Novel nanopolymer RNA therapeutics normalize human diabetic corneal wound healing and epithelial stem cells

Diabetic cornea; Gene therapy; Limbal stem cells; Nanobioconjugate; RNA therapeutics; Wound healing; miRNA.
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Andrei A Kramerov, Ruchi Shah, Hui Ding, Eggehard Holler, Sue Turjman, Yaron S Rabinowitz, Sean Ghiam, Ezra Maguen, Clive N Svendsen, Mehrnoosh Saghizadeh, Julia Y Ljubimova, Alexander V Ljubimov

  • Nanomedicine
  • 4.282
  • 2021 Feb:32:102332.
  • Mouse
  • 流式
  • 免疫/内分泌
  • 干细胞
  • 糖尿病
  • CD115 (CSF-1R)

Abstract

Human diabetic corneas develop delayed wound healing, epithelial stem cell dysfunction, recurrent erosions, and keratitis. Adenoviral gene therapy modulating c-Met, cathepsin F and MMP-10 normalized wound healing and epithelial stem cells in organ-cultured diabetic corneas but showed toxicity in stem cell-enriched cultured limbal epithelial cells (LECs). For a safer treatment, we engineered a novel nanobiopolymer (NBC) that carried antisense oligonucleotide (AON) RNA therapeutics suppressing cathepsin F or MMP-10, and miR-409-3p that inhibits c-Met. NBC was internalized by LECs through transferrin receptor (TfR)-mediated endocytosis, inhibited cathepsin F or MMP-10 and upregulated c-Met. Non-toxic NBC modulating c-Met and cathepsin F accelerated wound healing in diabetic LECs and organ-cultured corneas vs. control NBC. NBC treatment normalized levels of stem cell markers (keratins 15 and 17, ABCG2, and ΔNp63), and signaling mediators (p-EGFR, p-Akt and p-p38). Non-toxic nano RNA therapeutics thus present a safe alternative to viral gene therapy for normalizing diabetic corneal cells.Keywords: Diabetic cornea; Gene therapy; Limbal stem cells; Nanobioconjugate; RNA therapeutics; Wound healing; miRNA.
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