Uncovering Regeneration-Permissive Cues in Lower Vertebrate Retina to Inform Retinal Regenerative Medicine

[1606128]视网膜变性是一种普遍且日益严重的健康问题，尤其是与衰老相关的视网膜变性，目前影响着美国210万人，给经济带来了沉重的负担（每年约20亿美元）。它是全世界失明的主要原因。细胞移植策略显示出巨大的前景，但受到细胞存活率和整合水平低（1%）的显著限制。提出的研究旨在阐明使移植细胞存活和整合所需的化学和物理线索。这种基本的定量理解将为指导未来的视网膜再生策略和材料设计提供一个框架，以作为有效的细胞递送载体。拟议的项目还将培育一个有凝聚力的教育计划，包括从K-12到研究生的各个教育水平的教学。虽然一些低等脊椎动物的视网膜可以再生，但人类的视网膜却不能。下脊椎动物基质组成、结构和机械特性在组织再生中的作用尚未被探索，但可能是重要的，可以利用来实现再生医学的重大进展。PI最近的初步结果表明，哺乳动物视网膜祖细胞以一种类似于细胞移植的方式被引入低等脊椎动物（蝾螈）的视网膜时，表现出与哺乳动物视网膜截然不同的行为。具体来说，下部脊椎动物视网膜中的RPCs表现出明显增强的细胞存活率和融入视网膜的过程，这是再生的核心。本项目将分析下部脊椎动物允许再生视网膜微环境与成年哺乳动物视网膜微环境的化学和物理特征。该研究将包括深入研究人类视网膜祖细胞对下脊椎动物视网膜基质的反应，包括附着、存活、分化和3D迁移，这些过程对成功的细胞移植策略至关重要。该研究将分析细胞对完整组织和脱细胞基质的反应，在存在和不存在组织条件培养基的情况下，分析细胞与非细胞（基质）成分和可溶性与不可溶性线索的影响。重要的是，综合系统生物学方法将定量地联系细胞对组织/基质/可溶性因子线索和细胞信号事件的反应。这种定量机制的理解将为指导未来的视网膜再生策略和生物材料设计提供一个框架。该奖项由CBET部门的生物技术和生化工程项目颁发，由材料研究部的生物材料项目共同资助。

1606128 Carrier, Rebecca L. Retinal degeneration, particularly associated with aging, is a widespread and increasing health problem currently affecting 2.1 million people in the U.S. and imposing a substantial burden on the economy (~$2 billion annually). It is the leading cause of blindness worldwide. Cell transplantation strategies have shown tremendous promise, but are significantly limited by low levels of cell survival and integration (1%). The proposed studies aim to elucidate the chemical and physical cues required to enable survival and integration of transplanted cells. This fundamental, quantitative understanding will provide a framework to guide future retinal regeneration strategies and design of materials to serve as effective cell delivery vehicles. The proposed project will also nurture a cohesive education plan including teaching and learning across education levels spanning from K-12 to postgraduate. While the retinas of some lower vertebrate species do regenerate, the human retina does not. The role of lower vertebrate matrix composition, structure, and mechanical properties in tissue regeneration has not been explored, yet is likely significant and could be exploited to enable significant advances in regenerative medicine. The PI's recent preliminary results indicate that mammalian retinal progenitor cells display strikingly different behavior when introduced to the lower vertebrate (axolotl) retina, in a manner analogous to cell transplantation, relative to a mammalian retina. Specifically, the RPCs in the lower vertebrate retina exhibit markedly enhanced cell survival and integration into the retina - processes central to regeneration. This project will analyze the chemical and physical features of the lower vertebrate regeneration-permissive retinal microenvironment compared to that of adult mammalian retina. The research will include an in-depth study of human retinal progenitor cell response to lower vertebrate retinal matrix, including attachment, survival, differentiation and 3D migration, processes essential to successful cell transplantation strategies. The study will analyze cell response to both intact tissue and decellularized matrix, in the presence and absence of tissue-conditioned medium, to parse effects of cellular vs. acellular (matrix) components and soluble vs. insoluble cues. Importantly, an integrated systems-biology approach will quantitatively relate cell response to tissue/matrix/soluble factor cues and cell signaling events. This quantitative mechanistic understanding will provide a framework to guide future retinal regeneration strategies and biomaterial design.This award by the Biotechnology and Biochemical Engineering Program of the CBET Division is co-funded by the Biomaterials Program of the Division of Materials Research.