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DICER1 deficit induces Alu RNA toxicity in age-related macular degeneration

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Abstract

Geographic atrophy (GA), an untreatable advanced form of age-related macular degeneration, results from retinal pigmented epithelium (RPE) cell degeneration. Here we show that the microRNA (miRNA)-processing enzyme DICER1 is reduced in the RPE of humans with GA, and that conditional ablation of Dicer1, but not seven other miRNA-processing enzymes, induces RPE degeneration in mice. DICER1 knockdown induces accumulation of Alu RNA in human RPE cells and Alu-like B1 and B2 RNAs in mouse RPE. Alu RNA is increased in the RPE of humans with GA, and this pathogenic RNA induces human RPE cytotoxicity and RPE degeneration in mice. Antisense oligonucleotides targeting Alu/B1/B2 RNAs prevent DICER1 depletion-induced RPE degeneration despite global miRNA downregulation. DICER1 degrades Alu RNA, and this digested Alu RNA cannot induce RPE degeneration in mice. These findings reveal a miRNA-independent cell survival function for DICER1 involving retrotransposon transcript degradation, show that Alu RNA can directly cause human pathology, and identify new targets for a major cause of blindness.

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Figure 1: DICER1 deficit in GA induces RPE degeneration.
Figure 2: Alu RNA accumulation in GA triggered by DICER reduction.
Figure 3: DICER1 degrades Alu RNA.
Figure 4: DICER1 protects RPE cells from Alu RNA cytotoxicity.
Figure 5: DICER1 dysregulation induces RPE cell death via Alu RNA accumulation.

Accession codes

Data deposits

The Alu sequences have been deposited in GenBank under the accession numbers HN176584 and HN176585.

Change history

  • 16 March 2011

    In the paragraph beginning, ‘Subretinal injection delivered Alu RNA to RPE cells...’, two figure citations were corrected.

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Acknowledgements

We thank M. Chrenek, J. Garcia-Perez, T. Heidmann, C. Kanellopoulou, D. M. Livingston, J. V. Moran, R. F. Mullins, J. M. Nickerson, E. A. Pearce, A. Tarakhovsky, B. Vogelstein, V. E. Velculescu and D. J. Zack for providing mice, reagents or tissues; R. King, L. Xu, M. McConnell, C. Payne, G. R. Pattison, G. J. Jaffe, S. Medearis and C. Spee for technical assistance; and A. Sinai, R. Mohan, T. S. Khurana, R. A. Brekken, P. L. Deininger, S. Bondada, P. A. Pearson, A. M. Rao, G. S. Rao and K. Ambati for discussions. J.A. was supported by National Eye Institute (NEI)/National Institutes of Health (NIH) grants R01EY015422, R01EY018350, R01EY018836, R01EY020672, R21EY019778, RC1EY020442, the Doris Duke Distinguished Clinical Scientist Award, the Burroughs Wellcome Fund Clinical Scientist Award in Translational Research, and the Dr E. Vernon Smith and Eloise C. Smith Macular Degeneration Endowed Chair. Research to Prevent Blindness Senior Scientist Investigator Awards or departmental unrestricted grants supported J.A., H.E.G. and W.W.H.; J.Z.B. was supported by University of Kentucky Physician Scientist Award, International Retinal Research Foundation, and American Health Assistance Foundation; B.K.A. by VA Merit Award and Department of Defense; D.-k.L. by Global Research Laboratory program by MEST, Korea; D.R.H. by Arnold and Mabel Beckman Foundation; W.W.H. by Macular Vision Research Foundation and Foundation Fighting Blindness; J.M.P. by ARC Centres of Excellence Grant CE0561903; M.C.M. by Sydney Foundation for Medical Research. B.K.A was supported by NIH R01EY017182 and R01EY017950; R.E.B. by NIH R01HD027215; G.C. by NIH R21AI076757; H.E.G. by NIH P30EY06360; W.W.H. by NIH U10EY013729, R01EY011123, and P30EY008571; J.F.K. and J.A.G. by NIH R01GM068414; J.L.D. by NIH R01EY015240; D.R.H. by NIH P30EY003040 and R01EY001545; M.E.K. and S.B. by NIH T32HL091812. P.P. is a Senior Scholar from the Fonds de la Recherche en Santé du Québec (FRSQ). M.M.W.C. is a QEII Fellow of the Australian Research Council and is supported by National Health and Medical Research Council, Australia Project Grant 637228. E.F. and D.R.L. are investigators of the Howard Hughes Medical Institute.

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H.K., S.D., V.T., W.G.C., B.J.F., M.E.K., S.L.P., J.F.K., J.A.G., K.K., N.L.J., B.D.G., Y.H., R.J.C.A., A.D.B., S.B, J.W., M.H, Y.S. and J.Z.B. performed experiments. W.W.H., V.A.C, D.-k.L., J.W.Y., C.M.R, D.R.H., H.E.G., Q.Z., J.M.P., M.C.M., A.H.M., M.M.W.C., D.R.L., E.F., P.P., F.W.B., R.E.B., S.M., G.C. and J.L.D. provided tissues or reagents. J.A. conceived and directed the project, and wrote the paper with assistance from P.P., C.M.R., K.K., J.F.K., J.A.G., E.F., M.M.W.C., B.J.F., B.D.G. and B.K.A. All authors had the opportunity to discuss the results and comment on the manuscript.

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Correspondence to Jayakrishna Ambati.

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J.A. is named as an inventor on patent applications filed by his employer, the University of Kentucky, on technologies described in this paper. W.W.H. and the University of Florida have a financial interest in the use of AAV therapies, and own equity in a company (AGTC Inc.) that might, in the future, commercialize some aspects of this work.

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Kaneko, H., Dridi, S., Tarallo, V. et al. DICER1 deficit induces Alu RNA toxicity in age-related macular degeneration. Nature 471, 325–330 (2011). https://doi.org/10.1038/nature09830

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