LncRNA-1810058I24Rik及其编码多肽在视网膜发育和维持中的调控作用

Retinal development is a complex process regulated by various intrinsic and extrinsic factors. Recent studies have showed that long non-coding RNA (LncRNA) is an important intrinsic factor regulating retinal development. Among the LncRNAs with high expressions in retina, 1810058I24Rik is highly expressed during retinal embryonic development, but gradually down-regulated after birth, indicating that 1810058I24Rik plays an important role in retinal development. Our preliminary results showed that overexpression of 1810058I24Rik transcript was sufficient to promote the generation of amacrine, bipolar and Müller cells. Further in vitro experiments demonstrated that a short open reading frame V2sORF (144 bp) in 1810058I24Rik encoded a conserved mitochondrial peptide (47aa), and overexpression of V2sORF also promoted the differentiation of amacrine, bipolar and Müller cells. Therefore, we hypothesize that 1810058I24Rik regulates retinal development mainly mediated by V2sORF encoded peptide. In this project, by using gene overexpression, shRNA interference, gene knockout, immunostaining and other techniques, we aim to verify our hypothesis, and confirm the effect of 1810058I24Rik in the maintenance of retinal function in knockout mice. This project is expected to reveal the new mechanism of LncRNA in regulating retinal development, and provide a scientific basis for establishing LncRNA as a new target for the treatment of retinal diseases.

视网膜发育是一个受到多种因子调控的复杂过程，长链非编码RNA（LncRNA）作为一类基因元件，在视网膜发育过程中发挥重要作用。在视网膜高表达的LncRNA中，1810058I24Rik（18R）在胚胎期高表达，出生后下调，提示18R在视网膜发育中可能起重要作用。我们发现过表达18R促进视网膜无长突细胞、双极细胞和Müller细胞分化。进一步体外实验证明18R序列中短开放阅读框V2sORF编码保守线粒体功能多肽（47aa），过表达V2sORF产生与过表达18R相同的细胞命运改变效应。我们提出假设：18R对视网膜发育的调控主要通过V2sORF编码多肽介导。本项目拟采用基因过表达、shRNA干扰、基因敲除和免疫染色等方法，验证我们的假设，以及在基因敲除小鼠中验证18R在视网膜功能维持中的作用。本项目有望揭示LncRNA调控视网膜发育的新机制，为确立LncRNA作为视网膜疾病治疗靶点提供科学依据。

视网膜发育和损伤是受到多种因子调控的复杂过程，长链非编码RNA（LncRNA）作为一类基因元件，在视网膜发育和损伤调控过程中发挥重要作用。我们通过对胚胎到成年小鼠视网膜进行系统的RNA-seq分析，鉴定出一个LncRNA-1810058I24Rik (181Rik)参与调控视网膜发育和损伤过程。181Rik在胚胎期高表达，出生后下调，可编码一个47 氨基酸的线粒体短肽Stmp1。在视网膜中过表达181Rik和Stmp1都能够促进双极细胞、无长突细胞和Müller细胞的分化，而减少感光细胞的分化，表明181Rik可调控视网膜发育过程。作为许多眼部疾病（如糖尿病视网膜病变和青光眼）的常见病理，视网膜缺血/再灌注（IR）会引发炎症和小胶质细胞的激活，导致不可逆的视网膜损伤。然而，目前对视网膜IR损伤的详细分子机制仍知之甚少。在我们构建的181Rik 敲除小鼠（181Rik-KO）中，我们发现181Rik的缺失可减轻IR模型中损伤视网膜的凋亡，提高IR模型损伤视网膜的神经节细胞（RGC）生存率，抑制IR模型损伤视网膜的炎症反应，以及小胶质细胞的增值、激活和吞噬作用。即181Rik在小鼠视网膜中的缺失通过减弱小胶质细胞和Nlrp3炎性小体通路的激活来保护RGC免受IR的损伤。 这一作用可能是通过抑制代谢传感器Ucp2表达以及激活Ca2+传感器S100a8/a9表达来控制线粒体和代谢功能，从而触发Nlrp3炎症小体的激活。我们的发现为视网膜发育机制和IR损伤的分子发病机制提供了新的启示，为确立LncRNA作为IR相关的神经退行性疾病提供一个新的治疗目标。