Biodistribution of adeno-associated virus type 2 carrying multi-characteristic opsin in dogs following intravitreal injection
© 2021 Nanoscope Technologies LLC. Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd..
Gene therapy of retinal diseases using recombinant adeno-associated virus (rAAV) vector-based delivery has shown clinical success, and clinical trials based on rAAV-based optogenetic therapies are currently in progress. Recently, we have developed multi-characteristic opsin (MCO), which has been shown to effectively re-photosensitize photoreceptor-degenerated retina in mice leading to vision restoration at ambient light environment. Here, we report the biodistribution of the rAAV2 carried MCO (vMCO-I) in live samples and post-mortem organs following intraocular delivery in wild-type dogs. Immunohistochemistry showed that the intravitreal injection of vMCO-I resulted in gene transduction in the inner nuclear layer (INL) but did not induce detectable inflammatory or immune reaction in the dog retina. Vector DNA analysis of live body wastes and body fluids such as saliva and nasal secretions using quantitative polymerase chain reaction (qPCR) showed no correlative increase of vector copy in nasal secretions or saliva, minimal increase of vector copy in urine in the low-dose group 13 weeks after injection and in the faeces of the high-dose group at 3-13 weeks after injection suggesting clearance of the virus vector via urine and faeces. Further analysis of vector DNA extracted from faeces using PCR showed no transgene after 3 weeks post-injection. Intravitreal injection of vMCO-I resulted in few sporadic off-target presences of the vector in the mesenteric lymph node, liver, spleen and testis. This study showed that intravitreal rAAV2-based delivery of MCO-I for retinal gene therapy is safe.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2021 |
|---|---|
| Erschienen: |
2021 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:25 |
|---|---|
| Enthalten in: |
Journal of cellular and molecular medicine - 25(2021), 18 vom: 21. Sept., Seite 8676-8686 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Tchedre, Kissaou T [VerfasserIn] |
|---|
| Links: |
|---|
| Themen: |
Biodistribution |
|---|
| Anmerkungen: |
Date Completed 21.02.2022 Date Revised 03.04.2024 published: Print-Electronic Citation Status MEDLINE |
|---|
| doi: |
10.1111/jcmm.16823 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM329616927 |
|---|
| LEADER | 01000caa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM329616927 | ||
| 003 | DE-627 | ||
| 005 | 20240403232925.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231225s2021 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1111/jcmm.16823 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1362.xml |
| 035 | |a (DE-627)NLM329616927 | ||
| 035 | |a (NLM)34418301 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Tchedre, Kissaou T |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Biodistribution of adeno-associated virus type 2 carrying multi-characteristic opsin in dogs following intravitreal injection |
| 264 | 1 | |c 2021 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 21.02.2022 | ||
| 500 | |a Date Revised 03.04.2024 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © 2021 Nanoscope Technologies LLC. Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd. | ||
| 520 | |a Gene therapy of retinal diseases using recombinant adeno-associated virus (rAAV) vector-based delivery has shown clinical success, and clinical trials based on rAAV-based optogenetic therapies are currently in progress. Recently, we have developed multi-characteristic opsin (MCO), which has been shown to effectively re-photosensitize photoreceptor-degenerated retina in mice leading to vision restoration at ambient light environment. Here, we report the biodistribution of the rAAV2 carried MCO (vMCO-I) in live samples and post-mortem organs following intraocular delivery in wild-type dogs. Immunohistochemistry showed that the intravitreal injection of vMCO-I resulted in gene transduction in the inner nuclear layer (INL) but did not induce detectable inflammatory or immune reaction in the dog retina. Vector DNA analysis of live body wastes and body fluids such as saliva and nasal secretions using quantitative polymerase chain reaction (qPCR) showed no correlative increase of vector copy in nasal secretions or saliva, minimal increase of vector copy in urine in the low-dose group 13 weeks after injection and in the faeces of the high-dose group at 3-13 weeks after injection suggesting clearance of the virus vector via urine and faeces. Further analysis of vector DNA extracted from faeces using PCR showed no transgene after 3 weeks post-injection. Intravitreal injection of vMCO-I resulted in few sporadic off-target presences of the vector in the mesenteric lymph node, liver, spleen and testis. This study showed that intravitreal rAAV2-based delivery of MCO-I for retinal gene therapy is safe | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, N.I.H., Extramural | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a MCO | |
| 650 | 4 | |a biodistribution | |
| 650 | 4 | |a qPCR | |
| 650 | 4 | |a rAAV | |
| 650 | 4 | |a retinal gene therapy | |
| 700 | 1 | |a Batabyal, Subrata |e verfasserin |4 aut | |
| 700 | 1 | |a Galicia, Melissa |e verfasserin |4 aut | |
| 700 | 1 | |a Narcisse, Darryl |e verfasserin |4 aut | |
| 700 | 1 | |a Mustafi, Sourajit Mitra |e verfasserin |4 aut | |
| 700 | 1 | |a Ayyagari, Ananta |e verfasserin |4 aut | |
| 700 | 1 | |a Chavala, Sai |e verfasserin |4 aut | |
| 700 | 1 | |a Mohanty, Samarendra K |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Journal of cellular and molecular medicine |d 2000 |g 25(2021), 18 vom: 21. Sept., Seite 8676-8686 |w (DE-627)NLM118801236 |x 1582-4934 |7 nnns |
| 773 | 1 | 8 | |g volume:25 |g year:2021 |g number:18 |g day:21 |g month:09 |g pages:8676-8686 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1111/jcmm.16823 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 25 |j 2021 |e 18 |b 21 |c 09 |h 8676-8686 | ||