Reserpine maintains photoreceptor survival in retinal ciliopathy by resolving proteostasis imbalance and ciliogenesis defects.
Reserpine maintains photoreceptor survival in retinal ciliopathy by resolving proteostasis imbalance and ciliogenesis defects.
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DOI：
10.7554/elife.83205
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发表时间：
2023-03-28
期刊：
影响因子：
7.7
通讯作者：
Swaroop A
中科院分区：
文献类型：
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作者：
Chen HY;Swaroop M;Papal S;Mondal AK;Song HB;Campello L;Tawa GJ;Regent F;Shimada H;Nagashima K;de Val N;Jacobson SG;Zheng W;Swaroop A
纤毛病表现为感觉异常到多器官病变综合征，其中视网膜变性是一种高度渗透的表型。感光细胞死亡是视网膜纤毛病导致无法治愈失明的主要原因。为了确定维持光感受器存活的候选药物，我们使用从 rd16 小鼠的诱导多能干细胞 (iPSC) 分化而来的视网膜类器官对 6000 多种生物活性小分子进行了公正、高通量的筛选，rd16 小鼠是由纤毛中心体基因 CEP290 突变引起的 Leber 先天性黑蒙 (LCA) 10 型模型。我们确定了五个无毒的积极作用，包括主导分子利血平，它维持了 rd16 类器官中光感受器的发育和存活。利血平还改善了来自 LCA10 患者诱导多能干细胞的视网膜类器官和体内 rd16 小鼠视网膜中的光感受器。利血平治疗的患者类器官揭示了与细胞存活/死亡、代谢和蛋白质稳态相关的信号通路的调节。进一步的研究发现，自噬失调与患者类器官和 rd16 小鼠视网膜中初级纤毛生物发生受损相关。利血平至少部分通过增加货物接头 p62 来部分恢复自噬和泛素蛋白酶体系统之间的平衡，从而改善初级纤毛组装。我们的研究通过使用 iPSC 衍生的类器官进行跨物种药物发现，确定了 CEP290 视网膜纤毛病临床前研究中的有效候选药物，强调了蛋白质稳态在纤毛病发病机制中的影响，并为视网膜神经变性的治疗提供了新的见解。莱伯先天性黑蒙 (LCA) 是一种遗传性疾病，会影响眼睛并导致儿童早期视力丧失，并且通常会随着时间的推移而恶化。患有这种疾病的个体存在基因突变，导致眼睛后部视网膜区域的光敏感细胞（称为光感受器）死亡。携带 CEP290 基因突变的患者占所有 LCA 的 20-25%。目前，治疗方案仅适用于有限数量的 LCA 患者。一种选择是使用小分子作为药物，可以靶向或绕过眼睛内的错误过程，帮助光感受器在许多不同形式的 LCA 和其他视网膜疾病中存活。然而，超过90%的候选新药未能通过针对人类疾病的第一阶段临床试验。部分原因是候选药物是使用细胞培养物或动物模型开发的，不能忠实地反映人体的运作方式。细胞和发育生物学的最新进展使研究人员能够使用源自人类的干细胞在实验室的培养皿中培养视网膜组织。这些组织被称为视网膜类器官，与传统动物模型的视网膜相比，其行为方式更类似于人眼中的视网膜。然而，制造和维护人类视网膜类器官的方法既耗时又费力，迄今为止限制了它们在寻找新疗法中的应用。为了应对这一挑战，陈等人。开发了一种大规模方法，从 rd16 突变小鼠干细胞（这是 CEP290 突变引起的 LCA 的良好模型）培养视网膜类器官，并使用这些类器官的光感受器筛选 6,000 多种现有药物，以确定它们促进光感受器存活的能力。实验发现，先前被批准用于治疗高血压的药物利血平也有助于光感受器在患病的类器官中存活。利血平在人类 LCA 患者和 rd16 小鼠的视网膜类器官中也有类似的作用。进一步的实验表明，利血平可以通过部分恢复身体破坏和回收细胞中旧的和受损的蛋白质的过程来帮助 LCA 患者。这项工作的下一步将是进行进一步的测试，以证明利血平的这种用途可以安全地进入临床试验，作为 LCA 和其他类似眼病的治疗方法。
Ciliopathies manifest from sensory abnormalities to syndromic disorders with multi-organ pathologies, with retinal degeneration a highly penetrant phenotype. Photoreceptor cell death is a major cause of incurable blindness in retinal ciliopathies. To identify drug candidates to maintain photoreceptor survival, we performed an unbiased, high-throughput screening of over 6000 bioactive small molecules using retinal organoids differentiated from induced pluripotent stem cells (iPSC) of rd16 mouse, which is a model of Leber congenital amaurosis (LCA) type 10 caused by mutations in the cilia-centrosomal gene CEP290. We identified five non-toxic positive hits, including the lead molecule reserpine, which maintained photoreceptor development and survival in rd16 organoids. Reserpine also improved photoreceptors in retinal organoids derived from induced pluripotent stem cells of LCA10 patients and in rd16 mouse retina in vivo. Reserpine-treated patient organoids revealed modulation of signaling pathways related to cell survival/death, metabolism, and proteostasis. Further investigation uncovered dysregulation of autophagy associated with compromised primary cilium biogenesis in patient organoids and rd16 mouse retina. Reserpine partially restored the balance between autophagy and the ubiquitin-proteasome system at least in part by increasing the cargo adaptor p62, resulting in improved primary cilium assembly. Our study identifies effective drug candidates in preclinical studies of CEP290 retinal ciliopathies through cross-species drug discovery using iPSC-derived organoids, highlights the impact of proteostasis in the pathogenesis of ciliopathies, and provides new insights for treatments of retinal neurodegeneration. Leber congenital amaurosis (LCA) is an inherited disease that affects the eyes and causes sight loss in early childhood, which generally gets worse over time. Individuals with this condition have genetic mutations that result in the death of light-sensitive cells, known as photoreceptors, in a region called the retina at the back of the eye. Patients carrying a genetic change in the gene CEP290 account for 20-25% of all LCA. At present, treatment options are only available for a limited number of patients with LCA. One option is to use small molecules as drugs that may target or bypass the faulty processes within the eye to help the photoreceptors survive in many different forms of LCA and other retinal diseases. However, over 90% of new drug candidates fail the first phase of clinical trials for human diseases. This in part due to the candidates having been developed using cell cultures or animal models that do not faithfully reflect how the human body works. Recent advances in cell and developmental biology are now enabling researchers to use stem cells derived from humans to grow retina tissues in a dish in the laboratory. These tissues, known as retinal organoids, behave in a more similar way to retinas in human eyes than those of traditional animal models. However, the methods for making and maintaining human retinal organoids are time-consuming and labor-intensive, which has so far limited their use in the search for new therapies. To address this challenge, Chen et al. developed a large-scale approach to grow retinal organoids from rd16 mutant mice stem cells (which are a good model for LCA caused by mutations to CEP290) and used the photoreceptors from these organoids to screen over 6,000 existing drugs for their ability to promote the survival of photoreceptors. The experiments found that the drug reserpine, which was previously approved to treat high blood pressure, also helped photoreceptors to survive in the diseased organoids. Reserpine also had a similar effect in retinal organoids derived from human patients with LCA and in the rd16 mice themselves. Further experiments suggest that reserpine may help patients with LCA by partially restoring a process by which the body destroys and recycles old and damaged proteins in the cells. The next steps following on from this work will be to perform further tests to demonstrate that this use of reserpine is safe to enter clinical trials as a treatment for LCA and other similar eye diseases.
TSC，但在色素性视网膜炎小鼠的饥饿锥中的PTEN损失不会导致自噬缺陷和MTORC1与溶酶体的分离。
影响因子：
9
作者：
通讯作者：
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