Metabonomics A New Potential Diagnostic Tool David Huffman Western 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

Goals Early diagnosis when clinical manifestions are mild Predict neurological and liver function changes Metabonomics

Definition of terms METABOLITE 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 terms METABOLOMICS 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

“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 Tests For 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  M

Metabotypes genes 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 The metabotype consists of a variable part (environment) and an invariant part (genetics + environment) The invariant part persists for at least two-three years biomedical potential 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 Bertini Claudio Luchinat

Huffman Group Summer 2010