Method Profile
Preprint method combining a 17-compound mouse retinal organoid screen with selected drug tests in one affected human CEP290 family model.
Prioritize compounds that preserve photoreceptor-associated features, then compare response, toxicity and unsuccessful combinations in human disease-associated organoids.
Cep290 rd16 mouse retinal organoids for discovery; human iPSC-derived retinal organoids from one affected compound-heterozygous CEP290 child and the unaffected mother for follow-up.
Three-day mouse treatment beginning at day 22 with day 29 rhodopsin staining; human treatment beginning at day 90 with immunofluorescent rod, ciliary and other retinal markers.
Discovery and human exposures
17 discovery compounds. Human reserpine 30 μM; halofantrine (A3), C2 and F2 each 10 μM. Selected lower-dose halofantrine combinations were also compared.
Readouts and controls
Rhodopsin fluorescence, ARL13B cilia staining and cone/bipolar/synaptic markers; vehicle and unaffected-family controls. Animal functional and exposure follow-up are separate validation arms.
Matched disease and family-control organoids are compared under the same treatments. Organoid and experimental-group counts describe replication within one affected family background. Rod-marker preservation in organoids and visual-function measurements in animals are separate endpoints.
Sequential mouse discovery and human disease-associated retesting, followed by separate retinal-degeneration animal models.
One affected human donor and one unaffected mother; Figure 2 reports at least three independent groups with at least four organoids per group.
Vehicle, reserpine, other prioritized candidates, unaffected-family organoids and lower-dose combinations.
Reserpine and halofantrine significantly increased human rhodopsin staining. F2 had a nonsignificant trend; C2 was toxic. Tested halofantrine combinations added no further augmentation. Ciliary rescue was partial.
A staged screen identified halofantrine as a candidate photoreceptor protectant. Human follow-up used a CEP290-associated retinal degeneration line and an unaffected family control; animal studies added functional signals, route-dependent exposure and unresolved dose effects.
