Differentiated neuroprogenitor cells incubated with human or canine adenovirus, or lentiviral vectors have distinct transcriptome profiles.
Several studies have demonstrated the potential for vector-mediated gene transfer to the brain. Helper-dependent (HD) human (HAd) and canine (CAV-2) adenovirus, and VSV-G-pseudotyped self-inactivating HIV-1 vectors (LV) effectively transduce human brain cells and their toxicity has been partly analysed. However, their effect on the brain homeostasis is far from fully defined, especially because of the complexity of the central nervous system (CNS). With the goal of dissecting the toxicogenomic signatures of the three vectors for human neurons, we transduced a bona fide human neuronal system with HD-HAd, HD-CAV-2 and LV. We analysed the transcriptional response of more than 47,000 transcripts using gene chips. Chip data showed that HD-CAV-2 and LV vectors activated the innate arm of the immune response, including Toll-like receptors and hyaluronan circuits. LV vector also induced an IFN response. Moreover, HD-CAV-2 and LV vectors affected DNA damage pathways--but in opposite directions--suggesting a differential response of the p53 and ATM pathways to the vector genomes. As a general response to the vectors, human neurons activated pro-survival genes and neuron morphogenesis, presumably with the goal of re-establishing homeostasis. These data are complementary to in vivo studies on brain vector toxicity and allow a better understanding of the impact of viral vectors on human neurons, and mechanistic approaches to improve the therapeutic impact of brain-directed gene transfer.
Dipartimento di Biologia e Biotecnologie Chrales Darwin, Sapienza, Università di Roma, Roma, Italy.
Stefania
S
Piersanti
Letizia
L
Astrologo
Valerio
V
Licursi
Rossella
R
Costa
Enrica
E
Roncaglia
Aurelie
A
Gennetier
Sandy
S
Ibanes
Miguel
M
Chillon
Rodolfo
R
Negri
Enrico
E
Tagliafico
Eric J
EJ
Kremer
Isabella
I
Saggio
D006516:Adenoviruses, Canine / Q000502:physiology*; D000260:Adenoviruses, Human / Q000502:physiology*; D000818:Animals; D064007:Ataxia Telangiectasia Mutated Proteins / Q000378:metabolism; D002453:Cell Cycle / Q000235:genetics; D002454:Cell Differentiation / Q000235:genetics*; D004249:DNA Damage / Q000235:genetics; D004285:Dogs; D015536:Down-Regulation / Q000235:genetics; D004705:Endocytosis / Q000235:genetics; D020869:Gene Expression Profiling; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D007109:Immunity / Q000235:genetics; D007372:Interferons / Q000235:genetics / Q000378:metabolism; D016086:Lentivirus; D008636:Mesencephalon / Q000166:cytology; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000821:virology; D009474:Neurons / Q000166:cytology / Q000821:virology; D015398:Signal Transduction / Q000235:genetics; D051193:Toll-Like Receptors / Q000235:genetics / Q000378:metabolism; D015533:Transcriptional Activation; D059467:Transcriptome / Q000235:genetics*; D014161:Transduction, Genetic; D051153:Wnt Proteins / Q000235:genetics / Q000378:metabolism
D051193:Toll-Like Receptors; D051153:Wnt Proteins; D007372:Interferons; D064007:Ataxia Telangiectasia Mutated Proteins