A5013075360- Retinal Development and Disorders
- Photoreceptor and optogenetics research
- Retinal Diseases and Treatments
- Virus-based gene therapy research
- Neuroscience and Neuropharmacology Research
- CRISPR and Genetic Engineering
- Neuroscience and Neural Engineering
- Connexins and lens biology
- Epigenetics and DNA Methylation
- Retinal and Optic Conditions
- Neuroinflammation and Neurodegeneration Mechanisms
- RNA modifications and cancer
- RNA regulation and disease
- Olfactory and Sensory Function Studies
- Ion channel regulation and function
- Viral Infectious Diseases and Gene Expression in Insects
- Neurobiology and Insect Physiology Research
- Nicotinic Acetylcholine Receptors Study
- Advanced biosensing and bioanalysis techniques
- Phosphodiesterase function and regulation
- RNA Interference and Gene Delivery
- RNA and protein synthesis mechanisms
- Receptor Mechanisms and Signaling
- Advanced Fluorescence Microscopy Techniques
- bioluminescence and chemiluminescence research
Ludwig-Maximilians-Universität München
2016-2025
LMU Klinikum
2014-2025
Center for Integrated Protein Science Munich
2012-2021
5–Hydroxymethylcytosine (hmC) was recently detected as the sixth base in mammalian tissue at so far controversial levels. The function of modified is currently unknown, but it certain that generated from 5-methylcytosine (mC). This fuels hypothesis represents an intermediate active demethylation process, which could involve further oxidation hydroxymethyl group to a formyl or carboxyl followed by either deformylation decarboxylation. Here, we use ultra-sensitive and accurate isotope based...
Mind over matter: LC-MS has allowed the amount of post-replicatively formed DNA base 5-hydroxymethylcytosine (see picture; left) to be quantified in brain tissue. The nucleoside is most abundant areas that are associated with higher cognitive functions, and its content mouse hippocampi seems increase age. new method enables hydroxymethylcytosine unprecedented accuracy.
Cell death in neurodegenerative diseases is often thought to be governed by apoptosis; however, an increasing body of evidence suggests the involvement alternative cell mechanisms neuronal degeneration. We studied retinal neurodegeneration using 10 different animal models, covering all major groups hereditary human blindness (rd1, rd2, rd10, Cngb1 KO, Rho S334ter, P23H, Cnga3 cpfl1, Rpe65 KO), investigating metabolic processes relevant for forms death. show that apoptosis plays only a minor...
Achromatopsia linked to variations in the CNGA3 gene is associated with day blindness, poor visual acuity, photophobia, and involuntary eye movements owing lack of cone photoreceptor function. No treatment currently available.To assess safety vision outcomes supplemental therapy adeno-associated virus (AAV) encoding (AAV8.CNGA3) patients CNGA3-linked achromatopsia.This open-label, exploratory nonrandomized controlled trial tested vector AAV8.CNGA3 administered by subretinal injection at a...
The mammalian main olfactory epithelium (MOE) recognizes and transduces cues through a G protein-coupled, cAMP-dependent signaling cascade. Additional chemosensory transduction mechanisms have been suggested but remain controversial. We show that subset of MOE neurons expressing the orphan receptor guanylyl cyclase GC-D cyclic nucleotide-gated channel subunit CNGA3 employ an excitatory cGMP-dependent mechanism for chemodetection. By combining gene targeting Gucy2d, which encodes GC-D, with...
Congenital absence of cone photoreceptor function is associated with strongly impaired daylight vision and loss color discrimination in human achromatopsia. Here, we introduce viral gene replacement therapy as a potential treatment for this disease the CNGA3−/− mouse model. We show that such can restore cone-specific visual processing central nervous system even if photoreceptors had been nonfunctional from birth. The restoration was assessed at different stages along pathway. Treated mice...
Abstract The absolute levels of 5‐hydroxymethylcytosine (hmC) and 5‐methylcytosine (mC) in human brain tissues at various ages were determined. Additionally, 5‐formylcytosine (fC) adult individuals cytosine modification sorted neurons quantified. These data compared with age‐related fC, hmC, mC mouse samples. For an initial steady increase is observed, which off age to a final steady‐state value 1.2 % tissue. This level nearly twice as high cerebral cortex. In contrast, fC declines rapidly...
Neuronal differentiation involves major epigenetic changes, such as accumulation of 5hydroxymethylcytosine (5hmC).Perera et al. found that TET3 interacts with transcriptional regulators and histone writers.They demonstrate REST recruits for context-specific 5hmC formation NSD3 H3K36 trimethylation.
Abstract Until recently, it was believed that the genomes of higher organisms contain, in addition to four canonical DNA bases, only 5‐methyl‐dC (m 5 dC) as a modified base control epigenetic processes. In recent years, this view has changed dramatically with discovery 5‐hydroxymethyl‐dC (hmdC), 5‐formyl‐dC (fdC), and 5‐carboxy‐dC (cadC) from stem cells brain tissue. N 6 ‐methyldeoxyadenosine dA) is most reported be present genome various eukaryotic organisms. This base, together 4...
Purpose: To investigate shedding and biodistribution characteristics of recombinant adeno-associated virus serotype 8 (rAAV8) after single-dose subretinal or intravitreal injection in nonhuman primates (NHP, Macaca fascicularis) as a surrogate for environmental hazard patient safety. Methods: In study regulatory submission, 22 NHP were divided into four cohorts receiving either single injections vehicle clinical grade rAAV8 (1 × 1011 1 1012 vector genomes [vg]) versus application vg. Viral...
Article22 February 2021Open Access Source DataTransparent process Novel AAV capsids for intravitreal gene therapy of photoreceptor disorders Marina Pavlou Department Ophthalmology, Ludwig-Maximilians-University, Munich, Germany Centre Integrated Protein Science Munich (CIPSM) at the Pharmacy, Search more papers by this author Christian Schön Laurence M Occelli Small Animal Clinical Sciences, Michigan State University, East Lansing, MI, USA Axel Rossi Laboratory Infection Biology and Gene...
To determine long-term safety and efficacy outcomes of a subretinal gene therapy for CNGA3-associated achromatopsia. We present data from an open-label, nonrandomised controlled trial (NCT02610582).Details the study design have been previously described. Briefly, nine patients were treated in three escalating dose groups with AAV8.CNGA3 between November 2015 October 2016. After first year, seen on yearly basis. Safety assessment constituted primary endpoint. On secondary level, multiple...
Cyclic nucleotide-gated (CNG) channels are important mediators in the transduction pathways of rod and cone photoreceptors. Native CNG heterotetramers composed homologous A B subunits. In heterologous expression systems, subunits alone cannot form functional channels, but they confer a number channel properties when coexpressed with To investigate importance CNGB vivo, we deleted CNGB1 gene mice. absence CNGB1, only trace amounts CNGA1 subunit were found on outer segment. As consequence,...
Hyperpolarization-activated cation channels of the HCN gene family are crucial for regulation cell excitability. Importantly, these play a pivotal role in control cardiac and neuronal pacemaker activity. Dysfunction has been associated with human diseases, including arrhythmia, epilepsy, neuropathic pain. The properties three channel isoforms (HCN1, HCN2, HCN4) have extensively investigated. By contrast, due to lack an efficient heterologous expression system, functional characteristics HCN3...
purpose. To characterize molecular and cellular changes in the mouse retina caused by genetic deletion of cone cyclic nucleotide-gated channel (CNG) subunit CNGA3. methods. Retinas wild-type CNGA3-deficient (CNGA3−/−) mice from 9 days up to 22 months age were analyzed immunohistochemistry, electron microscopy, biological results. CNGA3−/− cones failed transport opsins into outer segments, downregulated various proteins phototransduction cascade, induced apoptotic death. Loss CNGA3 did not...
The rd1 natural mutant is one of the first and probably most commonly studied mouse model for retinitis pigmentosa (RP), a severe frequently blinding human retinal degeneration. In several decades research, link between increase in photoreceptor cGMP levels extremely rapid cell death gave rise to number hypotheses. Here, we provide clear evidence that presence cyclic nucleotide gated (CNG) channels outer segment membrane key rod loss. Cngb1−/−× double mutants devoid regular CNG channels, are...
Retinitis pigmentosa (RP) is a group of genetically heterogeneous, severe retinal diseases commonly leading to legal blindness. Mutations in the CNGB1a subunit rod cyclic nucleotide-gated (CNG) channel have been found cause RP patients. Here, we demonstrate efficacy gene therapy as potential treatment for by means recombinant adeno-associated viral (AAV) vectors CNGB1 knockout (CNGB1−/−) mouse model. To enable efficient packaging and rod-specific expression relatively large cDNA (∼4 kb),...
Purpose: Preclinical studies on photoreceptor transplantation provided evidence for restoration of visual function with pluripotent stem cells considered as a potential source sufficient amounts donor material. Adequate preclinical models representing retinal disease conditions future patients are needed translation research. Here we compared transplant integration in mouse mild (prominin1-deficient; Prom1−/−) or severe (cone loss 1/rhodopsin-deficient double-mutant; Cpfl1/Rho−/−) cone-rod...
Purpose: To study longitudinal changes of anti-drug antibody (ADA) titers to recombinant adeno-associated virus serotype 8 (rAAV8) capsid epitopes in nonhuman primates (NHP) and patients. Methods: Three groups six NHP each received subretinal injections (high dose: 1 × 1012 vector genomes [vg], low 1011 vg, or vehicle only). Four additional animals intravitreal the high dose (1 vg). patients 1010 vg as injections. ELISA quantified ADA levels at baseline 1, 2, 3, 7, 28, 90 days after surgery...