A Methodological Approach Using rAAV Vectors Encoding Nanobody-Based Biologics to Evaluate ARTC2.2 and P2X7 In Vivo
On murine T cells, mono-ADP ribosyltransferase ARTC2.2 catalyzes ADP-ribosylation of various surface proteins when nicotinamide adenine dinucleotide (NAD+) is released into the extracellular compartment. Covalent ADP-ribosylation of the P2X7 receptor by ARTC2.2 thereby represents an additional mecha...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2021-08-01
|
| Series: | Frontiers in Immunology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2021.704408/full |
| _version_ | 1818362983512801280 |
|---|---|
| author | Henri Gondé Henri Gondé Mélanie Demeules Romain Hardet Allan Scarpitta Marten Junge Carolina Pinto-Espinoza Rémi Varin Rémi Varin Friedrich Koch-Nolte Olivier Boyer Olivier Boyer Sahil Adriouch |
| author_facet | Henri Gondé Henri Gondé Mélanie Demeules Romain Hardet Allan Scarpitta Marten Junge Carolina Pinto-Espinoza Rémi Varin Rémi Varin Friedrich Koch-Nolte Olivier Boyer Olivier Boyer Sahil Adriouch |
| author_sort | Henri Gondé |
| collection | DOAJ |
| description | On murine T cells, mono-ADP ribosyltransferase ARTC2.2 catalyzes ADP-ribosylation of various surface proteins when nicotinamide adenine dinucleotide (NAD+) is released into the extracellular compartment. Covalent ADP-ribosylation of the P2X7 receptor by ARTC2.2 thereby represents an additional mechanism of activation, complementary to its triggering by extracellular ATP. P2X7 is a multifaceted receptor that may represents a potential target in inflammatory, and neurodegenerative diseases, as well as in cancer. We present herein an experimental approach using intramuscular injection of recombinant AAV vectors (rAAV) encoding nanobody-based biologics targeting ARTC2.2 or P2X7. We demonstrate the ability of these in vivo generated biologics to potently and durably block P2X7 or ARTC2.2 activities in vivo, or in contrast, to potentiate NAD+- or ATP-induced activation of P2X7. We additionally demonstrate the ability of rAAV-encoded functional heavy chain antibodies to elicit long-term depletion of T cells expressing high levels of ARTC2.2 or P2X7. Our approach of using rAAV to generate functional nanobody-based biologics in vivo appears promising to evaluate the role of ARTC2.2 and P2X7 in murine acute as well as chronic disease models. |
| first_indexed | 2024-12-13T21:41:15Z |
| format | Article |
| id | doaj.art-cbcdab012e614b578183a1da39767dad |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-12-13T21:41:15Z |
| publishDate | 2021-08-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-cbcdab012e614b578183a1da39767dad2022-12-21T23:30:32ZengFrontiers Media S.A.Frontiers in Immunology1664-32242021-08-011210.3389/fimmu.2021.704408704408A Methodological Approach Using rAAV Vectors Encoding Nanobody-Based Biologics to Evaluate ARTC2.2 and P2X7 In VivoHenri Gondé0Henri Gondé1Mélanie Demeules2Romain Hardet3Allan Scarpitta4Marten Junge5Carolina Pinto-Espinoza6Rémi Varin7Rémi Varin8Friedrich Koch-Nolte9Olivier Boyer10Olivier Boyer11Sahil Adriouch12Normandie University, UNIROUEN, INSERM U1234, Pathophysiology, Autoimmunity, Neuromuscular Diseases and Regenerative THERapies, Rouen, FranceRouen University Hospital, Department of Pharmacy, Rouen, FranceNormandie University, UNIROUEN, INSERM U1234, Pathophysiology, Autoimmunity, Neuromuscular Diseases and Regenerative THERapies, Rouen, FranceNormandie University, UNIROUEN, INSERM U1234, Pathophysiology, Autoimmunity, Neuromuscular Diseases and Regenerative THERapies, Rouen, FranceNormandie University, UNIROUEN, INSERM U1234, Pathophysiology, Autoimmunity, Neuromuscular Diseases and Regenerative THERapies, Rouen, FranceInstitute of Immunology, University Medical Center Hamburg-Eppendorf, Hamburg, GermanyInstitute of Immunology, University Medical Center Hamburg-Eppendorf, Hamburg, GermanyNormandie University, UNIROUEN, INSERM U1234, Pathophysiology, Autoimmunity, Neuromuscular Diseases and Regenerative THERapies, Rouen, FranceRouen University Hospital, Department of Pharmacy, Rouen, FranceInstitute of Immunology, University Medical Center Hamburg-Eppendorf, Hamburg, GermanyNormandie University, UNIROUEN, INSERM U1234, Pathophysiology, Autoimmunity, Neuromuscular Diseases and Regenerative THERapies, Rouen, FranceRouen University Hospital, Department of Immunology and Biotherapy, Rouen, FranceNormandie University, UNIROUEN, INSERM U1234, Pathophysiology, Autoimmunity, Neuromuscular Diseases and Regenerative THERapies, Rouen, FranceOn murine T cells, mono-ADP ribosyltransferase ARTC2.2 catalyzes ADP-ribosylation of various surface proteins when nicotinamide adenine dinucleotide (NAD+) is released into the extracellular compartment. Covalent ADP-ribosylation of the P2X7 receptor by ARTC2.2 thereby represents an additional mechanism of activation, complementary to its triggering by extracellular ATP. P2X7 is a multifaceted receptor that may represents a potential target in inflammatory, and neurodegenerative diseases, as well as in cancer. We present herein an experimental approach using intramuscular injection of recombinant AAV vectors (rAAV) encoding nanobody-based biologics targeting ARTC2.2 or P2X7. We demonstrate the ability of these in vivo generated biologics to potently and durably block P2X7 or ARTC2.2 activities in vivo, or in contrast, to potentiate NAD+- or ATP-induced activation of P2X7. We additionally demonstrate the ability of rAAV-encoded functional heavy chain antibodies to elicit long-term depletion of T cells expressing high levels of ARTC2.2 or P2X7. Our approach of using rAAV to generate functional nanobody-based biologics in vivo appears promising to evaluate the role of ARTC2.2 and P2X7 in murine acute as well as chronic disease models.https://www.frontiersin.org/articles/10.3389/fimmu.2021.704408/fullP2X7 (purino) receptorAAV vectorsnanobodies (VHH)animal modelsextracellular ATP (eATP)extracellular NAD+ |
| spellingShingle | Henri Gondé Henri Gondé Mélanie Demeules Romain Hardet Allan Scarpitta Marten Junge Carolina Pinto-Espinoza Rémi Varin Rémi Varin Friedrich Koch-Nolte Olivier Boyer Olivier Boyer Sahil Adriouch A Methodological Approach Using rAAV Vectors Encoding Nanobody-Based Biologics to Evaluate ARTC2.2 and P2X7 In Vivo Frontiers in Immunology P2X7 (purino) receptor AAV vectors nanobodies (VHH) animal models extracellular ATP (eATP) extracellular NAD+ |
| title | A Methodological Approach Using rAAV Vectors Encoding Nanobody-Based Biologics to Evaluate ARTC2.2 and P2X7 In Vivo |
| title_full | A Methodological Approach Using rAAV Vectors Encoding Nanobody-Based Biologics to Evaluate ARTC2.2 and P2X7 In Vivo |
| title_fullStr | A Methodological Approach Using rAAV Vectors Encoding Nanobody-Based Biologics to Evaluate ARTC2.2 and P2X7 In Vivo |
| title_full_unstemmed | A Methodological Approach Using rAAV Vectors Encoding Nanobody-Based Biologics to Evaluate ARTC2.2 and P2X7 In Vivo |
| title_short | A Methodological Approach Using rAAV Vectors Encoding Nanobody-Based Biologics to Evaluate ARTC2.2 and P2X7 In Vivo |
| title_sort | methodological approach using raav vectors encoding nanobody based biologics to evaluate artc2 2 and p2x7 in vivo |
| topic | P2X7 (purino) receptor AAV vectors nanobodies (VHH) animal models extracellular ATP (eATP) extracellular NAD+ |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2021.704408/full |
| work_keys_str_mv | AT henrigonde amethodologicalapproachusingraavvectorsencodingnanobodybasedbiologicstoevaluateartc22andp2x7invivo AT henrigonde amethodologicalapproachusingraavvectorsencodingnanobodybasedbiologicstoevaluateartc22andp2x7invivo AT melaniedemeules amethodologicalapproachusingraavvectorsencodingnanobodybasedbiologicstoevaluateartc22andp2x7invivo AT romainhardet amethodologicalapproachusingraavvectorsencodingnanobodybasedbiologicstoevaluateartc22andp2x7invivo AT allanscarpitta amethodologicalapproachusingraavvectorsencodingnanobodybasedbiologicstoevaluateartc22andp2x7invivo AT martenjunge amethodologicalapproachusingraavvectorsencodingnanobodybasedbiologicstoevaluateartc22andp2x7invivo AT carolinapintoespinoza amethodologicalapproachusingraavvectorsencodingnanobodybasedbiologicstoevaluateartc22andp2x7invivo AT remivarin amethodologicalapproachusingraavvectorsencodingnanobodybasedbiologicstoevaluateartc22andp2x7invivo AT remivarin amethodologicalapproachusingraavvectorsencodingnanobodybasedbiologicstoevaluateartc22andp2x7invivo AT friedrichkochnolte amethodologicalapproachusingraavvectorsencodingnanobodybasedbiologicstoevaluateartc22andp2x7invivo AT olivierboyer amethodologicalapproachusingraavvectorsencodingnanobodybasedbiologicstoevaluateartc22andp2x7invivo AT olivierboyer amethodologicalapproachusingraavvectorsencodingnanobodybasedbiologicstoevaluateartc22andp2x7invivo AT sahiladriouch amethodologicalapproachusingraavvectorsencodingnanobodybasedbiologicstoevaluateartc22andp2x7invivo AT henrigonde methodologicalapproachusingraavvectorsencodingnanobodybasedbiologicstoevaluateartc22andp2x7invivo AT henrigonde methodologicalapproachusingraavvectorsencodingnanobodybasedbiologicstoevaluateartc22andp2x7invivo AT melaniedemeules methodologicalapproachusingraavvectorsencodingnanobodybasedbiologicstoevaluateartc22andp2x7invivo AT romainhardet methodologicalapproachusingraavvectorsencodingnanobodybasedbiologicstoevaluateartc22andp2x7invivo AT allanscarpitta methodologicalapproachusingraavvectorsencodingnanobodybasedbiologicstoevaluateartc22andp2x7invivo AT martenjunge methodologicalapproachusingraavvectorsencodingnanobodybasedbiologicstoevaluateartc22andp2x7invivo AT carolinapintoespinoza methodologicalapproachusingraavvectorsencodingnanobodybasedbiologicstoevaluateartc22andp2x7invivo AT remivarin methodologicalapproachusingraavvectorsencodingnanobodybasedbiologicstoevaluateartc22andp2x7invivo AT remivarin methodologicalapproachusingraavvectorsencodingnanobodybasedbiologicstoevaluateartc22andp2x7invivo AT friedrichkochnolte methodologicalapproachusingraavvectorsencodingnanobodybasedbiologicstoevaluateartc22andp2x7invivo AT olivierboyer methodologicalapproachusingraavvectorsencodingnanobodybasedbiologicstoevaluateartc22andp2x7invivo AT olivierboyer methodologicalapproachusingraavvectorsencodingnanobodybasedbiologicstoevaluateartc22andp2x7invivo AT sahiladriouch methodologicalapproachusingraavvectorsencodingnanobodybasedbiologicstoevaluateartc22andp2x7invivo |