Nuclear Gene Research Articles

Recessive MECR pathogenic variants cause an LHON-like optic neuropathy

BackgroundLeber’s hereditary optic neuropathy (LHON) is a mitochondrial disorder characterised by complex I defect leading to sudden degeneration of retinal ganglion cells. Although typically associated with pathogenic variants in mitochondrial...

Revisiting gene typing and phylogeny of Trypanosoma cruzi reference strains: Comparison of the relevance of mitochondrial DNA, single-copy nuclear DNA, and the intergenic region of mini-exon gene

Chagas disease is a widespread neglected disease in Latin America. Trypanosoma cruzi, the causative agent of the disease, is currently subdivided into six DTUs (discrete typing units) named TcI-TcVI, and although no clear association has been found between parasite genetics and different clinical outcomes of the disease or different transmission cycles, genetic characterization of T. cruzi strains remains crucial for integrated epidemiological studies. Numerous markers have been used for this purpose, although without consensus. These include mitochondrial genes, single or multiple-copy nuclear genes, ribosomal RNA genes, and the intergenic region of the repeated mini-exon gene. To increase our knowledge of these gene sequences and their usefulness for strain typing, we sequenced fragments of three mitochondrial genes, nine single-copy nuclear genes, and the repeated intergenic part of the mini-exon gene by Next Generation Sequencing (NGS) on a sample constituted of 16 strains representative of T. cruzi genetic diversity, to which we added the corresponding genetic data of the 38 T. cruzi genomes fully sequenced until 2022.Our results show that single-copy nuclear genes remain the gold standard for characterizing T. cruzi strains; the phylogenetic tree from concatenated genes (3959 bp) confirms the six DTUs previously recognized and provides additional information about the alleles present in the hybrid strains. In the tree built from the three mitochondrial concatenated genes (1274 bp), three main clusters are identified, including one with TcIII, TcIV, TcV, and TcVI DTUs which are not separated. Nevertheless, mitochondrial markers remain necessary for detecting introgression and heteroplasmy. The phylogenetic tree built from the sequence alignment of the repeated mini-exon gene fragment (327 bp) displayed six clusters, but only TcI was associated with a single cluster. The sequences obtained from strains belonging to the other DTUs were scattered into different clusters. Therefore, while the mini-exon marker may bring, for some biological samples, some advantages in terms of sensibility due to its repeated nature, mini-exon sequences must be used with caution and, when possible, avoided for T. cruzi typing and phylogenetic studies.

Revolutionizing Gene Therapy: Biomaterials as Enabling Tools for Targeted Delivery and Enhanced Efficacy

Bioengineered smart nano/biomaterials have emerged as promising tools for efficient and targeted gene delivery in gene therapy. Gene therapy has the potential to treat various genetic disorders, cancers, and infectious diseases by replacing, repairing, or regulating the expression of disease-causing genes. However, the success of gene therapy critically depends on the efficient delivery of therapeutic genes into target cells or tissues while minimizing off-target effects and toxicity. Bioengineered smart nano/biomaterials offer several advantages over conventional gene delivery vehicles, such as viral vectors and cationic lipids, including improved stability, biocompatibility, and tunable properties. This review provides an overview of bioengineered smart nano/biomaterials for gene delivery, including their design, fabrication, mechanisms of action, and applications in gene therapy. The design and fabrication of smart nano/biomaterials involve the synthesis and functionalization of various materials, such as polymers, lipids, peptides, and inorganic nanoparticles, to optimize their physicochemical properties and biological interactions. Mechanistically, smart nano/biomaterials can efficiently deliver DNA/RNA cargo into target cells by overcoming biological barriers, such as cellular membranes and endosomes, and facilitating nuclear entry and gene expression. Applications of smart nano/biomaterials in gene therapy include the treatment of genetic disorders, cancers, and infectious diseases, as well as targeted gene therapy for specific cell types or tissues. However, several challenges remain to be addressed before the clinical translation of smart nano/biomaterials for gene therapy, including safety, toxicity, and immunogenicity issues, optimization of delivery efficiency and specificity, and regulatory and ethical considerations. In conclusion, bioengineered smart nano/biomaterials hold great promise for the development of safe and effective gene therapies that can revolutionize the treatment of various diseases.

Mitochondrially mediated RNA interference, a retrograde signaling system affecting nuclear gene expression.

Several functional classes of short noncoding RNAs are involved in manifold regulatory processes in eukaryotes, including, among the best characterized, miRNAs. One of the most intriguing regulatory networks in the eukaryotic cell is the mito-nuclear crosstalk: recently, miRNA-like elements of mitochondrial origin, called smithRNAs, were detected in a bivalve species, Ruditapes philippinarum. These RNA molecules originate in the organelle but were shown in vivo to regulate nuclear genes. Since miRNA genes evolve easily de novo with respect to protein-coding genes, in the present work we estimate the probability with which a newly arisen smithRNA finds a suitable target in the nuclear transcriptome. Simulations with transcriptomes of 12 bivalve species suggest that this probability is high and not species specific: one in a hundred million (1 × 10-8) if five mismatches between the smithRNA and the 3' mRNA are allowed, yet many more are allowed in animals. We propose that novel smithRNAs may easily evolve as exaptation of the pre-existing mitochondrial RNAs. In turn, the ability of evolving novel smithRNAs may have played a pivotal role in mito-nuclear interactions during animal evolution, including the intriguing possibility of acting as speciation trigger.

Abstract PO-011: Analysis of nuclear receptor expression in head and neck cancer

Abstract Background: The nuclear receptor (NR) superfamily contains a wide array of transcription factors responsible for cellular metabolism, inflammation, differentiation, and hormonal functions. For 45 years these receptors have been of interest in head and neck squamous cell carcinoma (HNSCC) treatment and prevention, especially retinoic acid receptor (RAR), peroxisome proliferator-activated receptor (PPAR), and vitamin D receptor (VDR) families. In the current study we examined expression patterns of these receptors in The Cancer Genome Atlas (TCGA). Methods: TCGA RNA-Seq data within the cBioportal platform comparing HNSCC samples (n=515 patients/samples) to normal tissue (n=82 patients/samples) was interrogated for significant differences in nuclear receptor expression. Genes with a z-score relative to normal samples (log RNA-Seq V2 RSEM normalized) greater than +2 or less than -2 in ≥50% of cases were considered significant. Unpaired, two-tailed t-tests were used to analyze comparisons in nuclear receptor genes’ z-scores across multiple clinical and pathologic parameters, p<0.05 was deemed significant. Statistical stringency for differential gene expression conformed to cBioportal and the University of Alabama at Birmingham’s Cancer Data Analysis Portal (UALCAN). Results: Of the 46 NR genes and 19 NR cofactors examined, ninety nine percent of tumor samples in the TCGA had some form of NR gene ‘alteration’ compared to normal tissue. These alterations predominantly encompass expression up- or down-regulation. NR genes Peroxisome Proliferator Activated Receptor Gamma (PPARγ) and Retinoic Acid Receptor Related Orphan Receptor C (RORC) (both in the thyroid hormone receptor-like family), and the NR cofactor, Nuclear Receptor Coactivator 1 (NCOA1), were differentially expressed and downregulated in tumors compared to normal tissue. Differences in specific NR expression varied significantly with HPV status (30 NRs), organ site (oral cavity vs hypopharynx & larynx; 20 NRs), tumor size (T1/2 vs. T3/4; 21 NRs), gender (male vs. female; 7 NRs); tumor staging (stage I/II vs. stage III/IV; 6 NRs); lymph node status (N0 vs. N+; 5 NRs), smoking history (smokers vs. non-smokers; 3 NRs), and patient race (White vs. Black/African American; 2 NRs). Conclusions: In prior studies of nuclear receptors in aerodigestive cancer, the retinoid receptor axis was downregulated, and a subsequent goal was stimulation and restoration of activity with natural or synthetic retinoids as a rationale for treatment. Presently, we have discovered significant decreases in PPARγ expression and are targeting this receptor with thiazolidinediones in a series of preclinical and human trials in a similar rationale to promote restoration of normal signaling and events like maturation. Here, we observed significant differences in a variety of parameters of several receptors associated with risk factors, stage, race, and gender that are observed and may become the focus of interest in future targeting efforts. Citation Format: Lindsey Mortensen, Beverly R. Wuertz, Frank G. Ondrey. Analysis of nuclear receptor expression in head and neck cancer [abstract]. In: Proceedings of the AACR-AHNS Head and Neck Cancer Conference: Innovating through Basic, Clinical, and Translational Research; 2023 Jul 7-8; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2023;29(18_Suppl):Abstract nr PO-011.

Wing pattern diversity in Eunica butterflies (Nymphalidae: Biblidinae): phylogenetic analysis implies decoupled adaptive trends in dorsal sexual dimorphism and ventral eyespot evolution.

Butterfly eyespots are wing patterns reminiscent of vertebrate eyes, formed by concentric rings of contrastingly coloured scales. Eyespots are usually located close to the wing margin and often regarded as the single most conspicuous pattern element of butterfly wing colour displays. Recent efforts to understand the processes involved in the formation of eyespots have been driven mainly by evo-devo approaches focused on model species. However, patterns of change implied by phylogenetic relationships can also inform hypotheses about the underlying developmental mechanisms associated with the formation or disappearance of eyespots, and the limits of phenotypic diversity occurring in nature. Here we present a combined evidence phylogenetic hypothesis for the genus Eunica, a prominent member of diverse Neotropical butterfly communities, that features notable variation among species in eyespot patterns on the ventral hind wing surface. The data matrix consists of one mitochondrial gene region (COI), four nuclear gene regions (GAPDH, RPS5, EF1a and Wingless) and 68 morphological characters. A combined cladistic analysis with all the characters concatenated produced a single most parsimonious tree that, although fully resolved, includes many nodes with modest branch support. The phylogenetic hypothesis presented corroborates a previously proposed morphological trend leading to the loss of eyespots, together with an increase in the size of the conserved eyespots, relative to outgroup taxa. Furthermore, wing colour pattern dimorphism and the presence of androconia suggest that the most remarkable instances of sexual dimorphism are present in the species of Eunica with the most derived eyespot patterns, and are in most cases accompanied by autapomorphic combinations of scent scales and "hair pencils". We discuss natural and sexual selection as potential adaptive explanations for dorsal and ventral wing patterns.

Origin of CMS-PET1 cytotype in cultivated sunflower: A new insight

The vast majority of commercial sunflower hybrids worldwide are produced using cytoplasmic male sterility (CMS) of the PET1 type, resulting from the interspecific hybridization of Helianthus petiolaris with Helianthus annuus. Due to the fact that CMS-PET1 was not previously detected in wild sunflower, it was believed that this cytotype could arise during interspecific hybridization and is specific solely for cultivated sunflower. In this study, the open reading frame, orfH522, associated with the CMS-PET1 phenotype, was revealed for the first time in the 3′-flanking region of the mitochondrial atpA gene in wild H. annuus. An analysis of whole genome data from 1089 accessions showed that the frequency of occurrence of CMS-orfH522 in wild H. annuus populations is 3.58%, while in wild H. petiolaris populations, it is 1.26%. In general, the analysis demonstrated that PET1-CMS is a natural cytotype of H. annuus, and the appearance of the CMS phenotype in cultivated sunflowers is associated with the loss of stabilizing nuclear genes of fertility restorers, which occurred during interspecific hybridization. These data can explain the patterns of differential cytoplasmic and nuclear introgression occurring in wild sunflower and are useful for further evolutionary studies.

Transcriptomic and genomic identification of spliceosomal genes from Euglena gracilis.

Diverse splicing types in nuclear and chloroplast genes of protist Euglena gracilis have been recognized for decades. However, the splicing machinery responsible for processing nuclear precursor messenger RNA introns, including trans-splicing of the 5' terminal outron and spliced leader (SL) RNA, remains elusive. Here, we identify 166 spliceosomal protein genes and two snRNA genes from E. gracilis by performing bioinformatics analysis from a combination of next-generation and full-length transcriptomic RNA sequencing (RNAseq) data as well as draft genomic data. With the spliceosomal proteins we identified in hand, the insensitivity of E. gracilis to some splicing modulators is revealed at the sequence level. The prevalence of SL RNA-mediated trans-splicing is estimated to be more than 70% from our full-length RNAseq data. Finally, the splicing proteomes between E. gracilis and its three evolutionary cousins within the same Excavata group are compared. In conclusion, our study characterizes the spliceosomal components in E. gracilis and provides the molecular basis for further exploration of underlying splicing mechanisms.

Nuclear pore complex and nucleocytoplasmic transport disruption in neurodegeneration.

Nuclear pore complexes (NPCs) play a critical role in maintaining the equilibrium between the nucleus and cytoplasm, enabling bidirectional transport across the nuclear envelope, and are essential for proper nuclear organization and gene regulation. Perturbations in the regulatory mechanisms governing NPCs and nuclear envelope homeostasis have been implicated in the pathogenesis of several neurodegenerative diseases. The ESCRT-III pathway emerges as a critical player in the surveillance and preservation of well-assembled, functional NPCs, as well as nuclear envelope sealing. Recent studies have provided insights into the involvement of nuclear ESCRT-III in the selective reduction of specific nucleoporins associated with neurodegenerative pathologies. Thus, maintaining quality control of the nuclear envelope and NPCs represents a pivotal element in the pathological cascade leading to neurodegenerative diseases. This review describes the constituents of the nuclear-cytoplasmic transport machinery, encompassing the nuclear envelope, NPC, and ESCRT proteins, and how their structural and functional alterations contribute to the development of neurodegenerative diseases.

AKIR-1 regulates proteasome subcellular function in Caenorhabditis elegans

Polyubiquitinated proteins are primarily degraded by the ubiquitin-proteasome system (UPS). Proteasomes are present both in the cytoplasm and nucleus. Here, we investigated mechanisms coordinating proteasome subcellular localization and activity in a multicellular organism. We identified the nuclear protein-encoding gene akir-1 as a proteasome regulator in a genome-wide Caenorhabditis elegans RNAi screen. We demonstrate that depletion of akir-1 causes nuclear accumulation of endogenous polyubiquitinated proteins in intestinal cells, concomitant with slower invivo proteasomal degradation in this subcellular compartment. Remarkably, akir-1 is essential for nuclear localization of proteasomes both in oocytes and intestinal cells but affects differentially the subcellular distribution of polyubiquitinated proteins. We further reveal that importin ima-3 genetically interacts with akir-1 and influences nuclear localization of a polyubiquitin-binding reporter. Our study shows that the conserved AKIR-1 is an important regulator of the subcellular function of proteasomes in a multicellular organism, suggesting a role for AKIR-1 in proteostasis maintenance.

Evolution of the expression and regulation of the nuclear hormone receptor ERR gene family in the chordate lineage

The Estrogen Related Receptor (ERR) nuclear hormone receptor genes have a wide diversity of roles in vertebrate development. In embryos, ERR genes are expressed in several tissues, including the central and peripheral nervous systems. Here we seek to establish the evolutionary history of chordate ERR genes, their expression and their regulation. We examine ERR expression in mollusc, amphioxus and sea squirt embryos, finding the single ERR orthologue is expressed in the nervous system in all three, with muscle expression also found in the two chordates. We show that most jawed vertebrates and lampreys have four ERR paralogues, and that vertebrate ERR genes were ancestrally linked to Estrogen Receptor genes. One of the lamprey paralogues shares conserved expression domains with jawed vertebrate ERRγ in the embryonic vestibuloacoustic ganglion, eye, brain and spinal cord. Hypothesising that conserved expression derives from conserved regulation, we identify a suite of pan-vertebrate conserved non-coding sequences in ERR introns. We use transgenesis in lamprey and chicken embryos to show that these sequences are regulatory and drive reporter gene expression in the nervous system. Our data suggest an ancient association between ERR and the nervous system, including expression in cells associated with photosensation and mechanosensation. This includes the origin in the vertebrate common ancestor of a suite of regulatory elements in the 3’ introns that drove nervous system expression and have been conserved from this point onwards.

Genetic divergence of farmed blue mussels (Mytilus sp.) in Australian waters

As valuable ecosystem engineers, blue mussels, are one of the most ecologically and economically important bivalve species in Australia. The need for understanding levels of genetic diversity in both wild and farmed populations of Australian mussels is ever-growing due to the increased risk of disease and decreased fitness associated with the constantly changing global environment. This study, therefore, investigated the phylogenetic relationship, and the genetic variation of farmed blue mussels (Mytilus sp., previously noted as M. galloprovincialis, M. planulatus or a hybrid of both) at four sites along the south coast of Australia, Victoria, South Australia, Western Australia, and Tasmania. Analyses of two genes, Cytochrome c oxidase subunit 1 gene (mtCOI) and the nuclear gene Elongation factor 1 alpha (EF1a) was completed for all mussels sampled. The phylogenetic reconstruction of mtDNA sequences was not able to cluster the samples into species. The analyses of the nuclear marker (EF1a) returned similar results, except that we could conclude the Australian samples not being M. chilensis. Furthermore, the analyses of EF1a revealed the presence of hybridisation, a pattern commonly observed in mussels. The genetic diversity analysis of mtCOI demonstrated that samples from Victoria and Western Australia had the highest genetic diversity, while South Australia had the lowest. When analysing the nuclear EF1a sequences, mussels from Western Australian had the lowest genetic diversity, and the Victorian samples had the highest.The study extended our knowledge on the population genetic variation of farmed blue mussels (Mytilus sp.) along the south coast of Australia. The identified genetic differences between the four southern states of Australia could be interesting for the aquaculture industry when aiming for more adaptable and profitable stocks.

Molecular phylogeny of the land snail family Euconulidae in Thailand and its position in the superfamily Trochomorphoidea (Stylommatophora: Limacoidei), with description of a new genus

The Euconulidae is a globally distributed land snail family but there is no record of this family from Thailand. In this study, we describe a new genus, Siamoconus gen. nov., based on comparative studies of the shell characteristics, external morphology of animals, radula, genital and spermatophore structures, and molecular phylogeny. We performed phylogenetic analyses of two mitochondrial gene fragment (cytochrome c oxidase I, COI; and 16S rRNA) and one nuclear (28S rRNA) gene fragment to clarify the relationships with other euconulid genera and its position in the superfamily Trochomorphoidea. We also analysed shell morphometrics, male genitalia and mantle pigmentation of Siamoconus gen. nov., confirming the status of three new species with keeled shells (S. boreas sp. nov., S. coleus sp. nov. and S. geotrochoides sp. nov.) and one new species with a rounded shell (S. destitutus sp. nov.). These new species are restricted to limestone areas in northern and north-eastern Thailand. We also re-examined the status of other genera in the superfamily Trochomorphoidea based on shell features, body pigmentation, radula, genital anatomy, spermatophore morphology, and a phylogenetic analysis of all available trochomorphoidean DNA sequences. Our analysis suggests that the family Geotrochidae, previously synonymised with the Trochomorphidae, should be resurrected to represent the genus Geotrochus from Borneo, and this family is retrieved as the sister clade of the Dyakiidae. ZooBank: urn:lsid:zoobank.org:pub:C5205F4E-5DDE-42E6-A532-761CAFE099C3

Johansenicoccus eremophilus gen. et sp. nov., a novel evolutionary lineage in Chlorophyceae with unusual genomic features

Background – Green algae are a diverse group of photosynthetic eukaryotes, yet are still vastly understudied compared to land plants. For many years, green algae were characterized based on their morphology and life cycles. More recently, phylogenetic and genomic analyses have been added to the phycological toolkit for a better understanding of algal biodiversity and evolutionary history. Material and methods – A desert strain of green algae was isolated from Joshua Tree National Park (JTNP) in southern California as part of a larger biodiversity survey. The alga’s nuclear rRNA genes as well as the chloroplast genome were sequenced, annotated, and analysed in addition to a morphological assessment. Results – Morphologically this strain is especially similar to Pseudomuriella and Rotundella, and its lipid profile resembles that of other soil algae, but phylogenomic analyses demonstrate that it is a distinct evolutionary lineage in Chlorophyceae. The alga exhibits several unusual genomic features, the most remarkable being its highly derived yet apparently functional nuclear rRNA genes, 18S and 28S. Both genes are GC-rich and bear many compensatory base changes to maintain a similar secondary structure to that of other green algae. The chloroplast genome has a distinct gene order and repeat arrangement from other published green algal plastomes, but contains the expected genes and also provides phylogenetically informative data. Conclusion – We conclude that the strain be placed into a new species and genus in the class Chlorophyceae, and propose the name Johansenicoccus eremophilus for this new taxon. Johansenicoccus eremophilus exemplifies science’s insufficient understanding of the range of genomic variations among inconspicuous soil algae.

Calcarisporium yuanyangense sp. nov. (Calcarisporiaceae, Hypocreales), a fungal pathogen isolated from Ophiocordyceps nutans

In this study, a new species, Calcarisporium yuanyangense, was isolated from Ophiocordyceps nutans collected from Yuanyang County, Yunnan Province, China, and described by morphology and molecular phylogeny. Based on the combined dataset of five genes including the internal transcribed spacer gene region (ITS), the nuclear ribosomal small and large subunit gene regions (nrSSU and nrLSU), the gene encoding translation elongation factor 1-α (tef-1α), and the RNA polymerase II second largest subunit gene region (rpb2), phylogenetic analyses were performed by the maximum likelihood (ML) and the Bayesian inference (BI) methods to determine the phylogenetic position of C. yuanyangense in Calcarisporium. This species was clustered with C. cordycipiticola, C. xylariicola, and C. arbuscula to form a separate branch. Calcarisporium yuanyangense had the basic morphological characteristics of Calcarisporium, but still differed morphologically from the other species in the genus. This species was similar to C. arbuscula in conidia shape, but the conidia of C. yuanyangense were larger and had a higher length/width ratio than those of C. arbuscula. This study deepens the understanding of the host diversity and the distribution of this taxonomic unit, as well as provides important taxonomic data for further exploration of the genus Calcarisporium.

A Heterolepidoderma and Halichaetoderma gen. nov. (Gastrotricha: Chaetonotidae) riddle: integrative taxonomy and phylogeny of six new freshwater species from Central Europe

Abstract Heterolepidoderma is a widely distributed gastrotrich genus encompassing 22 freshwater and 15 marine species. In the present study, two new Heterolepidoderma and four new Heterolepidoderma-like species were discovered in the inland waters of Central Europe. Although Heterolepidoderma-like species exhibited all the morphological features typical of Heterolepidoderma, both nuclear and mitochondrial genes robustly showed their sister-group relationship to the marine genus Halichaetonotus. Most Heterolepidoderma species, including the new ones, were grouped with some Chaetonotus and Ichthydium species in a strongly supported cluster (so-called Heterolepidoderma s.l. clade). Since statistical tree topology tests robustly refuted the close affinity of Heterolepidoderma-like species to the Heterolepidoderma s.l. clade, a new genus, Halichaetoderma gen. nov., was proposed for them. Stochastic mapping suggested that the last common ancestor (LCA) of the Chaetonotidae‒Neogosseidae‒Dasydytidae clade had scales carrying keels with spines. Scales became oblong in the LCA of the Halichaetoderma + Halichaetonotus clade and three times independently within the Heterolepidoderma s.l. clade. Spines were lost in the LCAs of the Halichaetoderma + Halichaetonotus and Heterolepidoderma s.l. clades convergently. State-by-state associations of cuticular characters suggested that particular scale morphologies tend to be correlated with particular numbers of keels and the presence/absence of spines during the chaetonotid phylogeny.

Nightmare or delight: Taxonomic circumscription meets reticulate evolution in the phylogenomic era

Phylogenetic studies in the phylogenomics era have demonstrated that reticulate evolution greatly impedes the accuracy of phylogenetic inference, and consequently can obscure taxonomic treatments. However, the systematics community lacks a broadly applicable strategy for taxonomic delimitation in groups characterized by pervasive reticulate evolution. The red-fruit genus, Stranvaesia, provides an ideal model to examine the influence of reticulation on generic circumscription, particularly where hybridization and allopolyploidy dominate the evolutionary history. In this study, we conducted phylogenomic analyses integrating data from hundreds of single-copy nuclear (SCN) genes and plastomes, and interrogated nuclear paralogs to clarify the inter/intra-generic relationship of Stranvaesia and its allies in the framework of Maleae. Analyses of phylogenomic discord and phylogenetic networks showed that allopolyploidization and introgression promoted the origin and diversification of the Stranvaesia clade, a conclusion further bolstered by cytonuclear and gene tree discordance. With a well-inferred phylogenetic backbone, we propose an updated generic delimitation of Stranvaesia and introduce a new genus, Weniomeles. This new genus is distinguished by its purple-black fruits, thorns trunk and/or branches, and a distinctive fruit core anatomy characterized by multilocular separated by a layer of sclereids and a cluster of sclereids at the top of the locules. Through this study, we highlight a broadly-applicable workflow that underscores the significance of reticulate evolution analyses in shaping taxonomic revisions from phylogenomic data.

Establishing induced pluripotent stem cell lines from two dominant optic atrophy patients with distinct OPA1 mutations and clinical pathologies.

Dominant optic atrophy (DOA) is an inherited disease that leads to the loss of retinal ganglion cells (RGCs), the projection neurons that relay visual information from the retina to the brain through the optic nerve. The majority of DOA cases can be attributed to mutations in optic atrophy 1 (OPA1), a nuclear gene encoding a mitochondrial-targeted protein that plays important roles in maintaining mitochondrial structure, dynamics, and bioenergetics. Although OPA1 is ubiquitously expressed in all human tissues, RGCs appear to be the primary cell type affected by OPA1 mutations. DOA has not been extensively studied in human RGCs due to the general unavailability of retinal tissues. However, recent advances in stem cell biology have made it possible to produce human RGCs from pluripotent stem cells (PSCs). To aid in establishing DOA disease models based on human PSC-derived RGCs, we have generated iPSC lines from two DOA patients who carry distinct OPA1 mutations and present very different disease symptoms. Studies using these OPA1 mutant RGCs can be correlated with clinical features in the patients to provide insights into DOA disease mechanisms.

Mitochondrial Factors in the Cell Nucleus.

The origin of eukaryotic organisms involved the integration of mitochondria into the ancestor cell, with a massive gene transfer from the original proteobacterium to the host nucleus. Thus, mitochondrial performance relies on a mosaic of nuclear gene products from a variety of genomes. The concerted regulation of their synthesis is necessary for metabolic housekeeping and stress response. This governance involves crosstalk between mitochondrial, cytoplasmic, and nuclear factors. While anterograde and retrograde regulation preserve mitochondrial homeostasis, the mitochondria can modulate a wide set of nuclear genes in response to an extensive variety of conditions, whose response mechanisms often merge. In this review, we summarise how mitochondrial metabolites and proteins-encoded either in the nucleus or in the organelle-target the cell nucleus and exert different actions modulating gene expression and the chromatin state, or even causing DNA fragmentation in response to common stress conditions, such as hypoxia, oxidative stress, unfolded protein stress, and DNA damage.

Myopathy and Ophthalmologic Abnormalities in Association With Multiple Skeletal Muscle Mitochondrial DNA Deletions.

Establishing a molecular diagnosis of mitochondrial diseases due to pathogenic mitochondrial DNA (mtDNA) variants can be difficult because of varying levels of tissue heteroplasmy, and identifying these variants is important for clinical management. Here, we present clinical and molecular findings in 8 adult patients with classical features of mitochondrial ophthalmologic and/or muscle disease and multiple mtDNA deletions isolated to muscle. The patients were identified via a retrospective review of patients seen in both a tertiary ophthalmology center and a genetics clinic with a clinical diagnosis of chronic progressive external ophthalmoplegia, optic nerve abnormalities, and/or mitochondrial myopathy. Age at onset of symptoms ranged from 18 to 61 years. Ocular manifestations included bilateral optic neuropathy in one patient, bilateral optic disc cupping without optic neuropathy in 2 patients, ptosis in 4 patients, and ocular motility deficits in 2 patients. Five patients had generalized weakness. Pathogenic variants in mtDNA were not found in the blood or buccal sample from any patient, but 7 of 8 patients had multiple mtDNA deletions identified in muscle tissue. One patient had a single mtDNA deletion identified in the muscle. Heteroplasmy was less than 15% for all of the identified deletions, with the exception of one deletion that had a heteroplasmy of 50%-60%. None of the patients were found to have a nuclear gene variant known to be associated with mitochondrial DNA maintenance. mtDNA deletions were identified in adult patients with ophthalmologic and/or musle abnormalities and may underlie their clinical presentations.