PMID- 30021812
OWN - NLM
STAT- MEDLINE
DCOM- 20190328
LR  - 20190328
IS  - 1468-2079 (Electronic)
IS  - 0007-1161 (Linking)
VI  - 102
IP  - 10
DP  - 2018 Oct
TI  - Targeted therapy for the post-operative conjunctiva: SPARC silencing reduces
      collagen deposition.
PG  - 1460-1470
LID - 10.1136/bjophthalmol-2018-311937 [doi]
AB  - BACKGROUND: To develop targeted antifibrotic therapy for glaucoma filtration
      surgery; this study determines the effectiveness of small interfering RNA (siRNA)
      to reduce in vivo secreted protein acidic and rich in cysteine (SPARC) expression
      using the mouse model of conjunctival scarring. METHODS: Experimental surgery was
      performed as described for the mouse model of conjunctival scarring. Scrambled
      (siScram) or Sparc (siSparc) siRNAs, loaded on layer-by-layer (LbL)
      nanoparticles, were injected into the conjunctiva immediately after surgery.
      Expression of Sparc, Col1a1, Fn1 and Mmp14 was measured by real-time PCR and
      immunoblotting on days 7 and 14 postsurgery. Live imaging of the operated eyes
      was performed using slit lamp, anterior segment-optical coherence tomography and 
      confocal microscopy. Tissue pathology was evaluated by histochemical and
      immunofluorescent analyses of operated conjunctival cryosections. Tissue
      apoptosis was quantitated by annexin V assay. RESULTS : siSparc, delivered via
      expanded LbL nanoparticles, significantly inhibited Sparc transcription in both
      day 7 (2.04-fold) and day 14 (1.39-fold) treated tissues. Sparc suppression on
      day 7 was associated with a significant reduction of Col1a1 (2.52-fold), Fn1
      (2.89-fold) and Mmp14 (2.23-fold) mRNAs. At the protein level, both SPARC and
      collagen 1A1 (COL1A1) were significantly reduced at both time points with siSparc
      treatment. Nanoparticles were visualised within cell-like structures by confocal 
      microscopy, while overt tissue response or apoptosis was not observed.
      CONCLUSIONS : SPARC targeted therapy effectively reduced both SPARC and collagen 
      production in the operated mouse conjunctiva. This proof-of-concept study
      suggests that targeted treatment of fibrosis in glaucoma surgery is safe and
      feasible, with the potential to extend to a range of potential genes associated
      with fibrosis.
CI  - (c) Author(s) (or their employer(s)) 2018. Re-use permitted under CC BY-NC. No
      commercial re-use. See rights and permissions. Published by BMJ.
FAU - Seet, Li Fong
AU  - Seet LF
AD  - Ocular Therapeutics and Drug Delivery, Singapore Eye Research Institute,
      Singapore, Singapore.
AD  - Department of Ophthalmology, Yong Loo Lin School of Medicine, National University
      of Singapore, Singapore, Singapore.
AD  - Duke-NUS Medical School, Singapore, Singapore.
FAU - Tan, Yang Fei
AU  - Tan YF
AD  - School of Materials Science and Engineering, Nanyang Technological University,
      Singapore, Singapore.
FAU - Toh, Li Zhen
AU  - Toh LZ
AD  - Ocular Therapeutics and Drug Delivery, Singapore Eye Research Institute,
      Singapore, Singapore.
FAU - Chu, Stephanie Wl
AU  - Chu SW
AD  - Ocular Therapeutics and Drug Delivery, Singapore Eye Research Institute,
      Singapore, Singapore.
FAU - Lee, Ying Shi
AU  - Lee YS
AD  - Ocular Therapeutics and Drug Delivery, Singapore Eye Research Institute,
      Singapore, Singapore.
FAU - Venkatraman, Subbu S
AU  - Venkatraman SS
AD  - Ocular Therapeutics and Drug Delivery, Singapore Eye Research Institute,
      Singapore, Singapore tina.wong.t.l@singhealth.com.sg ASSubbu@ntu.edu.sg.
AD  - School of Materials Science and Engineering, Nanyang Technological University,
      Singapore, Singapore.
AD  - NTU - Northwestern University Institute for Nanomedicine, Singapore, Singapore.
AD  - MedTech, National Heart Centre, Singapore, Singapore.
FAU - Wong, Tina T
AU  - Wong TT
AD  - Ocular Therapeutics and Drug Delivery, Singapore Eye Research Institute,
      Singapore, Singapore tina.wong.t.l@singhealth.com.sg ASSubbu@ntu.edu.sg.
AD  - Department of Ophthalmology, Yong Loo Lin School of Medicine, National University
      of Singapore, Singapore, Singapore.
AD  - Duke-NUS Medical School, Singapore, Singapore.
AD  - School of Materials Science and Engineering, Nanyang Technological University,
      Singapore, Singapore.
AD  - Glaucoma Service, Singapore National Eye Centre, Singapore, Singapore.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20180718
PL  - England
TA  - Br J Ophthalmol
JT  - The British journal of ophthalmology
JID - 0421041
RN  - 0 (Osteonectin)
RN  - 0 (SPARC protein, mouse)
RN  - 63231-63-0 (RNA)
RN  - 9007-34-5 (Collagen)
SB  - IM
MH  - Animals
MH  - Cells, Cultured
MH  - Collagen/*metabolism
MH  - Conjunctiva/*pathology
MH  - Conjunctival Diseases/genetics/metabolism/*therapy
MH  - Cornea/*metabolism/pathology
MH  - Disease Models, Animal
MH  - Filtering Surgery/*adverse effects
MH  - Flow Cytometry
MH  - Gene Expression Regulation
MH  - Genetic Therapy/*methods
MH  - Glaucoma/surgery
MH  - Immunoblotting
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - Microscopy, Confocal
MH  - Osteonectin/biosynthesis/genetics/*therapeutic use
MH  - Postoperative Complications
MH  - RNA/genetics
MH  - Real-Time Polymerase Chain Reaction
MH  - Tomography, Optical Coherence
PMC - PMC6173823
OTO - NOTNLM
OT  - *Experimental - animal models
OT  - *conjunctiva
OT  - *treatment surgery
OT  - *wound healing
COIS- Competing interests: None declared.
EDAT- 2018/07/20 06:00
MHDA- 2019/03/29 06:00
CRDT- 2018/07/20 06:00
PHST- 2018/01/17 00:00 [received]
PHST- 2018/04/22 00:00 [revised]
PHST- 2018/05/24 00:00 [accepted]
PHST- 2018/07/20 06:00 [pubmed]
PHST- 2019/03/29 06:00 [medline]
PHST- 2018/07/20 06:00 [entrez]
AID - bjophthalmol-2018-311937 [pii]
AID - 10.1136/bjophthalmol-2018-311937 [doi]
PST - ppublish
SO  - Br J Ophthalmol. 2018 Oct;102(10):1460-1470. doi:
      10.1136/bjophthalmol-2018-311937. Epub 2018 Jul 18.