An eye-drop formulation of Fas-mediated apoptosis inhibitor attenuates dry AMD = Fas 매개 세포사멸 억제 치료제의 점안 투여를 통한 건성 황반변성 치료

다국어 초록 (Multilingual Abstract)

Age -related macular degeneration (AMD) is a degenerative disease that occurs in the macula of the retina by Fas-mediated apoptosis. While dry AMD accounts for ~90% of macular degeneration, wet AMD accounts for approximately 10% of people with macular degeneration and can be treated with anti-VEGF therapies. However, current pharmaceuticals are unresponsive in about 40% of patients and intravitreal injection treatments are inaccessible, complex to use, and prone to side effects. Furthermore, no treatments are available to patients with dry AMD.

Fas is a major regulator of the death signal. Fas induced apoptosis in the retina causes macular degeneration and retinal cell death results in loss of vision. In this study, we used Fas-binding peptide (FBP) to inhibit Fas-mediated apoptosis in retinal cells, inhibiting AMD progression in sodium iodate (NaIO3)-induced dry AMD mice models. Intravitreal inoculation of FBP successfully binds to Fas and inhibits Fas-mediated apoptosis in retinal inhibiting AMD progression in vivo. However, an eye-drop formulation of FBP could not deliver to the posterior segment of the retina. To address this limitation, we used an eye-drop formulation of FBP conjugated with hexa-arginine (6R-FBP) for ocular delivery of FBP to the retinal segment of the eye. An eye drop formulation of 6R-FBP delivered to the posterior ocular regions of the macula of the retina segment of the eye and attenuated AMD by blocking Fas-mediated apoptosis. Application of an eye-drop formulation of 6R-FBP attenuates Fas-mediated apoptosis in retinal cells and downregulates inflammation in dry AMD.

Our data demonstrates that the inhibition of Fas-mediated signaling results in a reduction of retinal cell death and provides a therapeutic treatment for dry AMD. Our study provides an eye-drop formulation of 6R-FBP is a convenient and patient-compliant route of drug administration as a novel therapeutic intervention in treating AMD.

목차 (Table of Contents)

• Introduction 1
• Material and Methods 6
• 1. Peptide 6
• 2. Cell culture studies 7
• 3. Evaluation of Fas expression in vitro 7
• 4. Cell viability test of Fas inhibition in vitro 8
• 5. Apoptosis inhibition analysis in vitro 8
• 6. CPP toxicity test in vitro 8
• 7. Animal studies 9
• 8. Evaluation for localization of delivered FBP in vivo 9
• 9. Evaluation for delivery of FBP in vivo 10
• 10. Evaluation of cytokine level in vivo 11
• 11. Evaluation of therapeutic efficacy in vivo 13
• 12. Statistical Analysis 13
• Results 14
• 1. FBP can reduce inflammation and apoptosis in ARPE-19 cells by binding to the Fas receptor 14
• 2. FBP in vivo delivery test in AMD mice model 20
• 3. FBP in vivo therapeutic test in AMD mice model 27
• 4. FBP in vivo therapeutic test in AMD rabbit model 32
• 5. FBP in vivo size dependent delivery test in AMD mice model 38
• Discussion & Conclusion 41
• References 43
• 국문 요지 50