Severe facial clefting in Insig-deficient mouse embryos caused by sterol accumulation and reversed by lovastatin.
Open accessAbstract
Insig-1 and Insig-2 are regulatory proteins that restrict the cholesterol biosynthetic pathway by preventing proteolytic activation of SREBPs and by enhancing degradation of HMG-CoA reductase. Here, we created Insig-double-knockout (Insig-DKO) mice that are homozygous for null mutations in Insig-1 and Insig-2. After 18.5 days of development, 96% of Insig-DKO embryos had defects in midline facial development, ranging from cleft palate (52%) to complete cleft face (44%). Middle and inner ear structures were abnormal, but teeth and skeletons were normal. The animals were lethargic and runted; they died within 1 day of birth. The livers and heads of Insig-DKO embryos overproduced sterols, causing a marked buildup of sterol intermediates. Treatment of pregnant mice with the HMG-CoA reductase inhibitor lovastatin reduced sterol synthesis in Insig-DKO embryos and reduced the pre-cholesterol intermediates. This treatment ameliorated the clefting syndrome so that 54% of Insig-DKO mice had normal faces, and only 7% had cleft faces. We conclude that buildup of pre-cholesterol sterol intermediates interferes with midline fusion of facial structures in mice. These findings have implications for the pathogenesis of the cleft palate component of Smith-Lemli-Opitz syndrome and other human malformation syndromes in which mutations in enzymes catalyzing steps in cholesterol biosynthesis produce a buildup of sterol intermediates.
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In mammals, Insig is a key regulator of SREBP function where it binds to SCAP and prevents SREBP cleavage in the presence of sterols by maintaining the SREBP-SCAP complex in the endoplasmic reticulum membrane.
“In mammals, Insig is a key regulator of SREBP function where it binds to SCAP and prevents SREBP cleavage in the presence of sterols by maintaining the SREBP-SCAP complex in the endoplasmic reticulum membrane [65],[66].”
From A Sterol-Regulatory Element Binding Protein Is Required for Cell Polarity, Hypoxia Adaptation, Azole Drug Resistance, and Virulence in Aspergillus fumigatus (2008) · cites this paper as [65]
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SREBPs: activators of the complete program of cholesterol and fatty acid synthesis in the liver
Horton, J. D., Goldstein, J. L., & Brown, M. S. (2002) · Journal of Clinical Investigation · doi:10.1172/jci15593 · Cited by 4,285 · see citing works
Self-citationBrown, M. S., & Goldstein, J. L. (1997) · Cell · doi:10.1016/s0092-8674(00)80213-5 · Cited by 3,834 · see citing works
Self-citationCyclopia and defective axial patterning in mice lacking Sonic hedgehog gene function
Chiang, C. C., Litingtung, Y., Lee, E., Young, K. E., Corden, J. L., Westphal, H., & Beachy, P. A. (1996) · Nature · doi:10.1038/383407a0 · Cited by 3,062 · see citing works
Douarin, N. M. L., & Kalcheim, C. (1999) · Cambridge University Press eBooks · doi:10.1017/cbo9780511897948 · Cited by 3,003 · see citing works
Protein Sensors for Membrane Sterols
Goldstein, J. L., DeBose‐Boyd, R. A., & Brown, M. S. (2006) · Cell · doi:10.1016/j.cell.2005.12.022 · Cited by 1,685 · see citing works
Self-citationCholesterol Modification of Hedgehog Signaling Proteins in Animal Development
Porter, J. A., Young, K. E., & Beachy, P. A. (1996) · Science · doi:10.1126/science.274.5285.255 · Cited by 1,360 · see citing works
Crucial Step in Cholesterol Homeostasis
Yang, T., Espenshade, P. J., Wright, M. E., Yabe, D., Gong, Y., Aebersold, R., Goldstein, J. L., & Brown, M. S. (2002) · Cell · doi:10.1016/s0092-8674(02)00872-3 · Cited by 987 · see citing works
Self-citationShimano, H., Horton, J. D., Hammer, R. E., Shimomura, I., Brown, M. S., & Goldstein, J. L. (1996) · Journal of Clinical Investigation · doi:10.1172/jci118951 · Cited by 810 · see citing works
Self-citationGene/environment causes of cleft lip and/or palate
Murray, J. (2002) · Clinical Genetics · doi:10.1034/j.1399-0004.2002.610402.x · Cited by 787 · see citing works
Yabe, D., Brown, M. S., & Goldstein, J. L. (2002) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.162488899 · Cited by 557 · see citing works
Self-citationBrown, M. S., Faust, J. R., Goldstein, J. L., Kaneko, I., & Endo, A. (1978) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(17)38120-6 · Cited by 556 · see citing works
Self-citationFerguson, M. W. J. (1988) · Development · doi:10.1242/dev.103.supplement.41 · Cited by 515 · see citing works
SREBPs: activators of the complete program of cholesterol and fatty acid synthesis in the liver
Horton, J. D., Goldstein, J. L., & Brown, M. S. (2002) · Journal of Clinical Investigation · doi:10.1172/jci200215593 · Cited by 481 · see citing works
Self-citationThe Smith-Lemli-Opitz syndrome
Kelley, R. I., & Hennekam, R. C. M. (2000) · Journal of Medical Genetics · doi:10.1136/jmg.37.5.321 · Cited by 480 · see citing works
The Sonic Hedgehog–Patched–Gli Pathway in Human Development and Disease
Villavicencio, E. H., Walterhouse, D. O., & Iannaccone, P. M. (2000) · The American Journal of Human Genetics · doi:10.1016/s0002-9297(07)62934-6 · Cited by 427 · see citing works
Stanier, P. (2004) · Human Molecular Genetics · doi:10.1093/hmg/ddh052 · Cited by 419 · see citing works
Sever, N., Yang, T., Brown, M. S., Goldstein, J. L., & DeBose‐Boyd, R. A. (2003) · Molecular Cell · doi:10.1016/s1097-2765(02)00822-5 · Cited by 372 · see citing works
Self-citationDefective cholesterol biosynthesis in Smith-Lemli-Opitz syndrome
Irons, M., Roy Elias, E., Salen, G., Tint, G. S., & Batta, A. (1993) · The Lancet · doi:10.1016/0140-6736(93)90983-n · Cited by 368 · see citing works
Pathogenesis of cleft palate in TGF-β3 knockout mice
Taya, Y., O’Kane, S., & Ferguson, M. W. J. (1999) · Development · doi:10.1242/dev.126.17.3869 · Cited by 326 · see citing works
Kita, T., Brown, M. S., & Goldstein, J. L. (1980) · Journal of Clinical Investigation · doi:10.1172/jci109938 · Cited by 306 · see citing works
Self-citationSever, N., Song, B., Yabe, D., Goldstein, J. L., Brown, M. S., & DeBose‐Boyd, R. A. (2003) · Journal of Biological Chemistry · doi:10.1074/jbc.m310053200 · Cited by 305 · see citing works
Self-citationDerry, J. M. J., Gormally, E., Means, G., Zhao, W., Meindl, A., Kelley, R. I., Boyd, Y., & Herman, G. E. (1999) · Nature Genetics · doi:10.1038/10350 · Cited by 244 · see citing works
Engelking, L. J. (2005) · Journal of Clinical Investigation · doi:10.1172/jci25614 · Cited by 210 · see citing works
Self-citationMeasurement of rates of cholesterol synthesis using tritiated water
Dietschy, J. M., & Spady, D. K. (1984) · Journal of Lipid Research · doi:10.1016/s0022-2275(20)34420-5 · Cited by 201 · see citing works
Disorders of cholesterol biosynthesis: prototypic metabolic malformation syndromes
Herman, G. E. (2003) · Human Molecular Genetics · doi:10.1093/hmg/ddg072 · Cited by 189 · see citing works
Generation of Viable Cholesterol-Free Mice
Wechsler, A. S., Brafman, A., Shafir, M., Heverin, M., Gottlieb, H., Damari, G. M., Gozlan-Kelner, S., Spivak, I., Moshkin, O., Fridman, E., et al. (2003) · Science · doi:10.1126/science.1090776 · Cited by 179 · see citing works
I NBORN E RRORS OF S TEROL B IOSYNTHESIS
Kelley, R. I., & Herman, G. E. (2001) · Annual Review of Genomics and Human Genetics · doi:10.1146/annurev.genom.2.1.299 · Cited by 168 · see citing works
Wassif, C. A. (2001) · Human Molecular Genetics · doi:10.1093/hmg/10.6.555 · Cited by 164 · see citing works
Krakowiak, P. A. (2003) · Human Molecular Genetics · doi:10.1093/hmg/ddg172 · Cited by 161 · see citing works
The gene mutated in bare patches and striated mice encodes a novel 3β-hydroxysteroid dehydrogenase
Liu, X. Y., Dangel, A. W., Kelley, R. I., Zhao, W., Denny, P., Botcherby, M., Cattanach, B. M., Peters, J., Hunsicker, P. R., Mallon, A., et al. (1999) · Nature Genetics · doi:10.1038/9700 · Cited by 153 · see citing works
Fitzky, B. U., Moebius, F. F., Asaoka, H., Waage-Baudet, H., Xu, L., Xu, G., Maeda, N., Kluckman, K. D., Hiller, S., Yu, H., et al. (2001) · Journal of Clinical Investigation · doi:10.1172/jci200112103 · Cited by 153 · see citing works
Fitzky, B. U., Moebius, F. F., Asaoka, H., Waage-Baudet, H., Xu, L., Xu, G., Maeda, N., Kluckman, K. D., Hiller, S., Yu, H., et al. (2001) · Journal of Clinical Investigation · doi:10.1172/jci12103 · Cited by 150 · see citing works
Dual roles for cholesterol in mammalian cells
Xu, F., Rychnovsky, S. D., Belani, J. D., Hobbs, H. H., Cohen, J. C., & Rawson, R. B. (2005) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0503590102 · Cited by 140 · see citing works
Treatment of Smith-Lemli-Opitz syndrome: Results of a multicenter trial
Irons, M., Elias, E. R., Abuelo, D. N., Bull, M. J., Greene, C. L., Johnson, V. P., Keppen, L. D., Schanen, C., Tint, G. S., & Salen, G. (1997) · American Journal of Medical Genetics · doi:10.1002/(sici)1096-8628(19970131)68:3 · Cited by 138 · see citing works
Nohturfft, A., Brown, M. S., & Goldstein, J. L. (1998) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.95.22.12848 · Cited by 135 · see citing works
Self-citationTint, G. S., Seller, M. J., Hughes-Benzie, R., Batta, A. K., Shefer, S., Genest, D., Irons, M., Elias, E. R., & Salen, G. (1995) · Journal of Lipid Research · doi:10.1016/s0022-2275(20)39757-1 · Cited by 131 · see citing works
Human malformation syndromes due to inborn errors of cholesterol synthesis
Porter, F. D. (2003) · Current Opinion in Pediatrics · doi:10.1097/00008480-200312000-00011 · Cited by 127 · see citing works
Belknap, W. M., & Dietschy, J. (1988) · Journal of Clinical Investigation · doi:10.1172/jci113829 · Cited by 114 · see citing works
Simvastatin: a new therapeutic approach for Smith-Lemli-Opitz syndrome
Jira, P. E., Wevers, R. A., de Jong, J. G. N., Rubio‐Gozalbo, M. E., Janssen-Zijlstra, F. S. M., van Heyst, A. F. J., Sengers, R. C. A., & Smeitink, J. A. M. (2000) · Journal of Lipid Research · doi:10.1016/s0022-2275(20)33442-8 · Cited by 113 · see citing works
Porter, F. D. (2000) · Molecular Genetics and Metabolism · doi:10.1006/mgme.2000.3069 · Cited by 108 · see citing works
Cholesterol supplementation does not improve developmental progress in Smith-Lemli-Opitz syndrome
Sikora, D. M., Ruggiero, M., Pettit-Kekel, K., Merkens, L. S., Connor, W. E., & Steiner, R. D. (2004) · The Journal of Pediatrics · doi:10.1016/j.jpeds.2004.02.036 · Cited by 88 · see citing works
Recent advances in hedgehog signalling
Tabin, C. J., & McMahon, A. P. (1997) · Trends in Cell Biology · doi:10.1016/s0962-8924(97)01159-8 · Cited by 85 · see citing works
RSH (so-called Smith-Lemli-Opitz) syndrome
Opitz, J. M. (1999) · Current Opinion in Pediatrics · doi:10.1097/00008480-199908000-00015 · Cited by 83 · see citing works
Genetic Disorders of Cholesterol Biosynthesis in Mice and Humans
Nwokoro, N. A., Wassif, C. A., & Porter, F. D. (2001) · Molecular Genetics and Metabolism · doi:10.1006/mgme.2001.3226 · Cited by 72 · see citing works
The Sonic Hedgehog–Patched–Gli Pathway in Human Development and Disease
Villavicencio, E. H., Walterhouse, D. O., & Iannaccone, P. M. (2000) · The American Journal of Human Genetics · doi:10.1086/321187 · Cited by 67 · see citing works
Genetic Defects in Postsqualene Cholesterol Biosynthesis
Moebius, F. F., Fitzky, B. U., & Glossmann, H. H. (2000) · Trends in Endocrinology and Metabolism · doi:10.1016/s1043-2760(00)00235-6 · Cited by 60 · see citing works
Simvastatin treatment in the SLO syndrome: A safe approach?
Starck, L., Lövgren‐Sandblom, A., & Björkhem, I. (2002) · American Journal of Medical Genetics · doi:10.1002/ajmg.10722 · Cited by 41 · see citing works
Starck, L., Lövgren‐Sandblom, A., & Björkhem, I. (2002) · Journal of Internal Medicine · doi:10.1046/j.1365-2796.2002.01037.x · Cited by 32 · see citing works
Effects of Dietary Cholesterol on Plasma Lipoproteins in Smith-Lemli-Opitz Syndrome
Merkens, L. S., Connor, W. E., Linck, L. M., Lin, D. S., Flavell, D. P., & Steiner, R. D. (2004) · Pediatric Research · doi:10.1203/01.pdr.0000141522.14177.4f · Cited by 27 · see citing works
Developmental Biology: Frontiers for Clinical Genetics
McInnes, R. R. (1998) · Clinical Genetics · doi:10.1111/j.1399-0004.1998.tb02667.x · Cited by 15 · see citing works
Palatal Dysmorphogenesis: Palate Organ Culture
Abbott, B. D. (2003) · Humana Press eBooks · doi:10.1385/1-59259-065-9:195 · Cited by 5 · see citing works
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Regulation of Sterol Synthesis in Eukaryotes
Espenshade, P. J., & Hughes, A. L. (2007) · Annual Review of Genetics · doi:10.1146/annurev.genet.41.110306.130315 · Cited by 593 · see citing works
Willger, S. D., Puttikamonkul, S., Kim, K.-H., Burritt, J. B., Grahl, N., Metzler, L. J., Barbuch, R., Bard, M., Lawrence, C. B., & Cramer, R. A. (2008) · PLoS Pathogens · doi:10.1371/journal.ppat.1000200 · Cited by 345 · see citing works
- Authors
- Engelking, L. J., Evers, B. M., Richardson, J. A., Goldstein, J. L., Brown, M. S., & Liang, G.
- Affiliations
- The University of Texas Southwestern Medical Center (US)
- ORCID
- Venue
- Journal of Clinical Investigation
- Published
- 2006-09-01
- Volume
- 116
- Issue
- 9
- Pages
- 2356-2365
- ISSN
- 0021-9738 (ISSN-L)
- OpenAlex
- W2110721130
- Cited by
- 82
- References
- 51
- Citations by year
- 2026: 1 · 2025: 2 · 2024: 1 · 2022: 3 · 2021: 4 · 2020: 5 · 2019: 2 · 2018: 2 · 2017: 4 · 2016: 2 · 2015: 5 · 2014: 1 · 2013: 6 · 2012: 5
- FWCI
- 5.49
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Journal of Clinical Investigation (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (HL20948)
- National Heart, Lung, and Blood Institute (HL-20948, P01HL020948)
- National Institute of General Medical Sciences (GM-08014, T32GM008014)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Cleft Lip and Palate Research
- Health Sciences › Medicine › Surgery › Cholesterol and Lipid Metabolism
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer, Lipids, and Metabolism
- Keywords
- INSIG1, INSIG2, cleft palate, facial clefting, sterol accumulation, cholesterol biosynthesis, HMG-CoA reductase, lovastatin, SREBP, Smith-Lemli-Opitz syndrome, embryonic development, congenital malformations
- MeSH terms
- Animals, Bone and Bones, Female, Lovastatin, Membrane Proteins, Mouth Abnormalities, Pregnancy, Sterols, Mice, Knockout, Hydroxymethylglutaryl-CoA Reductase Inhibitors, Mice
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