AAV8 Can Induce Innate and Adaptive Immune Response in the Primate Eye

Reichel, Felix Friedrich; Dauletbekov, Daniyar; Klein, Reinhild; Peters, Tobias; Ochakovski, G. Alex; Seitz, Immanuel P.; Wilhelm, Barbara; Ueffing, Marius; Biel, Martin; Wissinger, Bernd; Michalakis, Stylianos; Bartz‐Schmidt, Karl Ulrich; Fischer, Manuel

doi:10.1016/j.ymthe.2017.08.018

Cited by 106 publications

(156 citation statements)

References 25 publications

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“…Surprisingly, the AAV8-GFP construct also demonstrated some non-negligible effects on disease phenotypes of SMA mice (Fig 3). While we cannot be certain as to why that is, we speculate that it may be related to possible effects on the immune system, similarly to previous reports for AAV8 vectors 40,41 . Seeing as SMA mice have an altered immune response 42,43 and that inflammation can display both protective and adverse systemic properties 44 , including the CNS 45 and muscle 46 , it is possible that an activated immune response results in acute and/or intermittent benefits in AAV8-treated SMA animals.…”

Section: Discussionsupporting

confidence: 82%

Muscle overexpression of Klf15 via an AAV8-Spc5-12 construct does not provide benefits in spinal muscular atrophy mice

Ahlskog

Hayler

Krüger

et al. 2019

Preprint

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ABSTRACTSpinal muscular atrophy (SMA) is a neuromuscular disease caused by loss of the survival motor neuron (SMN) gene. While there are currently two approved gene-based therapies for SMA, availability, high cost, and differences in patient response indicate that alternative treatment options are needed. Optimal therapeutic strategies will likely be a combination of SMN-dependent and -independent treatments aimed at alleviating symptoms in the central nervous system and peripheral muscles. Krüppel-like factor 15 (KLF15) is a transcription factor that regulates key metabolic and ergogenic pathways in muscle. We have recently reported significant downregulation of Klf15 in muscle of pre-symptomatic SMA mice. Importantly, perinatal upregulation of Klf15 via transgenic and pharmacological methods resulted in improved disease phenotypes in SMA mice, including weight and survival. In the current study, we designed an adeno-associated virus serotype 8 (AAV8) vector to overexpress a codon-optimised Klf15 cDNA under the muscle-specific Spc5-12 promoter (AAV8-Klf15). Administration of AAV8-Klf15 to severe Taiwanese Smn−/−;SMN2 or intermediate Smn2B/− SMA mice significantly increased Klf15 expression in muscle. We also observed significant activity of the AAV8-Klf15 vector in liver and heart. AAV8-mediated Klf15 overexpression moderately improved survival in the Smn2B/− model but not in the Taiwanese mice. An inability to specifically induce Klf15 expression at physiological levels in a time- and tissue-dependent manner may have contributed to this limited efficacy. Thus, our work demonstrates that an AAV8-Spc5-12 vector induces high gene expression as early as P2 in several tissues including muscle, heart and liver, but highlights the challenges of achieving meaningful vector-mediated transgene expression of Klf15.

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Section: Discussionsupporting

confidence: 82%

Muscle overexpression of Klf15 via an AAV8-Spc5-12 construct does not provide benefits in spinal muscular atrophy mice

Ahlskog

Hayler

Krüger

et al. 2019

Preprint

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“…In non‐human primates, subretinal injection of AAV‐8 gene vectors triggers both the innate and the adaptive immune response (Reichel et al. ). In humans, cytotoxic T‐cells are found after subretinal injection of the same AAV‐gene vector when immunosuppression is stopped (Reichel et al.…”

Section: Discussionmentioning

confidence: 99%

“…In humans, cytotoxic T‐cells are found after subretinal injection of the same AAV‐gene vector when immunosuppression is stopped (Reichel et al. ). These findings emphasize that the idea of the eye as an immune‐privileged organ favoring ocular gene therapy needs to be reconsidered.…”

Section: Discussionmentioning

confidence: 99%

Subretinal surgery: functional and histological consequences of entry into the subretinal space

Sørensen

2019

Acta Ophthalmologica

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Background and objectives: Gene-therapy, stem-cell transplantation and surgical robots hold the potential for treatment of currently untreatable retinal degenerative diseases. All of the techniques require entry into the subretinal space, which is a potential space located between the retina and the retinal pigment epithelium (RPE). Knowledge about obstacles and critical steps in relation to subretinal procedures is therefore needed. This thesis explores the functional and histological consequences of separation of the retina from the RPE, extensive RPE damage, a large cut in the retina (retinotomy) and RPE phagocytosis in a porcine model. Methods: Experiments were performed in 106 female domestic pigs of Danish landrace distributed over five studies. Under general anesthesia, different procedures for expansion of the subretinal space were conducted. Outcomes were visual function measured electrophysiologically with multifocal electroretinogram (mfERG) and retinal morphology examined histologically. Study I: The effect of anesthesia on mfERG was examined by repeated recordings for 3 hr in isoflurane or propofol anesthesia. Outcome was mfERG amplitude. Study II: Consequences of a large separation of the photoreceptors from the RPE were examined by injecting a perfluorocarbon-liquid (decalin) into the subretinal space. Two weeks after, in a second surgery, decalin was withdrawn. Outcomes were mfERG and histology 4 weeks after decalin injection. Study III: Extensive RPE damage was examined by expanding the subretinal space with saline and removing large sheets of RPE-cells through a retinotomy. Outcomes were mfERG and histology 2, 4 and 6 weeks after the procedure. Study IV: Consequences of a large retinotomy were examined by similar procedures as in Study III, but in study IV only a few RPE cells were removed. Outcomes were mfERG and histology 2 and 6 weeks after surgery. Study V: Clearance of the subretinal space was examined by injecting fluorescent latex beads of various sizes into the subretinal space. Outcome was histologic location of the beads at different time intervals after the procedure. Results: Study I: MfERG amplitudes decreased linearly as a function of time in propofol or isoflurane anesthesia. Duration of mfERG recording could be decreased without compromising quality, and thereby could time in anesthesia be reduced. Study II: MfERG and histology remained normal after reattachment of a large and 2-week long separation of the photoreceptors and RPE. Repeated entry into the subretinal space was well tolerated. Fluid injection into the subretinal space constitutes a risk of RPE-damage. Study III: Removal of large sheets of retinal pigment epithelial cells triggered a widespread rhegmatogenous-like retinal detachment resulting in visual loss. Study IV: A large retinotomy with limited damage of the RPE was well tolerated, and visual function was preserved. Study V: Subretinal latex beads up to 4 lm were phagocytosed by the RPE and passed into the sub-RPE space. Beads up to 2 lm travelled further through...

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“…The primate-derived variant NHP#26 transduced the outer retina at greatly reduced titer. Increasingly, it is understood that immune response to high doses of AAV vectors represents a significant factor influencing the success of retinal and other gene therapies( 21, 22 ). This vector may enable safer gene therapies for retinal degeneration in patients as a result of the decreased vector load required for transgene expression and may reduce risk of vector-related toxicity or immunogenicity.…”

Section: Discussionmentioning

confidence: 99%

In vivo directed evolution of AAV in the primate retina

Visel

et al. 2019

Preprint

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AbstractEfficient AAV-mediated gene delivery remains a significant obstacle to effective retinal gene therapies. Here, we apply directed evolution – guided by deep sequencing and followed by direct in vivo secondary selection of high-performing vectors with a GFP-barcoded library – to create AAV viral capsids with new capabilities to deliver genes to the outer retina in primates. A replication incompetent library, produced via providing rep in trans, was created to mitigate risk of AAV propagation. Six rounds of in vivo selection with this library in primates – involving intravitreal library administration, recovery of genomes from outer retina, and extensive next generation sequencing of each round – resulted in vectors with redirected tropism to the outer retina and increased gene delivery efficiency to retinal cells. These new viral vectors expand the toolbox of vectors available for primate retina, and may enable less invasive delivery of therapeutic genes to patients, potentially offering retina-wide infection at a similar dosage to vectors currently in clinical use.

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AAV8 Can Induce Innate and Adaptive Immune Response in the Primate Eye

Cited by 106 publications

References 25 publications

Muscle overexpression of Klf15 via an AAV8-Spc5-12 construct does not provide benefits in spinal muscular atrophy mice

Muscle overexpression of Klf15 via an AAV8-Spc5-12 construct does not provide benefits in spinal muscular atrophy mice

Subretinal surgery: functional and histological consequences of entry into the subretinal space

In vivo directed evolution of AAV in the primate retina

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