Harnessing autophagy for cell fate control gene therapy. Autophagy 2013 Jul;9(7):1069-79
Date
05/02/2013Pubmed ID
23633667Pubmed Central ID
PMC3722316DOI
10.4161/auto.24639Scopus ID
2-s2.0-84880885997 (requires institutional sign-in at Scopus site) 7 CitationsAbstract
We hypothesized that rapamycin, through induction of autophagy and promotion of an antiapoptotic phenotype, would permit lentiviral (LV)-based transgene delivery to human T-Rapa cells, which are being tested in phase II clinical trials in the setting of allogeneic hematopoietic cell transplantation. Manufactured T-Rapa cells were exposed to supernatant enriched for a LV vector encoding a fusion protein consisting of truncated CD19 (for cell surface marking) and DTYMK/TMPKΔ, which provides "cell-fate control" due to its ability to phosphorylate (activate) AZT prodrug. LV-transduction in rapamycin-treated T-Rapa cells: (1) resulted in mitochondrial autophagy and a resultant antiapoptotic phenotype, which was reversed by the autophagy inhibitor 3-MA; (2) yielded changes in MAP1LC3B and SQSTM1 expression, which were reversed by 3-MA; and (3) increased T-Rapa cell expression of the CD19-DTYMKΔ fusion protein, despite their reduced proliferative status. Importantly, although the transgene-expressing T-Rapa cells expressed an antiapoptotic phenotype, they were highly susceptible to cell death via AZT exposure both in vitro and in vivo (in a human-into-mouse xenogeneic transplantation model). Therefore, rapamycin induction of T cell autophagy can be used for gene therapy applications, including the CD19-DTYMKΔ cell-fate control axis to improve the safety of T cell immuno-gene therapy.
Author List
Felizardo TC, Foley J, Steed K, Dropulic B, Amarnath S, Medin JA, Fowler DHAuthor
Jeffrey A. Medin PhD Professor in the Pediatrics department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AdenineAnimals
Antigens, CD19
Apoptosis
Autophagy
CD4-Positive T-Lymphocytes
Cell Lineage
Cell Proliferation
Cell Survival
Genetic Therapy
Humans
Lentivirus
Mice
Nucleoside-Phosphate Kinase
Phenotype
Sirolimus
Transduction, Genetic
Transgenes
Zidovudine
