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Biochemical Profiling of Lipids: Applications to Drug Development and Toxicology

Steven M. Watkins, Ph.D. President/CSO

Scientific Approach

• Building technology for understanding lipid metabolism in global and molecular detail. – Lipid metabolism in disease – Effects of foods and drugs on lipid metabolism – Lipids as central components of individual

health status

Metabolomics Integrates

MetabolismDrugs

Nutrition Health

Exercise Disease

Environment

Genes

Metabolic Assessment

• High-content analyses provide individual context

• Context allows interpretation

Metabolomics provides both

Tools for Enabling Metabolic Assessment

• High-content, quantitative analysis of metabolites

• Analysis of flux through metabolic pathways

Metabolomics provides both

Quantitative Analysis

PATIENT

Lipid Extract

SAMPLE

CLINICAL / HEALTH INFORMATION

Phosphatidylcholine

Phosphatidylserine

Phosphatidylethanolamine

Cardiolipin

Sphingomyelin

Phosphatidylinositol

Lyso-PLCholine

Triglycerides

Diacylglycerides

Monoacylglycerides

Free Fatty Acids

Cholesterol Esters

Sterols

DATABASE

Surveyor® Data Visualization

Steady-State Flux Analyses

ƒFlux

Experimental Quantitative Map Against Flux Models Analysis Pathway Signature

Building Diagnostics for Metabolic Disease Therapy

Edward Leiter The Jackson Laboratory

Profiling the Effects of Insulin-Sensitizing Drugs

• Investigated the effects of two diabetes drugs on lipid metabolism in a obese-diabetic model

• PPARγ agonist: Rosiglitazone • β-3 adrenergic agonist: CL316,243

PPAR-γ β3-Adrenergic 450 350

400 300

350

250 300

L 250 L 200

r d r de 200 e p p 150 s s

mg 150

mg

100 100

50 50

0 0 Cholesterol Glucose Triglycerides Cholesterol Glucose Triglycerides

Control Treatment

Clinical Biomarkers

PPAR-γ β3-Adrenergic 90 50

80 45

4070

3560

30

30 15

20 10

10 5

Gra

ms

0 0

Gra

ms 50

25 40

20

Body Weight post-treatment Body Weight post-treatment

Control Treatment

Body Weight Following Treatment

PPAR-γ Treatment

Hepatic Lipid Accumulation

PPAR-γ Treatment

β3-Adrenergic Agonist

Hepatic Lipid Synthesis

PPAR-γ β-3 Agonist

-100%

200%

-150% 150%

Hyp

o -lip

idem

icD

rug

A

-100%

200%

-150% 150%

Hyp

o -lip

idem

icD

rug

A

-100%

200%

-150% 150%

Hyp

o -lip

idem

icD

rug

AH

ypo -

lipid

emic

Dru

g A

De

De

De

Decccc

rrrreeeeaaaassss

eeeedddd w www

iiiitttthhhhIIIInnnn

crcrcrcreaeaeaea

sssseeeedddd

wwwwiiiitttthhhh

d dddrrrruuuu

gggg A AAA

drdrdrdrugugugug

A AAA

Plasma Metabolome Reveals Mechanism

Signature of lipogenesis

PPA

R-γ

Increased with drug BDecreased with drug B Increased with drug BDecreased with drug Increased with drug BDecreased with drug B

-100%

200%

-150% 150%

Increased with drug BDecreased with drug

All other metabolites

β3-Adrenergic 0

50

100

150

200

250

300

350

Cholesterol Glucose Triglycerides

mgs

per

dL

0

50

100

150

200

250

300

350

400

450

Cholesterol Glucose Triglycerides

mgs

per

dL

Must know both the identity and quantity of each metabolite

Metabolic Assessment

• High-content assays are required to elucidate metabolic basis for phenotype

• Want data on both metabolite concentrations and metabolic flux

Must know both the identity and quantity of each metabolite

Building Diagnostics

• Single chronic diseases have more than one metabolic (or environmental) cause

• Next generation diagnostics need to provide actionable information. – What dysregulation needs treatment?

• Metabolic assessment enables actionable, individual-specific diagnoses.

Delivering Health to the Individual

Individual Dx Map to Pathway Action

Biochemical Profiling of Lipids: Applications to Drug Development and Toxicology

Steven M. Watkins, Ph.D. President/CSO