Periodic Reporting for period 1 - I-SEE (Cell Rejuvenation Therapy for Age-related Macular Degeneration)
Periodo di rendicontazione: 2022-06-01 al 2023-11-30
Age-related macular degeneration (AMD) stands as a predominant cause of blindness, impacting the macula and resulting in central vision loss due to retinal damage. This affliction, prevalent in individuals over 55 years old, affected 196 million people globally in 2020, with an estimated economic burden of $343 billion, encompassing $255 billion in direct healthcare costs. The macula, a critical region for visual acuity, relies on the intricate interplay of photoreceptors and retinal pigmented epithelium (RPE) cells. In the normal eye, RPE cells play a pivotal role in supporting photoreceptor function, maintaining the health of the retina, and facilitating optimal visual perception.
However, in AMD, a disease that progressively debilitates vision, photoreceptors and RPE cells undergo degeneration during aging. The prevailing standard of care involves periodic injections of anti-VEGF medications like ranibizumab (Lucentis), aflibercept (Eylea), and brolucizumab (Beovu), administered every 1-3 months. While these treatments impede disease progression, they fall short in restoring lost vision and entail side effects such as ocular bleeding.
Within the context of an ERC Starting project, the principal investigator unveiled a novel method that can potentially delay AMD by enhancing the function of RPE cells—a breakthrough in AMD treatment. The proposed strategy involves rejuvenation through partial reprogramming, a technique the results of which indicate both safety and efficacy in rejuvenating mouse fibroblasts while improving their function.
In this application, our focus shifts to RPE studies to pave the way for the implementation of our innovative approach embodied in the envisioned product, "I-SEE." "I-SEE" leverages the discovery of a trophoblast stem cell-specific (TSC) combination of factors, proven successful in reprogramming cells, to drive the rejuvenation and improved function of aging RPE cells. Our plan includes meticulous calibration of the expression of TSC reprogramming factors in RPE cells, followed by testing for rejuvenation and improved function in AMD mouse models.
The culmination of our efforts, "I-SEE," is built upon a PCT patent Application No. 63/210,030, featuring a cocktail of mRNA molecules directly applied to the eye. This innovative approach aims to provide the first AMD treatment capable of delaying vision lost, marking a significant advancement in the field.
However, in AMD, a disease that progressively debilitates vision, photoreceptors and RPE cells undergo degeneration during aging. The prevailing standard of care involves periodic injections of anti-VEGF medications like ranibizumab (Lucentis), aflibercept (Eylea), and brolucizumab (Beovu), administered every 1-3 months. While these treatments impede disease progression, they fall short in restoring lost vision and entail side effects such as ocular bleeding.
Within the context of an ERC Starting project, the principal investigator unveiled a novel method that can potentially delay AMD by enhancing the function of RPE cells—a breakthrough in AMD treatment. The proposed strategy involves rejuvenation through partial reprogramming, a technique the results of which indicate both safety and efficacy in rejuvenating mouse fibroblasts while improving their function.
In this application, our focus shifts to RPE studies to pave the way for the implementation of our innovative approach embodied in the envisioned product, "I-SEE." "I-SEE" leverages the discovery of a trophoblast stem cell-specific (TSC) combination of factors, proven successful in reprogramming cells, to drive the rejuvenation and improved function of aging RPE cells. Our plan includes meticulous calibration of the expression of TSC reprogramming factors in RPE cells, followed by testing for rejuvenation and improved function in AMD mouse models.
The culmination of our efforts, "I-SEE," is built upon a PCT patent Application No. 63/210,030, featuring a cocktail of mRNA molecules directly applied to the eye. This innovative approach aims to provide the first AMD treatment capable of delaying vision lost, marking a significant advancement in the field.
In this grant proposal, our primary objective has been to propel our groundbreaking cell rejuvenation technology into the industry, specifically emphasizing its potential in rejuvenating retinal pigment epithelium (RPE) cells. Our overarching goals encompassed:
1. Identifying the optimal delivery system for RPE cells, both in vitro and in vivo, and showcasing the rejuvenation properties of RPE cells in vitro.
2. Demonstrating the rejuvenation capabilities in RPE cells in elderly wild-type mice and an AMD model mouse, FAM161.
The unfortunate unavailability of adult RPE cells, due to the cessation of commercialization by the supplier, prompted an immediate shift in focus. Consequently, we assessed the delivery system in vivo within the animal eye. These experiments utilized clinically-approved Adeno-Associated Viruses (AAVs) and unequivocally exhibited the expression of reprogramming factors using either a robust promoter (CAGGS) or an RPE-specific promoter (BEST1), combined with dox-dependent control.
Given the exigencies of lacking in vitro RPE cells, we adapted our strategy and conducted further validation using human fibroblasts instead of human RPE, as initially planned. Rejuvenated human fibroblasts displayed a significantly younger epigenetic age, enhanced DNA damage repair following gamma radiation exposure, accelerated proliferation, improved resistance to oxidative stress, and superior DNA compaction.
These findings are of paramount importance, propelling us a significant step closer to practical clinical applications.
As in vivo experiments demand substantial time and meticulous calibration, we are presently engaged in fine-tuning the optimal timing and levels of the reprogramming factors in both wild-type and FAM161 mutant mice.
1. Identifying the optimal delivery system for RPE cells, both in vitro and in vivo, and showcasing the rejuvenation properties of RPE cells in vitro.
2. Demonstrating the rejuvenation capabilities in RPE cells in elderly wild-type mice and an AMD model mouse, FAM161.
The unfortunate unavailability of adult RPE cells, due to the cessation of commercialization by the supplier, prompted an immediate shift in focus. Consequently, we assessed the delivery system in vivo within the animal eye. These experiments utilized clinically-approved Adeno-Associated Viruses (AAVs) and unequivocally exhibited the expression of reprogramming factors using either a robust promoter (CAGGS) or an RPE-specific promoter (BEST1), combined with dox-dependent control.
Given the exigencies of lacking in vitro RPE cells, we adapted our strategy and conducted further validation using human fibroblasts instead of human RPE, as initially planned. Rejuvenated human fibroblasts displayed a significantly younger epigenetic age, enhanced DNA damage repair following gamma radiation exposure, accelerated proliferation, improved resistance to oxidative stress, and superior DNA compaction.
These findings are of paramount importance, propelling us a significant step closer to practical clinical applications.
As in vivo experiments demand substantial time and meticulous calibration, we are presently engaged in fine-tuning the optimal timing and levels of the reprogramming factors in both wild-type and FAM161 mutant mice.
Thus far, we have successfully showcased the efficient and cell-type-specific expression of reprogramming factors in retinal pigment epithelium (RPE) cells in vivo, conducted within the eyes of both wild-type (WT) and FAM161 mutant mice. Our investigations have not only confirmed these expressions but also substantially broadened our understanding of the rejuvenation potential inherent in our reprogramming factors, as evidenced by a comprehensive series of assays in human fibroblasts. These findings are of paramount importance, propelling us a significant step closer to practical clinical applications.
Currently, our focus remains on in vivo studies designed to rigorously evaluate the ability of the reprogramming factors to rejuvenate and enhance the functionality of RPE cells. This ongoing phase of calibration is of utmost importance, as it plays a pivotal role in ensuring the precision and efficacy of our technology within the complex environment of a living organism. These efforts are integral to the seamless translation of our innovative approach into practical clinical applications, marking a crucial step toward the realization of our therapeutic goals.
Currently, our focus remains on in vivo studies designed to rigorously evaluate the ability of the reprogramming factors to rejuvenate and enhance the functionality of RPE cells. This ongoing phase of calibration is of utmost importance, as it plays a pivotal role in ensuring the precision and efficacy of our technology within the complex environment of a living organism. These efforts are integral to the seamless translation of our innovative approach into practical clinical applications, marking a crucial step toward the realization of our therapeutic goals.