rAAV-CFTRΔR Rescues the Cystic Fibrosis Phenotype in Human Intestinal Organoids and Cystic Fibrosis Mice.
BACKGROUND
Gene therapy holds promise for a curative mutation-independent treatment applicable to all patients with cystic fibrosis (CF). The various viral vector-based clinical trials conducted in the past have demonstrated safety and tolerance of different vectors, but none have led to a clear and persistent clinical benefit. Recent clinical breakthroughs in recombinant adeno-associated viral vector (rAAV)-based gene therapy encouraged us to reexplore an rAAV approach for CF.


OBJECTIVE
We evaluated the preclinical potential of rAAV gene therapy for CF to restore chloride and fluid secretion in two complementary models: intestinal organoids derived from subjects with CF and a CF mouse model, an important milestone toward the development of a clinical rAAV candidate for CF gene therapy.


METHODS
We engineered an rAAV vector containing a truncated CF transmembrane conductance regulator (CFTRΔR) combined with a short promoter (CMV173) to ensure optimal gene expression. A rescue in chloride and fluid secretion after rAAV-CFTRΔR treatment was assessed by forskolin-induced swelling in CF transmembrane conductance regulator (CFTR)-deficient organoids and by nasal potential differences in ΔF508 mice.


RESULTS
rAAV-CFTRΔR transduction of human CFTR-deficient organoids resulted in forskolin-induced swelling, indicating a restoration of CFTR function. Nasal potential differences demonstrated a clear response to low chloride and forskolin perfusion in most rAAV-CFTRΔR-treated CF mice.


CONCLUSIONS
Our study provides robust evidence that rAAV-mediated gene transfer of a truncated CFTR functionally rescues the CF phenotype across the nasal mucosa of CF mice and in patient-derived organoids. These results underscore the clinical potential of rAAV-CFTRΔR in offering a cure for all patients with CF in the future.
1 Molecular Virology and Gene Therapy.
Dragana
D
Vidović
1 Molecular Virology and Gene Therapy.
Marianne S
MS
Carlon
2 INSERM U1151, University Paris Descartes, Faculté de Médecine Necker Enfants-Malades, Paris, France.
Mélanie F
MF
da Cunha
3 Department of Pediatric Pulmonology, Wilhelmina Children's Hospital, and.|4 Laboratory of Translational Immunology, University Medical Center, Utrecht, the Netherlands.
Johanna F
JF
Dekkers
2 INSERM U1151, University Paris Descartes, Faculté de Médecine Necker Enfants-Malades, Paris, France.
Monika I
MI
Hollenhorst
5 Department of Gastroenterology and Hepatology, Erasmus University Medical Center, Rotterdam, the Netherlands.
Marcel J C
MJ
Bijvelds
6 Stem Cell Biology and Embryology.
Anabela S
AS
Ramalho
7 Neurobiology and Gene Therapy.|8 Leuven Viral Vector Core, and.
Chris
C
Van den Haute
9 Translational Research in Gastrointestinal Disorders, KU Leuven, Flanders, Belgium.
Marc
M
Ferrante
7 Neurobiology and Gene Therapy.
Veerle
V
Baekelandt
10 Department of Pediatric Pulmonology, Erasmus University Medical Centre/Sophia Children's Hospital, Rotterdam, the Netherlands; and.
Hettie M
HM
Janssens
11 University Hospital Leuven, Flanders, Belgium.
Kris
K
De Boeck
2 INSERM U1151, University Paris Descartes, Faculté de Médecine Necker Enfants-Malades, Paris, France.
Isabelle
I
Sermet-Gaudelus
5 Department of Gastroenterology and Hepatology, Erasmus University Medical Center, Rotterdam, the Netherlands.
Hugo R
HR
de Jonge
1 Molecular Virology and Gene Therapy.|8 Leuven Viral Vector Core, and.
Rik
R
Gijsbers
3 Department of Pediatric Pulmonology, Wilhelmina Children's Hospital, and.|4 Laboratory of Translational Immunology, University Medical Center, Utrecht, the Netherlands.
Jeffrey M
JM
Beekman
2 INSERM U1151, University Paris Descartes, Faculté de Médecine Necker Enfants-Malades, Paris, France.
Aleksander
A
Edelman
1 Molecular Virology and Gene Therapy.
Zeger
Z
Debyser
D000818:Animals; D001826:Body Fluids / Q000378:metabolism; D018118:Chloride Channels / Q000235:genetics; D002712:Chlorides / Q000378:metabolism; D005576:Colforsin / Q000494:pharmacology; D003550:Cystic Fibrosis / Q000235:genetics / Q000628:therapy*; D000229:Dependovirus; D004195:Disease Models, Animal; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D005838:Genotype; D006367:HeLa Cells; D006801:Humans; D007422:Intestines; D051379:Mice; D009940:Organoids / Q000378:metabolism; D014161:Transduction, Genetic
D018118:Chloride Channels; D002712:Chlorides; D005576:Colforsin
airways; gene therapy; nasal potential difference; patient-derived organoid cultures; viral vectors