Regulating the expression of therapeutic transgenes by controlled intake of dietary essential amino acids.
Widespread application of gene therapy will depend on the development of simple methods to regulate the expression of therapeutic genes. Here we harness an endogenous signaling pathway to regulate therapeutic gene expression through diet. The GCN2-eIF2α signaling pathway is specifically activated by deficiencies in any essential amino acid (EAA); EAA deficiency leads to rapid expression of genes regulated by ATF4-binding cis elements. We found that therapeutic genes under the control of optimized amino acid response elements (AAREs) had low basal expression and high induced expression. We applied our system to regulate the expression of TNFSF10 (TRAIL) in the context of glioma therapy and found that intermittent activation of this gene by EEA-deficient meals retained its therapeutic efficacy while abrogating its toxic effects on normal tissue. The GCN2-eIF2α pathway is expressed in many tissues, including the brain, and is highly specific to EAA deficiency. Our system may be particularly well suited for intermittent regulation of therapeutic transgenes over short or long time periods.
Institut National de la Recherche Agronomique (INRA), UMR-1019 Nutrition Humaine, Genes-Nutrients team, Centre de Clermont-Theix, Université Clermont-Ferrand 1, 63122 Saint Genès Champanelle, France.
Cédric
C
Chaveroux
Institut National de la Recherche Agronomique (INRA), UMR-1019 Nutrition Humaine, Genes-Nutrients team, Centre de Clermont-Theix, Université Clermont-Ferrand 1, 63122 Saint Genès Champanelle, France.
Alain
A
Bruhat
Institut National de la Recherche Agronomique (INRA), UMR-1019 Nutrition Humaine, Genes-Nutrients team, Centre de Clermont-Theix, Université Clermont-Ferrand 1, 63122 Saint Genès Champanelle, France.
Valérie
V
Carraro
Institut National de la Recherche Agronomique (INRA), UMR-1019 Nutrition Humaine, Genes-Nutrients team, Centre de Clermont-Theix, Université Clermont-Ferrand 1, 63122 Saint Genès Champanelle, France.
Céline
C
Jousse
Institut National de la Recherche Agronomique (INRA), UMR-1019 Nutrition Humaine, Genes-Nutrients team, Centre de Clermont-Theix, Université Clermont-Ferrand 1, 63122 Saint Genès Champanelle, France.
Julien
J
Averous
Institut National de la Recherche Agronomique (INRA), UMR-1019 Nutrition Humaine, Genes-Nutrients team, Centre de Clermont-Theix, Université Clermont-Ferrand 1, 63122 Saint Genès Champanelle, France.
Anne-Catherine
AC
Maurin
Institut National de la Recherche Agronomique (INRA), UMR-1019 Nutrition Humaine, Genes-Nutrients team, Centre de Clermont-Theix, Université Clermont-Ferrand 1, 63122 Saint Genès Champanelle, France.
Laurent
L
Parry
Institut National de la Recherche Agronomique (INRA), UMR-1019 Nutrition Humaine, Genes-Nutrients team, Centre de Clermont-Theix, Université Clermont-Ferrand 1, 63122 Saint Genès Champanelle, France.
Florent
F
Mesclon
Institut National de la Recherche Agronomique (INRA), UMR-1019 Nutrition Humaine, Genes-Nutrients team, Centre de Clermont-Theix, Université Clermont-Ferrand 1, 63122 Saint Genès Champanelle, France.
Yuki
Y
Muranishi
CRCT, Université de Toulouse, Inserm, UPS, Toulouse, France.
Pierre
P
Cordelier
CRCT, Université de Toulouse, Inserm, UPS, Toulouse, France.
Aline
A
Meulle
Centre de Biophysique Moléculaire, Centre National de la Recherche Scientifique (CNRS) UPR4301, Université d'Orléans and Institut National de la Santé Et de la Recherche Médicale (INSERM), Orléans, France.
Patrick
P
Baril
CNRS UMR7225, INSERM U1127, Université Pierre et Marie Curie, Institut du Cerveau et de la Moelle (ICM), Biotechnology and Biotherapy Team, Paris, France.
Anh
A
Do Thi
CNRS UMR7225, INSERM U1127, Université Pierre et Marie Curie, Institut du Cerveau et de la Moelle (ICM), Biotechnology and Biotherapy Team, Paris, France.
Philippe
0000-0002-0393-9262
P
Ravassard
CNRS UMR7225, INSERM U1127, Université Pierre et Marie Curie, Institut du Cerveau et de la Moelle (ICM), Biotechnology and Biotherapy Team, Paris, France.|Department of Neurological Surgery, University of California San Francisco, San Francisco, California, USA.
Jacques
J
Mallet
Institut National de la Recherche Agronomique (INRA), UMR-1019 Nutrition Humaine, Genes-Nutrients team, Centre de Clermont-Theix, Université Clermont-Ferrand 1, 63122 Saint Genès Champanelle, France.|Centre de Biophysique Moléculaire, Centre National de la Recherche Scientifique (CNRS) UPR4301, Université d'Orléans and Institut National de la Santé Et de la Recherche Médicale (INSERM), Orléans, France.
Pierre
P
Fafournoux
D000284:Administration, Oral; D000601:Amino Acids, Essential / Q000008:administration & dosage* / Q000493:pharmacokinetics; D000818:Animals; D001932:Brain Neoplasms / Q000235:genetics* / Q000628:therapy*; D019587:Dietary Supplements; D004305:Dose-Response Relationship, Drug; D004435:Eating / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods*; D008297:Male; D051379:Mice; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000235:genetics* / Q000627:therapeutic use*; D019076:Transgenes / Q000235:genetics; D016896:Treatment Outcome
D000601:Amino Acids, Essential; D053221:TNF-Related Apoptosis-Inducing Ligand; C505654:Tnfsf10 protein, mouse