Noninvasive tracking of gene transcript and neuroprotection after gene therapy

Ren, Jun; Chen, Yi; Liu, CH; Pc, Chen; Prentice, Howard; Wu, Jianzhong; Liu, PK

doi:10.1038/gt.2015.81

Cited by 21 publications

(21 citation statements)

References 41 publications

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“…After 30 min of ischemia, the aneurysm clips were removed from both CCAs and the incision was sutured closed. Post-surgery analgesics were administered, and animals could wake up within 10 to 20 min after surgery [26]. All mice subjected to BCAO exhibited a significant reduction in regional cerebral blood flow (RCBF) to 51% pre-BCAO values and the RCBF retuned to 91% of the pre-BCAO level at the beginning of reperfusion as monitored with a Laser Doppler Flowmeter (LDF).…”

Section: Bilateral Common Carotid Artery Occlusion (Bcao)mentioning

confidence: 99%

“…Restoration of blood flow (reoxygenation) was also observed directly under the microscope. Sham-operated controls were subjected to the same surgical procedures except that CCAs were not occluded [26]. The body temperature was monitored and maintained at 37°C ± 0.5°C during surgery and during the immediate postoperative period until the animals recovered fully from anesthesia.…”

Section: Bilateral Common Carotid Artery Occlusion (Bcao)mentioning

confidence: 99%

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Mode of action of granulocyte-colony stimulating factor (G-CSF) as a novel therapy for stroke in a mouse model

Modi

Menzie-Suderam

et al. 2020

J Biomed Sci

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Background: The FDA approved drug granulocyte-colony stimulating factor (G-CSF) displays anti-apoptotic and immunomodulatory properties with neurogenesis and angiogenic functions. It is known to demonstrate neuroprotective mechanisms against ischemic global stroke. Autophagy is a method for the degradation of intracellular components and in particular, unrestrained autophagy may lead to uncontrolled digestion of affected neurons as well as neuronal death in cerebral ischemia. Mitochondrial dynamics is vital for the regulation of cell survival and death after cerebral ischemia and an early upstream event in neuronal death is mitochondrial fission. We examined the pro-survival mechanisms of G-CSF against apoptosis resulting from autophagy, mitochondrial stress and endoplasmic reticulum (ER) stress. Methods: Male Swiss Webster mice (20 weeks of age) were subjected to bilateral common carotid artery occlusion (BCAO) for 30 min. After occlusion, mice were injected with G-CSF (50 μg/kg) subcutaneously for 4 days. Behavioral analysis was carried out using the corner test and locomotor activity test before animals were sacrificed on day 4 or day 7. Key proteins in ER stress, autophagy and mitochondrial stress induced apoptosis were analyzed by immunoblotting.Results: G-CSF improved neurological deficits and improved behavioral performance on corner and locomotor test. G-CSF binds to G-CSF receptors and its activation leads to upregulation of Akt phosphorylation (P-Akt) which in turn decreases levels of the ER stress sensor, GRP 78 and expression of proteins involved in ER stress apoptosis pathway; ATF6, ATF4, eIF2α, XBP1, Caspase 12 and CHOP. G-CSF treatment significantly decreased Beclin-1, an autophagy marker, and decreased mitochondrial stress biomarkers DRP1 and P53. G-CSF also up-regulated the mitochondrial fusion protein, OPA1 and anti-apoptotic protein Bcl-2 while down-regulating the pro-apoptotic proteins Bax, Bak and PUMA. Conclusions: G-CSF is an endogenous ligand in the CNS that has a dual activity that is beneficial both in reducing acute neuronal degeneration and adding to long-term plasticity after cerebral ischemia. G-CSF treatment exerts neuroprotective effects on damaged neurons through the suppression of the ER stress and mitochondrial stress and maintains cellular homeostasis by decreasing pro-apoptotic proteins and increasing of anti-apoptotic proteins.

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Section: Bilateral Common Carotid Artery Occlusion (Bcao)mentioning

confidence: 99%

Section: Bilateral Common Carotid Artery Occlusion (Bcao)mentioning

confidence: 99%

Mode of action of granulocyte-colony stimulating factor (G-CSF) as a novel therapy for stroke in a mouse model

Modi

Menzie-Suderam

et al. 2020

J Biomed Sci

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“…Based on subtraction R2* maps that identified regional repair/remodeling [23, 29, 31], we are able to also identify changes in HDAC5 expression in the chronic paradigm. Figure 11a shows R2* elevation in the LS of three consecutive MR sections (a total of 1.5 mm in the dorsal LS); however, we observed several ROIs in the NAc/Striatal (arrows).…”

Section: Resultsmentioning

confidence: 99%

Epigenetics of amphetamine-induced sensitization: HDAC5 expression and microRNA in neural remodeling

Liu

2016

J Biomed Sci

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BackgroundHistone deacetylase (HDAC) activities modify chromatin structure and play a role in learning and memory during developmental processes. Studies of adult mice suggest HDACs are involved in neural network remodeling in brain repair, but its function in drug addiction is less understood. We aimed to examine in vivo HDAC5 expression in a preclinical model of amphetamine-induced sensitization (AIS) of behavior. We generated specific contrast agents to measure HDAC5 levels by in vivo molecular contrast-enhanced (MCE) magnetic resonance imaging (MRI) in amphetamine-naïve mice as well as in mice with AIS. To validate the MRI results we used ex vivo methods including in situ hybridization, RT-PCR, immunohistochemistry, and transmision electron microscopy.MethodsWe compared the expression of HDAC5 mRNA in an acute exposure paradigm (in which animals experienced a single drug exposure [A1]) and in a chronic-abstinence-challenge paradigm (in which animals were exposed to the drug once every other day for seven doses, then underwent 2 weeks of abstinence followed by a challenge dose [A7WA]). Control groups for each of these exposure paradigms were given saline. To delineate how HDAC5 expression was related to AIS, we compared the expression of HDAC5 mRNA at sequences where no known microRNA (miR) binds (hdac5AS2) and at sequences where miR-2861 is known to bind (miD2861). We synthesized and labeled phosphorothioated oligonucleic acids (sODN) of hdac5AS2 or miD2861 linked to superparamagentic iron oxide nanoparticles (SPION), and generated HDAC5-specific contrast agents (30 ± 20 nm, diameter) for MCE MRI; the same sequences were used for primers for TaqMan® analysis (RT-qPCR) in ex vivo validation. In addition, we used subtraction R2* maps to identify regional HDAC5 expression.ResultsNaïve C57black6 mice that experience acute exposure to amphetamine (4 mg/kg, by injection intraperitoneally) show expression of both total and phosphorylated (S259) HDAC5 antigens in GFAP+ and GFAP− cells, but the appearance of these cells was attenuated in the chronic paradigm. We found that MCE MRI reports HDAC5 mRNA with precision in physiological conditions because the HDAC5 mRNA copy number reported by TaqMan analysis was positively correlated (with a linear coefficient of 1.0) to the ΔR2* values (the frequency of signal reduction above background, 1/s) measured by MRI. We observed SPION-mid2861 as electron dense nanoparticles (EDNs) of less than 30 nm in the nucleus of the neurons, macrophages, and microglia, but not in glia and endothelia. We found no preferential distribution in any particular type of neural cells, but observed scattered EDNs of 60–150 nm (dia) in lysosomes. In the acute paradigm, mice pretreated with miD2861 (1.2 mmol/kg, i.p./icv) exhibited AIS similar to that exibited by mice in the chronic exposure group, which exhibited null response to mid2861 pretreatment. Moreover, SPION-miD2861 identified enhanced HDAC5 expression in the lateral septum and the striatum after amphetamine, where we found neurprogenitor cel...

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“…Given that the composition of NVU is the same in the CNS and the retina (1) and that our previous studies demonstrated that BCAO compromises the BRB (7), we delivered chimeric anti‐Rho‐SPION‐Ran (a T2 MR‐CA) through an eyedrop solution administered to the left eye (2 μg Fe in 1.5 μl) of C57black6 mice ( n =3) 1 d after BCAO (2). We acquired MR images of the eyes at 6 time points (each mouse received sequential MRI twice, with an interval of 1.5 h awakeness) and analyzed the R2∗ values using the right eye of the same mouse as the negative control.…”

Section: Resultsmentioning

confidence: 99%

“…Throughout the operation we maintained the animals' body temperature at 36 ± 1 °C using a heating blanket, and we monitored temperature with a rectal thermosensor. We used Doppler analysis to assess the blood flow in the middle cerebral artery in mice that underwent surgical procedures, positioning the Doppler probes to the skull during surgery (7). By the third day after BCAO‐60, all of the surviving postischemic mice exhibited spontaneous activity in their home cages, were responsive to handling, and engaged in normal self‐grooming behavior.…”

Section: Methodsmentioning

confidence: 99%

Imaging rhodopsin degeneration in vivo in a new model of ocular ischemia in living mice

et al. 2015

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Delivery of antibodies to monitor key biomarkers of retinopathy in vivo represents a significant challenge because living cells do not take up immunoglobulins to cellular antigens. We met this challenge by developing novel contrast agents for retinopathy, which we used with magnetic resonance imaging (MRI). Biotinylated rabbit polyclonal to chick IgY (rIgPxcIgY) and phosphorylthioate-modified oligoDNA (sODN) with random sequence (bio-sODN-Ran) were conjugated with NeutrAvidin-activated superparamagnetic iron oxide nanoparticles (SPION). The resulting Ran-SPION-rIgPxcIgY carries chick polyclonal to microtubule-associated protein 2 (MAP2) as Ran-SPIONrIgP/cIgY-MAP2, or to rhodopsin (Rho) as anti-Rho-SPIONRan. We examined the uptake of Ran-SPION-rIgP/ cIgY-MAP2 or SPION-rIgP/cIgY-MAP2 in normal C57black6 mice (n = 3 each, 40 mg/kg, i.c.v.); we found retention of Ran-SPION-rIgP/cIgY-MAP2 using molecular contrastenhanced MRI in vivo and validated neuronal uptake using Cy5-goat IgPxcIgY ex vivo. Applying this novel method to monitor retinopathy in a bilateral carotid artery occlusion-induced ocular ischemia, we observed pericytes (at d 2, using Gd-nestin, by eyedrop solution), significant photoreceptor degeneration (at d 20, using anti-Rho-SPION-Ran, eyedrops, P = 0.03, Student's t test), and gliosis in Müller cells (at 6 mo, using SPION-glial fibrillary acidic protein administered by intraperitoneal injection) in surviving mice (n ‡ 5). Molecular contrastenhanced MRI results were confirmed by optical and electron microscopy. We conclude that chimera and molecular contrast-enhanced MRI provide sufficient sensitivity for monitoring retinopathy and for theranostic applications.-Ren, J., Chen, Y. I., Mackey, A. M., Liu, P. K. Imaging rhodopsin degeneration in vivo in a new model of ocular ischemia in living mice. FASEB J. 30, 612-623 (2016). www.fasebj.org

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Noninvasive tracking of gene transcript and neuroprotection after gene therapy

Cited by 21 publications

References 41 publications

Mode of action of granulocyte-colony stimulating factor (G-CSF) as a novel therapy for stroke in a mouse model

Mode of action of granulocyte-colony stimulating factor (G-CSF) as a novel therapy for stroke in a mouse model

Epigenetics of amphetamine-induced sensitization: HDAC5 expression and microRNA in neural remodeling

Imaging rhodopsin degeneration in vivo in a new model of ocular ischemia in living mice

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