1. AMD Genetic Pathogenesis Peng ZHOU, M.D.

2. Aging population increasing  Elderly population over 143 million  Will reach 200 million by 2014 0% 5% 10% 15% 20% 25% 30% 35% 40% 45% 50% 195019701990 201020302050 Abobe 50 years old 0% 5% 10% 15% 20% 25% 50 岁 -60 岁 -70 岁 - 80 岁 - Prevalence of AMD 5.7 13.5 20.2 23.5 上图： Population division of the dept of the United Nations Secretariat, world population prospects,2004 下图：邹海东, 等. 中华眼科杂志.2005;41:15-19 Klein R, et.al Am J Ophthalmol.2004; 137:486-495 2 Prevalence of AMD increased with age  About 20% in Shanghai over 70 y/o  25% in the United States over 70 y/o Age-related Macular Degeneration (AMD)

3. Becoming No. 1 cause of blindness in China  7.6-15.5% in China over the age of 50  21.4 million AMD patients in China 3 黄晓波, 等. 中华眼科杂志.2009;45:786-92 No. 1 cause of blindness in U.S. No. 1 irreversible blindness developed world Congdon N.Arch Ophthalmol. 2004:122:477-85. Klein R, et.al Am J Ophthalmol.2004; 137:486-495 Age-related Macular Degeneration (AMD)

4. Oxidative stress Neovascularization Immune Genetics Environment (smoking) Previous research: Fragmented 1. No complete theory for AMD pathogenesis 2. Can not explain the different phenotypes Pathogenesis of AMD

5. AMD pathogenesis hypothesis Genetics Immunology NeovascularizationOxidativeStress Scar The sequence of different mechanisms :

6. AMD pathogenesis of the research strategy  Previous Strategy has its limitations  Population genetics provides an effective way  Theoretically, in a large population, the disease- causing gene polymorphism will lead to changes of AMD susceptibility  Our research strategy: ： Exon sequencing  AMD susceptibility genes Gene function  the pathogenesis of AMD 遗传与环境

7. 一、 Exon sequencing  Exon sequencing found many new genes  Needs more samples  HERPUD1 and COL15A1 related to PCV  An important way for early screening of PCV 遗传与环境

8. 二、 Functional Studies of Four genes  ARMS2  CFH  HTRA1  TLR3 遗传与环境

9. 1 、 ARMS2 遗传与环境 ◆ Background:  ARMS2 is associated with AMD ◆ In our study ：  Associated with AMD in Chinese Population  New CNV associated SNPs  New PCV associated SNPs  Difference between CNV and PCV Cheng Y, Li XX, et.al. Genetic and Functional Dissection of ARMS2 in Age-Related Macular Degeneration and Polypoidal Choroidal Vasculopathy. PloS ONE

10. 1 、 ARMS2 ◆ Background:  The mechanism of ARMS2 in unclear ◆ In our study ：  ARMS2 mutation plasmid affect the biological behavior of the vascular endothelial cells and RPE cell proliferation, adhesion and migration 遗传与环境 增殖能力：突变型＞野生型＞对照组，有统计学 差异 M 对照 野生型 突变型 55k Da 43k Da 34k Da ARMS2 野生型 、 突 变 型基因蛋白表达 升高

11. 2 、 CFH 遗传与环境 ◆ Background:  CFH is associated with AMD ◆ In our study ：  Associated with AMD in Chinese Population  Published in 2012 IOVS Tian J, Li XX. et.al Association of Genetic Polymorphisms and Age-Related Macular Degeneration in Chinese Population. Invest Ophthalmol Vis Sci. 2012;53:4262–4269

12. 2 、 CFH 遗传与环境 ◆ Background:  The mechanism of CFH in unclear ◆ In our study ：  H402Y significantly increases binding capacity of CFH to oxidizing lipids  Link genetic - oxidative stress - immune - neovascularization together  Support our proposal  PNAS in press CFH genotypes impact risk of AMD by interaction with oxidized phospholipids PNAS 2012

13. 3 、 HTRA1 遗传与环境 ◆ Background:  HTRA1 is associated with AMD ◆ In our study ：  Associated with AMD in Chinese Population  New CNV associated SNPs  New PCV associated SNPs  Difference between CNV and PCV Jiang JJ, Li XX, et.al. Overexpression of HTRA1 Leads to Down-regulation of Fibronectin and Functional Changes in RF/6A Cells and HUVECs. PloS ONE

14. 3 、 HTRA1 遗传与环境 ◆ Background:  HTRA1 is a serine protease  The mechanism of HTRA1 in unclear ◆ In our study ：  Destruct the vessel wall, lead to PCV like lesions  Published in 2011 PNAS Jones A, Zhang K, et.al. Increased expression of multifunctional serine protease, HTRA1, in retinal pigment epithelium induces polypoidal choroidal vasculopathy in mice. PNAS 2011:108:14578– 14583

15. 3 、 HTRA1 遗传与环境 ◆ Furthermore ：  HTRA1through Differentiation Factor 6 lead to CNV  Published in 2011 JBC Zhang L, Zhang K, et.al. High Temperature Requirement Factor A1 (HTRA1) Gene Regulates Angiogenesis through Transforming Growth Factor- Family Member Growth Differentiation Factor 6. JBC. 2012: 287:1520–1526

16. 4 、 TLR3 遗传与环境 ◆ Background:  TLR3 is NOT associated with wet AMD ◆ In our study ：  TLR3 Exon sequencing  Associated with wet AMD in Chinese Population Sun YY, Li XX, et.al. Role of TLR3 C1234T Variant on Choroidal Neovascularization is Controversial between Meta Analysis and Experimental Study. IOVS

17. 4 、 TLR3 ◆ Background:  The mechanism of TLR3 in GA in unclear ◆ In our study ：  TLR3 rs5743391decreases the binding capacity of TLR3 to dsRNA  Protects against from GA 遗传与环境

18. Molecular Pathogenesis of AMD Genetics Multi-gene diseases Each gene mutation may lead to AMD Genetic - environment interactions lead to different phenotypes Four genes play an important role in the pathogenesis of AMD HTRA1 Elastase CFH PCV CNV TRL3 Geographic Atrophy ARMS2 Upstream needs further investigation Oxidation Immunology Neovascularization Scar GDF6

19. Thank s ！