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Cells Aug 2023Cardiovascular diseases (CVDs) are the prevalent cause of mortality worldwide. A combination of environmental and genetic effectors modulates the risk of developing...
Cardiovascular diseases (CVDs) are the prevalent cause of mortality worldwide. A combination of environmental and genetic effectors modulates the risk of developing them. Thus, it is vital to identify candidate genes and elucidate their role in the manifestation of the disease. Large-scale human studies have revealed the implication of Craniofacial Development Protein 1 (CFDP1) in Coronary Artery Disease (CAD). CFDP1 belongs to the evolutionary conserved Bucentaur (BCNT) family, and to date, its function and mechanism of action in Cardiovascular Development are still unclear. We utilized zebrafish to investigate the role of in the developing heart due to the high genomic homology, similarity in heart physiology, and ease of experimental manipulations. We showed that was expressed during development, and we tested two morpholinos and generated a mutant line. The embryos developed arrhythmic hearts and exhibited defective cardiac performance, which led to a lethal phenotype. Findings from both knockdown and knockout experiments showed that abrogation of leads to downregulation of Wnt signaling in embryonic hearts during valve development but without affecting Notch activation in this process. The zebrafish mutant line provides a valuable tool for unveiling the novel mechanism of regulating cardiac physiology and function. is essential for cardiac development, a previously unreported phenotype most likely due to early lethality in mice. The detected phenotype of bradycardia and arrhythmias is an observation with potential clinical relevance for humans carrying heterozygous CFDP1 mutations and their risk of developing CAD.
Topics: Animals; Humans; Cardiovascular Diseases; Heart; Nuclear Proteins; Phenotype; Wnt Signaling Pathway; Zebrafish
PubMed: 37566073
DOI: 10.3390/cells12151994 -
Drugs of Today (Barcelona, Spain : 1998) Oct 2019Duchenne muscular dystrophy is the most common lethal X-linked genetic disorder, characterized by progressive muscle loss, with cardiac and respiratory complications. It... (Review)
Review
Duchenne muscular dystrophy is the most common lethal X-linked genetic disorder, characterized by progressive muscle loss, with cardiac and respiratory complications. It is caused by a lack of dystrophin protein due to mutations in the DMD gene, which can disrupt the reading frame of the dystrophin primary transcript. Antisense oligonucleotides such as phosphorodiamidate morpholino oligomers (PMOs) can induce exon skipping during pre-mRNA splicing and restore the reading frame of the DMD primary transcript. The resulting dystrophin protein is internally deleted but partially functional. Viltolarsen, also known as NS-065/NCNP-01, is a PMO developed through comprehensive sequence optimization and is designed to skip exon 53 on the DMD primary transcript. Exclusion of exon 53 from the DMD primary transcript can treat 8-10% of DMD patients worldwide. This review paper summarizes the mechanism of action, pharmacokinetics and safety of viltolarsen from preclinical and clinical trials.
Topics: Clinical Trials as Topic; Dystrophin; Exons; Humans; Morpholinos; Muscular Dystrophy, Duchenne; Oligonucleotides; Oligonucleotides, Antisense; RNA Precursors
PubMed: 31720560
DOI: 10.1358/dot.2019.55.10.3045038 -
International Journal of Molecular... Oct 2021Animal models of human neurodegenerative disease have been investigated for several decades. In recent years, zebrafish () and medaka () have become popular in... (Review)
Review
Animal models of human neurodegenerative disease have been investigated for several decades. In recent years, zebrafish () and medaka () have become popular in pathogenic and therapeutic studies about human neurodegenerative diseases due to their small size, the optical clarity of embryos, their fast development, and their suitability to large-scale therapeutic screening. Following the emergence of a new generation of molecular biological technologies such as reverse and forward genetics, morpholino, transgenesis, and gene knockout, many human neurodegenerative disease models, such as Parkinson's, Huntington's, and Alzheimer's, were constructed in zebrafish and medaka. These studies proved that zebrafish and medaka genes are functionally conserved in relation to their human homologues, so they exhibit similar neurodegenerative phenotypes to human beings. Therefore, fish are a suitable model for the investigation of pathologic mechanisms of neurodegenerative diseases and for the large-scale screening of drugs for potential therapy. In this review, we summarize the studies in modelling human neurodegenerative diseases in zebrafish and medaka in recent years.
Topics: Animals; Disease Models, Animal; Humans; Neurodegenerative Diseases; Oryzias; Species Specificity; Zebrafish
PubMed: 34639106
DOI: 10.3390/ijms221910766 -
Journal of Biosciences 2023Duchenne muscular dystrophy (DMD) is an X-linked genetic disease primarily affecting boys causing loss of the dystrophin protein, ultimately leading to muscle wastage... (Review)
Review
Duchenne muscular dystrophy (DMD) is an X-linked genetic disease primarily affecting boys causing loss of the dystrophin protein, ultimately leading to muscle wastage and death by cardiac or respiratory failure. The genetic mutation involved can be overcome with antisense oligonucleotides which bind to a pre-mRNA and results in reading frame restoration by exon skipping. Phosphorodiamidate morpholino oligonucleotides (PMOs) are a class of antisense agents with a neutral backbone derived from RNA which can induce effective exon skipping. In this review, the evolution of PMOs in exon skipping therapy for the last two decades has been detailed with the gradual structural and functional advancements. Even though the success rate of PMObased therapy has been high with four FDA approved drugs, several key challenges are yet to overcome, one being the dystrophin restoration in cardiac muscle. The current scenario in further improvement of PMOs has been discussed along with the future perspectives that have the potential to revolutionize the therapeutic benefits in DMD.
Topics: Male; Humans; Morpholinos; Dystrophin; Muscular Dystrophy, Duchenne; Oligonucleotides, Antisense; Exons
PubMed: 37846020
DOI: No ID Found -
Journal of Personalized Medicine Sep 2020With the development of novel targeted therapies, including exon skipping/inclusion and gene replacement therapy, the field of neuromuscular diseases has drastically...
With the development of novel targeted therapies, including exon skipping/inclusion and gene replacement therapy, the field of neuromuscular diseases has drastically changed in the last several years. Until 2016, there had been no FDA-approved drugs to treat Duchenne muscular dystrophy (DMD), the most common muscular dystrophy. However, several new personalized therapies, including antisense oligonucleotides eteplirsen for DMD exon 51 skipping and golodirsen and viltolarsen for DMD exon 53 skipping, have been approved in the last 4 years. We are witnessing the start of a therapeutic revolution in neuromuscular diseases. However, the studies also made clear that these therapies are still far from a cure. Personalized genetic medicine for neuromuscular diseases faces several key challenges, including the difficulty of obtaining appropriate cell and animal models and limited its applicability. This Special Issue "Molecular Diagnosis and Novel Therapies for Neuromuscular/Musculoskeletal Diseases" highlights key areas of research progress that improve our understanding and the therapeutic outcomes of neuromuscular diseases in the personalized medicine era.
PubMed: 32947786
DOI: 10.3390/jpm10030129 -
Journal of Genetics and Genomics = Yi... Jul 2019Gene knockdown approaches using antisense oligo nucleotides or analogs such as siRNAs and morpholinos have been widely adopted to study gene functions although the... (Review)
Review
Gene knockdown approaches using antisense oligo nucleotides or analogs such as siRNAs and morpholinos have been widely adopted to study gene functions although the off-target issue has been always a concern in these studies. On the other hand, classic genetic analysis relies on the availability of loss-of-function or gain-of-function mutants. The fast development of genome editing technologies such as TALEN and CRISPR/Cas9 has greatly facilitated the generation of null mutants for the functional studies of target genes in a variety of organisms such as zebrafish. Surprisingly, an unexpected discrepancy was observed between morphant phenotype and mutant phenotype for many genes in zebrafish, i.e., while the morphant often displays an obvious phenotype, the corresponding null mutant appears relatively normal or only exhibits a mild phenotype due to gene compensation. Two recent reports have partially answered this intriguing question by showing that a pre-mature termination codon and homologous sequence are required to elicit the gene compensation and the histone modifying complex COMPASS is involved in activating the expression of the compensatory genes. Here, I summarize these exciting new progress and try to redefine the concept of genetic compensation and gene compensation.
Topics: Animals; Dosage Compensation, Genetic; Gene Expression Regulation; Gene Knockdown Techniques; Genotype; Models, Genetic; Morpholinos; Multigene Family; Mutation; Nonsense Mediated mRNA Decay; Phenotype; RNA, Messenger; Zebrafish
PubMed: 31377237
DOI: 10.1016/j.jgg.2019.07.001 -
Drugs of Today (Barcelona, Spain : 1998) Dec 2021Duchenne muscular dystrophy (DMD) is a genetic disorder affecting 1 in 5,000 males which causes progressive muscle deterioration, loss of mobility and eventual death,...
Duchenne muscular dystrophy (DMD) is a genetic disorder affecting 1 in 5,000 males which causes progressive muscle deterioration, loss of mobility and eventual death, with an average lifespan of around 25 years. While no cure currently exists for DMD, a novel treatment known as antisense-mediated exon skipping therapy has shown great promise. Exon skipping therapy induces the skipping of mutated exons, restoring the reading frame in dystrophin transcripts and resulting in a truncated but partially functional protein product. In February 2021, Sarepta Therapeutics received accelerated Food and Drug Administration (FDA) approval for their new antisense oligonucleotide, casimersen (brand name Amondys 45). Casimersen targets exon 45 of the dystrophin gene and is expected to treat ~8% of the DMD patient population. The continued approval of this drug will be dependent on satisfactory clinical results from an ongoing phase III trial. This article summarizes the preclinical and clinical data currently available for casimersen, emphasizing pharmacokinetics and safety.
Topics: Exons; Humans; Male; Muscular Dystrophy, Duchenne; Oligonucleotides; Oligonucleotides, Antisense
PubMed: 34909800
DOI: 10.1358/dot.2021.57.12.3352740 -
Methods in Molecular Biology (Clifton,... 2021Primordial germ cells (PGCs) are unique cells in an embryo. These cells contain all genetic information and therefore represent the best source to store maternal and...
Primordial germ cells (PGCs) are unique cells in an embryo. These cells contain all genetic information and therefore represent the best source to store maternal and paternal genomes until embryo cryopreservation is achieved. However, the number of these cells in an embryo is very low limiting their potential application in cryopreservation and surrogate production. However, it was assumed that the induction of fish PGCs in vitro is not possible because in vivo they inherit germ plasm. In this chapter, we describe a successful differentiation protocol explaining the crucial factors and steps for in vitro PGC generation.
Topics: Animals; Cell Differentiation; Cells, Cultured; Cryopreservation; Embryo, Nonmammalian; Female; Germ Cells; Male; Zebrafish
PubMed: 33606224
DOI: 10.1007/978-1-0716-0970-5_7 -
Advances in Experimental Medicine and... 2024Over the last few decades, the study of congenital heart disease (CHD) has benefited from various model systems and the development of molecular biological techniques... (Review)
Review
Over the last few decades, the study of congenital heart disease (CHD) has benefited from various model systems and the development of molecular biological techniques enabling the analysis of single gene as well as global effects. In this chapter, we first describe different models including CHD patients and their families, animal models ranging from invertebrates to mammals, and various cell culture systems. Moreover, techniques to experimentally manipulate these models are discussed. Second, we introduce cardiac phenotyping technologies comprising the analysis of mouse and cell culture models, live imaging of cardiogenesis, and histological methods for fixed hearts. Finally, the most important and latest molecular biotechniques are described. These include genotyping technologies, different applications of next-generation sequencing, and the analysis of transcriptome, epigenome, proteome, and metabolome. In summary, the models and technologies presented in this chapter are essential to study the function and development of the heart and to understand the molecular pathways underlying CHD.
Topics: Animals; Humans; Heart Defects, Congenital; Disease Models, Animal; Mice; Phenotype; High-Throughput Nucleotide Sequencing; Cell Culture Techniques
PubMed: 38884724
DOI: 10.1007/978-3-031-44087-8_22 -
Trends in Biotechnology Oct 2019Living organisms create life-sustaining macromolecular biocompounds including biopolymers. Artificial polymers can selectively recognize biocompounds and are more... (Review)
Review
Living organisms create life-sustaining macromolecular biocompounds including biopolymers. Artificial polymers can selectively recognize biocompounds and are more resistant to harsh physical, chemical, and physiological conditions than biopolymers are. Due to recognition at a molecular level, molecularly imprinted polymers (MIPs) provide powerful tools to correlate structure with biological functionality and are often used to build next-generation chemosensors. We envision an increasing emergence of nucleic acid analogs (NAAs) or biorelevant monomers built into nature-mimicking polymers. For example, if nucleobases bearing monomers arranged by a complementary template are polymerized to form NAAs, the resulting MIPs will open up novel perspectives for synthesizing NAAs. Despite their usefulness, it is still challenging to use MIPs to devise adaptive biomaterials and to implement them in point-of-care testing.
Topics: Aptamers, Nucleotide; Biopolymers; Biosensing Techniques; Humans; Molecular Imprinting; Nucleic Acids; Oligonucleotides; Polymorphism, Single Nucleotide
PubMed: 31109738
DOI: 10.1016/j.tibtech.2019.04.003