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Employing a Gain-of-Function Factor IX Variant R338L to Advance the Efficacy and Safety of Hemophilia B Human Gene Therapy: Preclinical Evaluation Supporting an Ongoing Adeno-Associated Virus Clinical Trial

Vector capsid dose-dependent inflammation of transduced liver has limited the ability of adeno-associated virus (AAV) factor IX (FIX) gene therapy vectors to reliably convert severe to mild hemophilia B in human clinical trials. These trials also identified the need to understand AAV neutralizing an...

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Autores principales: Monahan, Paul E., Sun, Junjiang, Gui, Tong, Hu, Genlin, Hannah, William B., Wichlan, David G., Wu, Zhijian, Grieger, Joshua C., Li, Chengwen, Suwanmanee, Thipparat, Stafford, Darrel W., Booth, Carmen J., Samulski, Jade J., Kafri, Tal, McPhee, Scott W.J., Samulski, R. Jude
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Mary Ann Liebert, Inc. 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4326268/
https://www.ncbi.nlm.nih.gov/pubmed/25419787
http://dx.doi.org/10.1089/hum.2014.106
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author Monahan, Paul E.
Sun, Junjiang
Gui, Tong
Hu, Genlin
Hannah, William B.
Wichlan, David G.
Wu, Zhijian
Grieger, Joshua C.
Li, Chengwen
Suwanmanee, Thipparat
Stafford, Darrel W.
Booth, Carmen J.
Samulski, Jade J.
Kafri, Tal
McPhee, Scott W.J.
Samulski, R. Jude
author_facet Monahan, Paul E.
Sun, Junjiang
Gui, Tong
Hu, Genlin
Hannah, William B.
Wichlan, David G.
Wu, Zhijian
Grieger, Joshua C.
Li, Chengwen
Suwanmanee, Thipparat
Stafford, Darrel W.
Booth, Carmen J.
Samulski, Jade J.
Kafri, Tal
McPhee, Scott W.J.
Samulski, R. Jude
author_sort Monahan, Paul E.
collection PubMed
description Vector capsid dose-dependent inflammation of transduced liver has limited the ability of adeno-associated virus (AAV) factor IX (FIX) gene therapy vectors to reliably convert severe to mild hemophilia B in human clinical trials. These trials also identified the need to understand AAV neutralizing antibodies and empty AAV capsids regarding their impact on clinical success. To address these safety concerns, we have used a scalable manufacturing process to produce GMP-grade AAV8 expressing the FIXR338L gain-of-function variant with minimal (<10%) empty capsid and have performed comprehensive dose–response, biodistribution, and safety evaluations in clinically relevant hemophilia models. The scAAV8.FIXR338L vector produced greater than 6-fold increased FIX specific activity compared with wild-type FIX and demonstrated linear dose responses from doses that produced 2–500% FIX activity, associated with dose-dependent hemostasis in a tail transection bleeding challenge. More importantly, using a bleeding model that closely mimics the clinical morbidity of hemophilic arthropathy, mice that received the scAAV8.FIXR338L vector developed minimal histopathological findings of synovitis after hemarthrosis, when compared with mice that received identical doses of wild-type FIX vector. Hemostatically normal mice (n=20) and hemophilic mice (n=88) developed no FIX antibodies after peripheral intravenous vector delivery. No CD8(+) T cell liver infiltrates were observed, despite the marked tropism of scAAV8.FIXR338L for the liver in a comprehensive biodistribution evaluation (n=60 animals). With respect to the role of empty capsids, we demonstrated that in vivo FIXR338L expression was not influenced by the presence of empty AAV particles, either in the presence or absence of various titers of AAV8-neutralizing antibodies. Necropsy of FIX(–/–) mice 8–10 months after vector delivery revealed no microvascular or macrovascular thrombosis in mice expressing FIXR338L (plasma FIX activity, 100–500%). These preclinical studies demonstrate a safety:efficacy profile supporting an ongoing phase 1/2 human clinical trial of the scAAV8.FIXR338L vector (designated BAX335).
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spelling pubmed-43262682015-02-24 Employing a Gain-of-Function Factor IX Variant R338L to Advance the Efficacy and Safety of Hemophilia B Human Gene Therapy: Preclinical Evaluation Supporting an Ongoing Adeno-Associated Virus Clinical Trial Monahan, Paul E. Sun, Junjiang Gui, Tong Hu, Genlin Hannah, William B. Wichlan, David G. Wu, Zhijian Grieger, Joshua C. Li, Chengwen Suwanmanee, Thipparat Stafford, Darrel W. Booth, Carmen J. Samulski, Jade J. Kafri, Tal McPhee, Scott W.J. Samulski, R. Jude Hum Gene Ther Research Articles Vector capsid dose-dependent inflammation of transduced liver has limited the ability of adeno-associated virus (AAV) factor IX (FIX) gene therapy vectors to reliably convert severe to mild hemophilia B in human clinical trials. These trials also identified the need to understand AAV neutralizing antibodies and empty AAV capsids regarding their impact on clinical success. To address these safety concerns, we have used a scalable manufacturing process to produce GMP-grade AAV8 expressing the FIXR338L gain-of-function variant with minimal (<10%) empty capsid and have performed comprehensive dose–response, biodistribution, and safety evaluations in clinically relevant hemophilia models. The scAAV8.FIXR338L vector produced greater than 6-fold increased FIX specific activity compared with wild-type FIX and demonstrated linear dose responses from doses that produced 2–500% FIX activity, associated with dose-dependent hemostasis in a tail transection bleeding challenge. More importantly, using a bleeding model that closely mimics the clinical morbidity of hemophilic arthropathy, mice that received the scAAV8.FIXR338L vector developed minimal histopathological findings of synovitis after hemarthrosis, when compared with mice that received identical doses of wild-type FIX vector. Hemostatically normal mice (n=20) and hemophilic mice (n=88) developed no FIX antibodies after peripheral intravenous vector delivery. No CD8(+) T cell liver infiltrates were observed, despite the marked tropism of scAAV8.FIXR338L for the liver in a comprehensive biodistribution evaluation (n=60 animals). With respect to the role of empty capsids, we demonstrated that in vivo FIXR338L expression was not influenced by the presence of empty AAV particles, either in the presence or absence of various titers of AAV8-neutralizing antibodies. Necropsy of FIX(–/–) mice 8–10 months after vector delivery revealed no microvascular or macrovascular thrombosis in mice expressing FIXR338L (plasma FIX activity, 100–500%). These preclinical studies demonstrate a safety:efficacy profile supporting an ongoing phase 1/2 human clinical trial of the scAAV8.FIXR338L vector (designated BAX335). Mary Ann Liebert, Inc. 2015-02-01 2014-11-24 /pmc/articles/PMC4326268/ /pubmed/25419787 http://dx.doi.org/10.1089/hum.2014.106 Text en Copyright 2015, Mary Ann Liebert, Inc. © Paul E. Monahan et al. 2015; Published by Mary Ann Liebert, Inc. This Open Access article is distributed under the terms of the Creative Commons License Attribution-Non-Commercial Share Alike (http://creativecommons.org/licenses/by-nc-sa/4.0/).
spellingShingle Research Articles
Monahan, Paul E.
Sun, Junjiang
Gui, Tong
Hu, Genlin
Hannah, William B.
Wichlan, David G.
Wu, Zhijian
Grieger, Joshua C.
Li, Chengwen
Suwanmanee, Thipparat
Stafford, Darrel W.
Booth, Carmen J.
Samulski, Jade J.
Kafri, Tal
McPhee, Scott W.J.
Samulski, R. Jude
Employing a Gain-of-Function Factor IX Variant R338L to Advance the Efficacy and Safety of Hemophilia B Human Gene Therapy: Preclinical Evaluation Supporting an Ongoing Adeno-Associated Virus Clinical Trial
title Employing a Gain-of-Function Factor IX Variant R338L to Advance the Efficacy and Safety of Hemophilia B Human Gene Therapy: Preclinical Evaluation Supporting an Ongoing Adeno-Associated Virus Clinical Trial
title_full Employing a Gain-of-Function Factor IX Variant R338L to Advance the Efficacy and Safety of Hemophilia B Human Gene Therapy: Preclinical Evaluation Supporting an Ongoing Adeno-Associated Virus Clinical Trial
title_fullStr Employing a Gain-of-Function Factor IX Variant R338L to Advance the Efficacy and Safety of Hemophilia B Human Gene Therapy: Preclinical Evaluation Supporting an Ongoing Adeno-Associated Virus Clinical Trial
title_full_unstemmed Employing a Gain-of-Function Factor IX Variant R338L to Advance the Efficacy and Safety of Hemophilia B Human Gene Therapy: Preclinical Evaluation Supporting an Ongoing Adeno-Associated Virus Clinical Trial
title_short Employing a Gain-of-Function Factor IX Variant R338L to Advance the Efficacy and Safety of Hemophilia B Human Gene Therapy: Preclinical Evaluation Supporting an Ongoing Adeno-Associated Virus Clinical Trial
title_sort employing a gain-of-function factor ix variant r338l to advance the efficacy and safety of hemophilia b human gene therapy: preclinical evaluation supporting an ongoing adeno-associated virus clinical trial
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4326268/
https://www.ncbi.nlm.nih.gov/pubmed/25419787
http://dx.doi.org/10.1089/hum.2014.106
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