Related Experiment Video
Updated: Jul 5, 2026
Combining Human Organoids and Organ-on-a-Chip Technology to Model Intestinal Region-Specific Functionality
Published on: May 5, 2022
Patient-derived enteroids provide a platform for the development of therapeutic approaches in microvillus inclusion
Meri Kalashyan1, Krishnan Raghunathan1, Haley Oller1
1Division of Gastroenterology, Hepatology and Nutrition, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts, USA.
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.
Insights
Microvillus inclusion disease (MVID) patient enteroids model MYO5B mutations. γ-Secretase inhibition with DAPT restored brush border structure and Na+/H+ exchange, offering therapeutic potential.
Area of Science:
- Gastroenterology
- Cell Biology
- Genetics
Background:
- Microvillus inclusion disease (MVID) is a severe infantile disorder caused by MYO5B mutations.
- It leads to diarrhea, malabsorption, and nutritional challenges, often requiring parenteral support.
- Patient-derived enteroids are valuable for studying monogenic epithelial disorders like MVID.
Purpose of the Study:
- To develop and characterize human enteroids modeling MVID with MYO5B loss-of-function variants.
- To investigate the structural and functional defects in MVID enterocytes.
- To explore potential therapeutic strategies for MVID.
Main Methods:
- Generation of human enteroids from MVID patients with MYO5B variants.
- Multiplex immunofluorescence imaging of duodenal tissues and enteroids.
- Functional analysis of electrolyte transport and fluid secretion.
- Treatment with γ-secretase inhibitor (DAPT) and transcriptomic analysis.
Main Results:
- MVID enteroids recapitulated the structural changes observed in patient enterocytes.
- Significant loss of Na+/H+ exchange (NHE) activity was detected in MVID enteroids.
- DAPT treatment restored apical brush border structure and functional NHE activity.
- Transcriptomic analysis identified SGK2 and NHERF3 as potential rescue pathways.
Conclusions:
- Patient-derived MVID enteroids serve as a robust model for studying the disease.
- γ-Secretase inhibition shows promise as a therapeutic approach for MVID.
- Understanding altered transporter function and identifying rescue pathways are crucial for MVID treatment.
