Perspective of the GEMSTONE Consortium on Current and Future Approaches to Functional Validation for Skeletal Genetic Disease Using Cellular, Molecular and Animal-Modeling Techniques

Frontiers in Endocrinology, Год журнала: 2021, Номер 12

Опубликована: Ноя. 30, 2021

The availability of large human datasets for genome-wide association studies (GWAS) and the advancement sequencing technologies have boosted identification genetic variants in complex rare diseases skeletal field. Yet, interpreting results from remains a challenge. To bridge gap between causality, systematic functional investigation is necessary. Multiple unknowns exist putative causal genes, including cellular localization molecular function. Intermediate traits (“endophenotypes”), e.g. quantitative trait loci (molQTLs), are needed to identify mechanisms underlying associations. Furthermore, index often reside non-coding regions genome, therefore challenging interpretation. Knowledge variance (e.g. ncRNAs), repetitive sequences, regulatory interactions enhancers their target genes central understanding conditions. Animal models with deep phenotyping cell culture already facilitated fine mapping some signals, elucidated gene mechanisms, revealed disease-relevant biology. However, accelerate research towards bridging current causality diseases, alternative vivo platforms need be used developed parallel -omics traditional resources. Therefore, we argue that as field establish resource-sharing standards collectively address questions. These will promote data integration various dissection traits. In this mission statement, review available resources group propose consensus facilitate resource sharing using existing future Such coordination efforts maximize acquisition knowledge different approaches thus reduce redundancy duplication measures help understand pathogenesis osteoporosis other defining new more efficient therapeutic targets.

Язык: Английский

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Zebrafish mutants reveal unexpected role of Lrp5 in osteoclast regulation

Frontiers in Endocrinology, Год журнала: 2022, Номер 13

Опубликована: Сен. 2, 2022

Low-density Lipoprotein Receptor-related Protein 5 ( LRP5 ) functions as a co-receptor for Wnt ligands, controlling expression of genes involved in osteogenesis. In humans, loss-of-function mutations cause Osteoporosis-Pseudoglioma syndrome, low bone mass disorder, while gain-of-function missense have been observed individuals with high mass. Zebrafish Danio rerio is popular model human disease research, genetic determinants that control formation are generally conserved between zebrafish and mammals. We generated lrp5- knock-out to study its role skeletogenesis homeostasis. Loss lrp5 leads craniofacial deformities mineral density (total body head) at adult ages. To understand the mechanism consequences phenotypes, we performed transcriptome analysis cranium mutants siblings. Enrichment revealed upregulation significantly associated hydrolase activity: mmp9, mmp13a, acp5a . encodes Tartrate-resistant acid phosphatase (TRAP) which commonly used an osteoclast marker, Matrix metalloprotease 9, Mmp9, known be secreted by osteoclasts stimulate resorption. These point changes differentiation regulated analyze these functionally, assessed dynamics increased TRAP staining, larger resorption areas, developmental skeletal dysmorphologies mutant, suggesting higher resorptive activity absence Lrp5 signaling. Our findings support maintaining unexpected insights into function homeostasis through moderation function.

Язык: Английский

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Disruption of the foxe1 gene in zebrafish reveals conserved functions in development of the craniofacial skeleton and the thyroid

Frontiers in Cell and Developmental Biology, Год журнала: 2023, Номер 11

Опубликована: Март 13, 2023

Introduction: Mutations in the FOXE1 gene are implicated cleft palate and thyroid dysgenesis humans. Methods: To investigate whether zebrafish could provide meaningful insights into etiology of developmental defects humans related to FOXE1, we generated a mutant that has disruption nuclear localization signal foxe1 gene, thereby restraining access transcription factor. We characterized skeletal development thyroidogenesis these mutants, focusing on embryonic larval stages. Results: Mutant larvae showed aberrant phenotypes ceratohyal cartilage had reduced whole body levels Ca, Mg P, indicating critical role for early development. Markers bone (precursor) cells were differentially expressed mutants post-migratory cranial neural crest pharyngeal arch at 1 dpf, induction chondrogenesis 3 dpf start endochondral formation 6 dpf. Foxe1 protein was detected differentiated follicles, suggesting factor thyroidogenesis, but follicle morphology or differentiation unaffected mutants. Discussion: Taken together, our findings highlight conserved show differential signaling osteogenic chondrogenic genes mutation.

Язык: Английский

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From multiallele fish to nonstandard environments, how ZFIN assigns phenotypes, human disease models, and gene expression annotations to genes

Genetics, Год журнала: 2023, Номер 224(1)

Опубликована: Март 2, 2023

Danio rerio is a model organism used to investigate vertebrate development. Manipulation of the zebrafish genome and resultant gene products by mutation or targeted knockdown has made good system for investigating function, providing resource genetic contributors phenotype human disease. Phenotypic outcomes can be result mutation, products, manipulation experimental conditions, any combination thereof. Zebrafish have been in various chemical screens identify environmental disease outcomes. The Information Network (ZFIN, zfin.org) central repository genetic, genomic, phenotypic data that from research using D. rerio. Here we describe how ZFIN annotates phenotype, expression, across designs, computationally determine wild-type gene, these results allow us propagate correct related entity.

Язык: Английский

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Crispant analysis in zebrafish as a tool for rapid functional screening of disease-causing genes for bone fragility

Опубликована: Окт. 7, 2024

Heritable Fragile Bone Disorders (FBDs) encompass a spectrum of conditions, from widespread multifactorial to rare monogenic diseases, all characterized by an elevated risk fractures. The process validating causative genes and elucidating their pathogenic mechanisms remains daunting resource-intensive task. In this study, we evaluated the feasibility semi-high throughput zebrafish screening platform for rapid validation in vivo functional testing candidate disease-causing wide range heritable FBDs. Six associated with severe recessive forms Osteogenesis Imperfecta (OI) four BMD, key osteoporosis indicator, identified through genome-wide association studies (GWAS) were selected. crispant approach, based on CRISPR/Cas9 technology, was used phenotype directly F0 mosaic founder zebrafish. Next-Generation Sequencing (NGS) analysis revealed mean indel efficiency 88% across ten different crispants, indicating high proportion knock-out alleles thus resembling stable models. We applied multiple techniques evaluate skeletal characteristics at 7, 14 90 days post-fertilization (dpf), including microscopy osteoblast reporter visualization mineralization Alizarin Red S staining, microCT quantitative analysis. While larval crispants exhibited variable differences osteoblast-positive mineralized surface areas, adult-stage displayed more pronounced consistent phenotypes. Notably, developed malformed neural haemal arches, majority presenting vertebral fractures fusions, some showing significant alterations bone volume density. addition, aldh7a1 mbtps2 experienced increased mortality due deformities. RT-qPCR differentiation formation markers stages indicated differential expression osteogenic bglap col1a1a substantial portion hinting utility as biomarkers FBD screening. summary, our findings demonstrate that offers viable efficient strategy assessment genes. advocate comprehensive approach integrates various evaluates distinct molecular profiles developmental adult stages. This methodology has potential provide new insights into role these biology.

Язык: Английский

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The genetic overlap between osteoporosis and craniosynostosis

Frontiers in Endocrinology, Год журнала: 2022, Номер 13

Опубликована: Сен. 26, 2022

Osteoporosis is the most prevalent bone condition in ageing population. This systemic disease characterized by microarchitectural deterioration of bone, leading to increased fracture risk. In past 15 years, genome-wide association studies (GWAS), have pinpointed hundreds loci associated with mineral density (BMD), helping elucidate underlying molecular mechanisms and genetic architecture However, challenge remains pinpointing causative genes driving GWAS signals as a pivotal step drawing translational therapeutic roadmap. Recently, skull BMD-GWAS uncovered an intriguing intersection craniosynostosis, congenital anomaly due premature suture fusion skull. Here, we recapitulate contribution both osteoporosis describing biological underpinnings this overlap using zebrafish models leverage functional investigation development skeletal homeostasis.

Язык: Английский

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Intermediate cells of in vitro cellular reprogramming and in vivo tissue regeneration require desmoplakin

Science Advances, Год журнала: 2022, Номер 8(43)

Опубликована: Окт. 28, 2022

Amphibians and fish show considerable regeneration potential via dedifferentiation of somatic cells into blastemal cells. In terms dedifferentiation, in vitro cellular reprogramming has been proposed to share common processes with vivo tissue regeneration, although the details are elusive. Here, we identified cytoskeletal linker protein desmoplakin (Dsp) as a factor mediating both regeneration. Our analysis revealed that Dsp expression is elevated distinct intermediate during reprogramming. Knockdown impedes induced pluripotent stem neural stem/progenitor well zebrafish fins. Notably, reduced impairs formation These findings suggest there Dsp-mediated evolutionary link between mammals lower vertebrates may provide alternative approaches for mammalian regenerative therapy.

Язык: Английский

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Animal Models of Tuberculosis

Springer eBooks, Год журнала: 2023, Номер unknown, С. 139 - 170

Опубликована: Янв. 1, 2023

Язык: Английский

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MOXD1 is a lineage-specific gene and a tumor suppressor in neuroblastoma

Science Advances, Год журнала: 2024, Номер 10(25)

Опубликована: Июнь 21, 2024

Neuroblastoma is a childhood developmental cancer; however, its embryonic origins remain poorly understood. Moreover, in-depth studies of early tumor-driving events are limited because the lack appropriate models. Herein, we analyzed RNA sequencing data obtained from human neuroblastoma samples and found that loss expression trunk neural crest–enriched gene MOXD1 associates with advanced disease worse outcome. Further, by using single-cell cells fetal adrenal glands creating in vivo models zebrafish, chick, mouse, show determinate tumor development. In addition, highly conserved restricted to mesenchymal Schwann cell precursors during healthy Our findings identify as lineage-restricted tumor-suppressor neuroblastoma, potentiating further stratification these tumors development novel therapeutic interventions.

Язык: Английский

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Identification of Rare LRP5 Variants in a Cohort of Males with Impaired Bone Mass

International Journal of Molecular Sciences, Год журнала: 2021, Номер 22(19), С. 10834 - 10834

Опубликована: Окт. 7, 2021

Osteoporosis is the most common bone disease characterized by reduced mass and increased fragility. Genetic contribution one of main causes primary osteoporosis; therefore, both genders are affected this skeletal disorder. Nonetheless, osteoporosis in men has received little attention, thus being underestimated undertreated. The aim study was to identify novel genetic variants a cohort 128 males with idiopathic low using next-generation sequencing (NGS) panel including genes whose mutations could result mineral density (BMD). analysis detected eleven patients ten rare heterozygous within LRP5 gene, which were categorized as VUS (variant uncertain significance), likely pathogenic benign according American College Medical Genetics Genomics (ACMG) guidelines. Protein structural Bayesian performed on identified pointed out p.R1036Q p.R1135C pathogenic, therefore suggesting association these two phenotype. In conclusion, expands our understanding importance functional protein formation highlights necessity sequence gene subjects BMD.

Язык: Английский

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