CRISPR/Cas9-Mediated Constitutive Loss of VCP (Valosin-Containing Protein) Impairs Proteostasis and Leads to Defective Striated Muscle Structure and Function In Vivo

Voisard, Philipp; Diofano, Federica; Glazier, Amelia; Rottbauer, Wolfgang; Just, Steffen

doi:10.3390/ijms23126722

Cited by 5 publications

(1 citation statement)

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“…Some studies have shown that vcp is the central mediator of lysosomal clearance and biogenesis in skeletal muscle [ 50 ]. Philipp [ 51 ] and other researchers showed that deletion of vcp in zebrafish damages protein homeostasis and leads to structural and functional defects in striated muscle in vivo. Real-time PCR showed that the expression level of the vcp gene in miR-196a-1 knockout zebrafish was significantly decreased, but there was no significant difference in the miR-196b knockout group.…”

Section: Discussionmentioning

confidence: 99%

Roles of miR-196a and miR-196b in Zebrafish Motor Function

et al. 2023

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Background: The exertion of motor function depends on various tissues, such as bones and muscles. miR-196 has been widely studied in cancer and other fields, but its effect on bone and skeletal muscle is rarely reported. In order to explore the role of miR-196 family in bone and skeletal muscle, we used the previously successfully constructed miR-196a-1 and miR-196b gene knockout zebrafish animal models for research. Methods: The behavioral trajectories of zebrafish from 4 days post-fertilization (dpf) to 7 dpf were detected to analyze the effect of miR-196a-1 and miR-196b on motor ability. Hematoxylin-eosin (HE) staining and transmission electron microscopy (TEM) were used to detect the dorsal muscle tissue of zebrafish. The bone tissue of zebrafish was detected by microcomputed tomography (micro-CT). Real-time PCR was used to detect the expression levels of related genes, including vcp, dpm1, acta1b, mylpfb, col1a1a, bmp8a, gdf6a, and fgfr3. Results: The behavioral test showed that the total behavioral trajectory, movement time, and movement speed of zebrafish larvae were decreased in the miR-196a-1 and miR-196b gene knockout lines. Muscle tissue analysis showed that the structure of muscle fibers in the zebrafish lacking miR-196a-1 and miR-196b was abnormal and was characterized by vacuolar degeneration of muscle fibers, intranuclear migration, melanin deposition, and inflammatory cell infiltration. Bone CT examination revealed decreased bone mineral density and trabecular bone number. The real-time PCR results showed that the expression levels of vcp, dpm1, gdf6a, fgfr3, and col1a1a were decreased in the miR-196b gene knockout group. The expression levels of dpm1, acta1b, mylpfb, gdf6a, and col1a1a were decreased, and the expression level of fgfr3 was increased in the miR-196b gene knockout group compared with the wild-type group. Conclusions: miR-196a-1 and miR-196b play an important role in muscle fiber structure, bone mineral density, and bone trabecular quantity by affecting the expression of vcp, dpm1, acta1b, mylpfb, gdf6a, fgfr3, and col1a1a and then affect the function of the motor system

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Section: Discussionmentioning

confidence: 99%

Roles of miR-196a and miR-196b in Zebrafish Motor Function

et al. 2023

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SMYD1-mediated Mono-Methylation of Lysine K35 of the sarcomeric Myosin Heavy Chain (MHC) is fundamental for thick filament assembly in zebrafish and human iPSC-derived cardiomyocytes

Diofano,

Amadi,

Gahr

et al. 2024

Preprint

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The SMYD family is a unique class of lysine methyltransferases (KMTases) known to methylate histones but also non-histone proteins. Among the five SMYD family members (1-5), SMYD1 was identified as a heart- and skeletal muscle-specific KMTase, which, together with Unc45b and Hsp90a, interacts with Myosin thereby regulating thick filament assembly. However, the process by which SMYD1 orchestrates Myosin assembly is largely unknown. Here, we found that SMYD1 physically interacts with Myosin heavy chain (Myh) at its N-terminus and that the Myh N-terminus specifically gets mono-methylated by SMYD1 at lysine 35 (K35). Accordingly, methylated Myh is properly integrated into functional sarcomeres, whereas unmethylated Myh molecules in Smyd1-deficient zebrafish are efficiently degraded by the Ubiquitin Proteasome System (UPS) leading to defective thick filament assembly. Although the inhibition of the UPS by MG132 is able to reconstitute Myosin levels in Smyd1-deficient zebrafish embryos, thick filament assembly is still blocked due to the lack of K35 Myh mono-methylation. Similar to the situation in zebrafish striated muscle cells, SMYD1-mediated MYH methylation is also critical for thick filament assembly in human cardiomyocytes, indicating cross-species conservation of this fundamental mechanism of Myosin methylation, which has been first described about 40 years ago. Further investigations will now be essential to explore the therapeutic potential of targeting this pathway in cardiomyopathies and skeletal muscle disorders.

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CRISPR/Cas9: implication for modeling and therapy of amyotrophic lateral sclerosis

Shi

Zhao

et al. 2023

Front. Neurosci.

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Amyotrophic lateral sclerosis (ALS) is a deadly neurological disease with a complicated and variable pathophysiology yet to be fully understood. There is currently no effective treatment available to either slow or terminate it. However, recent advances in ALS genomics have linked genes to phenotypes, encouraging the creation of novel therapeutic approaches and giving researchers more tools to create efficient animal models. Genetically engineered rodent models replicating ALS disease pathology have a high predictive value for translational research. This review addresses the history of the evolution of gene editing tools, the most recent ALS disease models, and the application of CRISPR/Cas9 against ALS disease.

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CRISPR/Cas9-Mediated Constitutive Loss of VCP (Valosin-Containing Protein) Impairs Proteostasis and Leads to Defective Striated Muscle Structure and Function In Vivo

Cited by 5 publications

References 44 publications

Roles of miR-196a and miR-196b in Zebrafish Motor Function

Roles of miR-196a and miR-196b in Zebrafish Motor Function

SMYD1-mediated Mono-Methylation of Lysine K35 of the sarcomeric Myosin Heavy Chain (MHC) is fundamental for thick filament assembly in zebrafish and human iPSC-derived cardiomyocytes

CRISPR/Cas9: implication for modeling and therapy of amyotrophic lateral sclerosis

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