Investigative Ophthalmology & Visual Science Cover Image for Volume 65, Issue 7
June 2024
Volume 65, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2024
Characterization of a broadband ambient light activatable multi-characteristic opsin supporting its superior performance in restoring vision
Author Affiliations & Notes
  • Samarendra Mohanty
    Nanoscope, Bedford, Texas, United States
  • Pratyajit Mohapatra
    Nanoscope, Bedford, Texas, United States
  • Amir Singh
    Nanoscope, Bedford, Texas, United States
  • Subrata Batabyal
    Nanoscope, Bedford, Texas, United States
  • Michael Carlson
    Nanoscope, Bedford, Texas, United States
  • Sanghoon Kim
    Nanoscope, Bedford, Texas, United States
  • Najam (Naj) Sharif
    Nanoscope, Bedford, Texas, United States
  • Footnotes
    Commercial Relationships   Samarendra Mohanty Nanoscope, Code E (Employment), Nanoscope, Code I (Personal Financial Interest), Nanoscope, Code P (Patent); Pratyajit Mohapatra Nanoscope, Code E (Employment); Amir Singh Nanoscope, Code E (Employment); Subrata Batabyal Nanoscope, Code E (Employment), Nanoscope, Code I (Personal Financial Interest); Michael Carlson Nanoscope, Code E (Employment), Nanoscope, Code I (Personal Financial Interest); Sanghoon Kim Nanoscope, Code E (Employment), Nanoscope, Code I (Personal Financial Interest); Najam Sharif Nanoscope, Code E (Employment), Nanoscope, Code I (Personal Financial Interest)
  • Footnotes
    Support  NIH Grant EY025905
Investigative Ophthalmology & Visual Science June 2024, Vol.65, 6078. doi:
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      Samarendra Mohanty, Pratyajit Mohapatra, Amir Singh, Subrata Batabyal, Michael Carlson, Sanghoon Kim, Najam (Naj) Sharif; Characterization of a broadband ambient light activatable multi-characteristic opsin supporting its superior performance in restoring vision. Invest. Ophthalmol. Vis. Sci. 2024;65(7):6078.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Intravitreal multi-characteristic opsin (MCO) gene therapy has demonstrated improvement in patients with severe vision loss due to retinitis pigmentosa and Stargardt disease. Here, we present structure and functioning of individual subunits of MCO that imparts broadband light sensitivity with high dynamic range and fast kinetics. We also present comparison of key characteristics of MCO with other opsins that are being developed for vision restoration.

Methods : Studies were conducted on HEK293 cells and retinal explants from rd10 mice (animal model of retinal degeneration) which had been transduced with an adeno-associated virus-2 carried MCO (MCO-010). A combination of predictive in-silico protein modeling, Western blot-based protease protection assays, immunostaining as well as electrophysiological techniques were utilized to gather structural and functional data.

Results : Our multi-disciplinary studies revealed that the first subunit of MCO-010 acts as an ion-channel, while the second subunit is partially embedded in the membrane extending to the third intracellular cytoplasmic subunit. Patch-clamp recordings showed that MCO-010 expressing cells get depolarized upon light activation with large dynamic range across the visible light spectrum. Deletion of transmembrane subunit 1 or intracellular subunit 3 led to either hyperpolarization in response to broadband visible light or significant decrease in photosensitivity. MCO-010 sensitized cells were activated with fast on-off kinetics (time-constants of ~ 2 to 3 ms). Furthermore, multi-electrode array recordings confirmed multi-color light-activation of MCO-010 expressing bipolar neurons in the photoreceptor degenerated mouse retina.

Conclusions : The unique structure and functioning of MCO-010 represent key characteristics needed to restore vision in subjects with degenerative retinal conditions. Broadband light sensitivity with high dynamic range of MCO has opened new vistas in restoring vision at ambient lighting environments, without the need for a light stimulation device.

This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.

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