Impaired healing of skin wounds can lead to fibrotic changes, which in turn result in a violation of the mechanical properties of the tissue. The YAP signaling is actively involved in the regeneration process, which makes it possible to use this protein as a target for therapy. The investigation is focused on the potential of utilizing a suspension of primary keratinocytes (KC) and human dermal papilla (DP) cells to stimulate scar-free regeneration of human skin. The effect was then compared with that of the YAP inhibitor verteporfin (VP). The results of the study indicate the important role of YAP in the regenerative process, since its activation was associated with the formation of a fibrotic phenotype in the skin, while pronounced inhibition of the signaling led to impaired regeneration and the inability to restore normal tissue morphology. The findings demonstrate that the suspension of KC + DP cells contributes to the inhibition of YAP activity, leading to a decrease in the expression of extracellular matrix (ECM) by scar fibroblasts in vitro. The injection of cell suspension in the in vivo model into the dermis of xenografted human skin also led to a decrease in activity of the YAP, but did not cause its complete suppression, which contributed to the approximation of skin morphology to a normal state. Concurrently, the expression of ECM elements remained unchanged, while inflammatory changes exhibited a decrease in intensity. Furthermore, the epidermis underwent a process of restoration, and graft vascularization demonstrated a marked improvement. The study indicates the potential of using KC + DP cells to stimulate normal skin regeneration by regulating the activity of the YAP signaling.
Spinocerebellar ataxia type 27B (SCA27B) is a hereditary ataxia caused by a pathogenic expansion of GAA repeats (>250) in the first intron of the FGF14 gene, which encodes fibroblast growth factor 14. The clinical presentation includes slowly progressive cerebellar ataxia, often combined with sensory or sensorimotor neuropathy, bilateral vestibulopathy, nystagmus, paroxysmal diplopia, dizziness, and characteristic atrophy of the cerebellar vermis and hemispheres. To study the pathogenic mechanisms of SCA27B in a cellular model, we generated induced pluripotent stem cell (iPSC) (lines RCPCMi016-A and RCPCMi016-B) from fibroblasts of a patient with an expansion of GAA repeats in the FGF14 gene. The iPSCs exhibited typical pluripotent stem cell morphology and expression of pluripotency markers. Pluripotency was confirmed by differentiation into derivates of all three germ layers. The iPSC lines displayed normal karyotype (46,XX).
The endometrium of mammals is characterized by a high degree of physiological plasticity and undergoes cyclic remodeling, which requires precise coordination of hormonal signals and the activity of stromal progenitor cells. In the stromal tissue of the mouse endometrium, CD90+ cells are identified, which possess progenitor properties and are involved in the implantation process. One of the proposed regulators of their functional state is local thyroid signaling mediated by type 2 deiodinase (DIO2). Additionally, the Notch signaling pathway is considered a key mediator that controls cell differentiation and maintains the progenitor status of cells. There is reason to believe that thyroid hormones can modulate the activity of the Notch signaling pathway; however, the relationship between thyroid activation, Notch, and the regulation of CD90+ stromal cells remains unexplored. The objective of the study was to determine the mechanisms regulating endometrial stromal cells, specifically the influence of sex steroid hormones on the CD90+ population and the role of thyroid signaling. Using molecular biology methods, it was established that Dio2+ cells, predominantly represented by CD90+ fibroblasts, exhibit increased expression of decidualization genes. In vitro and in vivo analyses showed that the Notch signaling pathway acts as a necessary intermediary for the DIO2-mediated effects of thyroid hormones on uterine tissue. The findings deepen the understanding of hormonal regulation of stromal progenitor cells and the plasticity of the endometrium during the implantation period.
Induced pluripotent stem cell (iPSC) line FAMRCi016-A was generated by reprogramming peripheral blood mononuclear cells (PBMCs) from a patient with the genetic variant FLNC:c.3557C>T p.(Ala1186Val). The iPSC cell line was characterized according to the general standards: the cells had a typical morphology, a normal karyotype (46,XY), expressed pluripotent cell markers (OCT4, NANOG, SOX2, SSEA4), and demonstrated the ability to produce derivatives of three germ layers by directed differentiation. The iPSC line FAMRCi016-A can be used in studies investigating the pathogenic genetic variant FLNC Ala1186Val.
Developmental and epileptic encephalopathy-17 (DEE17) is a severe inherited neurological disorder caused by mutations in the GNAO1 gene. The GNAO1 gene encodes the alpha subunit of heterotrimeric G-protein complexes and is involved in signal transduction from G-protein coupled receptors in the brain. The precise pathogenesis of the disease remains unclear, and optimal treatment strategies depend on the specific mutation. Therefore, generating models for studying GNAO1 encephalopathy is highly relevant. In this work, using genetic reprogramming, we generated induced pluripotent stem cells (iPSCs) from dermal fibroblasts of a patient diagnosed with DEE17 caused by the c.155A>G (p.Q52R) mutation in the GNAO1 gene. The established iPSCs line (RCPCMi015-A) exhibits iPSCs morphology, a normal karyotype, and is free of bacterial contamination. RCPCMi015-A cells express pluripotency markers and are capable of differentiating into derivatives of the three germ layers. Thus, we have created a cell model for studying the molecular mechanisms of GNAO1 encephalopathy and for testing potential therapeutic approaches.
Over the past 6–8 million years, in the evolutionary line leading to modern humans, the brain has undergone significant changes. Its volume has tripled compared to that of other great apes, and the increased structural complexity has been associated with a marked prolongation of developmental stages. The emergence of these specific morphological features is linked to profound alterations in the genetic programs regulating brain development. Changes in regulatory regions of the human genome, which can modify the spatiotemporal patterns of gene expression, have a significant impact on this evolutionary process. One of the drivers of such evolutionary changes is human accelerated regions (HARs), which represent conserved DNA regions in mammals that have acquired specific mutations in humans. The overwhelming majority of these elements are non-coding DNA sequences, localized in intronic and intergenic regions near genes essential for brain development. Genetic variants within HARs have been associated with neurodevelopmental and psychiatric disorders and are known to alter gene expression. One gene whose regulation may be influenced by HARs is CNTN6. Two HARs have been identified within its introns and frequently overlap with CNTN6 copy number variations observed in patients with neurodevelopmental disorders. To further investigate the functional role of one of these elements, HARsv2_1747, in human neurodevelopment, we used the CRISPR/Cas9 system to generate two human induced pluripotent stem cell (iPSC) lines: one with a homozygous deletion and another with a compound heterozygous deletion of this region. Both iPSC lines fulfill the key criteria for pluripotency, as they form colonies with typical pluripotent cell morphology, maintain a normal diploid karyotype, express pluripotency markers, and retain the ability to differentiate into derivatives of all three germ layers.
Genomic changes, including the emergence of multiple paralogs of regulatory genes with diverging functions, form the foundation of phenotypic transformations in evolution. Such events are most likely to be traceable in phylogenetically basal groups, which among vertebrates include jawless vertebrates (cyclostomes) and ancient jawed vertebrates (gnathostomes)—cartilaginous fishes. In the present study, we investigated the phylogeny, local genomic synteny, and spatial expression patterns of the chordin-like2 gene in the river lamprey Lampetra fluviatilis, a representative of cyclostomes, and compared it with the chordin-like gene of the gray catshark, a representative of cartilaginous fishes, the basal clade of gnathostomes. We found that chordin-like genes in lampreys and cartilaginous fishes are not orthologous but instead illustrate a case of hidden paralogy. The chordin-like2 gene identified in lampreys represents an ancient Bilateria gene, being present in the genomes of some protostomes. In lamprey, chordin-like2 activity is observed at prelarval stages and is associated with the development of specific structures, including the endostyle and the tail bud, which markedly distinguishes it from the chordin-like1 gene of gnathostomes. Analysis of local genomic synteny indicates that chordin-like1, a novel gene of gnathostomes, may have arisen through duplication of chordin-like2.
CANVAS syndrome is a rare neurodegenerative disorder belonging to the group of ataxias with impaired DNA repair. It is characterized by degeneration of the sensory ganglia, posterior columns of the spinal cord, and the cerebellum. CANVAS is caused by a biallelic expansion of AAGGG pentanucleotide repeats in the second intron of the RFC1 gene, which encodes a subunit of the RFC complex involved in DNA replication and repair. To study the pathogenesis mechanisms of CANVAS in a cell model, we generated iPSC line RCPCMi017-A (IPSCANVAS1E6) from fibroblasts of a patient with a biallelic AAGGG expansion in the RFC1 gene and a diagnosis of CANVAS. The iPSCs exhibited typical stem cell morphology and expression of pluripotency markers. The pluripotency of the iPSCs was confirmed by their ability to differentiate into cells derived from the three germ layers. The iPSC line had a normal karyotype (46,XX).
Research on stem cell differentiation increasingly focuses on the vesicular paracrine component, which facilitates intercellular communication and defines cellular functional status. This study compares the content (mRNA and proteins) of extracellular vesicles (EVs) from human induced pluripotent stem cells (iPSCs) with the molecular profile of the cells themselves during directed neural differentiation. We found that changes in EV content mirrored the cell differentiation trajectory: expression of pluripotency marker genes (Oct4, Sox2, cMyc, Nanog) decreased, while markers of neural differentiation (Pax6, Nestin, Tubb, Map2, S100B, GFAP) appeared. Mass spectrometry analysis revealed a functional diversification in the EV proteome, reflecting various stages of differentiation—from regulatory processes to the functional specification of neural derivatives.
Genomic rearrangements, including the emergence of multiple paralogs of regulatory genes, may serve as the genetic foundation for the appearance of novel structures and traits during evolution. In the course of subsequent evolution, some paralogs arising through duplications may be lost due to functional redundancy. In the present study, we investigated chordin-like gene paralogs in cartilaginous and teleost fishes. It was found that, in contrast to most jawed vertebrates, whose genomes contain two chordin-like paralogs, cartilaginous and most teleost fishes have retained only a single paralog of this gene. At the same time, it turned out that the retained chordin-like genes in cartilaginous and teleost fishes are not orthologs but instead represent an example of hidden paralogy. The chordin-like1 gene is expressed in fins, as well as in jaw and branchial structures, in representatives of cartilaginous fishes and sturgeons, which constitute basal groups of jawed vertebrates. This may indicate the role of this gene in the emergence of these morphological innovations of jawed vertebrates at early stages of the group’s evolution. The secondary loss of chordin-like1 in teleosts, in turn, may be associated with the transformation of the endoskeletal organization of their fins and the loss of the metapterygial basal element.
Induced pluripotent stem cell lines (iPSC) FAMRCi013-A, FAMRCi013-B, and FAMRCi013-C were generated by reprogramming peripheral blood mononuclear cells from a patient with the genetic variant PKP2:c.355delT. The iPSC lines were characterized according to the general standards and showed the pluripotent cells’ properties: they had characteristic morphology, normal karyotype (46,XY), expressed markers typical for pluripotent cells (OCT4, NANOG, SOX2, SSEA4), and had the ability to produce derivatives of three germ layers during directed differentiation. These lines can be used to study the likely pathogenic variant PKP2:c.355delT associated with arrhythmogenic right ventricular cardiomyopathy.
West syndrome, first described in 1841, belongs to a group of infantile epileptic encephalopathies. To date, several missense and nonsense mutations in various genes have been associated with this disorder, including the missense variant p.Arg87Cys in the CYFIP2 gene. More than 50
For the first time, an analysis of the parasite and endosymbiont fauna of the larval forms of the moor frog (Rana arvalis Nilsson, 1842) at different stages of ontogenesis in natural and urban landscapes of the Middle Urals is presented. Six species of helminths of the phyla Nematoda and Platyhelminthes at different developmental stages and one species of endosymbiont (phylum Bigyra) were found. The greatest infestation of R. arvalis tadpoles and juveniles with the intestinal trematode O. ranae of different age groups was noted. Nematodes were found in metamorphosed amphibians. Along the urbanization gradient, parasite communities in tadpoles and froglets become simplified, which is likely associated not only with the biology of the developmental stages of R. arvalis but also with the specifics of urban ecosystems. Five types of anomalies were identified in R. arvalis larvae at different stages of ontogenesis, which apparently may be initiated by mass infection of amphibians with metacercariae of the trematode O. ranae.
To assess the reaction of trees, including conifers, which differ in the physical and mechanical properties of wood, to external factors, it is especially important to study the development of phloem and xylem cells and the deposition of biomass in tree trunks growing in the same exogenous conditions. The phenology of the cambium, the development of phloem and xylem cells and the deposition of biomass in wood annual layers in the trunks of Scots pine, Siberian larch and Siberian spruce, growing under the same exogenous conditions in Eastern Siberia, were studied. It was shown that during ontogenesis each species demonstrated its own seasonal dynamics of cambium activity in the production of phloem and xylem cells and the accumulation of biomass in tracheids walls. Of the three species studied, the largest number of cells produced by the cambium was found in larch. Spruce had the least activity in terms of these growth characteristics. During periods of maximum growth activity, the accumulation of biomass in the walls of larch xylem cells always exceeded these indicators in pine and especially spruce. The differences in xylogenesis of pine, larch and spruce are the consequence of species-specific features of their metabolism and its plasticity in response to changes in the external factors.
Hypertrophic cardiomyopathy (HCM) is one of the most common cardiovascular diseases and occurs with frequency 1 : 500. Up to 60
Neurodevelopmental disorders, particularly autism spectrum disorder (ASD) and intellectual disability (ID), comprise a heterogeneous group characterized by a range of clinical phenotypes that are influenced by both genetic and environmental factors. ASD is notable for its diverse behavioral manifestations and varying cognitive profiles, which complicate the processes of diagnosis and intervention. In this study, we reprogrammed blood mononuclear cells from a patient with ASD, ID and multiple structural variations into induced pluripotent stem cells (iPSCs) using episomal vector transfection technology. The resulting iPSC line, designated ICGi058-A, demonstrated robust proliferation as compact colonies of cells with large nuclei and maintained karyotype 46,XY,inv(2)(p21q23);t(3;7)(p13;q11.2). Characterization of these iPSCs revealed the expression of key pluripotency markers, including OCT4, SOX2, NANOG and TRA-1-60. Moreover, the functional pluripotency of ICGi058-A was confirmed through the formation of embryoid bodies and successful differentiation into the three germ layers: ectoderm, mesoderm, and endoderm. Comprehensive genetic analysis (STR) showed that the derived iPSC line is identical to the original cell material obtained from the patient.
The mouse digit tip regeneration model is one of the most phylogenetically close models to the human, and studying it at the cellular and transcriptomic levels may allow the development of novel approaches in regenerative medicine. Two papers with single-cell RNA sequencing applied to this model have been published, but none of them aimed to analyze differential gene expression between blastema and uninjured cells. In our study, we aim to conduct this analysis. All data processing stages, including cell filtering, clusterization, integration, and differential gene expression analysis were carried out in R with the aid of the Seurat package. Based on the presence of intermediate phenotypes after clusterization, we found out that epithelial-mesenchymal and endothelial-mesenchymal transitions may occur during digit tip regeneration, but not in uninjured samples. Differential gene expression analysis yielded about 1000 genes between stromal cells in blastema and uninjured digits in both analyzed datasets. These genes were associated both with morphogenetic processes: ossification, angiogenesis, extracellular matrix organization, and cell adhesion, and with a number of metabolic processes. We also selected from this differentially expressed gene set transcription and paracrine factors, among which putative regulators of the described processes were found. Further research on the presented here transcription and paracrine factors may elucidate the mechanisms of digit tip regeneration.
Тhis study represents the first stage of a research project investigating the potential of heat shock as a tool for purifying fish genomes of slightly deleterious mutations. Experimental offspring of common carp (Cyprinus carpio L.) were obtained by fertilizing eggs with sperm treated with the chemical mutagen N-ethyl-N-nitrosourea (ENU). The half-lethal dose (LD50) of ENU was determined to be 1.6 ± 0.8 mM. Optimal heat-shock conditions, resulting in 50
H2O2 exposure causes oxidative stress, which plays a role in oocyte aging. Folic acid (FA), which is regarded as an antioxidant, can effectively scavenge oxidising free radicals. MicroRNAs (miRNAs) play an important role in regulating gene expression during oocyte development and maturation and may be involved in the H2O2-exposure-induced oxidative stress in oocytes. In the present study, we explored if FA protects oocytes from reactive oxygen species (ROS) in vitro and the role of microRNAs (miRNAs) in this process. We compared the oocyte quality, ROS levels, mitochondrial membrane potential (MMP), 2 pronucleus (2PN) ratios, embryonic developmental potential, and miRNA expression in fresh mouse oocytes, H2O2-treated mouse oocytes, and FA and H2O2-treated mouse oocytes. The results demonstrated that FA could significantly reduce the ROS levels and fragments of H2O2-treated mouse oocytes and increase superoxide dismutase (SOD) levels, alleviating the H2O2-mediated oocyte 2PN ratio decreasing. H2O2-treated oocytes showed increased miR-10a expression, whereas FA treatment decreased miR-10a expression. In conclusion, miR-10a plays a vital role in the prevention of oxidative stress in oocytes.
The honey bee (Apis mellifera) and the silkworm (Bombyx mori) are not only important agricultural targets but also valuable model organisms for biomedical and genetic research. They have unique characteristics that allow the study of different biological processes, such as parthenogenesis and polyploidy, which contribute to the development of new technologies in genetics and breeding. The works of academician B.L. Astaurov on the creation of fertile tetraploid lines of silkworms using artificial parthenogenesis laid the foundation for further research in the field of insect genetics, opening new horizons for industrial silk production. Modern research confirms the possibility of using parthenogenesis and polyploidy in bees to create stable breeds, increase productivity and protect against disease. According to Vavilov’s law of homologous series, forms of hereditary variability can arise in closely related species, expanding the prospects for further use of parthenogenesis and polyploidy in subspecies of bees. This phenomenon can be of great importance for breeding programmes aimed at improving the efficiency of beekeeping, which is particularly important under conditions of mass hybridisation. The introduction of methods of artificial oocyte activation for parthenogenesis (teletokia, arrenotokia), developed on the basis of approaches of B.L. Astaurov, can significantly increase the stability of silkworm and honeybee populations, which will contribute to the development of agricultural sustainability, molecular genetic breeding and conservation of biodiversity.
