Protective effects of intrathecal injection of AAV9-RabGGTB-GFP+ in SOD1G93A mice

Gao, Tianchu; Xin, Cheng; Yang, Jianqi; Liu, Qi; Dong, Hui; Li, Rui; Liu, Yaling

doi:10.3389/fnagi.2023.1092607

Cited by 6 publications

(3 citation statements)

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“…The same result was obtained by delivering OPTN to SOD1 ‐G93A‐transfected cell lines through LV infection, as OPTN expression was observed in the cytoplasm of cells. These findings were consistent with the findings of Gao et al ., [30] who delivered AAV9‐ RabGGTB to mice by intrathecal injection and observed highly efficient gene expression in MNs. Rommereim et al .…”

Section: Discussionmentioning

confidence: 99%

OPTN gene therapy increases autophagy and protects mitochondria in SOD1‐G93A‐expressing transgenic mice and cells

Wen,

Ji,

et al. 2023

The FEBS Journal

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Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder characterized by progressive motor neuron (MN) death. Mutation of the superoxide dismutase 1 (SOD1) gene, which results in abnormal protein aggregation, is one of the causes of familial ALS. Autophagic dysfunction occurs in SOD1‐G93A mutant mice as the disease progresses, but the etiology of this disease is still unclear. Optineurin (OPTN) is an adaptor that is involved in autophagy and participates in aggrephagy and mitophagy. Previous studies have established that OPTN mutations contribute to diseases such as glaucoma and ALS. However, the function of OPTN in autophagy and mitophagy has not been intensively investigated in models of ALS. In this study, we assessed the beneficial effect of OPTN on autophagy and mitochondrial function by intrathecally injecting adeno‐associated virus 9 (AAV9)‐OPTN into SOD1‐G93A transgenic mice and by administering lentivirus (LV)‐OPTN to cells expressing the SOD1‐G93A mutant protein. The expression of voltage‐dependent anion channel 1 (VDAC1) was increased and autophagy was elevated after OPTN gene therapy, as shown by a lower level of p62 and a higher level of microtubule‐associated protein 1A/1B‐light chain 3 (LC3)‐II. Moreover, using electron microscopy, we observed a hyperpolarized mitochondrial transmembrane potential and reversal of mitochondrial morphological abnormalities. Furthermore, the protein level of TANK‐binding kinase 1 (TBK1) was increased, suggesting that mitophagy was increased. Our findings from both animal and cell line studies strongly suggest that OPTN gene therapy is a powerful strategy to increase autophagy and protect mitochondria to prevent the progression of ALS and could be effective in the treatment of ALS.

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Section: Discussionmentioning

confidence: 99%

OPTN gene therapy increases autophagy and protects mitochondria in SOD1‐G93A‐expressing transgenic mice and cells

Wen,

Ji,

et al. 2023

The FEBS Journal

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“…Autophagy obstacles have been confirmed to be one of the early pathological events of ALS [51,52], and autophagy plays an important role in inflammation [53]. In our previous studies, we found that the expression of RabGGTase (low RABGGTB expression and no change in RABGGTA) is lower in the lumbar and thoracic regions of spinal cord motoneurons in SOD1G93A mice compared with WT (wild-type) mice groups, and autophagy defects could be ameliorated by modulating RABGGTB in neurons [54]. However, it was difficult to obtain neurons in patients with ALS.…”

Section: Introductionmentioning

confidence: 88%

Rab Geranylgeranyltransferase Subunit Beta as a Potential Indicator to Assess the Progression of Amyotrophic Lateral Sclerosis

Yang,

Xin,

Huo

et al. 2023

Brain Sciences

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Background: Currently, there is no effective treatment for amyotrophic lateral sclerosis (ALS), a devastating neurodegenerative disorder. Many biomarkers have been proposed, but because ALS is a clinically heterogeneous disease with an unclear etiology, biomarker discovery for ALS has been challenging due to the lack of specificity of these biomarkers. In recent years, the role of autophagy in the development and treatment of ALS has become a research hotspot. In our previous studies, we found that the expression of RabGGTase (low RABGGTB expression and no change in RABGGTA) is lower in the lumbar and thoracic regions of spinal cord motoneurons in SOD1G93A mice compared with WT (wild-type) mice groups, and upregulation of RABGGTB promoted prenylation modification of Rab7, which promoted autophagy to protect neurons by degrading SOD1. Given that RabGGTase is associated with autophagy and autophagy is associated with inflammation, and based on the above findings, since peripheral blood mononuclear cells are readily available from patients with ALS, we proposed to investigate the expression of RabGGTase in peripheral inflammatory cells. Methods: Information and venous blood were collected from 86 patients diagnosed with ALS between January 2021 and August 2023. Flow cytometry was used to detect the expression of RABGGTB in monocytes from peripheral blood samples collected from patients with ALS and healthy controls. Extracted peripheral blood mononuclear cells (PBMCs) were differentiated in vitro into macrophages, and then the expression of RABGGTB was detected by immunofluorescence. RABGGTB levels in patients with ALS were analyzed to determine their impact on disease progression. Results: Using flow cytometry in monocytes and immunofluorescence in macrophages, we found that RABGGTB expression in the ALS group was significantly higher than in the control group. Age, sex, original location, disease course, C-reactive protein (CRP), and interleukin-6 (IL-6) did not correlate with the ALS functional rating scale—revised (ALSFRS-R), whereas the RABGGTB level was significantly correlated with the ALSFRS-R. In addition, multivariate analysis revealed a significant correlation between RABGGTB and ALSFRS-R score. Further analysis revealed a significant correlation between RABGGTB expression levels and disease progression levels (ΔFS). Conclusions: The RABGGTB level was significantly increased in patients with ALS compared with healthy controls. An elevated RABGGTB level in patients with ALS is associated with the rate of progression in ALS, suggesting that elevated RABGGTB levels in patients with ALS may serve as an indicator for tracking ALS progression.

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“…Overall, intrathecal injection has been employed to treat mouse models (Bailey et al., 2018; Calias et al., 2012; Gao et al., 2023; Georgiou et al., 2023; Habisch et al., 2007; Hinderer, Bell, Gurda, et al., 2014; Kagiava, Karaiskos, et al., 2021; Kagiava et al., 2018; Kagiava et al., 2019; Kagiava et al., 2016; Lee et al., 2019; Meyer et al., 2015; Miyake et al., 2021; Nagano et al., 2005; Passini et al., 2014; Schiza et al., 2019; Shyng et al., 2017; Stavrou et al., 2022), cats (Auclair et al., 2012; Farid et al., 2022; Genoni et al., 2016; Giroux et al., 2001; Hinderer, Bell, Gurda, et al., 2014), dogs (Bradbury et al., 2020; Genoni et al., 2016; Hinderer et al., 2018), pigs (Duque et al., 2015; Federici et al., 2012; Miyanohara et al., 2016; Pleticha et al., 2013) and humans (Cordts et al., 2020; Hache et al., 2016; Naveed & Calderon, 2021) (clinical trials NCT03381729 and NCT02362438) with neurological diseases. Lumbar intrathecal injection has also been tested in non‐human primates resulting in very encouraging biodistribution data (Beharry et al., 2022; Bey et al., 2020; Hinderer, Bell, Vite, et al., 2014; Hordeaux et al., 2019; Meseck et al., 2022; Meyer et al., 2015; Ohno et al., 2019).…”

Section: Introductionmentioning

confidence: 99%

Lumbar Intrathecal Injection in Adult and Neonatal Mice

Stavrou,

Georgiou,

Kleopa

2024

Current Protocols

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This article describes a step‐by‐step process of lumbar intrathecal injection of Evans blue dye and AAV9‐EGFP in adult (2‐month‐old) and neonatal (postnatal day 10) mice. Intrathecal injection is a clinically translatable technique that has already been extensively applied in humans. In mice, intrathecal injection is considered a challenging procedure that requires a trained and experienced researcher. For both adult and neonatal mice, lumbar intrathecal injection is directed into the L5‐L6 intervertebral space. Intrathecally injected material enters the cerebrospinal fluid (CSF) within the intrathecal space from where it can directly access the central nervous system (CNS) parenchyma. Simultaneously, intrathecally injected material exits the CSF with pressure gradient and enters the endoneurial fluid and ultimately the peripheral nerves. While in the CSF, the injectable material also enters the bloodstream and systemic circulation through the arachnoid villi. A successful lumbar intrathecal injection results in adequate biodistribution of the injectable material in the CNS, PNS, and peripheral organs. When correctly applied, this technique is considered as minimally invasive and non‐disruptive and can be used for the lumbar delivery of any solute. © 2024 Wiley Periodicals LLC.Basic Protocol 1: C57BL/6 adult and P10 mice lumbar intrathecal injectionBasic Protocol 2: Tissue collection and preparation for evaluating Evans blue dye diffusionBasic Protocol 3: Tissue collection and preparation for immunohistochemistry stainingBasic Protocol 4: Tissue collection and vector genome copy number analysis

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Protective effects of intrathecal injection of AAV9-RabGGTB-GFP+ in SOD1G93A mice

Cited by 6 publications

References 50 publications

OPTN gene therapy increases autophagy and protects mitochondria in SOD1‐G93A‐expressing transgenic mice and cells

OPTN gene therapy increases autophagy and protects mitochondria in SOD1‐G93A‐expressing transgenic mice and cells

Rab Geranylgeranyltransferase Subunit Beta as a Potential Indicator to Assess the Progression of Amyotrophic Lateral Sclerosis

Lumbar Intrathecal Injection in Adult and Neonatal Mice

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