1. 1 Pharmacogenetics for Genes Associated with Age-Related Macular Degeneration in the Comparison of AMD Treatments Trials (CATT) Hagstrom SA, Ying G-S, Pauer GJT, Sturgill-Short GM, Huang J, Callanan DG, Kim IK, Klein ML, Maguire MG Hagstrom SA, Ying G-S, Pauer GJT, Sturgill-Short GM, Huang J, Callanan DG, Kim IK, Klein ML, Maguire MG, Martin DF for the CATT Research Group Available through http://www.med.upenn.edu/cpob/publications_main.shtml http://www.med.upenn.edu/cpob/publications_main.shtml Hagstrom SA, Ying G-S, Pauer GJT, Sturgill-Short GM, Huang J, Callanan DG, Kim IK, Klein ML, Maguire MG Hagstrom SA, Ying G-S, Pauer GJT, Sturgill-Short GM, Huang J, Callanan DG, Kim IK, Klein ML, Maguire MG, Martin DF for the CATT Research Group Available through http://www.med.upenn.edu/cpob/publications_main.shtml http://www.med.upenn.edu/cpob/publications_main.shtml Supported by Cooperative Agreements from the National Eye Institute, National Institutes of Health, DHHS

2. Genotype Determination Each patient was genotyped for CFH, ARMS2, HTRA1, and C3. GeneSNPExonAmino Acid CFHrs10611709Y402H ARMS2rs104909241A69S HTRA1rs11200638promoter- C3rs22301993R80G InflammationLipid Metabolism Angiogenesis Oxidative Stress ECM Stasis AMD Biological Pathways VEGFA KDR CFI BF CFH C3 C2 ARMS2 ND2 HTRA1 TIMP3 COL8A1 FRK APOE LIPC CETP LPL ABCA1 Hagstrom et al. Pharmacogenetics of Anti-VEGF Therapy in CATT Ophthalmology 2013;120:593-599

3. Homozygous Allele 1 Homozygous Allele 2 Heterozygous Negative Controls Genotyping was performed using a custom made TaqMan OpenArray loaded with TaqMan SNP genotyping assays. Genotype Determination Hagstrom et al. Pharmacogenetics of Anti-VEGF Therapy in CATT Ophthalmology 2013;120:593-599

4. Outcome Variables All outcomes were determined following standardized protocols. VA assessed using eETDRS testing OCT outcomes determined by independent OCT Reading Center FA and photographic outcomes determined by independent Fundus Photographic Reading Center Hagstrom et al. Pharmacogenetics of Anti-VEGF Therapy in CATT Ophthalmology 2013;120:593-599

5. Comparison of baseline demographic and ocular characteristics between participants and non-participants in the genetic study (N=1149) Hagstrom et al. Pharmacogenetics of Anti-VEGF Therapy in CATT Ophthalmology 2013;120:593-599 Baseline Characteristics Subjects in genetic study (N=834) Alive subjects not in genetic study (N=315) P Value Age (years): Mean (SD)78.5 (7.5)80.9 (7.2)<0.0001 Female (%) 510 (61.2)204 (64.8)0.28 Former or current cigarette smoker (%)483 (57.9)169 (53.7)0.36 Presence of hypertension (%)563 (67.5)232 (73.7)0.045 Taking AREDS supplement (%)536 (64.3)189 (60.0)0.21 Baseline VA (letters): Mean (SD)61.3 (13.3)58.8 (13.7)0.005 Baseline area of CNV (DA): Mean (SD)1.70 (1.69)1.91 (1.90)0.096 Baseline total area of CNV lesion (DA): Mean (SD) 2.47 (2.55)2.49 (2.54)0.87 Presence of occult lesion (%)505 (60.6)169 (53.7)0.04 Presence of RAP lesion (%)80 (9.6)41 (13.0)0.12 Total foveal thickness (microns): Mean (SD)462 (190)456 (180)0.60 SD = standard deviation; AREDS = age-related eye disease study; VA = visual acuity; DA = disc area.

6. Visual Outcomes by Genotype 6GeneGenotypen Mean VA in letters Mean VA change in letters ≥ 15 letters increase CFH CC270707.928% TC391688.229% TT173698.634% P-Value0.300.610.22 ARMS2 TT170698.228% GT398698.331% GG266707.928% P-Value0.510.770.97 HTRA1 AA162698.429% AG398698.231% GG274697.929% P-Value0.680.690.99 C3 GG56718.128% CG318708.931% CC460687.629% P-Value0.030.340.72 # of Risk Alleles 0-1123687.029% 2141699.133% 3175697.927% 4170698.131% >=5225708.529% P-Value0.420.660.71

7. Genotypic Associations with Anatomic Outcome Measures on OCT at 1 Year (N=834) 7GeneGenotypen Retinal thinning <120 µm Retinal thickness >212 µm Mean change of total foveal thickness in microns Dry on OCT CFH CC270 17%12% -14227% TC391 22%9% -18829% TT173 21% 11%-17433% P-Value 0.62 0.030.18 ARMS2 TT17020%10%-18428% GT398 20% 9%-17633% GG266 21% 13%-15224% P-Value0.330.060.30 HTRA1 AA16221%10%-17828% AG39820%10%-17933% GG27420%12%-15225% P-Value0.640.100.23 C3 GG5620%11%-18235% CG31822%8%-16132% CC46019%12%-17427% P-Value0.510.670.07 # of Risk Alleles 0-112322%15%-15325% 214118%9%-15724% 317520%12%-20029% 417024%7%-17437% >=522517%11%-16029% P-Value0.290.680.30 Hagstrom et al. Pharmacogenetics of Anti-VEGF Therapy in CATT Ophthalmology 2013;120:593-599

8. FA and Fundus Outcomes by Genotype (N=834 ) 8GeneGenotypen Dye leakage on FA Mean change in lesion size in disc area CFH CC27046%0.2 TC39147%0.2 TT17344%0.4 P-Value0.710.48 ARMS2 TT17049%0.5 GT39844%0.0 GG26648%0.4 P-Value 0.890.80 HTRA1 AA16249%0.5 AG39843%0.1 GG27449%0.3 P-Value 0.840.42 C3 GG5650%0.0 CG31845%0.1 CC46046%0.3 P-Value 0.850.28 # of Risk Alleles 0-112355%0.2 214145%0.5 317538%0.1 417044%0.0 >=522550%0.3 P-Value 0.930.80 Hagstrom et al. Pharmacogenetics of Anti-VEGF Therapy in CATT Ophthalmology 2013;120:593-599

9. Number of Injections by Genotype 9GeneGenotypen Mean of total number of injections in PRN Groups CFH CC1257.4 TC1667.4 TT657.6 P-Value0.72 ARMS2 TT69 8.0 GT170 7.2 GG117 7.4 P-Value 0.35 HTRA1 AA678.0 AG1707.3 GG1197.3 P-Value 0.25 C3 GG27 7.0 CG138 7.9 CC1917.2 P-Value0.30 # of Risk Alleles 0-151 7.6 259 7.0 380 7.2 465 7.3 >=51017.9 P-Value 0.27 Hagstrom et al. Pharmacogenetics of Anti-VEGF Therapy in CATT Ophthalmology 2013;120:593-599

10. Conclusions 10 Largest pharmacogenetic analysis exploring the relationship between genotypes and response to anti-VEGF therapy with >90% power to detect a 1 line difference between the highest risk and lowest risk groups. The relationship between genotype and response to therapy did not vary by drug or dosing regimen. Although specific risk alleles for CFH, ARMS2, HTRA1 and C3 may predict the development of AMD, they did not predict response to anti-VEGF therapy. Further studies targeting SNPs in additional biological pathways are ongoing. Hagstrom et al. Pharmacogenetics of Anti-VEGF Therapy in CATT Ophthalmology 2013;120:593-599