Abstract |
Microvillus inclusion disease (MVID), caused by loss-of-function mutations in the motor protein myosin Vb (MYO5B), is a severe infantile disease characterized by diarrhea, malabsorption, and acid/base instability, requiring intensive parenteral support for nutritional and fluid management. Human patient-derived enteroids represent a model for investigation of monogenic epithelial disorders but are a rare resource from MVID patients. We developed human enteroids with different loss-of function MYO5B variants and showed that they recapitulated the structural changes found in native MVID enterocytes. Multiplex immunofluorescence imaging of patient duodenal tissues revealed patient-specific changes in localization of brush border transporters. Functional analysis of electrolyte transport revealed profound loss of Na+/H+ exchange (NHE) activity in MVID patient enteroids with near-normal chloride secretion. The chloride channel-blocking antidiarrheal drug crofelemer dose-dependently inhibited agonist-mediated fluid secretion. MVID enteroids exhibited altered differentiation and maturation versus healthy enteroids. γ- Secretase inhibition with DAPT recovered apical brush border structure and functional Na+/H+ exchange activity in MVID enteroids. Transcriptomic analysis revealed potential pathways involved in the rescue of MVID cells including serum/glucocorticoid-regulated kinase 2 (SGK2) and NHE regulatory factor 3 (NHERF3). These results demonstrate the utility of patient-derived enteroids for developing therapeutic approaches to MVID.
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Authors | Meri Kalashyan, Krishnan Raghunathan, Haley Oller, Marie-Theres Bayer, Lissette Jimenez, Joseph T Roland, Elena Kolobova, Susan J Hagen, Jeffrey D Goldsmith, Mitchell D Shub, James R Goldenring, Izumi Kaji, Jay R Thiagarajah |
Journal | The Journal of clinical investigation
(J Clin Invest)
Vol. 133
Issue 20
(10 16 2023)
ISSN: 1558-8238 [Electronic] United States |
PMID | 37643022
(Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
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Chemical References |
- Myosin Heavy Chains
- Myosin Type V
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Topics |
- Humans
- Microvilli
(genetics)
- Myosin Heavy Chains
(genetics)
- Myosin Type V
(genetics)
- Enterocytes
(metabolism)
- Malabsorption Syndromes
(genetics, therapy, metabolism)
- Mucolipidoses
(genetics, therapy, metabolism)
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