipid biomarkers of cancer: imaging MS 2. & intelligent knife Lipid biomarkers of diabetes Membrane lipid therapy New directions: towards single cell lipidomics Lipid metabolism - regulation New tools to investigate the regulation of lipid metabolism - genetic manipulations - lipidomics - systems biology (e.g. flux balance) Eugene P. Kennedy ( ) 60 years ago Yeast 8

10 Yeast Lipid synthesis (yeast) Global analysis of the yeast lipidome, Ejsing, PNAS, 2009 Lipid synthesis in Drosophila 9

11 Phospholipid synthesis and remodelling (mammalian) Lipid metabolism (mammalian) in context Sphingolipid metabolism (mammalian) 10

origin of lipid diversity The function of

Lipids in apoptosis Lipids as endogenous

Lipidomics: How to quantify several hundreds

Stress lipidomics Neurolipidomics & imaging

2. & intelligent knife Lipid biomarkers of

12 Modern view of biological membranes The complex world of lipids Lipid metabolism The origin of lipid diversity The function of microdomains Lipids in Calcium regulation Lipids in apoptosis Lipids as endogenous receptor agonists Lipids in inflammation Lipidomics: How to quantify several hundreds of lipid species in a couple of minutes? Stress lipidomics Neurolipidomics & imaging MS 1. Lipid biomarkers of cancer: imaging MS 2. & intelligent knife Lipid biomarkers of diabetes Membrane lipid therapy New directions: towards single cell lipidomics 11

The origin of lipid diversity Phylogenetic tree of life after Woese et al.

2009) The origin of lipid diversity temperature adaptation homeoviscous adaptation T= 60 C Alvinella pompejana 'Textbook biology

The alvinellids just didn't read the textbook.' S.C.

of microdomains Lipids in Calcium regulation Lipids in apoptosis Lipids as endogenous receptor agonists Lipids in inflammation

13 The origin of lipid diversity Phylogenetic tree of life after Woese et al with representative membrane lipids of the three domains of life: Bacteria, Archaea and Eukarya (modified after Schubotz, 2009) The origin of lipid diversity temperature adaptation homeoviscous adaptation T= 60 C Alvinella pompejana 'Textbook biology tells us that animals can be psychrophilic [cold loving] or thermophilic [heat loving] but not both. The alvinellids just didn't read the textbook.' S.C. Cary Modern view of biological membranes The complex world of lipids Lipid metabolism The origin of lipid diversity The function of microdomains Lipids in Calcium regulation Lipids in apoptosis Lipids as endogenous receptor agonists Lipids in inflammation Lipidomics: How to quantify several hundreds of lipid species in a couple of minutes? Stress lipidomics Neurolipidomics & imaging MS 1. Lipid biomarkers of cancer: imaging MS 2. & intelligent knife Lipid biomarkers of diabetes Membrane lipid therapy New directions: towards single cell lipidomics 12

14 Lateral segregation Seeger/Heimburg: Monte-Carlo simulation anullar lipids microdomains (e.g. rafts), anullar lipids protein Annular lipid-binding sites (Lee AG.) Cardiolipin is required for the proper respiratory supercomplex function membrane Cardiolipin is required for the proper respiratory supercomplex function Althoff et al.; Schlame M, Dowhan W 13

Membrane lipids such as cholesterol and sphingolipids influence

receptor (in light blue), with three bound cholesterol molecules

blue) with a bound partial inverse agonist carazolol (in green)

Cholesterol molecules between two receptor molecules are shown in

Jafurulla M & Chattopadhyay A 2013 Lateral segregation,

15 Membrane lipids such as cholesterol and sphingolipids influence the function of GPCRs The structure of the human A2A adenosine receptor (in light blue), with three bound cholesterol molecules (in yellow) The structure of the human 2-adrenergic receptor (in blue) with a bound partial inverse agonist carazolol (in green) embedded in a lipid bilayer. Cholesterol molecules between two receptor molecules are shown in orange. Jafurulla M & Chattopadhyay A 2013 Lateral segregation, microdomains (e.g. rafts), anullar lipids Seeger/Heimburg: Monte-Carlo simulation Membrane microdomain-mediated intracellular signaling pathways in colorectal cancer (Jahn KA et al. 2011) 14

Modern view of biological membranes The complex world of

function of microdomains Lipids in Calcium regulation

Lipids in inflammation Lipidomics: How to quantify several

Stress lipidomics Neurolipidomics & imaging MS 1.

& intelligent knife Lipid biomarkers of diabetes Membrane

16 Modern view of biological membranes The complex world of lipids Lipid metabolism The origin of lipid diversity The function of microdomains Lipids in Calcium regulation Lipids in apoptosis Lipids as endogenous receptor agonists Lipids in inflammation Lipidomics: How to quantify several hundreds of lipid species in a couple of minutes? Stress lipidomics Neurolipidomics & imaging MS 1. Lipid biomarkers of cancer: imaging MS 2. & intelligent knife Lipid biomarkers of diabetes Membrane lipid therapy New directions: towards single cell lipidomics Lipids in Calcium regulation: IP3 DAG 15

lipid diversity The function of microdomains

Lipids as endogenous receptor agonists Lipids in

hundreds of lipid species in a couple of minutes?

Lipid biomarkers of cancer: imaging MS 2.

Membrane lipid therapy New directions: towards

17 Modern view of biological membranes The complex world of lipids Lipid metabolism The origin of lipid diversity The function of microdomains Lipids in Calcium regulation Lipids in apoptosis Lipids as endogenous receptor agonists Lipids in inflammation Lipidomics: How to quantify several hundreds of lipid species in a couple of minutes? Stress lipidomics Neurolipidomics & imaging MS 1. Lipid biomarkers of cancer: imaging MS 2. & intelligent knife Lipid biomarkers of diabetes Membrane lipid therapy New directions: towards single cell lipidomics Lipids in apoptosis: Ceramide Membrane asymmetry 16

Apoptotic cells stained with annexin V (green) to show phosphatidylserine

externalization during apoptosis Modern view of biological membranes The

The function of microdomains Lipids in Calcium regulation Lipids in

in inflammation Lipidomics: How to quantify several hundreds of lipid

18 Apoptotic cells stained with annexin V (green) to show phosphatidylserine exposed on the outer membrane leaflet Oxidative signaling of PS externalization during apoptosis Modern view of biological membranes The complex world of lipids Lipid metabolism The origin of lipid diversity The function of microdomains Lipids in Calcium regulation Lipids in apoptosis Lipids as endogenous receptor agonists Endocannabinoids Lipids in inflammation Lipidomics: How to quantify several hundreds of lipid species in a couple of minutes? Stress lipidomics Neurolipidomics & imaging MS 1. Lipid biomarkers of cancer: imaging MS 2. & intelligent knife Lipid biomarkers of diabetes Membrane lipid therapy New directions: towards single cell lipidomics 17

Endogenous Cannabinoids N-arachidonoyl-ethanolamine O-arachidonoylethanolamine 2-arachidonyl glyceryl ether Myocardial infarction (ischaemia-reperfusion injury) (R, P, H) Heart failure,

(R, P, H) Obesity, non-alcoholic fatty liver disease, diabetic complications (R, H) Liver fibrosis, cirrhosis, alcohol-induced liver injury (R, H) Pancreatitis (R, H) Inflammatory bowel disease,

19 Endogenous Cannabinoids N-arachidonoyl-ethanolamine O-arachidonoylethanolamine 2-arachidonyl glyceryl ether Myocardial infarction (ischaemia-reperfusion injury) (R, P, H) Heart failure, cardiomyopathies (R, H) Atherosclerosis, restenosis (R, H) Stroke, spinal cord injury (R, H) Cirrhotic cardiomyopathy (R, H) Septic shock by live bacteria (R, H) Hepatic ischaemia-reperfusion injury (R, P, H) Obesity, non-alcoholic fatty liver disease, diabetic complications (R, H) Liver fibrosis, cirrhosis, alcohol-induced liver injury (R, H) Pancreatitis (R, H) Inflammatory bowel disease, colitis, diverticulitis (R, H) Nephropathy (R, H) Neurodegenerative/neuroinflammatory disorders (multiple sclerosis, Alzheimer's, Parkinson's and Huntington's disease, spinal cord injury) (R, H) Pain (R) Psychiatric disorders (anxiety and depression, schizophrenia) (R, H) Rheumatoid arthritis (H) Cancer (R, H) C, canine; H, human; P, pig; R, rodent. Modulating the endocannabinoid system in human health and disease successes and failures FEBS Journal pages , 22 APR

Modern view of biological membranes The complex world of lipids Lipid metabolism The origin of lipid diversity

agonists Lipids in inflammation Lipidomics: How to quantify several hundreds of lipid species in a couple of

& intelligent knife Lipid biomarkers of diabetes Membrane lipid therapy New directions: towards single cell

20 Modern view of biological membranes The complex world of lipids Lipid metabolism The origin of lipid diversity The function of microdomains Lipids in Calcium regulation Lipids in apoptosis Lipids as endogenous receptor agonists Lipids in inflammation Lipidomics: How to quantify several hundreds of lipid species in a couple of minutes? Stress lipidomics Neurolipidomics & imaging MS 1. Lipid biomarkers of cancer: imaging MS 2. & intelligent knife Lipid biomarkers of diabetes Membrane lipid therapy New directions: towards single cell lipidomics Lipids in inflammation: Eicosanoids and docosanoids Figure 1 Temporal changes in lipid mediators in the progression of acute inflammation. Yacoubian S and Serhan CN (2007) New endogenous anti-inflammatory and proresolving lipid mediators: implications for rheumatic diseases Nat Clin Pract Rheumatol 3: doi: /ncprheum

LTB 4 LIPID MAPS Modern view of biological membranes The complex world of lipids Lipid metabolism The origin of lipid diversity The function of microdomains Lipids in Calcium regulation

21 T M G. Balogh February 8, 2017 HETEs OH O OH O Prostaglandin s O OH 15(S)HETE Arachidonic Acid 15 LO HO O OH OH COX PGE 2 P450 Epoxyeicosatrienoids O OH H OH 5 LO Leukotrienes H OH O OH O 14,15-EET LTB 4 LIPID MAPS Modern view of biological membranes The complex world of lipids Lipid metabolism The origin of lipid diversity The function of microdomains Lipids in Calcium regulation Lipids in apoptosis Lipids as endogenous receptor agonists Lipids in inflammation Lipidomics: How to quantify several hundreds of lipid species in a couple of minutes? Stress lipidomics Neurolipidomics & imaging MS 1. Lipid biomarkers of cancer: imaging MS 2. & intelligent knife Lipid biomarkers of diabetes Membrane lipid therapy New directions: towards single cell lipidomics 20

22 Lipidome 21

23 Systems biology of lipids Technical development Lipid analysis in the 90s Large sample quantity TLC separation Fatty acid analysis Analysis time: days 22

Lipid analysis GC-MS Derivatisation FA composition (positional or cis/trans isomers) O O X FA1 FA2 (x10,000,000) Scan:TIC 1.50 SIM:TIC (1.19) MeO FA1 + MeO FA2 methyl esters 1.

25 14:0 14:1 15:0 15:1 16:0 16:1 n-7 17:0 17:1 18:0 18:1 n-9 trans 18:1 n-9 18:2 n-6 trans 18:2 n-6 18:3 n-6 18:3 n-3 20:0 20:1 20:2 21:0 20:3 n-6 20:4 n-6 20:3 n-3 22:0 22:1 20:5 n-3 22:2 + 23:0

24 Lipid analysis GC-MS Derivatisation FA composition (positional or cis/trans isomers) O O X FA1 FA2 (x10,000,000) Scan:TIC 1.50 SIM:TIC (1.19) MeO FA1 + MeO FA2 methyl esters 1.25 Positional isomers n-3 n :0 14:1 15:0 15:1 16:0 16:1 n-7 17:0 17:1 18:0 18:1 n-9 trans 18:1 n-9 18:2 n-6 trans 18:2 n-6 18:3 n-6 18:3 n-3 20:0 20:1 20:2 21:0 20:3 n-6 20:4 n-6 20:3 n-3 22:0 22:1 20:5 n-3 22:2 + 23:0 24:0 24: n ESI-MS lipidomics O 16:0 Ionisation Electrospray (ESI) O 20:4 lipid molecular species composition P Choline analysis time minutes high troughput PC(16:0/20:4) quantity low (hundreds of cells) (protein μg equivalent) Concentration nanomol (total) / picomol (species) Sample amount 5 µl Lipid(omic) analysis mass spectrometry Orbitrap Elite since Sept

3,000 100 1 P Lipidomics workflow Lipidomics workflow We

25 Lipid diversity structure, concentration and function PC Sphingomyelin Ceramide Sphingosine S1P 300,000 30,000 3, P Lipidomics workflow Lipidomics workflow We need all of the lipids extracted sample extraction lipid extract 24

Lipidomics workflow Triglycerides Phospholipids Gangliosides hexane water We need all of the lipids extracted sample extraction small sample size lipid extract Lipidomics

26 Lipidomics workflow Triglycerides Phospholipids Gangliosides hexane water We need all of the lipids extracted sample extraction small sample size lipid extract Lipidomics workflow Mass spectrometry We need IONS!!! Polarity + or - Proper solvent system, additives Lipidomics workflow Data analysis We need software LipidXplorer + In-House development 25

Lipidomics workflow Interpretation Statistics Bioinformatics Systems biology And again we

scan (MS1) High mass accuracy & resolution e.g.: 885.

8594 PI 18:0/20:4 PI 18:1/20:3 m/z Modern view of biological membranes The complex world

Lipids in Calcium regulation Lipids in apoptosis Lipids as endogenous receptor agonists

couple of minutes? Stress lipidomics Neurolipidomics & imaging MS 1.

27 Lipidomics workflow Interpretation Statistics Bioinformatics Systems biology And again we need brain Shotgun lipidomics identification of lipid molecular species Intensity Survey scan (MS1) High mass accuracy & resolution e.g.: (PI 38:4) m/z Intensity Fragmentation (MS2, MSn) Fatty acyl chains e.g.: PI 18:0/20:4 PI 18:1/20:3 m/z Modern view of biological membranes The complex world of lipids Lipid metabolism The origin of lipid diversity The function of microdomains Lipids in Calcium regulation Lipids in apoptosis Lipids as endogenous receptor agonists Lipids in inflammation Lipidomics: How to quantify several hundreds of lipid species in a couple of minutes? Stress lipidomics Neurolipidomics & imaging MS 1. Lipid biomarkers of cancer: imaging MS 2. & intelligent knife Lipid biomarkers of diabetes Membrane lipid therapy New directions: towards single cell lipidomics 26

Stress Heat shock proteins Membranes, lipid droplets Stress sensor

DEVELOPMENT 1998 Morimoto et al., 12:3788-3796 Vigh et al.

stress lipidomics Without stress there would be no life (Hans Selye)

28 Stress Heat shock proteins Membranes, lipid droplets Stress sensor models Unfolded Protein Membrane perturbation Morimoto, GENES & DEVELOPMENT 1998 Morimoto et al., 12: Vigh et al., TIBS (1998) Genes and Development (1998) Membranes are stress sensors stress lipidomics Without stress there would be no life (Hans Selye) Heat shock response (HSR) Lipid-related changes Composition Microdomain reorganization Signal lipids 27

69 65 61 49 53 57 13 17 21 25 29 33 37 41 45 9 1 5 G. Balogh February 8, 2017 Stress-induced lipid remodelling PCA ESI-MS/MS 4.50 4.00 3.50 ca.

29 G. Balogh February 8, 2017 Stress-induced lipid remodelling PCA ESI-MS/MS ca. 160 species (>1% within its class) 4 different conditions 4 replicates Unsupervised method Principal component analysis 0.00 S1 S5 S9 S13 S17 S21 PC1, stress vs control PC2, BA vs heat PC3, mild vs severe heat Stress-induced lipid remodelling Lipid classes C 41C 43C Phospholipase action Raft forming lipids 25 BA 20 mol% of total * ** *** 5 ** 2 1 * * *** 0 PC PC PE PE PS PI PG Cl SPM LPC Cer Chol CE diac. alk. diac. pl. Balogh, et al. BB A : 1036 Proposed involvement of phospholipases 28

Evolution of fever and heat stress signalling Kluger

(1975) Hasday and Singh (2000) Hypothesis: HS

vertebrates Lipid signals (cellular stress response)

FEBS Lett (2013) Stress-induced lipid remodelling

30 Evolution of fever and heat stress signalling Kluger et al. (1975) Hasday and Singh (2000) Hypothesis: HS sensor(s) evolved in parallel to the fever response in vertebrates Lipid signals (cellular stress response) Diabetes Neurodegenerative diseases Cancer Balogh et al. FEBS Lett (2013) Stress-induced lipid remodelling Microenvironmental changes cell culture conditions nutrition tumors insuline resistance Microenvironmental stress 29

pombe 1h mild (40 C) heat stress Membrane stabilization Signal, Lipid synthesis,?

31 Sequence of events during and after 1 h heat shock Balogh et al. FEBS Lett (2013) Stress Heat shock proteins Membranes, lipid droplets Lipid changes upon heat stress in fission yeast S. pombe 1h mild (40 C) heat stress Membrane stabilization Signal, Lipid synthesis,? Membrane stabilization Signal Signal, Lipid Membrane synthesis stabilization LD formation Regulation of protein translation, formation of mrna processing bodies during heat stress in budding yeast (Riezman group & Cowart Hannun) 30

32 DKO TG mutants in S. pombe Lipase activities FA synthase No FA uptake FA-CoA 14 30C_WT PA PE, PC DG DGA1 PLH1 PL TG mol % of membrane lipids C_DKO 4 2 WT DKO TG Specific lipid changes in the DKO TG mutant upon HS Membrane stabilization WT_30 WT_40 DKO_30 DKO_40 ESI-MS mol % of total Stress signalling 10 0 PC PE PI PS LPE pcer DG TG Dramatic increases in signal lipids at mol. spec. level 10 3 WT_30 WT_40 DKO_30 DKO_40 mol % of total 5 mol % of total 0 0 DG(34:1) DG(36:2) DG(36:1) pcer(38:1:2) pcer(38:0:2) pcer(38:0:3) pcer(44:0:3) pcer(44:0:4) Are they toxic? 31

DKO cells show cell cycle arrest due to mild HS

NO, only cell cycle arrest The engagement of

context-specific Protein denaturation Folded

hydrophobic surfaces Putative role of lipid

33 DKO cells show cell cycle arrest due to mild HS WT 30 C 40 C DKO 30 C 40 C HS: 40 C growth: 1 day 30 C Are they dead? WT 30 C 40 C DKO 30 C 40 C growth: 4 day 30 C NO, only cell cycle arrest The engagement of lipid signals in survival or death is highly context-specific Protein denaturation Folded state Heat Unfolded state Attack against hydrophobic surfaces Putative role of lipid droplets in stress management WT 30 C HS: 50 C 50 C DKO 30 C (no TG) 50 C TG TG holdase function 32

Modern view of biological membranes The complex world of lipids Lipid metabolism

regulation Lipids in apoptosis Lipids as endogenous receptor agonists Lipids in

& intelligent knife Lipid biomarkers of diabetes Membrane lipid therapy New

34 Modern view of biological membranes The complex world of lipids Lipid metabolism The origin of lipid diversity The function of microdomains Lipids in Calcium regulation Lipids in apoptosis Lipids as endogenous receptor agonists Lipids in inflammation Lipidomics: How to quantify several hundreds of lipid species in a couple of minutes? Stress lipidomics Neurolipidomics & imaging MS 1. Lipid biomarkers of cancer: imaging MS 2. & intelligent knife Lipid biomarkers of diabetes Membrane lipid therapy New directions: towards single cell lipidomics Neurolipidomics Neurolipidomics: lipid specificity in brain cerebellum pref.cortex striatum hippocampus 33

Neurolipidomics: lipid specificity in brain: imaging mass spectrometry Colsch et al.

origin of lipid diversity The function of microdomains Lipids in Calcium regulation Lipids in

quantify several hundreds of lipid species in a couple of minutes?

& intelligent knife Lipid biomarkers of diabetes Membrane lipid therapy New directions:

(colon carcinoma sample) Red m/z 861.5425 (PI 36:2) Blue 885.5495 (PI 38:4) Red 742.

35 Neurolipidomics: lipid specificity in brain: imaging mass spectrometry Colsch et al., 2011 Modern view of biological membranes The complex world of lipids Lipid metabolism The origin of lipid diversity The function of microdomains Lipids in Calcium regulation Lipids in apoptosis Lipids as endogenous receptor agonists Lipids in inflammation Lipidomics: How to quantify several hundreds of lipid species in a couple of minutes? Stress lipidomics Neurolipidomics & imaging MS 1. Lipid biomarkers of cancer: imaging MS 2. & intelligent knife Lipid biomarkers of diabetes Membrane lipid therapy New directions: towards single cell lipidomics Desorption electrospray ionization mass spectrometric imaging (colon carcinoma sample) Red m/z (PI 36:2) Blue (PI 38:4) Red (PE 36:2) Green ; Fatty acid dimer (40:6) Blue (Triacylglycerol fragment) Green (PS 36:1); Blue (PI 36:2) Zoltán Takáts 34

Lipidomics triumphant - Intelligent knife Zoltán Takáts Modern view of biological membranes The complex world

regulation Lipids in apoptosis Lipids as endogenous receptor agonists Lipids in inflammation Lipidomics: How to

Stress lipidomics Neurolipidomics & imaging MS 1. Lipid biomarkers of cancer: imaging MS 2.

36 Lipidomics triumphant - Intelligent knife Zoltán Takáts Modern view of biological membranes The complex world of lipids Lipid metabolism The origin of lipid diversity The function of microdomains Lipids in Calcium regulation Lipids in apoptosis Lipids as endogenous receptor agonists Lipids in inflammation Lipidomics: How to quantify several hundreds of lipid species in a couple of minutes? Stress lipidomics Neurolipidomics & imaging MS 1. Lipid biomarkers of cancer: imaging MS 2. & intelligent knife Lipid biomarkers of diabetes Membrane lipid therapy New directions: towards single cell lipidomics Plasma biomarkers of obesity related diseases 200 lipid species 35

Prediction of obesity PLS-DA obesity plasmamarkers 30 samples learning 10

Calcium regulation Lipids in apoptosis Lipids as endogenous receptor

New directions: towards single cell lipidomics Allo-network-type drugs:

37 Prediction of obesity PLS-DA obesity plasmamarkers 30 samples learning 10 unknown sample: prediction Neuronal network Multilayer perceptron Modern view of biological membranes The complex world of lipids Lipid metabolism The origin of lipid diversity The function of microdomains Lipids in Calcium regulation Lipids in apoptosis Lipids as endogenous receptor agonists Lipids in inflammation Lipidomics: How to quantify several hundreds of lipid species in a couple of minutes? Stress lipidomics Neurolipidomics & imaging MS 1. Lipid biomarkers of cancer: imaging MS 2. & intelligent knife Lipid biomarkers of diabetes Membrane lipid therapy New directions: towards single cell lipidomics Allo-network-type drugs: membrane lipid therapy Vigh et al TIBS (2007). Balogh et al. FEBS Lett (2013) Escribá, Busquets, Inokuchi, Balogh, Török, Horváth, Harwood, Vígh. Prog Lipid Res (2015) 36

An example for membrane lipid therapy Modulation of lipid metabolism by a specific fatty

Radiosensitizing Role of Gamma-Linolenic Acid in Glioma Cells Antal O, Péter M, Hackler L

BBA Molecular and Cell Biology of Lipids (2015) GLA supplementation reshapes the whole

38 An example for membrane lipid therapy Modulation of lipid metabolism by a specific fatty acid treatment Anticancer therapy by radiosensitisation Lipidomic Analysis Reveals a Radiosensitizing Role of Gamma-Linolenic Acid in Glioma Cells Antal O, Péter M, Hackler L Jr, Mán I, Szebeni G, Ayaydin F, Hideghéty K, Vigh L, Kitajka K, Balogh G, Puskás L.BBA Molecular and Cell Biology of Lipids (2015) GLA supplementation reshapes the whole lipidome Distinct patterns of lipid molecular species for the non-treated and GLA-supplemented cells 37

Membrane modulation by GLA and/or radiation stress - HSP

the protection by HSP expression Lipids Life Death

Zsolt Török Imre Gombos Begüm Peksel László Vígh Jr.

Crul Noémi Tóth Bálint Csoboz Burcin Güngör Ákos Hunya

39 Membrane modulation by GLA and/or radiation stress - HSP expression The susceptibility to oxidative stress overrides the protection by HSP expression Lipids Life Death Lipidomics Laboratory László Vígh Ibolya Horváth Mária Péter Zsolt Török Imre Gombos Begüm Peksel László Vígh Jr. Attila Glatz Lajos Németh Mónika Homolya Péter Gudmann Tim Crul Noémi Tóth Bálint Csoboz Burcin Güngör Ákos Hunya Ana-Maria Pilbat Erika Zukic Istvánné Bogdán Laboratory of Molecular Stress Biology 38

40 lipidlibrary.aocs.org CYBERLIPID CENTER lipidbank.jp

LABORATÓRIUMI GYAKORLAT SILLABUSZ SYLLABUS OF A PRACTICAL DEMONSTRATION. financed by the program

LABORATÓRIUMI GYAKORLAT SILLABUSZ SYLLABUS OF A PRACTICAL DEMONSTRATION. financed by the program TÁMOP-4.1.1.C-13/1/KONV-2014-0001 projekt Az élettudományi-klinikai felsőoktatás gyakorlatorientált és hallgatóbarát korszerűsítése a vidéki képzőhelyek nemzetközi versenyképességének erősítésére program