Reprogramming of iPSCs to NPCEC-like cells by biomimetic scaffolds for zonular fiber reconstruction

Ectopia lentis (EL), characterised by impaired zonular fibers originating from non-pigmented ciliary epithelial cells (NPCEC), presents formidable surgical complexities and potential risks of visual impairment. Cataract surgery is the only treatment method for EL, but it leads to the loss of accommodative power of the lens post-operatively. Furthermore, the challenge of repairing zonular ligaments in situ remains a significant global issue. Ocular tissue and aqueous humour samples from patients with EL were subjected to RNA sequencing and Olink high-throughput proteomic analysis, revealing the downregulation of pathogenic genes ( FBN1 , MFAP2 ) and upregulation of secretory proteins (IL-12, MMP-1). The high expression of FBN1 and MFAP2 in NPCECs suggests their potential as candidates for zonular fiber construction; however, the limited availability of donor sources restricts the feasibility of NPCEC transplantation therapy. The reprogramming and directional differentiation of induced pluripotent stem cells (iPSC) to NPCEC was successfully achieved using the developed biomimetic scaffolds that mimic the microstructures of natural radial zonular fibers. Excitingly, the single injection of induced NPCEC-like cells significantly contributed to restoring and enhancing mechanical properties in zonular fiber structures in a rabbit model with EL. This proposed in situ iPSC-based regeneration technique might serve as an innovative therapeutic strategy for clinical EL patients, reduce the cataract surgery rate, and retain the adjustment capacity of inherent lentis. • Identification of Pathogenic and Secretory Proteins: Through RNA sequencing and high-throughput proteomic analysis, our study identified the downregulation of pathogenic genes ( FBN1, MFAP2 ) and upregulation of secretory proteins (IL-12, MMP-1) in ocular tissues and aqueous humour samples from ectopia lentis patients. • Innovative iPSC Reprogramming and Differentiation Technique: We developed a novel biomaterials-based technique for the reprogramming and directed differentiation of induced pluripotent stem cells (iPSCs) into non-pigmented ciliary epithelial cells (NPCECs) using biomimetic scaffolds. This method effectively mimics the microstructures of natural radial zonular fibers. • Successful In Situ Regeneration in Animal Model: A single injection of induced NPCEC-like cells significantly restored and enhanced the mechanical properties of zonular fiber structures in a rabbit model with ectopia lentis, representing a groundbreaking advancement in in-situ repair of zonular ligaments through cell therapy.