June 2021
Volume 62, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2021
Extracellular vesicles (exosomes) derived from primary optic nerve head astrocytes enhance neurite outgrowth.
Author Affiliations & Notes
  • Simon Kaja
    Ophthalmology, Loyola University Chicago, Maywood, Illinois, United States
    Molecular Pharmacology and Neuroscience, Loyola University Chicago, Maywood, Illinois, United States
  • Harsh Nilesh Hariani
    Graduate Program in Neuroscience, Loyola University Chicago, Maywood, Illinois, United States
  • Anita K. Ghosh
    Graduate Program in Biochemistry and Molecular Biology, Loyola University Chicago, Maywood, Illinois, United States
  • Giedrius Kalesnykas
    Research & Development, Experimentica Ltd., Vilnius, Lithuania
  • Walter Keith Jones
    Molecular Pharmacology and Neuroscience, Loyola University Chicago, Maywood, Illinois, United States
  • Footnotes
    Commercial Relationships   Simon Kaja, Experimentica Ltd. (F), Experimentica Ltd. (I), Experimentica Ltd. (P), Experimentica Ltd. (R), Experimentica Ltd. (S), eyeNOS Inc. (P), K&P Scientific LLC (F), K&P Scientific LLC (I), K&P Scientific LLC (R), K&P Scientific LLC (S); Harsh Hariani, None; Anita Ghosh, Experimentica Ltd. (F), Experimentica Ltd. (R), Experimentica Ltd. (S), eyeNOS Inc. (I), eyeNOS Inc. (P), eyeNOS Inc. (S), K&P Scientific LLC (F), K&P Scientific LLC (C), K&P Scientific LLC (R); Giedrius Kalesnykas, Experimentica Ltd. (F), Experimentica Ltd. (I), Experimentica Ltd. (E), Experimentica Ltd. (P), Experimentica Ltd. (R), Experimentica Ltd. (S), Spouse - Experimentica Ltd. (I); Walter Jones, eyeNOS Inc. (I), eyeNOS Inc. (P), eyeNOS Inc. (S)
  • Footnotes
    Support  The Glaucoma Foundation (SK), Dr. John P. and Therese E. Mulcahy Endowed Professorship in Ophthalmology (SK), Illinois Society for Prevention of Blindness (HNH, AKG), “Smart FDI” (project no. 01.2.1-LVPA-T-848-01-005) under the Promotion of Scientific Research, Experimental Development and Innovations Priority of the Operational Programme of the European Union Structural Funds for 2014-2020 (GK).
Investigative Ophthalmology & Visual Science June 2021, Vol.62, 1651. doi:
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      Simon Kaja, Harsh Nilesh Hariani, Anita K. Ghosh, Giedrius Kalesnykas, Walter Keith Jones; Extracellular vesicles (exosomes) derived from primary optic nerve head astrocytes enhance neurite outgrowth.. Invest. Ophthalmol. Vis. Sci. 2021;62(8):1651.

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

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Abstract

Purpose : To determine the properties of primary optic nerve head astrocyte (ONHA)-derived extracellular vesicles (exosomes), their uptake into primary neurons, and to identify the functional effects of exosomes on neuronal survival and neurite outgrowth.

Methods : Exosomes were prepared by ultracentrifugation from primary rat ONHAs. To determine the functional effects of lysyl oxidase like-1 (Loxl1) knockdown, we transduced ONHA with lentivirus harboring either control or Loxl1-targeting small hairpin RNA. A stable line with ~50% Loxl1 knockdown was generated. Exosomes were characterized by nanoparticle tracking analysis and immunoblotting. Uptake of exosomes into primary cortical neurons was quantified by quantitative microscopy using RNA-based labeling. Functional effects of exosomes were determined by testing their antioxidative properties in primary cortical neuronal culture. To quantify effects on neuronal differentiation, exosomes were added to neuronal cultures on days 3 and 5 in vitro and neurite outgrowth was quantified on day 7.

Results : Exosomes derived from wildtype, control- and Loxl1 shRNA-transduced ONHAs were of similar size (119.8±4.3, 121.9±2.8 and 121.1±2.5 nm, respectively, n=3, P=0.89). ONHA exosomes expressed the prototypical markers Alix and CD63, but were deficient of CD81. Exosomes were taken up by primary neurons, and this uptake was time-dependent and peaked at 4 h after addition (n=3, P<0.05). Viability of rat primary cortical neuronal cultures in response to oxidative stress insult did not change in the presence of exosomes derived from wild-type, control- or Loxl1 shRNA-transduced ONHA cultures, suggesting lack of antioxidative properties. Wild-type ONHA exosomes increased neurite outgrowth of primary rat cortical neurons by 20.1±3.3% (from 57.6±4.4 to 77.6±6.5 µm, n=5, P<0.05); similarly, exosomes from control shRNA-transduced ONHAs increased neurite outgrowth by 18.7±2.0% (to 76.3±4.6 µm, n=5, P<0.05). In contrast, exosomes from Loxl1 shRNA-transduced ONHAs prevented any increase in neurite outgrowth (2.2±4.0% change; 59.8±3.3 µm, n=5, P=0.98).

Conclusions : Our data provide evidence that ONHA-derived exosomes can exert functional effects on neurons and identify Loxl1 as a putative mediator of these effects. Altered exosome-mediated neuron-glia signaling in exfoliation glaucoma, where LOXL1 expression is chronically reduced, may be of pathological importance.

This is a 2021 ARVO Annual Meeting abstract.

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