Stem Cell-Derived Photoreceptor Transplants Differentially Integrate Into Mouse Models of Cone-Rod Dystrophy.
Stem Cell-Derived Photoreceptor Transplants Differentially Integrate Into Mouse Models of Cone-Rod Dystrophy.
复制标题
DOI：
10.1167/iovs.16-19087
复制
发表时间：
2016-06
影响因子：
4.4
通讯作者：
T. Santos-Ferreira;Manuela Völkner;O. Borsch;Jochen Haas;P. Cimalla;P. Vasudevan;P. Carmeliet;D. Cor
中科院分区：
文献类型：
--
作者：
T. Santos-Ferreira;Manuela Völkner;O. Borsch;Jochen Haas;P. Cimalla;P. Vasudevan;P. Carmeliet;D. Cor
目的光感受器移植的临床前研究为利用多能干细胞恢复视功能提供了证据，多能干细胞被认为是获得足够供体材料的潜在来源。翻译研究需要足够的临床前模型来代表潜在未来患者的视网膜疾病状况。在这里，我们比较了轻度(Prominin1缺失；Prom1-/-)和重度(视锥细胞光感受器功能丧失1/视紫红质缺失双突变；Cpfl1/Rho-/-)视锥-杆变性小鼠模型中的移植整合。方法将小鼠胚胎干细胞视网膜器官系统与重组腺相关病毒(AAV)标记的视紫红质驱动的GFP细胞相结合，用于光感受器移植的生产。以CD73为基础的磁活化细胞分选(MACs)富集有机类光受体，并将其移植到野生型、Prom1-/-和Cpfl1/Rho-/-宿主体内。免疫组织化学方法分析供体细胞的存活、成熟和突触形成情况。结果视网膜有机体具有较高的光感受器数量，进一步富集光受体的纯度可达85%。移植的光感受器在所有模型小鼠的视网膜下空间存活下来。一些细胞整合到野生型和Prom1/-小鼠视网膜并获得成熟的形态，在靠近二级神经元的地方表达视杆和突触标记。相反，在光感受器完全变性的新型Cpfl1/Rho-/-模型中，移植细胞仍然局限于视网膜下空间，表达杆状特异的突触标志，但仅减少突触，并且没有获得成熟的形态。结论在临床前光感受器丢失或完全丢失的模型中，光感受器移植物的比较显示，移植的成功率不同，整合有效和受损。因此，Cpfl1/Rho-/-小鼠代表了一个潜在的基准模型，类似于严重的视网膜变性患者，以优化光感受器替代疗法。
PURPOSE Preclinical studies on photoreceptor transplantation provided evidence for restoration of visual function with pluripotent stem cells considered as a potential source for sufficient amounts of donor material. Adequate preclinical models representing retinal disease conditions of potential future patients are needed for translation research. Here we compared transplant integration in mouse models with mild (prominin1-deficient; Prom1-/-) or severe (cone photoreceptor function loss 1/rhodopsin-deficient double-mutant; Cpfl1/Rho-/-) cone-rod degeneration. METHODS For photoreceptor transplant production, we combined the mouse embryonic stem cell retinal organoid system with rhodopsin-driven GFP cell labeling by recombinant adeno-associated virus (AAV). Organoid-derived photoreceptors were enriched by CD73-based magnetic-activated cell sorting (MACS) and transplanted subretinally into wild-type, Prom1-/- and Cpfl1/Rho-/- hosts. The survival, maturation, and synapse formation of donor cells was analyzed by immunohistochemistry. RESULTS Retinal organoids yielded high photoreceptor numbers that were further MACS-enriched to 85% purity. Grafted photoreceptors survived in the subretinal space of all mouse models. Some cells integrated into wild-type as well as Prom1-/- mouse retinas and acquired a mature morphology, expressing rod and synaptic markers in close proximity to second-order neurons. In contrast, in the novel Cpfl1/Rho-/- model with complete photoreceptor degeneration, transplants remained confined to the subretinal space, expressed rod-specific but only reduced synaptic markers, and did not acquire mature morphology. CONCLUSIONS Comparison of photoreceptor grafts in preclinical models with incomplete or complete photoreceptor loss, showed differential transplant success with effective and impaired integration, respectively. Thus, Cpfl1/Rho-/- mice represent a potential benchmark model resembling patients with severe retinal degeneration to optimize photoreceptor replacement therapies.