Transduction of Systemically Administered Adeno-Associated Virus in the Colonic Enteric Nervous System and c-Kit Cells of Adult Mice

Wang, Lixin; Yuan, Pu‐Qing; Challis, Collin; Kumar, Sripriya Ravindra; Taché, Yvette

doi:10.3389/fnana.2022.884280

Cited by 3 publications

(7 citation statements)

References 69 publications

(104 reference statements)

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“…AAV9 has been used extensively to transduce cells of the sensory nervous system and has been proven more efficient than other AAV serotypes when delivered systemically in mice ( Gombash et al, 2014 ; Buckinx et al, 2016 ). AAV9 has also been effective at transducing motor, autonomic, and enteric neurons in mice ( Benkhelifa-Ziyyat et al, 2013 ; Ayers et al, 2015 ; Wang et al, 2022 ). PHP.S, engineered from AAV9, has achieved even higher transduction efficacy in both sensory and autonomic nervous systems compared to AAV9 in adult C57Bl/6J mice ( Chan et al, 2017 ; Asencor et al, 2021 ).…”

Section: Introductionmentioning

confidence: 99%

Evaluating the transduction efficiency of systemically delivered AAV vectors in the rat nervous system

Yang

Robilotto²,

Alom³

et al. 2023

Front. Neurosci.

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Gene delivery or manipulation with viral vectors is a frequently used tool in basic neuroscience studies. Adeno-associated viruses (AAV) are the most widely used vectors due to their relative safety and long-term efficacy without causing overt immunological complications. Many AAV serotypes have been discovered and engineered that preferentially transduce different populations of neurons. However, efficient targeting of peripheral neurons remains challenging for many researchers, and evaluation of peripheral neuron transduction with AAVs in rats is limited. Here, we aimed to test the efficiency of systemic AAVs to transduce peripheral neurons in rats. We administered AAV9-tdTomato, AAV-PHP.S-tdTomato, or AAV-retro-GFP systemically to neonatal rats via intraperitoneal injection. After 5 weeks, we evaluated expression patterns in peripheral sensory, motor, and autonomic neurons. No significant difference between the serotypes in the transduction of sensory neurons was noted, and all serotypes were more efficient in transducing NF200 + neurons compared to smaller CGRP + neurons. AAV-retro was more efficient at transducing motor neurons compared to other serotypes. Moreover, PHP.S was more efficient at transducing sympathetic neurons, and AAV-retro was more efficient at transducing parasympathetic neurons. These results indicate that specific AAV serotypes target peripheral neuron populations more efficiently than others in the neonatal rat.

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Section: Introductionmentioning

confidence: 99%

Evaluating the transduction efficiency of systemically delivered AAV vectors in the rat nervous system

Yang

Robilotto²,

Alom³

et al. 2023

Front. Neurosci.

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“…There are few reports of the tropism of specific AAV serotypes for cells in the gastric muscularis propria of mice 34,35 . Therefore, we used a previously reported method 38 for determining the pattern of transduction induced by intravenous delivery of scAAV9‐Cre to Gt(ROSA)26Sor tm4(ACTBtdTomato,‐EGFP)Luo/J mice.…”

Section: Resultsmentioning

confidence: 99%

“…There are few reports of the tropism of specific AAV serotypes for cells in the gastric muscularis propria of mice. 34,35 Therefore, we used a previously reported method 38 for determining the pattern of transduction induced by intravenous delivery of scAAV9-Cre to Gt( ROSA)26Sor tm4(ACTBtdTomato,-EGFP)Luo/J mice. Three weeks after injection of 1 × 10 12 vg/mouse of scAAV9-Cre, we observed expression of eGFP in smooth muscle cells of the circular and longitudinal muscles of the small intestine (Figure 1A) and stomach (Figure 1B).…”

Section: Efficiency Of Viral Deliverymentioning

confidence: 99%

“…However, only few studies to date use AAVs to deliver proteins to the gastrointestinal tract. One study screened different AAV serotypes to identify one that can achieve high transduction efficiency in small intestine, 34 while another study used AAV to investigate transduction of the ENS in the colon 35 . These studies indicate that use of AAV to deliver proteins to gastric tissue may be possible.…”

Section: Introductionmentioning

confidence: 99%

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Adeno‐associated virus‐9 reverses delayed gastric emptying of solids in diabetic mice

Li,

Ji,

Cipriani

et al. 2023

Neurogastroenterology Motil

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BackgroundGastroparesis is defined by delayed gastric emptying (GE) without obstruction. Studies suggest targeting heme oxygenase‐1 (HO1) may ameliorate diabetic gastroparesis. Upregulation of HO1 expression via interleukin‐10 (IL‐10) in the gastric muscularis propria is associated with reversal of delayed GE in diabetic NOD mice. IL‐10 activates the M2 cytoprotective phenotype of macrophages and induces expression of HO1 protein. Here, we assess delivery of HO1 by recombinant adeno‐associated viruses (AAVs) in diabetic mice with delayed GE.MethodsC57BL6 diabetic delayed GE mice were injected with 1 × 1012 vg scAAV9‐cre, scAAV9‐GFP, or scAAV9‐HO1 particles. Changes to GE were assessed weekly utilizing our [13C]‐octanoic acid breath test. Stomach tissue was collected to assess the effect of scAAV9 treatment on Kit, NOS1, and HO1 expression.Key ResultsDelayed GE returned to normal within 2 weeks of treatment in 7/12 mice receiving scAAV9‐cre and in 4/5 mice that received the scAAV9‐GFP, whereas mice that received scAAV9‐HO1 did not respond in the same manner and had GE that took significantly longer to return to normal (6/7 mice at 4–6 weeks). Kit, NOS1, and HO1 protein expression in scAAV9‐GFP‐treated mice with normal GE were not significantly different compared with diabetic mice with delayed GE.Conclusions and InferencesInjection of scAAV9 into diabetic C57BL6 mice produced a biological response that resulted in acceleration of GE independently of the cargo delivered by the AAV9 vector. Further research is needed to determine whether use of AAV mediated gene transduction in the gastric muscularis propria is beneficial and warranted.

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“…The macrophages in the periphery are equivalent to microglia in the brain. The colon has numerous macrophages labeled by ionized calcium-binding adaptor molecule 1 (Iba1), a microglia/macrophage-specific marker (Wang et al, 2022), and their distributions related to microvessels were also studied.…”

Section: Introductionmentioning

confidence: 99%

Vasculature in the mouse colon and spatial relationships with the enteric nervous system, glia, and immune cells

2023

Self Cite

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The distribution, morphology, and innervation of vasculature in different mouse colonic segments and layers, as well as spatial relationships of the vasculature with the enteric plexuses, glia, and macrophages are far from being complete. The vessels in the adult mouse colon were stained by the cardiovascular perfusion of wheat germ agglutinin (WGA)-Alexa Fluor 448 and by CD31 immunoreactivity. Nerve fibers, enteric glia, and macrophages were immunostained in the WGA-perfused colon. The blood vessels entered from the mesentery to the submucosa and branched into the capillary networks in the mucosa and muscularis externa. The capillary net formed anastomosed rings at the orifices of mucosa crypts, and the capillary rings surrounded the crypts individually in the proximal colon and more than two crypts in the distal colon. Microvessels in the muscularis externa with myenteric plexus were less dense than in the mucosa and formed loops. In the circular smooth muscle layer, microvessels were distributed in the proximal, but not the distal colon. Capillaries did not enter the enteric ganglia. There were no significant differences in microvascular volume per tissue volume between the proximal and distal colon either in the mucosa or muscularis externa containing the myenteric plexus. PGP9.5-, tyrosine hydroxylase-, and calcitonin gene-related peptide (CGRP)-immunoreactive nerve fibers were distributed along the vessels in the submucosa. In the mucosa, PGP9.5-, CGRP-, and vasoactive intestinal peptide (VIP)-immunoreactive nerves terminated close to the capillary rings, while cells and processes labeled by S100B and glial fibrillary acidic protein were distributed mainly in the lamina propria and lower portion of the mucosa. Dense Iba1 immunoreactive macrophages were closely adjacent to the mucosal capillary rings. There were a few macrophages, but no glia in apposition to microvessels in the submucosa and muscularis externa. In conclusion, in the mouse colon, (1) the differences in vasculature between the proximal and distal colon were associated with the morphology, but not the microvascular amount per tissue volume in the mucosa and muscle layers; (2) the colonic mucosa contained significantly more microvessels than the muscularis externa; and (3) there were more CGRP and VIP nerve fibers found close to microvessels in the mucosa and submucosa than in the muscle layers.

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Transduction of Systemically Administered Adeno-Associated Virus in the Colonic Enteric Nervous System and c-Kit Cells of Adult Mice

Cited by 3 publications

References 69 publications

Evaluating the transduction efficiency of systemically delivered AAV vectors in the rat nervous system

Evaluating the transduction efficiency of systemically delivered AAV vectors in the rat nervous system

Adeno‐associated virus‐9 reverses delayed gastric emptying of solids in diabetic mice

Vasculature in the mouse colon and spatial relationships with the enteric nervous system, glia, and immune cells

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