Small molecules facilitate single factor-mediated sweat gland cell reprogramming

Area of Science:

• Regenerative Medicine
• Dermatology
• Cell Biology

Background:

• Large skin defects can cause irreversible damage to sweat glands (SG), impairing skin function.
• Developing methods for functional skin repair and regeneration is crucial.
• Current strategies lack efficient ways to restore damaged SG.

Purpose of the Study:

• To develop a stepwise reprogramming strategy to convert fibroblasts into SG lineages.
• To generate functional induced SG-like cells (iSGCs) for skin regeneration.
• To evaluate the potential of iSGCs for in situ skin repair with SG restoration.

Main Methods:

• Fibroblast reprogramming using EDA overexpression in SG culture medium (SGM).
• Treatment with chemical cocktails to accelerate SG fate.
• Assessment of SG markers (CK5, CK10, CK18, CEA, AQP5) via qPCR, immunofluorescence, and flow cytometry.
• Functional evaluation using calcium activity analysis and in vivo mouse xenograft models (sweat tests, histology).

Main Results:

• EDA successfully drove fibroblast conversion into iSGCs, with increased SG marker expression.
• Chemical cocktails significantly enhanced SG marker expression (CK5, CK18, CK10, AQP5) and functional markers.
• iSGCs exhibited acetylcholine-induced calcium activity comparable to primary SG cells.
• In vivo studies showed regenerated SG structures and positive sweat tests in iSGC-treated mice.

Conclusions:

• A novel SG reprogramming strategy using EDA, SGM, and small molecules effectively generates functional iSGCs from fibroblasts.
• The generated iSGCs possess functional characteristics and demonstrate potential for in vivo skin regeneration.
• This approach holds significant implications for future skin regeneration therapies focused on restoring sweat gland function.