Abstract Body: Background: Retinal vasculopathy had been reported across the continuum of neurodegeneration. Retinal color fundus photography coupled with automated retinal vascular analysis offers potential to non-invazively screen for Alzheimer’s disease (AD) neurodegeneration. Amyloid positron emission tomography (PET) and apolipoprotein 4 (APOE4) carrier status are known biomarkers of AD risk. Herein, we examined the relationship between retinal vascular fractal dimensions in non-mydriatic color fundus photographs, amyloid-PET burden and APOE4 carrier status in a cohort of cognitively intact individuals. Methods: Our dataset included 91 macula-centered and 39 optic disc-centered images from 96 cognitively intact participants (29% male). 25 (26%) were amyloid-PET positive, defined as amyloid-PET standardized uptake value ratio centiloid cut-off > 20. AutoMorph software automatically determined retinal arteriolar and venular density, tortuosity and width, among other fractal dimensions. To test the effect of amyloid-PET status on the retinal vascular parameters, we used a generalized linear model compensated for age, sex and the interaction between APOE4 carrier and amyloid-PET status. We also compared amyloid-PET positive and negative subjects for vascular fractal dimensions correcting for age and gender. We adjusted for multiple comparisons using the False Discovery Rate correction and reported significant P values less than 0.05. Results: Compared to amyloid-PET negative, cognitively intact amyloid-PET positive cohort had greater mean age (71 vs 66 years, P=0.06), more males (60% vs 35%, P=0.02), more APOE4 carriers (P<0.001), similar vascular risk factors (hypertension, dyslipidemia, diabetes mellitus, smoking, P>0.05 for all) and significantly lower macular vessel tortuosity density (P=0.019). The amyloid-PET positive status had a significant effect on the retinal artery distance tortuosity (corrected P=8.75E-04, β=13.9, 95% CI [7.75-19.99]) and artery squared curvature tortuosity (corrected P=8.91E-10, β=464, 95% CI [359.10-569.15]). Conclusions: Automatically determined retinal vascular fractal dimension on non-mydriatic color fundus photographs predicts amyloid-PET burden in cognitively intact individuals. Future longitudinal studies should asssess the utility of automated quantitative retinal vasculopathy analysis to screen for presymptomatic AD.