PMID- 26854824 OWN - NLM STAT- MEDLINE DCOM- 20170613 LR - 20231123 IS - 1096-0007 (Electronic) IS - 0014-4835 (Print) IS - 0014-4835 (Linking) VI - 145 DP - 2016 Apr TI - Differential cytotoxic effects of 7-dehydrocholesterol-derived oxysterols on cultured retina-derived cells: Dependence on sterol structure, cell type, and density. PG - 297-316 LID - S0014-4835(16)30015-X [pii] LID - 10.1016/j.exer.2016.01.016 [doi] AB - Tissue accumulation of 7-dehydrocholesterol (7DHC) is a hallmark of Smith-Lemli-Opitz Syndrome (SLOS), a human inborn error of the cholesterol (CHOL) synthesis pathway. Retinal 7DHC-derived oxysterol formation occurs in the AY9944-induced rat model of SLOS, which exhibits a retinal degeneration characterized by selective loss of photoreceptors and associated functional deficits, Muller cell hypertrophy, and engorgement of the retinal pigment epithelium (RPE) with phagocytic inclusions. We evaluated the relative effects of four 7DHC-derived oxysterols on three retina-derived cell types in culture, with respect to changes in cellular morphology and viability. 661W (photoreceptor-derived) cells, rMC-1 (Muller glia-derived) cells, and normal diploid monkey RPE (mRPE) cells were incubated for 24 h with dose ranges of either 7-ketocholesterol (7kCHOL), 5,9-endoperoxy-cholest-7-en-3beta,6alpha-diol (EPCD), 3beta,5alpha-dihydroxycholest-7-en-6-one (DHCEO), or 4beta-hydroxy-7-dehydrocholesterol (4HDHC); CHOL served as a negative control (same dose range), along with appropriate vehicle controls, while staurosporine (Stsp) was used as a positive cytotoxic control. For 661W cells, the rank order of oxysterol potency was: EPCD > 7kCHOL >> DHCEO > 4HDHC approximately CHOL. EC50 values were higher for confluent vs. subconfluent cultures. 661W cells exhibited much higher sensitivity to EPCD and 7kCHOL than either rMC-1 or mRPE cells, with the latter being the most robust when challenged, either at confluence or in sub-confluent cultures. When tested on rMC-1 and mRPE cells, EPCD was again an order of magnitude more potent than 7kCHOL in compromising cellular viability. Hence, 7DHC-derived oxysterols elicit differential cytotoxicity that is dose-, cell type-, and cell density-dependent. These results are consistent with the observed progressive, photoreceptor-specific retinal degeneration in the rat SLOS model, and support the hypothesis that 7DHC-derived oxysterols are causally linked to that retinal degeneration as well as to SLOS. CI - Published by Elsevier Ltd. FAU - Pfeffer, Bruce A AU - Pfeffer BA AD - Research Service, VA Western New York Healthcare System, Buffalo, NY, USA; SUNY Eye Institute, Buffalo, NY, USA; Departments of Ophthalmology and Biochemistry, University at Buffalo, The State University of New York (SUNY), Buffalo, NY, USA. FAU - Xu, Libin AU - Xu L AD - Department of Medicinal Chemistry, University of Washington, Seattle, WA, USA. FAU - Porter, Ned A AU - Porter NA AD - Department of Chemistry and Vanderbilt Institute of Chemical Biology, Vanderbilt University, Nashville, TN, USA. FAU - Rao, Sriganesh Ramachandra AU - Rao SR AD - Research Service, VA Western New York Healthcare System, Buffalo, NY, USA; SUNY Eye Institute, Buffalo, NY, USA; Departments of Ophthalmology and Biochemistry, University at Buffalo, The State University of New York (SUNY), Buffalo, NY, USA. FAU - Fliesler, Steven J AU - Fliesler SJ AD - Research Service, VA Western New York Healthcare System, Buffalo, NY, USA; SUNY Eye Institute, Buffalo, NY, USA; Departments of Ophthalmology and Biochemistry, University at Buffalo, The State University of New York (SUNY), Buffalo, NY, USA. Electronic address: fliesler@buffalo.edu. LA - eng GR - P01 ES013125/ES/NIEHS NIH HHS/United States GR - R00 HD073270/HD/NICHD NIH HHS/United States GR - U54 HD083211/HD/NICHD NIH HHS/United States GR - R01 HD064727/HD/NICHD NIH HHS/United States GR - R01 EY007361/EY/NEI NIH HHS/United States GR - I01 BX002439/BX/BLRD VA/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20160213 PL - England TA - Exp Eye Res JT - Experimental eye research JID - 0370707 RN - 0 (Dehydrocholesterols) RN - 0 (Oxysterols) RN - BK1IU07GKF (7-dehydrocholesterol) SB - IM MH - Animals MH - Cell Count MH - Cell Death/drug effects MH - Cell Survival/drug effects MH - Cells, Cultured MH - Dehydrocholesterols/chemistry/metabolism/*pharmacology MH - Ependymoglial Cells/*drug effects MH - Epithelial Cells/*drug effects MH - Macaca mulatta MH - Models, Animal MH - Oxysterols/*pharmacology MH - Photoreceptor Cells, Vertebrate/*drug effects MH - Rats MH - Retina/*cytology/metabolism MH - Retinal Pigment Epithelium/cytology PMC - PMC5024725 MID - NIHMS814292 OTO - NOTNLM OT - Cell culture OT - Cell viability assay OT - Muller cell OT - Oxysterol OT - Photoreceptor OT - Retinal pigment epithelium EDAT- 2016/02/09 06:00 MHDA- 2017/06/14 06:00 PMCR- 2017/04/01 CRDT- 2016/02/09 06:00 PHST- 2015/09/17 00:00 [received] PHST- 2015/12/21 00:00 [revised] PHST- 2016/01/26 00:00 [accepted] PHST- 2016/02/09 06:00 [entrez] PHST- 2016/02/09 06:00 [pubmed] PHST- 2017/06/14 06:00 [medline] PHST- 2017/04/01 00:00 [pmc-release] AID - S0014-4835(16)30015-X [pii] AID - 10.1016/j.exer.2016.01.016 [doi] PST - ppublish SO - Exp Eye Res. 2016 Apr;145:297-316. doi: 10.1016/j.exer.2016.01.016. Epub 2016 Feb 13.