Identification of molecular targets and biomarkers in the mechanism and the treatment of obesity....

Identification of molecular targets and biomarkers in the mechanism and the

treatment of obesity. Introduction of the activity of the „OBEKON”

Tamás Pázmány, PhD

02. October 2009 CECON II Budapest 1

2nd Central European Congress on Obesity October 1-3, 2009 Budapest, Hungary

O B E K O NO B E K O NConsortium

Treatment of obesitySoranus - 2nd century – Alexandria and Rome

elixirs of laxatives and purgatives, heat, massage, and exercise.

Current strategies to treat obesityLifestyle changes in dietPhysical activityGastrointestinal surgeryPharmacological intervention

Suppression of the appetite Increase of the body's metabolism Interference with the body's ability to absorb

specific nutrients in food.

02. October 2009 CECON II Budapest


FDA approved prescription medication – for long-term weight loss

Sibutramine Appetite suppressant. Serotonin-norepinephrine reuptake inhibitor Meridia(US) – Reductil (EU) – prescription

Orlistat Reduces intestinal fat absorbtion by inhibiting pancreatic lipase Xenical - prescription Alli - OTC


…and for short term (up to 12 weeks):• Phentermine

• Appetite suppressant by inducing norepinephrine release in the hypothalamus


Arena Pharmaceutical’s Lorcaserin is an selective serotonin 2C receptor agonists.Phase III trials hase been completed: + results 7190 patients – 2009.09.18.

Merk’s Taranabant (MK 0364) is an selective inverse agonist to the human cannabinoid type 1 receptor. Stopped after phase III – 2008.10.02.

Orexigene Therapeutics’ Contrave is a combination of Wellbutrin ( antidepressant) and Naltrexone (addiction)Phase III trials hase been completed: + results 4500 patients – 2009.07.20.

Pfizer’s selective antagonist of the cannabinoid type 1 receptorStopped after phase III – 2008.11.06.

Merck’s MK 0557 Neuropeptid Y receptor (NPY5R) antagonist. Stopped after phase III – 2006.Oct.

Phase III

Pipeline of anti-obesity compounds (2006-)



OBEKON consortium – the start – 2006

Collaboration / alliance - consortium 7 members of academic institutions + university and industry – OBEKON 40 researcher - > 20 PhD and 4 members of the HAS

Submitted grant: „Identification of molecular targets and biomarkers in the mechanism and the treatment of obesity”

Start – 2006, summer with the significant financial support from the NKTH

Duration: 2.5 yrs – ends December, 2009



2006, spring - call for a grant application by the National Office for Research and Technology

Members of the OBEKON


•Richter Gedeon Plc. www.richter.hu

•Semmelweis University www.sote.hu Institute of Genetics, Cell- and Immunobiology

•Pázmány Péter Catholic University www.ppke.hu Faculty of Information Technology

•HAS, Chemical Research Center www.chemres.huStructural Chemistry Institute

•BioSystems International Ltd. www.biosys-intl.com

•TargetEx Research and Development Ltd. www.targetex.com

•QSX Quality Software Expert Ltd. www.qsx.hu


Our goals are:Using „systems biology” approach we intend to identify new molecular targets

and biomarkers by mapping of the molecular network of obesity. To collect large number of blood sample from obese and non-obese donors, prepare

plasma, DNA and RNA. Generate quality genomics and proteomics data. Analyse the data to identify new molecular targets and biomarkers by mapping of the

molecular network of obesity using bioinformatic tools.


…and the possible results - future• Initiate R & D project for the development of new obesity drug.

• Initiate the development of new diagnostic / prediction of the evolvement of obesity.


Work flow

Sample collection (human blood)

Data generation Clinical Genomics Proteomics

Data analysis - Bioinformatics

Sample and Data management – „Biobanking”



Collection of human samples Inclusion criteria

Age >18 yrsBMI > 30 kg/m2 - obeseBMI < 25 kg/m2 - control

Exclusion criteriaCancerPsychiatric disease Medication cause weight gain Endocrine diseaseChronic kidney disease Severe lung disease (e.g.asthma bronchiale, COPD III.st.)Severe heart diseaseLiver diseases (cirrhosis hepatis, hepatitis)GI surgerySport at competition level


8 clinical sites>1500 blood sample


Data generation - clinical data

Electronic questionnaireAnamnesis & family anamnesisPhysical conditionCurrent medicationLife-styleClinical laboratory data

Socio-demographic dataNumeric output: education, family status, economic

situation

Dietary data (3 days diary)Numeric output: energy, protein, fat, carbohydrates,

fatty acids, amino acids, etc….



Data generation - genomics


gene polymorphism: SNP

SNPstream® Genotyping System

Biomek FX Dual Arm System

Bioinformatics

gene expression profile: mRNA, miRNA, CGH micro-array


Data generation - proteomics


• Chromatography: ion-exchange, affinity, reverse phase,…

• Protein identification: HPLC-MS/MS

• Protein glycosylation pattern analysis: HPLC-MS

• Monoclonal antibodies production: hybridoma technology

• Epitop mapping: phage display

• Protein separation: one and two D gel-electrophoresis

• Cloning and expression


Data Generation summary



Source of data

Number of validated

sample

(obese/ nonob.)

Number of variable per

sample

Clinics

Anamnesis & laboratory & clinical data

1345

(836 / 509)≈ 100

Socio-demographic, life-style and dietary data

513 ≈ 100

Genomics

SNP, 55 genes-120SNP 1519 ≈ 100

CGH 15 ≈ 1000

Transcriptomics/mRNA

Whole Human Genome Microarray

32

(17 / 16)≈ 10000

Transcriptomics/ miRNA 32 ≈ 100

Proteomics

Biomarkers 3 x 50 ≈ 100

Glycosylation pattern analysis 3 x 50 ≈ 20

Data Analysis - Bioinformatics: Preparation Phase

Huge amount of dataToo many variables in some of the data sourcesReduction and modification using

Data evaluationStatistical analysisDiscretizationNormalizationStratification



Summary of Data Generation /Human



Source of data

Number of sample

(obese/ nonob.)

Number of variable per

sample

Derived number of

variable per sample for

further analysis

Clinics

Anamnesis & laboratory & clinical data

1345

(836 / 509)≈ 100 ≈ 10

Data evaluation

Socio-demographic, life-style and dietary data

519 ≈ 100 ≈ 10

Genomics

SNP, 55 genes-120SNP 1519 ≈ 100 ≈ 100CGH 15 ≈ 1000 ≈ 100

Transcriptomics/mRNA

Whole Human Genome Microarray

32

(17 / 16)≈ 10000

≈ 100Experimental

verification (qRT-PCR)

Transcriptomics/ miRNA 32 ≈ 100 ≈ 100

Proteomics Biomarkers 3 x 50 ≈ 100 ≈ 100

Glycosylation pattern analysis 3 x 50 ≈ 20 ≈ 20

Sample and data management: „OBEKON Biobank”Store samples

DNA, RNA, and plasmaand data

clinical, genomics, proteomicsContinuous maintenance

Clinical sampleData

Organized / regulated operation : SOP - in progressRegulated accessibilityMember of the National Biobank Network - www.biobank.hu



Summary

Large biobank: sample and data

Wide methodological arsenal – genomics, proteomics, and bioinformatics

Strong bases for further collaborative studies / analysis



Thank you forThank you for your attentionyour attention



1.Csaba Szalai: Investigation of the genomic background of obesity using single nucleotide polymorphism analysis in candidate genes

2.István Kurucz:

Application of proteomics methods in the identification of biomarkers, suitable for studying obesity and obesity related diseases.

3.Katalin Éder: Obesity related mRNA and miRNA profiling

4.Viola Tamasi: Obesity: genetic update by CGH analysis and its potential clinical implications



Further presentations: lectures

Further presentations: posters

1., Glycosylation pattern analysis with mass spectrometryLívia Budai, Ferenc Pollreisz, Olivér Ozohanics, Krisztina Ludányi, László Drahos, Károly Vékey

2., Epitope mapping of mABs recognizing protein markers of obesity: a phage display Flachner Beáta, Dobi Krisztina, Varga János, Lõrincz Zsolt, Cseh Sándor

3.,Pharmacologic therapy of obesity. Study of ligand-binding mode to lysophosphatidic acid receptors by molecular modelling methodsAndras Szilagyi, B Balogh, B Jojart, P Matyus



Identification of molecular targets and biomarkers in the mechanism and the treatment of obesity....

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