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Ontogenesis and Modulation of Intestinal Unesterified Cholesterol Sequestration in a Mouse Model of Niemann–Pick C1 Disease

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Abstract

Background

Mutations in the NPC1 gene result in sequestration of unesterified cholesterol (UC) and glycosphingolipids in most tissues leading to multi-organ disease, especially in the brain, liver, lungs, and spleen. Various data from NPC1-deficient mice suggest the small intestine (SI) is comparatively less affected, even in late stage disease.

Methods

Using the Npc1nih mouse model, we measured SI weights and total cholesterol (TC) levels in Npc1−/− versus Npc1+/+ mice as a function of age, and then after prolonged ezetimibe-induced inhibition of cholesterol absorption. Next, we determined intestinal levels of UC and esterified cholesterol (EC), and cholesterol synthesis rates in Npc1−/− and Npc1+/+ mice, with and without the cholesterol-esterifying enzyme SOAT2, following a once-only subcutaneous injection with 2-hydroxypropyl-β-cyclodextrin (2HPβCD).

Results

By ~ 42 days of age, intestinal TC levels averaged ~ 2.1-fold more (mostly UC) in the Npc1−/− versus Npc1+/+ mice with no further increase thereafter. Chronic ezetimibe treatment lowered intestinal TC levels in the Npc1−/− mice by only ~ 16%. In Npc1−/− mice given 2HPβCD 24 h earlier, UC levels fell, EC levels increased (although less so in mice lacking SOAT2), and cholesterol synthesis was suppressed equally in the Npc1−/−:Soat2+/+ and Npc1−/−:Soat2−/− mice.

Conclusions

The low and static levels of intestinal UC sequestration in Npc1−/− mice likely reflect the continual sloughing of cells from the mucosa. This sequestration is blunted by about the same extent following a single acute treatment with 2HPβCD as it is by a prolonged ezetimibe-induced block of cholesterol absorption.

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Acknowledgments

We thank Dr. Lawrence Rudel of Wake Forest University School of Medicine for helpful discussions about cholesterol-esterifying enzymes in the small intestine and Dr. Harry R. Davis Jr., previously at Merck & Co. Inc., for a gift of ezetimibe. Heather Waddell, Carolyn Crumpton, Monti Schneiderman, and Stephen Ostermann provided excellent technical assistance.

Funding

This research was supported by National Institutes of Health Grants R01 HL 009610 (SDT) and DK 078592 (JJR) and the Ara Parseghian Medical Research Foundation (JJR).

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Author notes

  1. Anna M. Taylor

    Present address: School of Behavioral and Brain Sciences, University of Texas at Dallas, Richardson, TX, 75080, USA

  2. Ryan D. Jones

    Present address: Department of Pathology, Northwestern University, Chicago, IL, 60611, USA

Authors and Affiliations

  1. Department of Internal Medicine, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd, Dallas, TX, 75390, USA

    Adam M. Lopez, Joyce J. Repa & Stephen D. Turley

  2. Department of Pediatrics, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd, Dallas, TX, 75390, USA

    Charina M. Ramirez

  3. Department of Physiology, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd, Dallas, TX, 75390, USA

    Anna M. Taylor, Ryan D. Jones & Joyce J. Repa

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  1. Adam M. Lopez
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Contributions

AML, CMR, AMT, RDJ, JJR, and SDT all had varying but significant roles in the conception and design of the research, performance of experiments, calculation and interpretation of the data, and the preparation of the initial draft of the manuscript. JJR performed statistical analyses, and SDT finalized the manuscript for submission. All authors approved the final manuscript.

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Correspondence to Stephen D. Turley.

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Lopez, A.M., Ramirez, C.M., Taylor, A.M. et al. Ontogenesis and Modulation of Intestinal Unesterified Cholesterol Sequestration in a Mouse Model of Niemann–Pick C1 Disease. Dig Dis Sci 65, 158–167 (2020). https://doi.org/10.1007/s10620-019-05736-x

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