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

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

doi:10.1016/j.nano.2020.102332

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

Nanomedicine. 2021 Feb:32:102332. doi: 10.1016/j.nano.2020.102332. Epub 2020 Nov 10.

Authors

Affiliations

¹ Eye Program, Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA; Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA; Biomedical Sciences, Cedars-Sinai Medical Center, Los Angeles, CA, USA.
² Neurosurgery, Cedars-Sinai Medical Center, Los Angeles, CA, USA.
³ Eye Program, Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA; Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA.
⁴ Eye Program, Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA; Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA; Surgery, Cedars-Sinai Medical Center, Los Angeles, CA, USA.
⁵ David Geffen School of Medicine at UCLA, Los Angeles, CA, USA.
⁶ American Eye Institute, Los Angeles, CA, USA.
⁷ Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA; Biomedical Sciences, Cedars-Sinai Medical Center, Los Angeles, CA, USA; David Geffen School of Medicine at UCLA, Los Angeles, CA, USA.
⁸ Eye Program, Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA; Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA; Biomedical Sciences, Cedars-Sinai Medical Center, Los Angeles, CA, USA; David Geffen School of Medicine at UCLA, Los Angeles, CA, USA.
⁹ Biomedical Sciences, Cedars-Sinai Medical Center, Los Angeles, CA, USA; Neurosurgery, Cedars-Sinai Medical Center, Los Angeles, CA, USA.
¹⁰ Eye Program, Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA; Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA; Biomedical Sciences, Cedars-Sinai Medical Center, Los Angeles, CA, USA; Neurosurgery, Cedars-Sinai Medical Center, Los Angeles, CA, USA; David Geffen School of Medicine at UCLA, Los Angeles, CA, USA. Electronic address: ljubimov@cshs.org.

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.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Adenoviridae / physiology
Adult
Aged
Aged, 80 and over
Biomarkers / metabolism
Cell Survival
Cells, Cultured
Cornea / drug effects
Cornea / pathology*
Diabetes Mellitus / pathology*
Epithelial Cells / drug effects
Epithelial Cells / pathology*
Epithelial Cells / virology
Female
Humans
Male
Middle Aged
Nanoparticles / chemistry*
Nanoparticles / ultrastructure
Oligonucleotides, Antisense / pharmacology
Polymers / chemistry*
RNA / pharmacology
RNA / therapeutic use*
Receptors, Cell Surface / metabolism
Signal Transduction / drug effects
Stem Cells / drug effects
Stem Cells / pathology*
Wound Healing* / drug effects

Substances

Biomarkers
Oligonucleotides, Antisense
Polymers
Receptors, Cell Surface
RNA

Abstract

Publication types

MeSH terms

Substances

Grants and funding