{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,22]],"date-time":"2026-06-22T03:07:26Z","timestamp":1782097646106,"version":"3.54.5"},"reference-count":44,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2023,1,18]],"date-time":"2023-01-18T00:00:00Z","timestamp":1674000000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Interdisciplinary Doctoral School in Health and Sciences (Doctoral School EDISS 205, Lyon, France)"},{"name":"University of Lyon"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Nutrients"],"abstract":"<jats:p>Abetalipoproteinemia (FHBL-SD1) and chylomicron retention disease (FHBL-SD3) are rare recessive disorders of lipoprotein metabolism due to mutations in MTTP and SAR1B genes, respectively, which lead to defective chylomicron formation and secretion. This results in lipid and fat-soluble vitamin malabsorption, which induces severe neuro-ophthalmic complications. Currently, treatment combines a low-fat diet with high-dose vitamin A and E supplementation but still fails in normalizing serum vitamin E levels and providing complete ophthalmic protection. To explore these persistent complications, we developed two knock-out cell models of FHBL-SD1 and FHBL-SD3 using the CRISPR\/Cas9 technique in Caco-2\/TC7 cells. DNA sequencing, RNA quantification and Western blotting confirmed the introduction of mutations with protein knock-out in four clones associated with i) impaired lipid droplet formation and ii) defective triglyceride (\u221257.0 \u00b1 2.6% to \u221283.9 \u00b1 1.6%) and cholesterol (\u221235.3 \u00b1 4.4% to \u221260.6 \u00b1 3.5%) secretion. A significant decrease in \u03b1-tocopherol secretion was also observed in these clones (\u221241.5 \u00b1 3.7% to \u221297.2 \u00b1 2.8%), even with the pharmaceutical forms of vitamin E: tocopherol-acetate and tocofersolan (\u03b1-tocopheryl polyethylene glycol succinate 1000). MTTP silencing led to a more severe phenotype than SAR1B silencing, which is consistent with clinical observations. Our cellular models thus provide an efficient tool to experiment with therapeutic strategies and will allow progress in understanding the mechanisms involved in lipid metabolism.<\/jats:p>","DOI":"10.3390\/nu15030505","type":"journal-article","created":{"date-parts":[[2023,1,19]],"date-time":"2023-01-19T04:19:40Z","timestamp":1674101980000},"page":"505","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Validation of Knock-Out Caco-2 TC7 Cells as Models of Enterocytes of Patients with Familial Genetic Hypobetalipoproteinemias"],"prefix":"10.3390","volume":"15","author":[{"given":"Claire","family":"Bordat","sequence":"first","affiliation":[{"name":"Aix-Marseille Universit\u00e9, INRAE, INSERM, C2VN, 13885 Marseille, France"},{"name":"CarMeN Laboratory, INSERM U1060, INRAE, UMR 1397, Universit\u00e9 Claude Bernard Lyon 1, 69495 Pierre-Benite, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Donato","family":"Vairo","sequence":"additional","affiliation":[{"name":"Aix-Marseille Universit\u00e9, INRAE, INSERM, C2VN, 13885 Marseille, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Charlotte","family":"Cuerq","sequence":"additional","affiliation":[{"name":"CarMeN Laboratory, INSERM U1060, INRAE, UMR 1397, Universit\u00e9 Claude Bernard Lyon 1, 69495 Pierre-Benite, France"},{"name":"Biochemistry Department, Hospices Civils de Lyon, 69495 Pierre-Benite, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Charlotte","family":"Halimi","sequence":"additional","affiliation":[{"name":"Aix-Marseille Universit\u00e9, INRAE, INSERM, C2VN, 13885 Marseille, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7198-0534","authenticated-orcid":false,"given":"Franck","family":"Peiretti","sequence":"additional","affiliation":[{"name":"Aix-Marseille Universit\u00e9, INRAE, INSERM, C2VN, 13885 Marseille, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3181-9564","authenticated-orcid":false,"given":"Armelle","family":"Penhoat","sequence":"additional","affiliation":[{"name":"CarMeN Laboratory, INSERM U1060, INRAE, UMR 1397, Universit\u00e9 Claude Bernard Lyon 1, 69495 Pierre-Benite, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aur\u00e9lie","family":"Vieille-Marchiset","sequence":"additional","affiliation":[{"name":"CarMeN Laboratory, INSERM U1060, INRAE, UMR 1397, Universit\u00e9 Claude Bernard Lyon 1, 69495 Pierre-Benite, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Teresa","family":"Gonzalez","sequence":"additional","affiliation":[{"name":"Aix-Marseille Universit\u00e9, INRAE, INSERM, C2VN, 13885 Marseille, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marie-Caroline","family":"Michalski","sequence":"additional","affiliation":[{"name":"CarMeN Laboratory, INSERM U1060, INRAE, UMR 1397, Universit\u00e9 Claude Bernard Lyon 1, 69495 Pierre-Benite, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marion","family":"Nowicki","sequence":"additional","affiliation":[{"name":"Aix-Marseille Universit\u00e9, INRAE, INSERM, C2VN, 13885 Marseille, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8359-5163","authenticated-orcid":false,"given":"No\u00ebl","family":"Peretti","sequence":"additional","affiliation":[{"name":"CarMeN Laboratory, INSERM U1060, INRAE, UMR 1397, Universit\u00e9 Claude Bernard Lyon 1, 69495 Pierre-Benite, France"},{"name":"Pediatric Hepato-Gastroenterology and Nutrition Unit, H\u00f4pital Femme M\u00e8re Enfant (HFME) de Lyon, Hospices Civils de Lyon, 69677 Bron, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4576-1992","authenticated-orcid":false,"given":"Emmanuelle","family":"Reboul","sequence":"additional","affiliation":[{"name":"Aix-Marseille Universit\u00e9, INRAE, INSERM, C2VN, 13885 Marseille, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,1,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1912","DOI":"10.1053\/j.gastro.2020.11.040","article-title":"Medical Advisory Panel New Classification and Management of Abetalipoproteinemia and Related Disorders","volume":"160","author":"Bredefeld","year":"2021","journal-title":"Gastroenterology"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1016\/B978-0-12-387025-4.00004-2","article-title":"Hypobetalipoproteinemia: Genetics, Biochemistry, and Clinical Spectrum","volume":"54","author":"Tarugi","year":"2011","journal-title":"Adv. Clin. Chem."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"423","DOI":"10.1007\/s11883-014-0423-3","article-title":"Update on Primary Hypobetalipoproteinemia","volume":"16","author":"Hooper","year":"2014","journal-title":"Curr. Atheroscler. Rep."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"999","DOI":"10.1126\/science.1439810","article-title":"Absence of Microsomal Triglyceride Transfer Protein in Individuals with Abetalipoproteinemia","volume":"258","author":"Wetterau","year":"1992","journal-title":"Science"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2243","DOI":"10.1161\/ATVBAHA.117.310121","article-title":"Understanding Chylomicron Retention Disease Through Sar1b Gtpase Gene Disruption: Insight From Cell Culture","volume":"37","author":"Seidman","year":"2017","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1038\/ng1145","article-title":"Mutations in a Sar1 GTPase of COPII Vesicles Are Associated with Lipid Absorption Disorders","volume":"34","author":"Jones","year":"2003","journal-title":"Nat. Genet."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2028","DOI":"10.1194\/jlr.M700207-JLR200","article-title":"Mechanisms Involved in Vitamin E Transport by Primary Enterocytes and in Vivo Absorption","volume":"48","author":"Anwar","year":"2007","journal-title":"J. Lipid Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1016\/0021-9150(95)05605-X","article-title":"Lipoproteins Containing Apolipoprotein B Isolated from Patients with Abetalipoproteinemia and Homozygous Hypobetalipoproteinemia: Identification and Characterization","volume":"118","author":"Aguie","year":"1995","journal-title":"Atherosclerosis"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1186\/1750-1172-5-24","article-title":"Guidelines for the Diagnosis and Management of Chylomicron Retention Disease Based on a Review of the Literature and the Experience of Two Centers","volume":"5","author":"Peretti","year":"2010","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1146\/annurev.nu.08.070188.002031","article-title":"Vitamin E Deficiency and Neurologic Disease","volume":"8","author":"Sokol","year":"1988","journal-title":"Annu. Rev. Nutr."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"915","DOI":"10.1093\/brain\/114.2.915","article-title":"Experimental Vitamin E Deficiency in Rats. Morphological and Functional Evidence of Abnormal Axonal Transport Secondary to Free Radical Damage","volume":"114","author":"Southam","year":"1991","journal-title":"Brain"},{"key":"ref_12","first-page":"451","article-title":"Myopathy in Vitamin E Deficient Rats: Muscle Fibre Necrosis Associated with Disturbances of Mitochondrial Function","volume":"183","author":"Thomas","year":"1993","journal-title":"J. Anat."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Cuerq, C., Bordat, C., Halimi, C., Blond, E., Nowicki, M., Peretti, N., and Reboul, E. (2020). Comparison of \u03b1-Tocopherol, \u03b1-Tocopherol Acetate, and \u03b1-Tocopheryl Polyethylene Glycol Succinate 1000 Absorption by Caco-2 TC7 Intestinal Cells. Nutrients, 13.","DOI":"10.3390\/nu13010129"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s10565-005-0085-6","article-title":"The Caco-2 Cell Line as a Model of the Intestinal Barrier: Influence of Cell and Culture-Related Factors on Caco-2 Cell Functional Characteristics","volume":"21","author":"Sambuy","year":"2005","journal-title":"Cell Biol. Toxicol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1242\/jcs.107.1.213","article-title":"Differential Expression of Sucrase-Isomaltase in Clones Isolated from Early and Late Passages of the Cell Line Caco-2: Evidence for Glucose-Dependent Negative Regulation","volume":"107","author":"Chantret","year":"1994","journal-title":"J. Cell Sci."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1186\/s13062-015-0035-z","article-title":"\u201cOff-Spotter\u201d: Very Fast and Exhaustive Enumeration of Genomic Lookalikes for Designing CRISPR\/Cas Guide RNAs","volume":"10","author":"Pliatsika","year":"2015","journal-title":"Biol. Direct."},{"key":"ref_17","unstructured":"Fortin, J., and Lun, A. (2022, July 22). On-Target and Off-Target Scoring Algorithms for CRISPR GRNAs. Issues Jfortin1\/CrisprScore R Package Version 1.0.0. Available online: https:\/\/github.com\/crisprVerse\/crisprScore."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1042\/BJ20040554","article-title":"Lutein Transport by Caco-2 TC-7 Cells Occurs Partly by a Facilitated Process Involving the Scavenger Receptor Class B Type I (SR-BI)","volume":"387","author":"Reboul","year":"2005","journal-title":"Biochem. J."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"126927","DOI":"10.1016\/j.foodchem.2020.126927","article-title":"Human Milk Pasteurisation Reduces Pre-Lipolysis but Not Digestive Lipolysis and Moderately Decreases Intestinal Lipid Uptake in a Combination of Preterm Infant in Vitro Models","volume":"329","author":"Vincent","year":"2020","journal-title":"Food Chem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1079\/BJN2001507","article-title":"Functional Characterization of Three Clones of the Human Intestinal Caco-2 Cell Line for Dietary Lipid Processing","volume":"87","author":"Salvini","year":"2002","journal-title":"Br. J. Nutr."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"e12018","DOI":"10.14814\/phy2.12018","article-title":"Chylomicrons Produced by Caco-2 Cells Contained ApoB-48 with Diameter of 80\u2013200 Nm","volume":"2","author":"Nauli","year":"2014","journal-title":"Physiol. Rep."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"816","DOI":"10.1126\/science.1225829","article-title":"A Programmable Dual-RNA\u2013Guided DNA Endonuclease in Adaptive Bacterial Immunity","volume":"337","author":"Jinek","year":"2012","journal-title":"Science"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1201","DOI":"10.1126\/science.1093131","article-title":"Niemann-Pick C1 Like 1 Protein Is Critical for Intestinal Cholesterol Absorption","volume":"303","author":"Altmann","year":"2004","journal-title":"Science"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"4075","DOI":"10.1074\/jbc.M510622200","article-title":"Compensatory Increase in Hepatic Lipogenesis in Mice with Conditional Intestine-Specific Mttp Deficiency","volume":"281","author":"Xie","year":"2006","journal-title":"J. Biol. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"30432","DOI":"10.1074\/jbc.M113.501247","article-title":"Lipid Absorption Defects in Intestine-Specific Microsomal Triglyceride Transfer Protein and ATP-Binding Cassette Transporter A1-Deficient Mice","volume":"288","author":"Iqbal","year":"2013","journal-title":"J. Biol. Chem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1016\/j.freeradbiomed.2017.04.013","article-title":"Cholesterol and Related Sterols Autoxidation","volume":"111","author":"Zerbinati","year":"2017","journal-title":"Free Radic. Biol. Med."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1016\/j.mce.2011.08.036","article-title":"Selective Liver X Receptor Modulators (SLiMs): What Use in Human Health?","volume":"351","author":"Viennois","year":"2012","journal-title":"Mol. Cell. Endocrinol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"693","DOI":"10.1016\/S0092-8674(00)81432-4","article-title":"Cholesterol and Bile Acid Metabolism Are Impaired in Mice Lacking the Nuclear Oxysterol Receptor LXR\u03b1","volume":"93","author":"Peet","year":"1998","journal-title":"Cell"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1259","DOI":"10.1016\/j.bbrc.2005.12.137","article-title":"Niemann\u2013Pick C1 like 1 Gene Expression Is down-Regulated by LXR Activators in the Intestine","volume":"340","author":"Duval","year":"2006","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1054","DOI":"10.1194\/jlr.M100358-JLR200","article-title":"LXR\/RXR Activation Enhances Basolateral Efflux of Cholesterol in CaCo-2 Cells","volume":"43","author":"Murthy","year":"2002","journal-title":"J. Lipid Res."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1103","DOI":"10.1194\/jlr.R084244","article-title":"Thematic Review Series: Lipid Transfer Proteins ABCG5 and ABCG8: More than a Defense against Xenosterols","volume":"59","author":"Patel","year":"2018","journal-title":"J. Lipid Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1007\/s004410051181","article-title":"Adipophilin Is a Specific Marker of Lipid Accumulation in Diverse Cell Types and Diseases","volume":"294","author":"Heid","year":"1998","journal-title":"Cell Tissue Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"499","DOI":"10.1042\/BC20110024","article-title":"The Proteome of Cytosolic Lipid Droplets Isolated from Differentiated Caco-2\/TC7 Enterocytes Reveals Cell-Specific Characteristics","volume":"103","author":"Bouchoux","year":"2011","journal-title":"Biol. Cell"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1173","DOI":"10.1016\/j.bbalip.2009.08.002","article-title":"Differential Association of Adipophilin and TIP47 Proteins with Cytoplasmic Lipid Droplets in Mouse Enterocytes during Dietary Fat Absorption","volume":"1791","author":"Lee","year":"2009","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"100085","DOI":"10.1016\/j.jlr.2021.100085","article-title":"Sar1b Mutant Mice Recapitulate Gastrointestinal Abnormalities Associated with Chylomicron Retention Disease","volume":"62","author":"Auclair","year":"2021","journal-title":"J. Lipid Res."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1023\/A:1020505512364","article-title":"New Insights into the Fatty Acid-Binding Protein (FABP) Family in the Small Intestine","volume":"239","author":"Besnard","year":"2002","journal-title":"Mol. Cell. Biochem."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"18532","DOI":"10.1074\/jbc.M301633200","article-title":"MGAT2, a Monoacylglycerol Acyltransferase Expressed in the Small Intestine","volume":"278","author":"Yen","year":"2003","journal-title":"J. Biol. Chem."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"603","DOI":"10.1177\/026119290503300618","article-title":"An Inter-Laboratory Study to Evaluate the Effects of Medium Composition on the Differentiation and Barrier Function of Caco-2 Cell Lines","volume":"33","author":"Zucco","year":"2005","journal-title":"Altern. Lab. Anim."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1146\/annurev.nu.16.070196.001541","article-title":"Vitamin E in Humans: Demand and Delivery","volume":"16","author":"Traber","year":"1996","journal-title":"Annu. Rev. Nutr."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1097\/MPG.0000000000003145","article-title":"Lipids Responsible for Intestinal or Hepatic Disorder: When to Suspect a Familial Intestinal Hypocholesterolemia?","volume":"73","author":"Sissaoui","year":"2021","journal-title":"J. Pediatr. Gastroenterol. Nutr."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1270","DOI":"10.1093\/ajcn\/59.6.1270","article-title":"Efficacy of Water-Soluble Vitamin E in the Treatment of Vitamin E Malabsorption in Short-Bowel Syndrome","volume":"59","author":"Traber","year":"1994","journal-title":"Am. J. Clin. Nutr."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"e23672","DOI":"10.1002\/jcla.23672","article-title":"A Novel p. Gly417Valfs* 12 Mutation in the MTTP Gene Causing Abetalipoproteinemia: Presentation of the First Patient in Mexico and Analysis of the Previously Reported Cases","volume":"35","year":"2021","journal-title":"J. Clin. Lab. Anal."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"554","DOI":"10.1016\/j.jacl.2019.05.013","article-title":"Novel Mutations of SAR1B Gene in Four Children with Chylomicron Retention Disease","volume":"13","author":"Simone","year":"2019","journal-title":"J. Clin. Lipidol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"919","DOI":"10.1007\/s10529-016-2064-9","article-title":"Improved Delivery of Cas9 Protein\/GRNA Complexes Using Lipofectamine CRISPRMAX","volume":"38","author":"Yu","year":"2016","journal-title":"Biotechnol. Lett."}],"container-title":["Nutrients"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6643\/15\/3\/505\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:09:28Z","timestamp":1760119768000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6643\/15\/3\/505"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,1,18]]},"references-count":44,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2023,2]]}},"alternative-id":["nu15030505"],"URL":"https:\/\/doi.org\/10.3390\/nu15030505","relation":{},"ISSN":["2072-6643"],"issn-type":[{"value":"2072-6643","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,1,18]]}}}