Glaucoma Epigenetic Treatment with Light Activated CRISPR-dCas9

青光眼是世界范围内导致不可逆转失明的主要原因。青光眼的视力丧失是由将视觉信息从视网膜传递到大脑的视网膜神经元的死亡引起的。目前，青光眼还没有治愈的方法。近年来，青光眼的大多数形式都是遗传性的，遗传因素在青光眼的发生和发展中起着关键作用。青光眼致病基因似乎是青光眼治疗的非常有前途的治疗靶点。最近还发现，视网膜神经元功能的丧失不仅可以减缓，而且还可以逆转。因此，似乎有希望不仅可以使用遗传和表观遗传学方法减缓青光眼的进展，而且还有可能通过视网膜神经元和视神经轴突再生来逆转青光眼的进展。研究人员提议开发一种基于可见光激活的CRISPR-dCas9的表观遗传学方法来治疗青光眼，并有可能恢复视力。CRISPR是一种用于多种基因组改变的革命性新技术，但CRISPR在预期靶点以外的位置诱导的高频率非靶点活动是一个主要问题，特别是在治疗和临床应用中。这个问题可以用一种光激活的CRISPR版本来解决，它可以进行局部的基因改变，将非靶标活动限制在只有受照细胞上。他们将开发和建造一种新的可见光局部激活CRISPR的仪器。拟议的基于光激活CRISPR-dCas9的青光眼治疗和视力恢复的新型表观遗传学方法具有变革性，因为该平台可能对为各种视网膜疾病开发高效眼科疗法产生深远影响。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

Glaucoma is the leading cause of irreversible blindness worldwide. Loss of vision in glaucoma is caused by the death of retinal neurons that convey visual information from the retina to the brain. Currently, there is no cure for glaucoma. In recent years it has become apparent that most forms of glaucoma are inherited and the genetic factors play a key role in its onset and development. Glaucoma-causative genes appear to be very promising therapeutic targets for glaucoma treatment. Recently, it has also been discovered that the loss of the retinal neuron function could not only be slowed down but could also be reversed. Thus, it seems hopeful that not only could the glaucoma’s progress be slowed down using genetic and epigenetic approaches, but it may also be possible to reverse its progression with retinal neuron and optic nerve axon regeneration.The investigators propose to develop a novel visible light activated CRISPR-dCas9 based epigenetic method for glaucoma treatment and the potential for vision restoration. CRISPR is a revolutionary new technology for versatile genome alterations but the high frequency of off-target activity induced by CRISPR at sites other than the intended on-target one is a major concern, especially for therapeutic and clinical applications. This problem could be solved with a light activatable version of CRISPR, which enables localized genetic alterations, limiting off-target activity only to the illuminated cells. They will develop and construct a new instrument for localized CRISPR activation with visible light. The proposed novel light activated CRISPR-dCas9 based epigenetic method for glaucoma treatment and vision restoration is transformative since the platform could have a profound impact on developing highly effective ophthalmic therapies for a wide range of retinal disorders.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.