Patient-derived enteroids provide a platform for the development of therapeutic approaches in microvillus inclusion disease.

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.

• 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)

Chemical References

• Myosin Heavy Chains
• Myosin Type V

Topics

• Humans
• Microvilli (genetics)
• Myosin Heavy Chains (genetics)
• Myosin Type V (genetics)
• Enterocytes (metabolism)
• Malabsorption Syndromes (genetics, therapy, metabolism)
• Mucolipidoses (genetics, therapy, metabolism)