Pharmacogenomics and Therapeutic Target Validation of Cardiovascular - - PowerPoint PPT Presentation

Pharmacogenomics and Therapeutic Target Validation of Cardiovascular Disease Models

Toshio Tanaka MD, PhD

Department of Molecular and Cellular Pharmacology Pharmacogenomics and Pharmacoinformatics Mie University Graduate School of Medicine February 25, 2009 UNAM, Mexico City, Mexico

Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics、Mie University Graduate School of Medicine

Drug Target

Medicine

Gene Gene Gene Drug Gene Disease Gene Disease Gene Drug Gene Therapeutic Gene Therapeutic Gene

Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics、Mie University Graduate School of Medicine

Pharmacogenomics Network Analysis of Therapeutic Genes

J Pharmacol Sci 107, 8 – 14 (2008)

Rat Cerebral Vasospasm Model

HE staining (x 100) Circulation (2004)110:1839-1846

Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics、Mie University Graduate School of Medicine

HSP72 Protein after SAH

Circulation (2004)110:1839-1846

Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics、Mie University Graduate School of Medicine

HSP72 Protein after SAH and Antisense

Saline SAH Antisense Sense

Circulation (2004)110:1839-1846

Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics、Mie University Graduate School of Medicine

Effect of Antisense HSP72 ODN

• n Cerebral Vasospasm

Circulation (2004)110:1839-1846

Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics、Mie University Graduate School of Medicine

Circulation (2004)110:1839-1846

Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics、Mie University Graduate School of Medicine

Effect of GGA on HSP72 Gene Expression

Circulation (2004)110:1839-1846

Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics、Mie University Graduate School of Medicine

Circulation (2004)110:1839-1846

Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics、Mie University Graduate School of Medicine

Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics、Mie University Graduate School of Medicine

Pharmacogenomics network analysis in treatment of cerebral vasospasm by GGA J Pharmacol Sci 107, 8 – 14 (2008)

20 16 12 8 4

pre-injection “1-saline” group “2-saline” group “1-hemorrhage” group “2-hemorrhage” group

HO-1/HO-2

10 min Day 12 10 min Day 12 10 min Day 12 10 min Day 12 10 min Day 12 10 min Day 12

Brain-stem Cerebellum Occipital Lobe Frontal Lobe Parietal Lobe Basilar Artery

Heme Oxygenase - 1 mRNA in Rat Cerebral Vasospasm

J.Clin. Invest. 104:59-66(1999)

Effect of Antisense HO-1 ODN on Angiographic Vasospasm in Rat SAH Model

• 5
• 10
• 15
• 20

5 Pre Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7 % Change in Vessel Diameter

a nti se ns e HO -1 O D N scr as mb le d O D N sen se H O-1 O DN

J.Clin. Invest. 104:59‐66(1999)

Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics、Mie University Graduate School of Medicine

Pharmacogenomics network analysis in treatment of cerebral vasospasm by AVS(nicaraven) J Pharmacol Sci 107, 8 – 14 (2008)

Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics、Mie University Graduate School of Medicine

Comparative Genomics of Disease Models Human(2003) Rat/Mouse Zebrafish (2005/2002) (2009) Low Throughput High Throughput

– 1. Short reproductive cycle. – 2. Large number of offspring per mating. – 3. Embryos are transparent allowing internal organs and neurons to be visualised. – 4. Strong correlation between human genetics, anatomy and disease. – 5. Rapid layout of body plan, allowing study of disease at early stages of embryonic development. – 6. Ethically preferred in the drive to reduce, refine and replace. – 7. Cost effective.

Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics、Mie University Graduate School of Medicine

Advantages of Zebrafish as Disease Model Animal

Zebrafish Heart Failure Models Zebrafish Heart Failure Models

Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics、Mie University Graduate School of Medicine

Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics、Mie University Graduate School of Medicine

Control Zebrafish Heart

Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics、Mie University Graduate School of Medicine

Zebrafish Heart Failure Model

Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics、Mie University Graduate School of Medicine

Carvedilol Treatment of Heart Failure Model

Carvedilol

Carvedilol Therapeutic Gene Discovery

Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics、Mie University Graduate School of Medicine

Laser-Captured Microdissection

HF Carvedilol HF

before after

Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics、Mie University Graduate School of Medicine

Therapeutic Gene Validation with Calvedilol

HF HF Carvedilol Control

Gene Expression

siRNA cocktail / Morpholino Antisense法

Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics、Mie University Graduate School of Medicine

赤：Up by PECI kd 緑：Down by PECI kd

Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics、Mie University Graduate School of Medicine

Metabolome Transcriptome

Genotype

Forward Pharmacogenomics Reverse Pharmacogenomics

Phenotype

Proteome

Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics、Mie University Graduate School of Medicine

J Pharmacol Sci 107, 8 – 14 (2008)