An Adeno-Associated Virus-Based Toolkit for Preferential Targeting and Manipulating Quiescent Neural Stem Cells in the Adult Hippocampus

Crowther, Andrew; Lim, Szu Aun; Asrican, Brent; Albright, Blake H.; Wooten, Josh; Yeh, Chia-Yu; Bao, Hechen; Cerri, Domenic H.; Hu, Junmei; Shih, Yen‐Yu Ian; Asokan, Aravind; Song, Juan

doi:10.1016/j.stemcr.2018.01.018

Cited by 12 publications

(13 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Interestingly, the total number of Sox2+ cells, composed of Type 1 and 2a NPCs, was relatively unaffected by rAAV, consistent with this population being largely quiescent (Figure 2 B,C). The fact that Sox2+ cells were mostly preserved in our experiments might explain why Type I cells are visualized in studies utilizing a large range of AAV titers, including titers greater than those reported here (Crowther et al, 2018;Kotterman et al, 2015;Ojala et al, 2018;Pulicherla et al, 2011;Song et al, 2012).…”

Section: A Developmental Window For Sensitivity To Raav-induced Toxicitymentioning

confidence: 71%

AAV ablates neurogenesis in the adult murine hippocampus

Johnston

Parylak

Kim

et al. 2021

eLife

View full text Add to dashboard Cite

Recombinant adeno-associated virus (rAAV) has been widely used as a viral vector across mammalian biology and has been shown to be safe and effective in human gene therapy. We demonstrate that neural progenitor cells (NPCs) and immature dentate granule cells (DGCs) within the adult murine hippocampus are particularly sensitive to rAAV-induced cell death. Cell loss is dose dependent and nearly complete at experimentally relevant viral titers. rAAV-induced cell death is rapid and persistent, with loss of BrdU-labeled cells within 18 hours post-injection and no evidence of recovery of adult neurogenesis at 3 months post-injection. The remaining mature DGCs appear hyperactive 4 weeks post-injection based on immediate early gene expression, consistent with previous studies investigating the effects of attenuating adult neurogenesis. In vitro application of AAV or electroporation of AAV2 inverted terminal repeats (ITRs) is sufficient to induce cell death. Efficient transduction of the dentate gyrus (DG)-without ablating adult neurogenesis-can be achieved by injection of rAAV2-retro serotyped virus into CA3. rAAV2-retro results in efficient retrograde labeling of mature DGCs and permits in vivo 2-photon calcium imaging of dentate activity while leaving adult neurogenesis intact. These findings expand on recent reports implicating rAAV-linked toxicity in stem cells and other cell types and suggest that future work using rAAV as an experimental tool in the DG and as a gene therapy for diseases of the central nervous system (CNS) should be carefully evaluated.

show abstract

Section: A Developmental Window For Sensitivity To Raav-induced Toxicitymentioning

confidence: 71%

AAV ablates neurogenesis in the adult murine hippocampus

Johnston

Parylak

Kim

et al. 2021

eLife

View full text Add to dashboard Cite

show abstract

“…An additional alternative method is to deliver transgenes plus cleavably-linked fluorescent reporters using viral vectors that target expression to specific cell populations based on viral serotype or cell-specific promoters. For example, certain adeno-associated viruses show preference for infecting specific NSPC subclasses and can be used to manipulate gene function in adulthood after stereotaxic delivery (Crowther et al, 2018).…”

Section: Discussionmentioning

confidence: 99%

Poor Concordance of Floxed Sequence Recombination in Single Neural Stem Cells: Implications for Cell Autonomous Studies

Dause

Kirby

2020

eNeuro

View full text Add to dashboard Cite

To manipulate target gene function in specific adult cell populations, tamoxifen (TAM)-dependent CreER T2 is widely used to drive inducible, site-specific recombination of loxP flanked sequences. In studies of cell autonomous target gene function, it is common practice to combine these CreER T2-lox systems with a ubiquitously expressed stop-floxed fluorescent reporter gene to identify single cells supposedly undergoing target gene recombination. Here, we studied the reliability of using Cre-induced recombination of one gene to predict recombination in another gene at the single-cell level in adult hippocampal neural stem and progenitor cells (NSPCs). Using both probabilistic predictions in a generic experimental paradigm, as well as a mouse model with two separate stop-floxed reporters plus a Nestin promoter-driven CreER T2 , we found that, in individual cells, recombination of one gene was a poor predictor of Significance Statement We investigate the reliability of a widely used transgenic mouse model in studies of adult neural stem and progenitor cells (NSPCs). Ligand-dependent Cre recombinases, such as the CreER T2 model, are a fundamental tool for inducible gene modification used to investigate gene function in many cell populations. It is common practice to combine NSPC-specific CreER T2-lox systems with a ubiquitously expressed stopfloxed fluorescent reporter gene to identify single cells undergoing target gene recombination in studies of cell autonomous gene function. Our probabilistic predictions and experimental data suggest that use of stop-floxed reporters to investigate cell autonomous gene function in NSPCs may lead to false conclusions because recombination in separate genes can show poor concordance in individual cells.

show abstract

“…2 B,C). The fact that Sox2+ cells were mostly preserved in our experiments might explain why Type I cells are visualized in studies utilizing a large range of AAV titers, including titers greater than those reported here (Crowther et al, 2018;Kotterman et al, 2015;Ojala et al, 2018;Pulicherla et al, 2011;Song et al, 2012).…”

Section: A Developmental Window For Sensitivity To Raav-induced Toxicitymentioning

confidence: 59%

AAV Ablates Neurogenesis in the Adult Murine Hippocampus

Johnston

Parylak

Kim

et al. 2020

Preprint

View full text Add to dashboard Cite

Recombinant adeno-associated virus (rAAV) has been widely used as a viral vector across mammalian biology and has been shown to be safe and effective in human gene therapy. We demonstrate that neural progenitor cells (NPCs) and immature dentate granule cells (DGCs) within the adult murine hippocampus are particularly sensitive to rAAV-induced cell death. Cell loss is dose dependent and nearly complete at experimentally relevant viral titers. rAAV-induced cell death is rapid and persistent, with loss of BrdU-labeled cells within 18 hours post-injection and no evidence of recovery of adult neurogenesis at 3 months post-injection. The remaining mature DGCs appear hyperactive 4 weeks post-injection based on immediate early gene expression, consistent with previous studies investigating the effects of attenuating adult neurogenesis. In vitro application of AAV or electroporation of AAV2 inverted terminal repeats (ITRs) is sufficient to induce cell death. Efficient transduction of the dentate gyrus (DG)-without ablating adult neurogenesis-can be achieved by injection of rAAV2-retro serotyped virus into CA3. rAAV2-retro results in efficient retrograde labeling of mature DGCs and permits in vivo 2photon calcium imaging of dentate activity while leaving adult neurogenesis intact. These findings expand on recent reports implicating rAAV-linked toxicity in stem cells and other cell types and suggest that future work using rAAV as an experimental tool in the DG and as a gene therapy for diseases of the central nervous system (CNS) should be carefully evaluated. et al., 2015). These stem cells and their immature progeny are sensitive to environmental stimuli; their proliferation, development, and survival are regulated by multiple intrinsic and extrinsic factors, including experience, stress, and inflammation (Gonçalves et al., 2016a;Kempermann et al., 2015;Monje et al., 2003;Snyder et al., 2009;Vivar et al.). Numerous studies demonstrate that abDGCs are critical for maintaining the physiological activity of mature DGCs and contribute to hippocampus-dependent behaviors (Akers et al. 17

show abstract

An Adeno-Associated Virus-Based Toolkit for Preferential Targeting and Manipulating Quiescent Neural Stem Cells in the Adult Hippocampus

Cited by 12 publications

References 31 publications

AAV ablates neurogenesis in the adult murine hippocampus

AAV ablates neurogenesis in the adult murine hippocampus

Poor Concordance of Floxed Sequence Recombination in Single Neural Stem Cells: Implications for Cell Autonomous Studies

AAV Ablates Neurogenesis in the Adult Murine Hippocampus

Contact Info

Product

Resources

About