Supergene Research Articles

Insights into lncRNA-mediated regulatory networks in Hevea brasiliensis under anthracnose stress

In recent years, long non-coding RNAs (lncRNAs) and microRNAs (miRNAs) have emerged as critical regulators in plant biology, governing complex gene regulatory networks. In the context of disease resistance in Hevea brasiliensis, the rubber tree, significant progress has been made in understanding its response to anthracnose disease, a serious threat posed by fungal pathogens impacting global rubber tree cultivation and latex quality. While advances have been achieved in unraveling the genetic and molecular foundations underlying anthracnose resistance, gaps persist in comprehending the regulatory roles of lncRNAs and miRNAs under such stress conditions. The specific contributions of these non-coding RNAs in orchestrating molecular responses against anthracnose in H. brasiliensis remain unclear, necessitating further exploration to uncover strategies that increase disease resistance. Here, we integrate lncRNA sequencing, miRNA sequencing, and degradome sequencing to decipher the regulatory landscape of lncRNAs and miRNAs in H. brasiliensis under anthracnose stress. We investigated the genomic and regulatory profiles of differentially expressed lncRNAs (DE-lncRNAs) and constructed a competitive endogenous RNA (ceRNA) regulatory network in response to pathogenic infection. Additionally, we elucidated the functional roles of HblncRNA29219 and its antisense hbr-miR482a, as well as the miR390-TAS3-ARF pathway, in enhancing anthracnose resistance. These findings provide valuable insights into plant-microbe interactions and hold promising implications for advancing agricultural crop protection strategies. This comprehensive analysis sheds light on non-coding RNA-mediated regulatory mechanisms in H. brasiliensis under pathogen stress, establishing a foundation for innovative approaches aimed at enhancing crop resilience and sustainability in agriculture.

Construction of the porcine genome mobile element variations and investigation of its role in population diversity and gene expression

BackgroundMobile element variants (MEVs) have a significant and complex impact on genomic diversity and phenotypic traits. However, the quantity, distribution, and relationship with gene expression and complex traits of MEVs in the pig genome remain poorly understood.ResultsWe constructed the most comprehensive porcine MEV library based on high-depth whole genome sequencing (WGS) data from 747 pigs across 59 breeds worldwide. This database identified a total of 147,993 polymorphic MEVs, including 121,099 short interspersed nuclear elements (SINEs), 26,053 long interspersed nuclear elements (LINEs), 802 long terminal repeats (LTRs), and 39 other transposons, among which 54% are newly discovered. We found that MEVs are unevenly distributed across the genome and are strongly influenced by negative selection effects. Importantly, we identified 514, 530, and 584 candidate MEVs associated with population differentiation, domestication, and breed formation, respectively. For example, a significantly differentiated MEV is located in the ATRX intron between Asian and European pigs, whereas ATRX is also differentially expressed between Asian and European pigs in muscle tissue. In addition, we identified 4,169 expressed MEVs (eMEVs) significantly associated with gene expression and 6,914 splicing MEVs (sMEVs) associated with gene splicing based on RNA-seq data from 266 porcine liver tissues. These eMEVs and sMEVs explain 6.24% and 9.47%, respectively, of the observed cis-heritability and highlight the important role of MEVs in the regulation of gene expression. Finally, we provide a high-quality SNP–MEV reference haplotype panel to impute MEV genotypes from genome-wide SNPs. Notably, we identified a candidate MEV significantly associated with total teat number, demonstrating the functionality of this reference panel.ConclusionsThe present investigation demonstrated the importance of MEVs in pigs in terms of population diversity, gene expression and phenotypic traits, which may provide useful resources and theoretical support for pig genetics and breeding.

PTPN23-dependent ESCRT machinery functions as a cell death checkpoint

Cell death plasticity is crucial for modulating tissue homeostasis and immune responses, but our understanding of the molecular components that regulate cell death pathways to determine cell fate remains limited. Here, a CRISPR screen of acute myeloid leukemia cells identifies protein tyrosine phosphatase non-receptor type 23 (PTPN23) as essential for survival. Loss of PTPN23 activates nuclear factor-kappa B, apoptotic, necroptotic, and pyroptotic pathways by causing the accumulation of death receptors and toll-like receptors (TLRs) in endosomes. These effects are recapitulated by depletion of PTPN23 co-dependent genes in the endosomal sorting complex required for transport (ESCRT) pathway. Through proximity-dependent biotin labeling, we show that NAK-associated protein 1 interacts with PTPN23 to facilitate endosomal sorting of tumor necrosis factor receptor 1 (TNFR1), sensitizing cells to TNF-α-induced cytotoxicity. Our findings reveal PTPN23-dependent ESCRT machinery as a cell death checkpoint that regulates the spatiotemporal distribution of death receptors and TLRs to restrain multiple cell death pathways.

TSC2-mTORC1 axis regulates morphogenesis and neurological function of Gli1+ adult-born dentate granule cells.

Aberrant adult hippocampal neurogenesis is implicated in neurological and mood disorders associated with dysregulation of the mechanistic target of rapamycin (mTOR). Understanding how the mTOR pathway shapes the functional development of different subpopulations of adult-born hippocampal neural stem cells will enable insight into potential therapeutic pathways for these disorders. Here we study how loss of TSC2, a regulator of mTOR pathway and a causal gene for Tuberous Sclerosis Complex (TSC), affects dentate gyrus granule cell (dGC) morphogenesis and hippocampal-dependent function. We found that Tsc2KO mice with TSC2 specifically ablated from Gli1+ adult-born neural stem cells showed neuronal hypertrophy, reduced NEUN expression, increased dendritic arborization, premature cellular senescence, and hypervascularization of the dentate gyrus (DG). Neurologically, Tsc2KO mice showed altered exploratory behavior, impaired spatial learning, abnormal contextual recall, and hypersensitivity to kainic acid-induced seizures. Importantly, genetic reduction of Raptor, essential for mTORC1 signaling, rebalanced mTORC1 signaling and mitigated molecular, cellular, and neurological deficits in Tsc2KO mice. This study uncovered functions of TSC2 in Gli1+ adult-born neural stem cells and highlights RAPTOR as a potential therapeutic target for reversing disease features associated with TSC2 mutations.

Gnas Knockdown Induces Obesity and AHO Features in Early Zebrafish Larvae

GNAS (Guanine Nucleotide-Binding Protein, Alpha Stimulating) is a complex gene that encodes the alpha subunit of the stimulatory G protein (Gsα), critical for signaling through various G protein-coupled receptors. Inactivating genetic and epigenetic changes in GNAS, resulting in Gsα deficiency, cause different variants of pseudohypoparathyroidism, which may manifest features of Albright hereditary osteodystrophy (AHO, a syndrome characterized by early-onset obesity and other developmental defects). Recent findings have linked Gsα deficiency with isolated, severe, early-onset obesity, suggesting it as a potential, underrecognized cause of monogenic, non-syndromic obesity. This study was prompted by identifying several GNAS variants of uncertain significance (VUSs) in pediatric patients presenting with unexplained, severe, early-onset obesity at Sidra Medicine in Qatar. To functionally characterize these variants, we developed the first zebrafish model of Gsα deficiency, offering numerous advantages over other model systems. This was achieved by knockdown of the ortholog through microinjection of translation-blocking Morpholino antisense oligonucleotides into the yolks of 1-8-cell-stage zebrafish embryos. The morphant larvae displayed an obese phenotype, marked by significantly enlarged yolk sacs, increased neutral lipid accumulation, and reduced metabolic rates, among other developmental abnormalities resembling those in AHO. This zebrafish model lays the foundation for efficient functional characterization of GNAS VUSs and paves the way for enhancing our understanding of Gsα deficiency-associated early-onset obesity.

Heterozygote advantage can explain the extraordinary diversity of immune genes.

The majority of highly polymorphic genes are related to immune functions and with over 100 alleles within a population, genes of the major histocompatibility complex (MHC) are the most polymorphic loci in vertebrates. How such extraordinary polymorphism arose and is maintained is controversial. One possibility is heterozygote advantage (HA), which can in principle maintain any number of alleles, but biologically explicit models based on this mechanism have so far failed to reliably predict the coexistence of significantly more than ten alleles. We here present an eco-evolutionary model showing that evolution can result in the emergence and maintenance of more than 100 alleles under HA if the following two assumptions are fulfilled: first, pathogens are lethal in the absence of an appropriate immune defence; second, the effect of pathogens depends on host condition, with hosts in poorer condition being affected more strongly. Thus, our results show that HA can be a more potent force in explaining the extraordinary polymorphism found at MHC loci than currently recognized.

Allele Diversity of the DRB1 Gene of the Major Histocompatibility Complex in Two Species of Hamsters of the Genus Allocricetulus Argyropulo 1932

Allele Diversity of the DRB1 Gene of the Major Histocompatibility Complex in Two Species of Hamsters of the Genus Allocricetulus Argyropulo 1932

A comprehensive analysis of gene expression and the immune landscape in gastric cancer through single-cell and multi-omics approaches

Gastric cancer (GC) is a common malignant tumor worldwide, characterized by complex biological processes. The distribution of various cell types and gene expression profiles in the GC microenvironment remains unclear. This study uses single-cell RNA sequencing to explore gene expression patterns and identify differentially expressed genes in GC samples, offering new insights into cellular diversity and potential molecular mechanisms. We conducted temporal and clustering analyses with single-cell sequencing, followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses to clarify their functions. Using machine learning, we identified relevant genes to create highly accurate prediction models. Additionally, ssGSEA analysis provided detailed insights into the immunosuppressive tumor microenvironment, revealing complex gene expression interactions and diverse immune infiltrates in cancer. Correlation analysis highlighted TIMP1 as having significant prognostic value across different immune cell subtypes. Single-cell RNA sequencing revealed the cellular landscape and gene expression profiles of the GC microenvironment, offering crucial data on how cell heterogeneity is regulated in relation to the tumor microenvironment. Moreover, new insights into the expression levels of AGT, INHBA, and TIMP1 showed distinct sex-biased gene functions within the tumor microenvironment. These findings enhance our understanding of the molecular mechanisms associated with gastric cancer development and may lay the groundwork for identifying novel therapeutic targets and diagnostic strategies.

Val66Met Polymorphism of the BDNF Gene and Work Intellectual Complexity: Associations with Speed Characteristics of Cognitive Activity in Aging

The high heterogeneity of cognitive aging is explained by the influence of both genetic and environmental factors. It has been shown that increasing cognitive reserve prevents the development of aging-related cognitive impairment. Education and level of professional activity are considered external factors in the formation of cognitive reserve. BDNF (brain-derived neurotrophic factor) is a neurotrophic factor involved in the processes of plasticity of the mature brain. A polymorphism (Val66Met) of the BDNF gene is associated with differential expression of BDNF, suggesting its potential role in the on cognitive training outcomes. The associations between Val66Met polymorphism and the effectiveness of mental training caused by and work intellectual complexity throughout adult life (in our study, a comparison of scientists - SA and people not associated with professional scientific activities — NSA remain unstudied. The objective of the study was to assess the modulating effect of these factors in relation to aging-related changes in the processes of attention and figurative creativity in models that allow us to consider both the efficiency and speed aspects of activity. The study involved 257 healthy young and 162 elderly Caucasians belonging to the groups of SA and NSA. It has been shown that the Val66Met polymorphism of the BDNF gene is associated with the influence of the level of professional activity on the efficiency of cognitive functions only in elderly subjects. At a higher intensity of intellectual activity (SA), Val/Val, but not Val/Met carriers, showed a reduction in the average reaction time in the Attention network test and an increase in fluency when testing figurative creativity compared to similar indicators of elderly subjects in the NSA group. The results obtained indicate greater plasticity of cognitive functions in carriers of the Val/Val genotype and may be used for prediction and development of methods for differentiated correction of age-related cognitive decline.

Combined transcriptomic, metabolomic and physiological analysis reveals the key role of nitrogen, but not phosphate and potassium in regulating anthocyanin biosynthesis induced by nutrient deficiency in Eucalyptus

Anthocyanin biosynthesis in Eucalyptus plants can be flexibly and rapidly modulated in response to hormones or environmental stimuli, including nutrient deprivation (ND). However, the underlying mechanism of ND in inducing anthocyanin biosynthesis in plants remains largely elusive. In this study, we discovered that anthocyanin levels in leaves and stems could well reflect nitrogen availability in Eucalyptus. Supplementation with nitrogen, but not with phosphate or potassium, effectively inhibited ND-induced anthocyanin biosynthesis. To further investigate how nitrogen regulates this process, comprehensive time-resolved transcriptomic and targeted metabolomic analyses were conducted. The results revealed that 3405, 2706, and 3153 genes were differentially regulated by nitrogen treatment at 1, 2, and 3 d, respectively. Pathway analysis indicated the majority of the enriched KEGG pathways were associated with cellular metabolism, such as amino acid and flavonoid biosynthesis. Moreover, metabolomic analysis showed that the most abundantly accumulated anthocyanins, cyanidin-3-O-galactoside and cyanidin-3-O-glucoside, along with key intermediates in the flavonoid pathway, were downregulated by nitrogen treatment. Furthermore, nitrogen-responsive MYB-bHLH-WDR complex genes were identified, including six EgrMYB113 family members. Overexpression of one of the EgrMYB113-like genes induced anthocyanin biosynthesis in the transgenic hairy roots of Eucalyptus. Interestingly, mutation of AtMYB113 inhibited nitrogen deficiency-induced anthocyanin biosynthesis in Arabidopsis, suggesting that MYB113 is a novel N-responsive MYB transcriptional regulator of anthocyanin biosynthesis. Additionally, nitrogen treatment upregulated a few GA biosynthesis genes while simultaneously downregulated the expression of several GA2ox genes. Exogenous application of GA3 decreased ND-induced anthocyanin biosynthesis. Conclusively, this study provides novel insights into the molecular mechanism of nitrogen in the regulation of anthocyanin biosynthesis in Eucalyptus.

Mechanisms of nano zero-valent iron in enhancing dibenzofuran degradation by a Rhodococcus sp.: Trade-offs between ATP production and protection against reactive oxygen species

Nano zero-valent iron (nZVI) can enhance pollutants biodegradation, but it displays toxicity towards microorganisms. Gram-positive (G+) bacteria exhibit greater resistance to nZVI than Gram-negative bacteria. However, mechanisms of nZVI accelerating pollutants degradation by G+ bacteria remain unclear. Herein, we explored effects of nZVI on a G+ bacterium, Rhodococcus sp. strain p52, and mechanisms by which nZVI accelerates biodegradation of dibenzofuran, a typical polycyclic aromatic compound. Electron microscopy and energy dispersive spectroscopy analysis revealed that nZVI could penetrate cell membranes, which caused damage and growth inhibition. nZVI promoted dibenzofuran biodegradation at certain concentrations, while higher concentration functioned later due to the delayed reactive oxygen species (ROS) mitigation. Transcriptomic analysis revealed that cells adopted response mechanisms to handle the elevated ROS induced by nZVI. ATP production was enhanced by accelerated dibenzofuran degradation, providing energy for protein synthesis related to antioxidant stress and damage repair. Meanwhile, electron transport chain (ETC) was adjusted to mitigate ROS accumulation, which involved downregulating expression of ETC complex I-related genes, as well as upregulating expression of the genes for the ROS-scavenging cytochrome bd complex and ETC complex II. These findings revealed the mechanisms underlying nZVI-enhanced biodegradation by G+ bacteria, offering insights into optimizing bioremediation strategies involving nZVI.

Network hub gene detection using the entire solution path information.

Gene co-expression networks typically comprise modules and their associated hub genes, which are regulating numerous downstream interactions within the network. Methods for hub screening, as well as data-driven estimation of hub co-expression networks using graphical models, can serve as useful tools for identifying these hubs. Graphical model-based penalization methods typically have one or multiple regularization terms, each of which encourages some favorable characteristics (e.g., sparsity, hubs, power-law) to the estimated complex gene network. It is common practice to find a single optimal graphical model corresponding to a specific value of the regularization parameter(s). However, instead of doing this, one could aggregate information across several graphical models, all of which depend on the same data set, along the solution path in the hub gene detection process. We propose a novel method for detecting hub genes that utilizes the information available in the solution path. Our procedure is related to stability selection, but we replace resampling with a simple statistic. This procedure amalgamates information from each node of the data-driven graphical models into a single influence statistic, similar to Cook's distance. We call this statistic the Mean Degree Squared Distance (MDSD). Our simulation and empirical studies demonstrate that the MDSD statistic maintains a good balance between false positive and true positive hubs. An R package MDSD is publicly available on GitHub under the General Public License https://github.com/markkukuismin/MDSD.

The Bi-Steric Inhibitor RMC-5552 Reduces mTORC1 Signaling and Growth in Lymphangioleiomyomatosis.

Mutations in the Tuberous Sclerosis Complex (TSC) genes result in the hyperactivation of the mechanistic/mammalian target of rapamycin 1 (mTORC1) growth pathway in mesenchymal pulmonary cells. Rapamycin (SirolimusTM), a naturally occurring macrolide, is the only therapeutic approved for women with lymphangioleiomyomatosis (LAM), a progressive, destructive lung disease caused by TSC gene mutations and mTORC1 hyperactivation. However, on cessation of the drug, lung function decline continues. We demonstrated here that pulmonary LAM cancer stem-like cells (SLS) most highly expressed the eukaryotic translation initiation factor 4E (eIF4E)-dependent translation initiation genes. We also showed that the eukaryotic initiation factor 4E-binding protein 1 (4E-BP1) gene has the lowest expression in these cells, indicating that the 4E-BP1/eIF4E ratio in LAM SLS cells favors unrestrained eIF4E oncogenic mRNA translation. The bi-steric mTORC1-selective compound RMC-5552 prevented growth of LAM-associated fibroblasts (LAFs) and phosphorylation of proteins in the ribosomal protein S6K1/ribosomal protein S6 (S6K1/S6) and 4E-BP1/eIF4E translation mTORC1-driven pathways, whereas rapamycin only blocked the S6K/S6 axis. Rapamycin inhibition of LAF growth was rapidly reversed, but RMC-5552 inhibition was more durable. RMC-5552, through its potential to eradicate LAM cancer SLS cells, may have therapeutic benefit in LAM and other diseases with mTORC1 hyperactivity.

Evolution of 3D Chromatin Folding.

Studies examining the evolution of genomes have focused mainly on sequence conservation. However, the inner working of a cell implies tightly regulated crosstalk between complex gene networks controlled by small dispersed regulatory elements of physically contacting DNA regions. How these different levels of chromatin organization crosstalk in different species underpins the potential for genome evolutionary plasticity. We review the evolution of chromatin organization across the Animal Tree of Life. We introduce general aspects of the mode and tempo of genome evolution to later explore the multiple layers of genome organization. We argue that both genome and chromosome size modulate patterns of chromatin folding and that chromatin interactions facilitate the formation of lineage-specific chromosomal reorganizations, especially in germ cells. Overall, analyzing the mechanistic forces involved in the maintenance of chromatin structure and function of the germ line is critical for understanding genome evolution, maintenance, and inheritance.

WebTWAS 2.0: update platform for identifying complex disease susceptibility genes through transcriptome-wide association study.

Transcriptome-wide association study (TWAS) has successfully identified numerous complex disease susceptibility genes in the post-genome-wide association study (GWAS) era. Over the past 3 years, the focus of TWAS algorithms has shifted from merely identifying associations to understanding how single nucleotide polymorphisms(SNPs)regulate gene expression, with a growing emphasis on incorporating fine-mapping techniques. Additionally, the rapid increase in GWAS summary statistics, driven largely by the UK Biobank and other consortia, has made it essential to update our webTWAS resource. To address these challenges and meet the growing needs of researchers, we developed webTWAS 2.0, an updated platform for identifying susceptibility genes for human complex diseases using TWAS. Additionally, webTWAS 2.0 provides an online TWAS analysis tool that simplifies conducting TWAS analyses. The updated resource includes 7247 GWAS summary statistics covering 1588 complex human diseases from 192 publications. It also incorporates multiple TWAS methods, such as sTF-TWAS, 3'aTWAS and GIFT, along with an updated interactive visualization tool that allows users to easily explore significant associations across different methods. Other upgrades include a personalized online analysis tool for user-submitted GWAS data and a refined search function that makes it easier to identify relevant associations and meet diverse user needs more efficiently. webTWAS 2.0 is freely accessible at http://www.webtwas.net.

EXTH-04. COMBINATION OF BEVACIZUMAB ENHANCES THE ANTI-TUMOR EFFICACY OF AD-SGE-REIC FOR GLIOMA

Abstract Reduced expression in immortalized cells/Dickkopf-3 (REIC/Dkk-3) is a tumor suppressor gene and its overexpression has been shown to exert anti-tumor effects as a therapeutic target gene in many human cancers. Recently, we revealed the anti-glioma effects of an adenoviral vector carrying REIC/Dkk-3 with the super gene expression system (Ad-SGE-REIC) and conducted the phase Ⅰ/Ⅱ a clinical trial for recurrent malignant glioma (jRCT2063190013). Anti-vascular endothelial growth factor treatments such as bevacizumab have demonstrated convincing therapeutic efficacy in patients with glioma. In this study, we examined the effects of Ad-SGE-REIC on glioma treated with bevacizumab in vitro and in vivo. Ad-SGE-REIC treatment resulted in a significant reduction in the number of invasion cells treated with bevacizumab. Western blot analyses revealed the increased expression of several endoplasmic reticulum stress markers in cells treated with both bevacizumab and Ad-SGE-REIC, as well as decreased β-catenin protein levels. In glioma-bearing mouse models, survival was extended in the combination therapy group. These results suggest that the combination therapy of Ad-SGE-REIC and bevacizumab showed anti-glioma effects by suppressing the angiogenesis and invasion of tumor cells. Combined Ad-SGE-REIC and bevacizumab might be a promising strategy for the treatment of malignant glioma.

Symmetry and Complexity in Gene Association Networks Using the Generalized Correlation Coefficient

High-dimensional gene expression data cause challenges for traditional statistical tools, particularly when dealing with non-linear relationships and outliers. The present study addresses these challenges by employing a generalized correlation coefficient (GCC) that incorporates a flexibility parameter, allowing it to adapt to varying levels of symmetry and asymmetry in the data distribution. This adaptability is crucial for analyzing gene association networks, where the GCC demonstrates advantages over traditional measures such as Kendall, Pearson, and Spearman coefficients. We introduce two novel adaptations of this metric, enhancing its precision and broadening its applicability in the context of complex gene interactions. By applying the GCC to relevance networks, we show how different levels of the flexibility parameter reveal distinct patterns in gene interactions, capturing both linear and non-linear relationships. The maximum likelihood and Spearman-based estimators of the GCC offer a refined approach for disentangling the complexity of biological networks, with potential implications for precision medicine. Our methodology provides a powerful tool for constructing and interpreting relevance networks in biomedicine, supporting advancements in the understanding of biological interactions and healthcare research.

Polygenic risk for alcohol use disorder affects cellular responses to ethanol exposure in a human microglial cell model.

Polygenic risk scores (PRSs) assess genetic susceptibility to alcohol use disorder (AUD), yet their molecular implications remain underexplored. Neuroimmune interactions, particularly in microglia, are recognized as notable contributors to AUD pathophysiology. We investigated the interplay between AUD PRS and ethanol in human microglia derived from iPSCs from individuals with AUD high-PRS (diagnosed with AUD) or low-PRS (unaffected). Ethanol exposure induced elevated CD68 expression and morphological changes in microglia, with differential responses between high-PRS and low-PRS microglial cells. Transcriptomic analysis revealed expression differences in MHCII complex and phagocytosis-related genes following ethanol exposure; high-PRS microglial cells displayed enhanced phagocytosis and increased CLEC7A expression, unlike low-PRS microglial cells. Synapse numbers in cocultures of induced neurons with microglia after alcohol exposure were lower in high-RPS cocultures, suggesting possible excess synapse pruning. This study provides insights into the intricate relationship between AUD PRS, ethanol, and microglial function, potentially influencing neuronal functions in developing AUD.

Evidence of a Lytic Pathway in an Invertebrate Complement System: Identification of a Terminal Complement Complex Gene in a Colonial Tunicate and Its Evolutionary Implications.

The complement system is a pivotal component of innate immunity, extensively studied in vertebrates but also present in invertebrates. This study explores the existence of a terminal complement pathway in the tunicate Botryllus schlosseri, aiming to understand the evolutionary integration of innate and adaptive immunity. Through transcriptome analysis, we identified a novel transcript, BsITCCP, encoding a protein with both MACPF and LDLa domains-a structure resembling that of vertebrate C9 but with a simpler organization. Phylogenetic reconstruction positions BsITCCP between invertebrate perforins and vertebrate terminal complement proteins, suggesting an evolutionary link. Localization studies confirmed that bsitccp is transcribed in cytotoxic morula cells (MCs), which are also responsible for producing other complement components like BsC3, BsMBL, BsMASP, and BsBf. Functional assays demonstrated that bsitccp transcription is upregulated in response to nonself challenges and is dependent on BsC3 activity; inhibition of BsC3 led to a significant reduction in BsITCCP expression. Electron microscopy revealed that MCs form contact with perforated yeast cells, indicating a possible mechanism of cell lysis similar to the immunological synapse observed in vertebrates. These findings suggest that a C3-governed lytic complement pathway exists in B. schlosseri, challenging the assumption that a C5 ortholog is necessary for such a pathway. This work enhances our understanding of the evolution of the complement system and suggests that invertebrates possess a terminal complement complex capable of mediating cell lysis, regulated by C3. Future studies will focus on confirming the pore-forming ability of BsITCCP and its role in the immunological synapse.

Variants in MICOS10 Identified by Whole Genome Sequencing and RNA Sequencing in a New Type of Hepatocerebral Mitochondrial DNA Depletion Syndrome.

The mitochondrial contact site and cristae organising system (MICOS) complex is required for cristae formation and is composed of seven proteins. Among the genes of MICOS complex, variants of MICOS13, IMMT and APOO have been reported to cause diseases. We report a case in which whole genome sequencing identified a variant of the MICOS10 gene associated with mitochondrial hepatopathy along with mitochondrial DNA depletion. We identified the deletion g.19596826_19601303del and the single nucleotide variant c.173G>C (p.Cys58Ser). The deletion including exon 1 might have caused complete loss of gene expression, indicating monoallelic expression from RNA sequencing. MIC10 was lost at the protein level in the patient's fibroblasts, and mitochondrial oxygen consumption was impaired. These were restored by overexpression of MICOS10 in the patient's fibroblasts. Taken together, these findings indicate that MICOS10 is a causative gene for hepatopathy and neuropathy, a disease very similar to that associated with MICOS13.