Retinal Circular RNA hsa_circ_0087207 Expression Promotes Apoptotic Cell Death in Induced Pluripotent Stem Cell-Derived Leber’s Hereditary Optic Neuropathy-like Models

Abstract

Backgrounds: Leber’s hereditary optic neuropathy (LHON) is known as an inherited retinal disorder characterized by the bilateral central vision loss and degeneration of retinal ganglion cells (RGCs). Unaffected LHON carriers are generally asymptomatic, suggesting that certain factors may contribute to the disease manifestations between carriers and patients who carry the same mutated genotypes. Methods: We first aimed to establish the iPSC-differentiated RGCs from the normal healthy subject, the carrier, and the LHON patient and then compared the differential expression profile of circular RNAs (CircRNAs) among RGCs from these donors in vitro. We further overexpressed or knocked down the most upregulated circRNA to examine whether this circRNA contributes to the distinct phenotypic manifestations between the carrier- and patient-derived RGCs. Results: iPSCs were generated from the peripheral blood cells from the healthy subject, the carrier, and the LHON patient and successfully differentiated into RGCs. These RGCs carried equivalent intracellular reactive oxygen species, but only LHON-patient iPSC-derived RGCs exhibited remarkable apoptosis. Next-generation sequencing and quantitative real-time PCR revealed the circRNA hsa_circ_0087207 as the most upregulated circRNA in LHON-patient iPSC-derived RGCs. Overexpression of hsa_circ_0087207 increased the apoptosis in carrier iPSC-derived RGCs, while knockdown of hsa_circ_0087207 attenuated the apoptosis in LHON-patient iPSC-derived RGCs. Predicted by bioinformatics approaches, hsa_circ_0087207 acts as the sponge of miR-665 to induce the expression of a variety of apoptosis-related genes in LHON patient iPSC-derived RGCs. Conclusions: Our data indicated that hsa_circ_0087207 upregulation distinguishes the disease phenotype manifestations between iPSC-derived RGCs generated from the LHON patient and carrier. Targeting the hsa_circ_0087207/miR-665 axis might hold therapeutic promises for the treatment of LHON.

Recognizing the Differentiation Degree of Human Induced Pluripotent Stem Cell-Derived Retinal Pigment Epithelium Cells Using Machine Learning and Deep Learning-Based Approaches

“…Nevertheless, the optic vesicles, form which the neurites projected, were heterogeneously consisted of somas and other cell structures (Figure 1). On the other hand, can be individually separated manually, however, it requires significant manpower [9,10]. The optic nerve is well known not to regenerate, and the repair of damaged optic nerves using iPSC-RGCs remains a hurdle in clinical transplantation [49].…”

“…The serial passaging was used to select MSC-like cells which were more homogenous and exhibited pronounced fibroblast morphology. For the differentiation of iPSCs into iPSC-RPEs and iPSC-RGCs, the differentiation following the procedures reported by Regent et al [28], Yang et al [8] and the protocols for the routine generation of iPSC-RGCs in our laboratory [9,10] were used, respectively.…”

“…In addition, following the stimuli by defined factors, iPSCs can undergo differentiation into different lineages and exhibit robust phenotypic and genotypic changes [2,7]. For instance, iPSCs can be differentiated into various retinal cell types, including retinal pigment epithelial cells (iPSC-RPEs) [8], retinal ganglion cells (iPSC-RGCs) [9,10], and retinal organoids [11,12]. These iPSC-derived retinal cell lineages have been utilized for disease modeling [9], pathogenesis investigation [9], and regeneration medicine [13].…”

Features of generation and differentiation of iPSCs into retinal cells for modeling human hereditary diseases

“…У ганглиозных клеток также нарушалась биологическая активность и изменялся профиль экспрессии генов по сравнению с нормальными клетками [13]. Кроме того, в работе [14] показано, что у ганглиозных клеток наблюдается повышенный уровень апоптоза, связанный с гиперэкспрессией циркулирующей РНК. В ещё одном исследовании, посвящённом получению ганглиозных клеток из ИПСК пациентов с LHON, показано снижение уровня экспрессии генов, связанных с глутаматергической синаптической передачей сигналов [21].…”

Leber’s hereditary optic neuropathy: Update on the novel genes and therapeutic options

“…Thus, the mutations impair ATP synthesis and increase the formation of reactive oxygen species (ROS), resulting in the apoptosis of retinal ganglion cells. [2][3][4] These pathophysiological mechanisms contribute to the development of the LHON phenotype. While the disease is primarily associated with maternal mtDNA mutation, the higher proportion of male patients suggests a potential involvement of nuclear DNA in hereditary "mitochondrial" optic neuropathy.…”