MiRNA Expression analysis from genomic form of Rhodopsin DNA Nanoparticles treated P23H Knock-in Heterozygous Mouse (P23H+/-)
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE153053 | GEO | 2020/06/24
REPOSITORIES: GEO
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Project description:miRNA Expression analysis from genomic form of Rhodopsin DNA Nanoparticles treated P23H Knock-in Heterozygous Mouse (P23H+/-)
| PRJNA641357 | ENA
Project description:Rhodopsin is essential for phototransduction, and many rhodopsin mutations cause heritable retinal degenerations. The P23H rhodopsin variant generates a misfolded rhodopsin protein that photoreceptors quickly target for degradation by mechanisms that are incompletely understood. To gain insight into how P23H rhodopsin is removed from rods, we used mass spectrometry to identify protein interaction partners of P23H rhodopsin immunopurified from RhoP23H/P23H mice and to compare with protein interaction partners of wild-type rhodopsin from Rho+/+ mice. We identified 286 proteins associated with P23H rhodopsin and 276 proteins associated with wild-type rhodopsin. Only 113 proteins were shared between wild-type and mutant rhodopsin protein interactomes. In the P23H rhodopsin protein interactome, we saw loss of phototransduction, retinal cycle, and rhodopsin protein trafficking proteins but gain of ubiquitin-related proteins when compared with the wild-type rhodopsin protein interactome. In the P23H rhodopsin protein interactome, we saw significant enrichment of gene ontology terms related to ER-associated protein degradation, ER stress, and translation. Protein-protein interaction network analysis revealed that translational and ribosomal quality control proteins were the most significant regulators in the P23H rhodopsin protein interactome. The protein partners identified in our study may provide new insights into how photoreceptors recognize and clear mutant rhodopsin and may offer novel targets involved in retinal degeneration pathogenesis.
2022-08-24 | MSV000090197 | MassIVE
Project description:Purpose: The goals of this study is to analyze the transcriptome change during retinal explant culture treated with pharmacological chaperone of rhodopsin (YC-001). Methods: The WT and RhoP23H/+ mouse eyes were enucleated at post natal day 15 and retinal explant were cultured for 24 hours followed by the 24 hours treatment with pharmacological chaperone of rhodopsin (YC-001) and dmso vehicle control. Another group of P23H/+ retinal explant was treated further till 9 days. We compare the transcript of RhoP23H/+ dmso with WT dmso at 24 hour treatment (1 days in vitro (1DIV)). RhoP23H/+ YC-001 were also compared with RhoP23H/+ dmso treated retinal explant at 1DIV and 9DIV. Results: A total of 338 differentially expressed genes (DEGs) were identified comparing the RhoP23H/+ vs. WT retinal explants with DMSO and 56 DEGs were identified comparing RhoP23H/+ YC-001 treated vs. DMSO treated retinal explant at 1 DIV. Further we observed an increased number of DEGs (8,597) comparing RhoP23H/+ vs dmso vehicle control at 9DIV. DEGs were identified using stringency with >1.5-fold change and P-value<0.05. Interestingly, we found 40 common genes when comparing RhoP23H/+ YC-001 treated vs. DMSO at 1DIV and 9DIV. Conclusions: This study used RNA-seq technology which represents the transcriptome of WT and RhoP23H/+ mouse retinal explant with and without pharmacological chaperone of rhodopsin (YC-001). Our NGS results show that YC-001 affect many biological pathways including visual transduction, protein homeostasis pathways and decreases the expression of immune response activation genes, to rescue the rhodospin homeostasis and retinal morphology in the retinal explant of P23H Rho mouse model of retinitis pigmentosa (RP).
2021-07-10 | GSE179754 | GEO
Project description:P23H is the most common mutation in the RHODOPSIN (RHO) gene leading to a dominant form of retinitis pigmentosa (RP), a rod photoreceptor degeneration that invariably causes vision loss. Specific disruption of the disease P23H RHO mutant while preserving the wild-type (WT) functional allele would be an invaluable therapy for this disease. However, various technologies tested in the past failed to achieve effective changes and consequently therapeutic benefits. We validated a CRISPR/Cas9 strategy to specifically inactivate the P23H RHO mutant, while preserving the WT allele in vitro. We, then, translated this approach in vivo by delivering the CRISPR/Cas9 components in murine Rho+/P23H mutant retinae. Targeted retinae presented a high rate of cleavage in the P23H but not WT Rho allele. This gene manipulation was sufficient to slow photoreceptor degeneration and improve retinal functions. To improve the translational potential of our approach, we tested intravitreal delivery of this system by means of adeno-associated viruses (AAVs). To this purpose, the employment of the AAV9-PHP.B resulted the most effective in disrupting the P23H Rho mutant. Finally, this approach was translated successfully in human cells engineered with the homozygous P23H RHO gene mutation. Overall, this is a significant proof-of-concept that gene allele specific targeting by CRISPR/Cas9 technology is specific and efficient and represents an unprecedented tool for treating RP and more broadly dominant genetic human disorders affecting the eye, as well as other tissues.
2019-03-20 | GSE128579 | GEO
Project description:The goal of this study focues on comparing the molecular signatures of immune cells, primarily microglia, in ectopically distinct mouse models of outer retinal degeneration. These models include the sodium iodate induced RPE injury model, rhodopsin-P23H knockin mice, an advanced aging model of 2-year-old mice and young (2-month old) wild-type control mice.
2024-01-07 | GSE195891 | GEO
Project description:G protein-coupled receptor (GPCR) kinases (GRKs) selectively phosphorylate agonist bound GPCRs, thereby switching the signal from G protein-dependent to arrestin-dependent pathways, including receptor internalization and downregulation. There are currently no high-resolution details known about how a GRK recognizes an activated receptor, but most functional studies indicate that its unique N-terminus is key to agonist-dependent phosphorylation. Herein, we report the 7.2 Å cryo-electron microscopy (EM) single particle reconstruction of the rhodopsin-rhodopsin kinase (GRK1) complex. The structure reveals a 1:1 assembly with multiple contact points between the GRK1 kinase domain and rhodopsin, the most prominent being the insertion of the GRK1 N-terminal helix directly into the cytoplasmic cleft formed by rhodopsin in its active state. Cross-linking coupled with mass spectrometry, along with functional studies, confirmed the observed interface between the proteins and revealed regions of the complex that are dynamic.
2021-07-15 | PXD019215 | Pride
Project description:Retinitis pigmentosa (RP) is a group of inherited retinal degenerative conditions and a leading cause of irreversible blindness. 25%-30% of RP cases are caused by inherited autosomal dominant (ad) mutations in the rhodopsin (Rho) protein of the retina, which impose a barrier for developing therapeutic treatments for this genetically heterogeneous disorder, as simple gene replacement is not sufficient to overcome dominant disease alleles. Previously, we have explored using the genomic short-form of Rho (sgRho) for gene augmentation therapy of RP in a Rho knockout mouse model. We have shown improved gene expression and fewer epigenetic modifications compared with the use of a Rho cDNA expression construct. In the current study, we altered our strategy by delivering a codon-optimized genomic form of Rho (co-sgRho) (for gene replacement) in combination with an RNAi-based inactivation of endogenous Rho alleles (gene suppression of both mutant Rho alleles, but mismatched with the co-sgRho) into a homozygous RhoP23H/P23H knock-in (KI) RP mouse model, which has a severe phenotype of adRP. In addition, we have conjugated a cell penetrating TAT peptide sequence to our previously established CK30PEG10 diblock co-polymer. The DNAs were compacted with CK30PEG10-TAT diblock co-polymer to form DNA nanoparticles (NPs). These NPs were injected into the sub-retinal space of the KI mouse eyes. As a proof of concept, we demonstrated the efficiency of this strategy in the partial improvement of visual function in the RhoP23H/P23H KI mouse model.
| S-EPMC5752119 | biostudies-literature
Project description:The study investigates the effect of a heterozygous MYBPC3 mutation combined with Western diet feeding on protein expression in a HCM mouse model. Hypertrophic cardiomyopathy (HCM) is the most common inherited genetic heart disease, characterized by asymmetric left ventricular hypertrophy and diastolic dysfunction. It is generally accepted that HCM is caused by mutations in genes encoding sarcomere proteins (e.g. MYBPC3, MYH7). However, not all mutation carriers will develop HCM and, moreover, phenotypical variation in terms of clinical presentation is large. The phenotypical expression of HCM thus requires additional disease modifiers on top of a sarcomere gene mutation. Clinical reports identify obesity and related comorbidities (diabetes, hyperlipidemia, hypertension) as a major factor contributing to disease expression and severity. We aimed to mimic this situation in a HCM mouse model by feeding a Western diet (8 weeks) to mice carrying a heterozygous mutation in MYBPC3. These heterozygous mice are phenotype negative under normal conditions, but developed cardiac dysfunction and hypertrophy upon Western diet feeding. The main goal of this proteomic screening is to identify clusters of proteins and pathways that are down/upregulated specifically in the left ventricle of mice suffering from a mutation and Western diet feeding. Also, we are interested in protein expression differences caused solely by mutation or Western diet. Group 1 are Wild type mice that received normal chow; group 2 are heterozygous mice that received normal chow; group 3 are Wild type mice that received a Western diet; group 4 are heterozygous mice that received a Western diet.
2023-10-01 | PXD043345 | Pride
Project description:We report RNAseq analysis of the transcriptome of retinas from mature rhodopsin knockout mice
2016-12-31 | GSE61239 | GEO
Project description:Neuronal plasticity of the inner retina has been observed in response to photoreceptor degeneration. Typically, this phenomenon has been considered maladaptive and may preclude vision restoration in the blind. However, several recent studies utilizing triggered photoreceptor ablation have shown adaptive responses in bipolar cell dendrites expected to support normal vision. Whether such homeostatic plasticity occurs during progressive photoreceptor degenerative disease to help maintain normal visual behavior is unknown. We addressed these issues in an established mouse model of Retinitis Pigmentosa caused by the P23H mutation in rhodopsin. We show robust modulation of the retinal transcriptomic network reminiscent of the neurodevelopmental state as well as potentiation of rod – rod bipolar cell signaling following rod photoreceptor degeneration. Additionally, we found highly sensitive night vision in P23H mice even when more than half of the rod photoreceptors were lost. The results implicate retinal adaptation leading to persistent visual function during photoreceptor degenerative disease.
2020-06-16 | GSE152474 | GEO