Potent spinal parenchymal AAV9-mediated gene delivery by subpial injection in adult rats and pigs

Miyanohara, Atsushi; Kamizato, Kota; Juhás, Štefan; Juhásová, Jana; Navarro, Michael; Marsala, Silvia; Lukáčová, Nadežda; Hruška-Plocháň, Marián; Curtis, Erik; Gabel, Brandon C; Ciacci, Joseph D.; Ahrens, Eric T.; Kaspar, Brian K.; Cleveland, Don W.; Maršala, Martin

doi:10.1038/mtm.2016.46

Cited by 40 publications

(26 citation statements)

References 18 publications

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“…The volume of vector that was injected is enough to bathe the entire cord at this early stage, which may explain the efficient vector spread. This efficiency also suggests that the spinal pia membrane barrier to transduction observed in adult animal models [42] may not be such an obstacle at E16 in mice.…”

Section: Discussionmentioning

confidence: 82%

High-efficiency transduction of spinal cord motor neurons by intrauterine delivery of integration-deficient lentiviral vectors

Ahmed

Waddington

Boza-Morán

et al. 2018

Journal of Controlled Release

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Integration-deficient lentiviral vectors (IDLVs) are promising gene delivery tools that retain the high transduction efficiency of standard lentiviral vectors, yet fail to integrate as proviruses and are instead converted into episomal circles. These episomes are metabolically stable and support long-term expression of transgenes in non-dividing cells, exhibiting a decreased risk of insertional mutagenesis. We have embarked on an extensive study to compare the transduction efficiency of IDLVs pseudotyped with different envelopes (vesicular stomatitis, Rabies, Mokola and Ross River viral envelopes) and self-complementary adeno-associated viral vectors, serotype-9 (scAAV-9) in spinal cord tissues after intraspinal injection of mouse embryos (E16). Our results indicate that IDLVs can transduce motor neurons (MNs) at extremely high efficiency regardless of the envelope pseudotype while scAAV9 mediates gene delivery to ~ 40% of spinal cord motor neurons, with other non-neuronal cells also transduced. Long-term expression studies revealed stable gene expression at 7 months post-injection. Taken together, the results of this study indicate that IDLVs may be efficient tools for in utero cord transduction in therapeutic strategies such as for treatment of inherited early childhood neurodegenerative diseases.

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Section: Discussionmentioning

confidence: 82%

High-efficiency transduction of spinal cord motor neurons by intrauterine delivery of integration-deficient lentiviral vectors

Ahmed

Waddington

Boza-Morán

et al. 2018

Journal of Controlled Release

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“…In our previous studies, we have described the technique of subpial AAV9 vector delivery and identified the pia mater as the major barrier which limits the penetration of the AAV9 vector into the deep spinal parenchyma in mice, rats and pigs. 12,13 Our original technique used subpial placement of a PE-5 or PE-10 catheter after the pia was punctured using a manually positioned 30G pia-penetrating needle. Our current technique was slightly modified with two XYZ manipulators mounted firmly on a spinal immobilization frame.…”

Section: Technical Requirements To Perform Effective and Safe Subpimentioning

confidence: 99%

“…Recently, we have described a novel subpial AAV9 delivery technique and demonstrated robust multisegmental transgene expression in adult pigs, rats, and mice . We have also demonstrated that the pia mater represents the primary barrier that limits vector penetration into the deep spinal parenchyma.…”

Section: Introductionmentioning

confidence: 97%

Spinal parenchymal occupation by neural stem cells after subpial delivery in adult immunodeficient rats

Maršala

Kamizato

Tadokoro

et al. 2019

Stem Cells Translational Medicine

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Neural precursor cells (NSCs) hold great potential to treat a variety of neurodegenerative diseases and injuries to the spinal cord. However, current delivery techniques require an invasive approach in which an injection needle is advanced into the spinal parenchyma to deliver cells of interest. As such, this approach is associated with an inherent risk of spinal injury, as well as a limited delivery of cells into multiple spinal segments.Here, we characterize the use of a novel cell delivery technique that employs single bolus cell injections into the spinal subpial space. In immunodeficient rats, two subpial injections of human NSCs were performed in the cervical and lumbar spinal cord, respectively. The survival, distribution, and phenotype of transplanted cells were assessed 6-8 months after injection. Immunofluorescence staining and mRNA sequencing analysis demonstrated a near-complete occupation of the spinal cord by injected cells, in which transplanted human NSCs (hNSCs) preferentially acquired glial phenotypes, expressing oligodendrocyte (Olig2, APC) or astrocyte (GFAP) markers. In the outermost layer of the spinal cord, injected hNSCs differentiated into glia limitans-forming astrocytes and expressed human-specific superoxide dismutase and laminin. All animals showed normal neurological function for the duration of the analysis. These data show that the subpial cell delivery technique is highly effective in populating the entire spinal cord with injected NSCs, and has a potential for clinical use in cell replacement therapies for the treatment of ALS, multiple sclerosis, or spinal cord injury. K E Y W O R D S glia limitans formation from grafted neural precursors, human-specific mRNA sequencing, immunodeficient rat, neuraxial neural precursor migration, subpial stem cell injection 1 | BACKGROUND The use of spinally targeted cell-replacement therapies for the treatment of a variety of neurodegenerative disorders, including amyotrophic lateral sclerosis (ALS) and spinal cord injury (SCI), has recently reached the clinical trial stage with several phase I or phase II trials underway.Extensive preclinical studies in rodent and large animal models of ALS and SCI have tested several spinal cell delivery techniques to define the most effective and safe cell delivery approach to be used in human clinical trials.

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“…A third option for gene delivery to the CNS is to inject vectors into the cerebral spinal fluid (CSF). Several access points can be used: the lateral ventricle (intracerebroventricular, ICV), the cisterna magna (CM), subpial (Miyanohara et al, 2016) or the intrathecal (IT) space along the spinal cord. When performed in neonates, ICV AAV administration can provide widespread gene delivery.…”

Section: Csf Deliverymentioning

confidence: 99%

Adeno-Associated Virus Technologies and Methods for Targeted Neuronal Manipulation

Haery

Deverman

Matho

et al. 2019

Preprint

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Cell-type-specific expression of molecular tools and sensors is critical to construct circuit diagrams and to investigate the activity and function of neurons within the nervous system. Strategies for targeted manipulation include combinations of classical genetic tools such as Cre/loxP and Flp/FRT, use of cis-regulatory elements, targeted knock-in transgenic mice, and gene delivery by AAV and other viral vectors. The combination of these complex technologies with the goal of precise neuronal targeting is a challenge in the lab. This report will discuss the theoretical and practical aspects of combining current technologies and establish best practices for achieving targeted manipulation of specific cell types. Novel applications and tools, as well as areas for development, will be envisioned and discussed.Selecting the Route of Administration and Capsid AAV tropism, as dictated by AAV capsid proteins, is an important factor affecting transduction efficiency and specificity across cell types. Since the mechanism of AAV transduction is through interaction of the AAV capsid with cell surface proteins and glycans, protein composition of the capsid (i.e., the AAV serotype) and the cell surface (i.e., based on cell type) determines transduction efficiency. Consequently, serotype and route of delivery should be carefully considered when designing experiments (Fig 1A, B). For an overview of the primary receptors for AAV serotypes, see (Schultz and Chamberlain, 2008). Direct intraparenchymal deliveryWhen injected directly into the brain, many of the naturally-occurring AAV capsids, which share homology ranging from 65-99% (Drouin and Agbandje-McKenna, 2013), have distinct but significantly overlapping tropisms and distribution characteristics. AAV1, AAV2, AAV5, AAV8,

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Potent spinal parenchymal AAV9-mediated gene delivery by subpial injection in adult rats and pigs

Cited by 40 publications

References 18 publications

High-efficiency transduction of spinal cord motor neurons by intrauterine delivery of integration-deficient lentiviral vectors

High-efficiency transduction of spinal cord motor neurons by intrauterine delivery of integration-deficient lentiviral vectors

Spinal parenchymal occupation by neural stem cells after subpial delivery in adult immunodeficient rats

Adeno-Associated Virus Technologies and Methods for Targeted Neuronal Manipulation

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