Expression of Normal or Mutated X-Linked BCOR Transcripts in OFCD iPSCs

Area of Science:

• Stem Cell Biology
• Genetics
• Developmental Biology

Background:

• Oculofaciocardiodental (OFCD) syndrome is a developmental disorder linked to mutations in the BCOR gene.
• Induced pluripotent stem cells (iPSCs) offer a model for studying genetic disorders and developing therapeutic strategies.
• Reprogramming patient-derived cells is crucial for creating disease-specific models.

Purpose of the Study:

• To reprogram stem cells from a patient with OFCD (SCAP-O) into transgene-free induced pluripotent stem cells (iPSCs).
• To generate isogenic iPSC lines with either normal (BCOR-WT) or mutated (BCOR-mut) BCOR transcripts for OFCD studies.
• To establish a foundation for studying OFCD mechanisms and potential gene correction therapies.

Main Methods:

• Isolation and subcloning of stem cells from tooth apical papilla (SCAP) from an OFCD patient.
• Reprogramming of SCAP-O cells and a BCOR-mutated subclone into iPSCs using an excisable lentiviral vector system (hSTEMCCA-loxP).
• Transgene removal via Cre-mediated excision and verification of BCOR transcript expression in iPSC clones by sequencing.

Main Results:

• Successfully generated transgene-free iPSCs from OFCD patient-derived stem cells (SCAP-O).
• Generated iPSC clones exhibited varying expression patterns of BCOR-WT and BCOR-mut transcripts.
• Established a method for creating isogenic iPSC lines, with some expressing solely BCOR-WT or BCOR-mut, and others expressing both.

Conclusions:

• The study successfully generated iPSCs from an OFCD patient, providing a valuable tool for disease modeling.
• The generation of isogenic iPSC lines with distinct BCOR expression profiles is a critical first step towards understanding OFCD pathogenesis.
• This work lays the groundwork for future research into OFCD mechanisms and the development of targeted therapeutic interventions.