Metabonomics

A New Potential Diagnostic Tool

David HuffmanWestern Michigan University

WDA 2010

Current Challenges• Difficulty in diagnosis

– Symptoms not always specific

– Confusion with other disorders

• Speed of diagnosis

– To minimize complications

• Monitoring treatment regimen

– Short and long term effects

Ala et al. Lancet 2007, 369, 397

http://www.nytimes.com/2009/08/18/health/18tran.html

Goals• Early diagnosis when clinical

manifestions are mild

• Predict neurological and liver function changes

Metabonomics

Definition of termsMETABOLITE is a small molecule

whose concentration changes as a function of something; usually not a protein or enzyme

METABOLOME is the repetoire of biochemicals present in cells, tissue, and body fluids

An example would be blood glucose

Metabonomics

Definition of termsMETABOLOMICS is the global

analysis of the type and quantity of all metabolites in biological fluids or tissue

METABONOMICS is the study of metabolic responses to pathophysiological stimuli

Metabonomics

Metabonomics“The relevance of metabonomics could be

greatly enhanced if it were possible to identify an invariant part of the individual metabolome of a ‘healthy’ subject with respect to, for example, pathological states, in such a way as to perform prediction, early diagnosis and prognosis of pathologies.”

Assfalg et al. Proc Natl Acad Sci 2008, 105, 1420

Metabonomics“. . . traditional biomedical/clinical

approaches are limited by the number of parameters as well as in their efficiency, and they provide only a fragmented perspective on the health status of an individual.”

Assfalg et al. Proc Natl Acad Sci 2008, 105, 1420

Current Biochemical TestsFor WD Typically don’t

measure small metabolites• AST and ALT (Aminotransferases)

• ALP (Alk Phosphatase)

• Bilirubin, Albumin

• INR, CBC

• Serum Ceruloplasmin

Roberts & Schilsky Hepatology 2003, 37, 1475

Current Testing Methods“However, most clinical chemistry tests

available today rely on old technologies that only measure a single chemical in blood, urine or other biofluid, and these tests are generally neither sensitive nor specific for any one particular disease. Comprehensive chemical analyses of urine or blood have simply never been done in a clinical setting before. ”

Slupsky, Biomarkers Med. 2010, 4, 195

Prospect of Metabonomics

• Measure a number of different metabolites simultaneously

• Obtain a unique fingerprint of a individual in metabolic space and time

• Develop new markers for WD diagnosis and monitoring

Metabonomics

Metabolites Monitored• amino acids

• amines

• lipids

• sugars and derivatives

• nucleic acids

• other metabolic intermediates

Metabonomics

Metabonomic Phenotypes

NMR Fingerprint from one individual*,‡

Multivariate Analysis of 21 Individuals

Assfalg et al. Proc Natl Acad Sci 2008, 105, 1420

*not actual data‡urine has over 2500 different metabolites at >1 mM

Metabotypesgenes

lifestyle etc.

Bernini, P.; Bertini, I.; Luchinat, C.; Nepi, S.; Saccenti, E.; Schäfer, H.; Schütz, B.; Spraul, M.; Tenori, L. Individual human phenotypes in metabolic space and time, J. Prot. Res. 2009

• The metabotype consists of a variable part (environment) and an invariant part (genetics + environment)• The invariant part persists for at least two-three years• The discovery of the existence of individual metabotypes has a strong biomedical potential

Celiac Disease Metabonomics

After gluten-free diet the metabolic fingerprint reverts to normal

Bertini, I.; Calabrò, A.; De Carli, V.; Luchinat, C.; Nepi, S.; Porfirio, B.; Renzi, D.; Saccenti, E.; Tenori, L. The metabonomic signature of celiac disease, J. Proteome Res. 2009, 8(1), 170

Metabonomic Applications

• New Markers for Disease– Facilitate rapid diagnosis

• Monitor Maintenance Therapy– Pre/Post Drug Treatment– Pre/Post Surgery

• Establish Parameters to Monitor Health– Routine Multivariate Analysis of body fluid

or tissue

Metabonomics

The Next Step

• Establish a metabonomics study of WD patients

Metabonomics

Current Funding

Metabonomics

National Science Foundation

Collaborators:Ivano BertiniClaudio Luchinat

Huffman Group Summer 2010