Effects of lentivirus-mediated C3 expression on trabecular meshwork cells and intraocular pressure

Junkai Tan, Ning Fan, Ningli Wang, Bingkai Feng, Ming Yang, Guo Liu, Yun Wang, Xianjun Zhu, Paul L. Kaufman, Iok Hou Pang, Xuyang Liu

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

PURPOSE. We evaluated the effects of lentivirus-mediated exoenzyme C3 transferase (C3) expression on cultured primary human trabecular meshwork (HTM) cells in vitro, and on rat intraocular pressure (IOP). METHODS. HTM cells were cultured and treated with lentivirus vectors expressing either green fluorescent protein (GFP) only (LV-GFP) or GFP and C3 together (LV-C3-GFP). Changes in cell morphology and actin stress fibers were assessed. The vectors were also injected into the anterior chamber of rats, and GFP expression was visualized by a Micron III Retinal Imaging Microscope in vivo and a fluorescence microscope ex vivo. Changes in rat IOP were monitored by using a rebound tonometer and the eyes were evaluated by slit lamp. RESULTS. LV-mediated C3 expression induced morphologic changes in HTM cells. The cells became retracted and rounded. GFP expression in the anterior chamber angle of rats was observed in vivo from 8 days to 48 days after injection of LV-C3-GFP or LV-GFP. IOP was significantly decreased in the LV-C3-GFP group starting 3 days post injection, and lasting for at least 40 days, when compared to either the contralateral control eyes (the LV-GFP group) or the ipsilateral baseline before injection (P < 0.05). No obvious inflammatory signs were observed in either the LV-C3-GFP or LV-GFP groups. CONCLUSIONS. LV-mediated C3 expression induced changes in morphology of cultured HTM cells. Intracameral injection of LV-C3-GFP lowered rat IOP for at least 40 days. No significant inflammatory reactions were observed in either the LV-C3-GFP or LV-GFP groups. This study supports the possible use of C3 gene therapy for the treatment of glaucoma.

Original languageEnglish
Pages (from-to)4937-4944
Number of pages8
JournalInvestigative Ophthalmology and Visual Science
Volume59
Issue number12
DOIs
StatePublished - Oct 2018

Fingerprint

Trabecular Meshwork
Lentivirus
Green Fluorescent Proteins
Intraocular Pressure
Injections
Anterior Chamber
Stress Fibers
Transferases

Keywords

  • C3 transferase
  • Intraocular pressure
  • Lentivirus
  • Rat
  • Trabecular meshwork

Cite this

Tan, Junkai ; Fan, Ning ; Wang, Ningli ; Feng, Bingkai ; Yang, Ming ; Liu, Guo ; Wang, Yun ; Zhu, Xianjun ; Kaufman, Paul L. ; Pang, Iok Hou ; Liu, Xuyang. / Effects of lentivirus-mediated C3 expression on trabecular meshwork cells and intraocular pressure. In: Investigative Ophthalmology and Visual Science. 2018 ; Vol. 59, No. 12. pp. 4937-4944.
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title = "Effects of lentivirus-mediated C3 expression on trabecular meshwork cells and intraocular pressure",
abstract = "PURPOSE. We evaluated the effects of lentivirus-mediated exoenzyme C3 transferase (C3) expression on cultured primary human trabecular meshwork (HTM) cells in vitro, and on rat intraocular pressure (IOP). METHODS. HTM cells were cultured and treated with lentivirus vectors expressing either green fluorescent protein (GFP) only (LV-GFP) or GFP and C3 together (LV-C3-GFP). Changes in cell morphology and actin stress fibers were assessed. The vectors were also injected into the anterior chamber of rats, and GFP expression was visualized by a Micron III Retinal Imaging Microscope in vivo and a fluorescence microscope ex vivo. Changes in rat IOP were monitored by using a rebound tonometer and the eyes were evaluated by slit lamp. RESULTS. LV-mediated C3 expression induced morphologic changes in HTM cells. The cells became retracted and rounded. GFP expression in the anterior chamber angle of rats was observed in vivo from 8 days to 48 days after injection of LV-C3-GFP or LV-GFP. IOP was significantly decreased in the LV-C3-GFP group starting 3 days post injection, and lasting for at least 40 days, when compared to either the contralateral control eyes (the LV-GFP group) or the ipsilateral baseline before injection (P < 0.05). No obvious inflammatory signs were observed in either the LV-C3-GFP or LV-GFP groups. CONCLUSIONS. LV-mediated C3 expression induced changes in morphology of cultured HTM cells. Intracameral injection of LV-C3-GFP lowered rat IOP for at least 40 days. No significant inflammatory reactions were observed in either the LV-C3-GFP or LV-GFP groups. This study supports the possible use of C3 gene therapy for the treatment of glaucoma.",
keywords = "C3 transferase, Intraocular pressure, Lentivirus, Rat, Trabecular meshwork",
author = "Junkai Tan and Ning Fan and Ningli Wang and Bingkai Feng and Ming Yang and Guo Liu and Yun Wang and Xianjun Zhu and Kaufman, {Paul L.} and Pang, {Iok Hou} and Xuyang Liu",
year = "2018",
month = "10",
doi = "10.1167/iovs.18-24978",
language = "English",
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Tan, J, Fan, N, Wang, N, Feng, B, Yang, M, Liu, G, Wang, Y, Zhu, X, Kaufman, PL, Pang, IH & Liu, X 2018, 'Effects of lentivirus-mediated C3 expression on trabecular meshwork cells and intraocular pressure', Investigative Ophthalmology and Visual Science, vol. 59, no. 12, pp. 4937-4944. https://doi.org/10.1167/iovs.18-24978

Effects of lentivirus-mediated C3 expression on trabecular meshwork cells and intraocular pressure. / Tan, Junkai; Fan, Ning; Wang, Ningli; Feng, Bingkai; Yang, Ming; Liu, Guo; Wang, Yun; Zhu, Xianjun; Kaufman, Paul L.; Pang, Iok Hou; Liu, Xuyang.

In: Investigative Ophthalmology and Visual Science, Vol. 59, No. 12, 10.2018, p. 4937-4944.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Effects of lentivirus-mediated C3 expression on trabecular meshwork cells and intraocular pressure

AU - Tan, Junkai

AU - Fan, Ning

AU - Wang, Ningli

AU - Feng, Bingkai

AU - Yang, Ming

AU - Liu, Guo

AU - Wang, Yun

AU - Zhu, Xianjun

AU - Kaufman, Paul L.

AU - Pang, Iok Hou

AU - Liu, Xuyang

PY - 2018/10

Y1 - 2018/10

N2 - PURPOSE. We evaluated the effects of lentivirus-mediated exoenzyme C3 transferase (C3) expression on cultured primary human trabecular meshwork (HTM) cells in vitro, and on rat intraocular pressure (IOP). METHODS. HTM cells were cultured and treated with lentivirus vectors expressing either green fluorescent protein (GFP) only (LV-GFP) or GFP and C3 together (LV-C3-GFP). Changes in cell morphology and actin stress fibers were assessed. The vectors were also injected into the anterior chamber of rats, and GFP expression was visualized by a Micron III Retinal Imaging Microscope in vivo and a fluorescence microscope ex vivo. Changes in rat IOP were monitored by using a rebound tonometer and the eyes were evaluated by slit lamp. RESULTS. LV-mediated C3 expression induced morphologic changes in HTM cells. The cells became retracted and rounded. GFP expression in the anterior chamber angle of rats was observed in vivo from 8 days to 48 days after injection of LV-C3-GFP or LV-GFP. IOP was significantly decreased in the LV-C3-GFP group starting 3 days post injection, and lasting for at least 40 days, when compared to either the contralateral control eyes (the LV-GFP group) or the ipsilateral baseline before injection (P < 0.05). No obvious inflammatory signs were observed in either the LV-C3-GFP or LV-GFP groups. CONCLUSIONS. LV-mediated C3 expression induced changes in morphology of cultured HTM cells. Intracameral injection of LV-C3-GFP lowered rat IOP for at least 40 days. No significant inflammatory reactions were observed in either the LV-C3-GFP or LV-GFP groups. This study supports the possible use of C3 gene therapy for the treatment of glaucoma.

AB - PURPOSE. We evaluated the effects of lentivirus-mediated exoenzyme C3 transferase (C3) expression on cultured primary human trabecular meshwork (HTM) cells in vitro, and on rat intraocular pressure (IOP). METHODS. HTM cells were cultured and treated with lentivirus vectors expressing either green fluorescent protein (GFP) only (LV-GFP) or GFP and C3 together (LV-C3-GFP). Changes in cell morphology and actin stress fibers were assessed. The vectors were also injected into the anterior chamber of rats, and GFP expression was visualized by a Micron III Retinal Imaging Microscope in vivo and a fluorescence microscope ex vivo. Changes in rat IOP were monitored by using a rebound tonometer and the eyes were evaluated by slit lamp. RESULTS. LV-mediated C3 expression induced morphologic changes in HTM cells. The cells became retracted and rounded. GFP expression in the anterior chamber angle of rats was observed in vivo from 8 days to 48 days after injection of LV-C3-GFP or LV-GFP. IOP was significantly decreased in the LV-C3-GFP group starting 3 days post injection, and lasting for at least 40 days, when compared to either the contralateral control eyes (the LV-GFP group) or the ipsilateral baseline before injection (P < 0.05). No obvious inflammatory signs were observed in either the LV-C3-GFP or LV-GFP groups. CONCLUSIONS. LV-mediated C3 expression induced changes in morphology of cultured HTM cells. Intracameral injection of LV-C3-GFP lowered rat IOP for at least 40 days. No significant inflammatory reactions were observed in either the LV-C3-GFP or LV-GFP groups. This study supports the possible use of C3 gene therapy for the treatment of glaucoma.

KW - C3 transferase

KW - Intraocular pressure

KW - Lentivirus

KW - Rat

KW - Trabecular meshwork

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U2 - 10.1167/iovs.18-24978

DO - 10.1167/iovs.18-24978

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