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Transcriptional Response of Human Neurospheres to Helper-Dependent CAV-2 Vectors Involves the Modulation of DNA Damage Response, Microtubule and Centromere Gene Groups.

Piersanti S, Burla R, Licursi V, Brito C, La Torre M, Alves PM, Simao D, Mottini C, Salinas S, Negri R, Tagliafico E, Kremer EJ, Saggio I - PLoS ONE (2015)

Bottom Line: We found that differentiated neurospheres are readily transduced by HD-CAV-2 and that transduction generates two main transcriptional responses: a DNA damage response and alteration of centromeric and microtubule probes.Future investigations on the biochemistry of processes highlighted by probe modulations will help defining the implication of HD-CAV-2 and CAR receptor binding in enchaining these functional pathways.We suggest here that the modulation of DNA damage genes is related to viral DNA, while the alteration of centromeric and microtubule probes is possibly enchained by the interaction of the HD-CAV-2 fibre with CAR.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology and Biotechnology "C. Darwin", Sapienza University of Rome, Rome, Italy.

ABSTRACT
Brain gene transfer using viral vectors will likely become a therapeutic option for several disorders. Helper-dependent (HD) canine adenovirus type 2 vectors (CAV-2) are well suited for this goal. These vectors are poorly immunogenic, efficiently transduce neurons, are retrogradely transported to afferent structures in the brain and lead to long-term transgene expression. CAV-2 vectors are being exploited to unravel behavior, cognition, neural networks, axonal transport and therapy for orphan diseases. With the goal of better understanding and characterizing HD-CAV-2 for brain therapy, we analyzed the transcriptomic modulation induced by HD-CAV-2 in human differentiated neurospheres derived from midbrain progenitors. This 3D model system mimics several aspects of the dynamic nature of human brain. We found that differentiated neurospheres are readily transduced by HD-CAV-2 and that transduction generates two main transcriptional responses: a DNA damage response and alteration of centromeric and microtubule probes. Future investigations on the biochemistry of processes highlighted by probe modulations will help defining the implication of HD-CAV-2 and CAR receptor binding in enchaining these functional pathways. We suggest here that the modulation of DNA damage genes is related to viral DNA, while the alteration of centromeric and microtubule probes is possibly enchained by the interaction of the HD-CAV-2 fibre with CAR.

No MeSH data available.


Related in: MedlinePlus

Biological processes, cellular components and molecular functions modulated by HD-CAV-2 in human neurospheres.The bioinformatic online software g:Profiler gGOSt set as significant only, hierarchical sorting and no hierarchical filtering was used to classify the genes transcriptionally modulated upon transduction of human neurospheres at both 2 h and 5 days time points. Among the biological processes the g:Profiler analysis identified cell cycle, mitosis, DNA damage, microtubule and centromere-related processes. The centromeric and microtubule aspects were revealed also by the cellular component category, as much as by the molecular function grouping. The full list of probes modulated in the groups is detailed on S1 Table.
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pone.0133607.g003: Biological processes, cellular components and molecular functions modulated by HD-CAV-2 in human neurospheres.The bioinformatic online software g:Profiler gGOSt set as significant only, hierarchical sorting and no hierarchical filtering was used to classify the genes transcriptionally modulated upon transduction of human neurospheres at both 2 h and 5 days time points. Among the biological processes the g:Profiler analysis identified cell cycle, mitosis, DNA damage, microtubule and centromere-related processes. The centromeric and microtubule aspects were revealed also by the cellular component category, as much as by the molecular function grouping. The full list of probes modulated in the groups is detailed on S1 Table.

Mentions: To interpret the biological significance of global gene modulation, the full list of probes modulated by HD-CAV-2 was uploaded into the bioinformatic online software g:Profiler. By g:GOSt filtering we identified groups including biological processes, cellular components and molecular functions (Fig 3 and S1 Table). Among the biological processes in silico analysis identified cell cycle, mitosis, DNA damage, microtubule and centromere-related processes. The centromere and microtubule aspects were revealed also by the cellular component category, and by the molecular function grouping. To highlight single genes involved in these processes, g:GOSt was performed with greater stringency (i.e. g:GOST hierarchical filtering parameter: best per parent group, strong). Single genes and relative groups identified through this analysis are reported on Fig 4. The cell cycle and DNA damage-related processes modulated by HD-CAV-2 in neurospheres are defined by the transcriptional induction of genes including BIRC5, BRCA2 and FANCD2. On the other hand, the positive modulation of the centromeric genes CENPE, CENPH, CENPK, CENPM, CENPN, CENPW suggests that HD-CAV-2 induces the modulation of the centromeric-related processes. Interestingly, several microtubule-associated molecular motors were part of the transcriptomic picture of HD-CAV-2-neurospheres. Specifically, transduction with HD-CAV-2 induced at day 5 the modulation of the kinesins KIF14, KIF15, KIF18A and KIF23.


Transcriptional Response of Human Neurospheres to Helper-Dependent CAV-2 Vectors Involves the Modulation of DNA Damage Response, Microtubule and Centromere Gene Groups.

Piersanti S, Burla R, Licursi V, Brito C, La Torre M, Alves PM, Simao D, Mottini C, Salinas S, Negri R, Tagliafico E, Kremer EJ, Saggio I - PLoS ONE (2015)

Biological processes, cellular components and molecular functions modulated by HD-CAV-2 in human neurospheres.The bioinformatic online software g:Profiler gGOSt set as significant only, hierarchical sorting and no hierarchical filtering was used to classify the genes transcriptionally modulated upon transduction of human neurospheres at both 2 h and 5 days time points. Among the biological processes the g:Profiler analysis identified cell cycle, mitosis, DNA damage, microtubule and centromere-related processes. The centromeric and microtubule aspects were revealed also by the cellular component category, as much as by the molecular function grouping. The full list of probes modulated in the groups is detailed on S1 Table.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4514711&req=5

pone.0133607.g003: Biological processes, cellular components and molecular functions modulated by HD-CAV-2 in human neurospheres.The bioinformatic online software g:Profiler gGOSt set as significant only, hierarchical sorting and no hierarchical filtering was used to classify the genes transcriptionally modulated upon transduction of human neurospheres at both 2 h and 5 days time points. Among the biological processes the g:Profiler analysis identified cell cycle, mitosis, DNA damage, microtubule and centromere-related processes. The centromeric and microtubule aspects were revealed also by the cellular component category, as much as by the molecular function grouping. The full list of probes modulated in the groups is detailed on S1 Table.
Mentions: To interpret the biological significance of global gene modulation, the full list of probes modulated by HD-CAV-2 was uploaded into the bioinformatic online software g:Profiler. By g:GOSt filtering we identified groups including biological processes, cellular components and molecular functions (Fig 3 and S1 Table). Among the biological processes in silico analysis identified cell cycle, mitosis, DNA damage, microtubule and centromere-related processes. The centromere and microtubule aspects were revealed also by the cellular component category, and by the molecular function grouping. To highlight single genes involved in these processes, g:GOSt was performed with greater stringency (i.e. g:GOST hierarchical filtering parameter: best per parent group, strong). Single genes and relative groups identified through this analysis are reported on Fig 4. The cell cycle and DNA damage-related processes modulated by HD-CAV-2 in neurospheres are defined by the transcriptional induction of genes including BIRC5, BRCA2 and FANCD2. On the other hand, the positive modulation of the centromeric genes CENPE, CENPH, CENPK, CENPM, CENPN, CENPW suggests that HD-CAV-2 induces the modulation of the centromeric-related processes. Interestingly, several microtubule-associated molecular motors were part of the transcriptomic picture of HD-CAV-2-neurospheres. Specifically, transduction with HD-CAV-2 induced at day 5 the modulation of the kinesins KIF14, KIF15, KIF18A and KIF23.

Bottom Line: We found that differentiated neurospheres are readily transduced by HD-CAV-2 and that transduction generates two main transcriptional responses: a DNA damage response and alteration of centromeric and microtubule probes.Future investigations on the biochemistry of processes highlighted by probe modulations will help defining the implication of HD-CAV-2 and CAR receptor binding in enchaining these functional pathways.We suggest here that the modulation of DNA damage genes is related to viral DNA, while the alteration of centromeric and microtubule probes is possibly enchained by the interaction of the HD-CAV-2 fibre with CAR.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology and Biotechnology "C. Darwin", Sapienza University of Rome, Rome, Italy.

ABSTRACT
Brain gene transfer using viral vectors will likely become a therapeutic option for several disorders. Helper-dependent (HD) canine adenovirus type 2 vectors (CAV-2) are well suited for this goal. These vectors are poorly immunogenic, efficiently transduce neurons, are retrogradely transported to afferent structures in the brain and lead to long-term transgene expression. CAV-2 vectors are being exploited to unravel behavior, cognition, neural networks, axonal transport and therapy for orphan diseases. With the goal of better understanding and characterizing HD-CAV-2 for brain therapy, we analyzed the transcriptomic modulation induced by HD-CAV-2 in human differentiated neurospheres derived from midbrain progenitors. This 3D model system mimics several aspects of the dynamic nature of human brain. We found that differentiated neurospheres are readily transduced by HD-CAV-2 and that transduction generates two main transcriptional responses: a DNA damage response and alteration of centromeric and microtubule probes. Future investigations on the biochemistry of processes highlighted by probe modulations will help defining the implication of HD-CAV-2 and CAR receptor binding in enchaining these functional pathways. We suggest here that the modulation of DNA damage genes is related to viral DNA, while the alteration of centromeric and microtubule probes is possibly enchained by the interaction of the HD-CAV-2 fibre with CAR.

No MeSH data available.


Related in: MedlinePlus