Exploring the human lacrimal gland using organoids and single-cell sequencing

Area of Science:

• Ophthalmology and Vision Science
• Regenerative Medicine
• Developmental Biology

Background:

• The lacrimal gland is crucial for ocular surface health, and its dysfunction leads to dry eye disease.
• Current models for studying lacrimal gland physiology and pathology are limited.

Purpose of the Study:

• To establish and characterize long-term 3D organoid cultures of mouse and human lacrimal glands.
• To investigate the role of Pax6 in lacrimal gland development and differentiation.
• To create a platform for studying lacrimal gland (patho-)physiology and tear secretion.

Main Methods:

• Development of long-term 3D organoid culture conditions for lacrimal glands.
• CRISPR-Cas9 genome editing to assess the function of Pax6.
• Single-cell RNA sequencing to analyze cellular heterogeneity.
• Neurotransmitter stimulation assays and orthotopic transplantation in mice.

Main Results:

• Established expandable 3D lacrimal gland organoids recapitulating ductal morphology and transcriptional profiles.
• Demonstrated that the transcription factor Pax6 is essential for adult lacrimal gland cell differentiation.
• Generated a single-cell atlas of human lacrimal gland tissue and organoids.
• Showcased that human organoids mimic neurotransmitter-induced tear secretion and can produce tear products after transplantation.

Conclusions:

• 3D lacrimal gland organoids provide a robust experimental system for studying gland function and disease.
• Pax6 is identified as a key regulator in lacrimal gland development.
• This platform facilitates research into dry eye disease and lacrimal gland disorders.