TY - JOUR
T1 - Preclinical Development of an AAV8-hUGT1A1 Vector for the Treatment of Crigler-Najjar Syndrome
AU - Collaud, Fanny
AU - Bortolussi, Giulia
AU - Guianvarc'h, Laurence
AU - Aronson, Sem J.
AU - Bordet, Thierry
AU - Veron, Philippe
AU - Charles, Severine
AU - Vidal, Patrice
AU - Sola, Marcelo Simon
AU - Rundwasser, Stephanie
AU - Dufour, Delphine G.
AU - Lacoste, Florence
AU - Luc, Cyril
AU - Wittenberghe, Laetitia v.
AU - Martin, Samia
AU - le Bec, Christine
AU - Bosma, Piter J.
AU - Muro, Andres F.
AU - Ronzitti, Giuseppe
AU - Hebben, Matthias
AU - Mingozzi, Federico
PY - 2019
Y1 - 2019
N2 - Adeno-associated viruses (AAVs) are among the most efficient vectors for liver gene therapy. Results obtained in the first hemophilia clinical trials demonstrated the long-term efficacy of this approach in humans, showing efficient targeting of hepatocytes with both self-complementary (sc) and single-stranded (ss) AAV vectors. However, to support clinical development of AAV-based gene therapies, efficient and scalable production processes are needed. In an effort to translate to the clinic an approach of AAV-mediated liver gene transfer to treat Crigler-Najjar (CN) syndrome, we developed an (ss)AAV8 vector carrying the human UDP-glucuronosyltransferase family 1-member A1 (hUGT1A1) transgene under the control of a liver-specific promoter. We compared our construct with similar (sc)AAV8 vectors expressing hUGT1A1, showing comparable potency in vitro and in vivo. Conversely, (ss)AAV8-hUGT1A1 vectors showed superior yields and product homogeneity compared with their (sc) counterpart. We then focused our efforts in the scale-up of a manufacturing process of the clinical product (ss)AAV8-hUGT1A1 based on the triple transfection of HEK293 cells grown in suspension. Large-scale production of this vector had characteristics identical to those of small-scale vectors produced in adherent cells. Preclinical studies in animal models of the disease and a good laboratory practice (GLP) toxicology-biodistribution study were also conducted using large-scale preparations of vectors. These studies demonstrated long-term safety and efficacy of gene transfer with (ss)AAV8-hUGT1A1 in relevant animal models of the disease, thus supporting the clinical translation of this gene therapy approach for the treatment of CN syndrome.
AB - Adeno-associated viruses (AAVs) are among the most efficient vectors for liver gene therapy. Results obtained in the first hemophilia clinical trials demonstrated the long-term efficacy of this approach in humans, showing efficient targeting of hepatocytes with both self-complementary (sc) and single-stranded (ss) AAV vectors. However, to support clinical development of AAV-based gene therapies, efficient and scalable production processes are needed. In an effort to translate to the clinic an approach of AAV-mediated liver gene transfer to treat Crigler-Najjar (CN) syndrome, we developed an (ss)AAV8 vector carrying the human UDP-glucuronosyltransferase family 1-member A1 (hUGT1A1) transgene under the control of a liver-specific promoter. We compared our construct with similar (sc)AAV8 vectors expressing hUGT1A1, showing comparable potency in vitro and in vivo. Conversely, (ss)AAV8-hUGT1A1 vectors showed superior yields and product homogeneity compared with their (sc) counterpart. We then focused our efforts in the scale-up of a manufacturing process of the clinical product (ss)AAV8-hUGT1A1 based on the triple transfection of HEK293 cells grown in suspension. Large-scale production of this vector had characteristics identical to those of small-scale vectors produced in adherent cells. Preclinical studies in animal models of the disease and a good laboratory practice (GLP) toxicology-biodistribution study were also conducted using large-scale preparations of vectors. These studies demonstrated long-term safety and efficacy of gene transfer with (ss)AAV8-hUGT1A1 in relevant animal models of the disease, thus supporting the clinical translation of this gene therapy approach for the treatment of CN syndrome.
UR - https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85059833634&origin=inward
UR - https://www.ncbi.nlm.nih.gov/pubmed/30705921
U2 - https://doi.org/10.1016/j.omtm.2018.12.011
DO - https://doi.org/10.1016/j.omtm.2018.12.011
M3 - Article
C2 - 30705921
SN - 2329-0501
VL - 12
SP - 157
EP - 174
JO - Molecular therapy. Methods & clinical development
JF - Molecular therapy. Methods & clinical development
ER -