bims-mitdis Biomed News
on Mitochondrial disorders
Issue of 2025–08–31
fifty-nine papers selected by
Catalina Vasilescu, Helmholz Munich



  1. EMBO J. 2025 Aug 26.
      A biochemical deficiency of mitochondrial complex I (CI) underlies approximately 30% of cases of primary mitochondrial disease, yet the inventory of molecular machinery required for CI assembly remains incomplete. We previously characterised patients with isolated CI deficiency caused by segregating variants in RTN4IP1, a gene that encodes a mitochondrial NAD(P)H oxidoreductase. Here, we demonstrate that RTN4IP1 deficiency causes a CI assembly defect in both patient fibroblasts and knockout cells, and report that RTN4IP1 is a bona fide CI assembly factor. Complexome profiling revealed accumulation of unincorporated ND5-module and impaired N-module production. RTN4IP1 patient fibroblasts also exhibited defective coenzyme Q biosynthesis, substantiating a second function of RTN4IP1. Thus, our data reveal RTN4IP1 plays necessary and independent roles in both the terminal stages of CI assembly and in coenzyme Q metabolism, and that pathogenic RTN4IP1 variants impair both functions in patients with mitochondrial disease.
    Keywords:  Coenzyme Q; Complex I Assembly; Complexome Profiling; Mitochondria; RTN4IP1
    DOI:  https://doi.org/10.1038/s44318-025-00533-x
  2. Sci Adv. 2025 Aug 29. 11(35): eady0240
      The PINK1/Parkin pathway targets damaged mitochondria for degradation via mitophagy. Genetic evidence implicates impaired mitophagy in Parkinson's disease, making its pharmacological enhancement a promising therapeutic strategy. Here, we characterize two mitophagy activators: a novel Parkin activator, FB231, and the reported PINK1 activator MTK458. Both compounds lower the threshold for mitochondrial toxins to induce PINK1/Parkin-mediated mitophagy. However, global proteomics revealed that FB231 and MTK458 independently induce mild mitochondrial stress, resulting in impaired mitochondrial function and activation of the integrated stress response, effects that result from PINK1/Parkin-independent off-target activities. We find that these compounds impair mitochondria by distinct mechanisms and synergistically decrease mitochondrial function and cell viability in combination with classical mitochondrial toxins. Our findings support a model whereby weak or "silent" mitochondrial toxins potentiate other mitochondrial stressors, enhancing PINK1/Parkin-mediated mitophagy. These insights highlight important considerations for therapeutic strategies targeting mitophagy activation in Parkinson's disease.
    DOI:  https://doi.org/10.1126/sciadv.ady0240
  3. Biochem Soc Trans. 2025 Aug 26. pii: BST20253044. [Epub ahead of print]
      Intrinsic protein quality control (QC) mechanisms are essential in maintaining mitochondrial health and function. These sophisticated molecular machineries govern protein trafficking and import, processing, folding, maturation and degradation, ensuring the organelle's health. Disruption in mitochondrial protein QC can lead to severe, multisystem disorders with variable age of onset and progression. In this review, we provide a snapshot of the intrinsic molecular protein QC machineries in mitochondria detailing their function, localisation and substrate specificity. We also highlight how dysfunction of these molecular machines contributes to mitochondrial disease. Ultimately, elucidating the consequences of proteostatic failure offers critical insights into the pathogenesis of complex mitochondrial disorders.
    Keywords:  AAA+; chaperone; disaggregase; extractase; mitochondria; mitochondrial disease; protease; protein quality control
    DOI:  https://doi.org/10.1042/BST20253044
  4. Neuromuscul Disord. 2025 Aug 13. pii: S0960-8966(25)00931-9. [Epub ahead of print]54 106204
      Primary mitochondrial disease refers to a group of genetic disorders caused by pathogenic variants in either the nuclear or mitochondrial genomes, leading to an impairment of oxidative phosphorylation. We present a young female with a prominent myopathic phenotype associated with an episode of cardiac decompensation. MRI of lower limb musculature revealed a selective pattern of fatty infiltration and muscle oedema. Skeletal muscle biopsy confirmed significant evidence of mitochondrial histopathological abnormalities characterised by multiple respiratory chain deficiencies whilst complete sequencing of the entire mitochondrial genome identified a rare, likely pathogenic m.8362T>G MT-TK gene (NC_012920.1) variant at high levels of heteroplasmy, which we confirmed to be maternally-inherited.
    Keywords:  MERRF; Multi-system disorder; Muscle MRI; Primary mitochondrial disorder
    DOI:  https://doi.org/10.1016/j.nmd.2025.106204
  5. Nat Commun. 2025 Aug 20. 16(1): 7757
      Proximity labeling with engineered ascorbate peroxidase (APEX) has been widely used to identify proteomes within various membrane-enclosed subcellular organelles. However, constructing protein distribution maps between two non-partitioned proximal spaces remains challenging with the current proximity labeling tools. Here, we introduce a proximity labeling approach using isotope-coded phenol probes for APEX labeling (ICAX) that enables the quantitative analysis of the spatial proteome at nanometer resolution between two distinctly localized APEX enzymes. Using this technique, we identify the spatial proteomic architecture of the mitochondrial intracristal space (ICS), which is not physically separated from the peripheral space. ICAX analysis further reveals unexpected dynamics of the mitochondrial spatiome under mitochondrial contact site and cristae organizing system (MICOS) complex inhibition and mitochondrial uncoupling, respectively. Overall, these findings highlight the importance of ICS for mitochondrial quality control under dynamic stress conditions.
    DOI:  https://doi.org/10.1038/s41467-025-62756-0
  6. Mitochondrion. 2025 Aug 18. pii: S1567-7249(25)00077-7. [Epub ahead of print]85 102080
      The diagnosis of disorders associated with mitochondrial DNA (mtDNA) variants presents substantial complexity due to their genetic and clinical heterogeneity, which is largely influenced by mtDNA heteroplasmy. However, the level of heteroplasmy alone is often not sufficient to predict the clinical phenotype including its severity and progression. This study concerns the characterization of the m.8357T > C variant in the MT-TK gene, encoding for mt-tRNA-Lys found in two pediatric siblings. Both had symptoms suggestive of a mitochondrial disease, including severe hearing loss, easy fatigability, decreased activity of mitochondrial complex I in muscle samples, epilepsy, metabolic acidosis with hyperkalemia, and mild kidney impairment. The m.8357T > C mtDNA variant was homoplasmic in muscle, blood, urine and fibroblasts. Immortalized fibroblasts from the patients showed reduced activity of mitochondrial complexes I, III and IV, decreased mitochondrial respiration, and abnormal depolarization of the mitochondrial membrane potential. The mt-tRNA-Lys levels were reduced as compared to the mt-tRNA-Leu (UUR) or the snRNA encoded by RNU6B nuclear gene; the level of three mitochondrial DNA encoded proteins was decreased, altogether suggesting a defective translation machinery in cells carrying the variant. Consistently, fibroblasts from the mother, who had only mild hearing loss, despite high level of heteroplasmy, showed some biochemical abnormalities, however milder than in her daughter and son. Contrariwise, their maternal aunt, who showed intellectual disability, mild hearing loss, easy fatigability and weakness was also virtually homoplasmic for the m.8357T > C in blood and urinary sediment cells. These findings suggest the pathogenicity of the m.8357T > C variant but only in condition of homoplasmy.
    Keywords:  Heteroplasmy; Mitochondrial disease; mt-tRNA-Lys
    DOI:  https://doi.org/10.1016/j.mito.2025.102080
  7. Biochim Biophys Acta Mol Cell Res. 2025 Aug 25. pii: S0167-4889(25)00156-9. [Epub ahead of print] 120051
      The Saccharomyces cerevisiae mitoribosome synthesizes eight mitochondrial DNA-encoded proteins essential for oxidative phosphorylation. Mitoribosome large subunit (mtLSU) biogenesis involves the conserved DEAD-box helicase Mrh4 and the GTPases Mtg1/GTPBP7 and Mtg2/GTPBP5. Here, we have employed genetic, biochemical, in vitro reconstitution, and cryo-EM approaches to elucidate their hierarchical action during the late stages of mtLSU assembly. We show that Mrh4-mediated bL33m incorporation precedes Mtg1 recruitment to the 21S rRNA. Cryo-EM structures of mitoribosome assembly intermediates accumulating in the absence of Mtg1 or uL16m reveal that Mtg1 restructures the 21S rRNA H73-75 and H93 domains to their mature fold. This subsequently allows the structuring of neighboring peptidyl transfer center region helices and the incorporation of uL6m, uL16m, bL35m, and bL36m during late mtLSU maturation. Unexpectedly, monosomes containing immature mtLSU assemble in Mrh4-, bL33m-, uL16m-, Mtg1-, and Mtg2-depleted mitochondria, at levels that increase with the maturation state of the mtLSU particle. Our data have shed light on the rRNA folding events and the structuring of the MRPs that occur during the late stages of assembly. They have provided insight into the roles of assembly factors Mrh4, Mtg1, and Mtg2 during the process and revealed evolutionarily conserved mechanisms underlying mitochondrial ribosome assembly.
    Keywords:  Cryo-EM; DEAD-box helicase; GTPase; Mitochondrial ribosome assembly; Mrh4; Mtg1; Oxidative phosphorylation; Yeast; bL33m; uL16m
    DOI:  https://doi.org/10.1016/j.bbamcr.2025.120051
  8. Nat Commun. 2025 Aug 26. 16(1): 7972
      Heme is an iron-containing cofactor generated in mitochondria that must leave this organelle to reach protein targets in other cell compartments. Because mitochondrial heme binding by cytosolic GAPDH enables its distribution in cells, we sought to uncover how heme reaches GAPDH. Experiments utilizing two human cell lines and a GAPDH reporter protein whose heme binding can be followed by fluorescence reveal that the mitochondrial protein FLVCR1b provides heme to GAPDH in concert with a rise and fall in their association. An absence of FLVCR1b diminishes GAPDH association with mitochondria and prevents GAPDH and cell hemeproteins from receiving heme. GAPDH heme procurement also requires the TANGO2 protein, which interacts with FLVCR1b to presumably support heme export. In isolated mitochondria, GAPDH associates with FLVCR1b to trigger heme release and delivery to client hemeproteins. Identifying FLVCR1b as the source of mitochondrial heme for GAPDH reveals a path by which this essential cofactor can reach multiple protein targets within eukaryotic cells.
    DOI:  https://doi.org/10.1038/s41467-025-62819-2
  9. J Lipid Res. 2025 Aug 25. pii: S0022-2275(25)00150-6. [Epub ahead of print] 100888
      Mitochondria are fundamental in energy homeostasis and undergo dynamic changes in brown and beige fat. Mitochondrial dysfunctions impair thermogenic capacity and cause obesity-associated metabolic diseases. The phospholipid composition is crucial for maintaining mitochondrial function and fission-fusion processes. Here, we described EBF2, a transcription factor pivotal for brown fat development and function that regulates the integrity of mitochondrial membrane composition, function and dynamics in brown adipocytes. Strikingly, Myf5Cre-driven targeted deletion of Ebf2 in brown adipose tissue (BAT) drastically reduces cardiolipin and phosphatidylethanolamine abundance and alters acyl chain remodeling of major phospholipids. BAT mitochondria of Ebf2-KO neonates exhibit a severe reduction of DRP1 and OPA1, key regulators of mitochondrial fission-fusion dynamics; further, Ebf2 deletion impairs fragmentation-fusion events in brown adipose tissue. Mechanistically, EBF2 directly binds to key genes, including Srebf1, which are involved in membrane lipid metabolism and differentially regulate their expression. Also, the deletion of Ebf2 downregulates CL and PE-synthesizing genes and accumulates phosphatidylserine and sphingomyelin levels in mitochondria. Thus, the deletion of Ebf2 perturbs the acyl chain remodeling of mitochondrial lipids and affects the fission-fusion cycle in neonatal brown adipocytes. To conclude, Ebf2 is crucial for regulating the levels and remodeling of bilayer and non-bilayer-forming lipids to conserve mitochondrial metabolism.
    Keywords:  Brown adipocytes; Cardiolipin; Ebf2-BKO; Lipid metabolism; Membrane lipids; Mitochondria; Mitochondrial lipidome; OXPHOS; Reactive oxygen species; Thermogenesis
    DOI:  https://doi.org/10.1016/j.jlr.2025.100888
  10. J Cardiovasc Dev Dis. 2025 Aug 20. pii: 318. [Epub ahead of print]12(8):
      Combined oxidative phosphorylation deficiency type 8 (COXPD8) is an autosomal recessive mitochondrial disorder caused by a mutation of the nuclear encoded mitochondrial alanyl-tRNA synthetase gene (AARS2). Clinical manifestations of COXPD8 include lethal infantile hypertrophic cardiomyopathy, pulmonary hypoplasia, generalized muscle weakness, and neurological involvement. We report a patient with COXPD8 caused by two mutations in the AARS2 gene. The c.1738 C>G mutation has not been previously reported, while the c.2872 C>T mutation has been associated with pulmonary hypoplasia and hypertrophic cardiomyopathy. Cardiac tissue, obtained through the LungMAP program, showed that, compared to other patients of similar ages, these two mutations affect not only the assembly of functional monomeric complexes (Cx) I and IV of the electron transport chain (ETC) but also limit the formation of respiratory supercomplexes. This patient had altered expression of some ETC proteins but normal expression of several enzymes of the tricarboxylic acid cycle. We also show that one of the control/comparison patients had an undiagnosed ETC Cx IV deficiency. In conclusion, our data demonstrate that the two mutations of the AARS2 gene are associated with failed assembly of Cx I and Cx IV and reduced formation of respiratory supercomplexes of the ETC, likely leading to acute bioenergetic stress.
    Keywords:  COXPD8; electron transport chain; hypertrophic cardiomyopathy; mitochondrial disease; mitochondrial supercomplexes
    DOI:  https://doi.org/10.3390/jcdd12080318
  11. Biomolecules. 2025 Aug 13. pii: 1159. [Epub ahead of print]15(8):
      Efficient mitochondrial matrix protein quality control (mPQC), regulated by the mitochondrial matrix protease LONP1, is essential for preserving cardiac bioenergetics, particularly in post-mitotic cardiomyocytes, which are highly susceptible to mitochondrial dysfunction. While cardiac mPQC defects could impair heart function, it remains unclear whether such defects can be mitigated through inter-organ crosstalk by modulating mPQC in extra-cardiac tissues, a potentially valuable strategy given the challenges of directly targeting the heart. To investigate this, we examined two mouse models of Lonp1 haploinsufficiency at young adulthood: a cardiomyocyte-specific heterozygous knockout (Lonp1CKO-HET) and a whole-body heterozygous knockout (Lonp1GKO-HET). Despite similar reductions in Lonp1 mRNA expression in the hearts, Lonp1GKO-HET mice exhibited no cardiac dysfunction, whereas Lonp1CKO-HET mice showed mild cardiac dysfunction accompanied by activation of the mitochondrial stress response, including induction of genes such as Clpx, Spg7, Hspa9, and Hspd1, increased mitochondrial dynamics (Pink1, Dnm1l), reduced mitochondrial biogenesis, and compensatory upregulation of the mtDNA transcriptional regulator Tfam, all occurring without overt structural remodeling. These alterations were absent in Lonp1GKO-HET hearts. Our findings reveal a novel adaptive mechanism in which systemic mPQC deficiency can buffer mitochondrial dysfunction in the heart through inter-organ communication that is lost with cardiomyocyte-specific mPQC disruption. This study identifies systemic modulation of Lonp1-mediated mitochondrial stress pathways as a promising strategy to promote cardiac resilience through protective inter-organ signaling.
    Keywords:  LONP1; cardiac dysfunction; heart; mitochondria; mitochondrial dysfunction; mitochondrial matrix; protein quality control
    DOI:  https://doi.org/10.3390/biom15081159
  12. J Biol Chem. 2025 Aug 19. pii: S0021-9258(25)02465-2. [Epub ahead of print] 110614
      An essential aspect of protein quality control is enzymatic removal of membrane proteins from the lipid bilayer. Failures in this critical cellular process are associated with neurodegenerative diseases and cancer. Msp1 is a AAA+ (ATPases Associated with diverse cellular Activities) ATPase that removes mistargeted membrane proteins from the outer mitochondrial membrane (OMM). How Msp1 selectively recognizes and extracts substrates within the complex OMM ecosystem, and how the lipid bilayer impacts these processes is unknown. Here, we describe the development of a fully defined, rapid, and quantitative extraction assay that retains physiological substrate selectivity. Using this new assay, we systematically modified both the model substrate and the lipid environment to demonstrate that Msp1 can recognize substrates by a hydrophobic mismatch between the substrate TMD and the lipid bilayer. We further demonstrate that the rate-limiting step in Msp1 activity is extraction of the TMD from the lipid bilayer. Together, these results provide foundational insights into how the lipid bilayer influences AAA+ mediated membrane protein extraction.
    Keywords:  ATPase Associated with diverse cellular Activities (AAA+); lipid bilayer; membrane protein; mitochondria; proteostasis
    DOI:  https://doi.org/10.1016/j.jbc.2025.110614
  13. Children (Basel). 2025 Aug 21. pii: 1102. [Epub ahead of print]12(8):
       BACKGROUND: Given the limited research on mitochondrial diseases in our area, specifically regarding their genetic variability and diverse clinical manifestations, and considering the significant number of consanguineous marriages in our region, we aimed to investigate the clinical and molecular characteristics of patients with mitochondrial disorders in Saudi Arabia.
    METHODS: This retrospective cross-sectional cohort study involved a chart review of patients diagnosed with mitochondrial disorders at the Ministry of National Guard Health Affairs tertiary health care centers in Saudi Arabia between 2013 and 2022.
    RESULTS: The study population comprised 116 patients with a mean age of 10 years (±7 SD). Among the study cohort, 34.5% (n = 40) had died. The primary cause of death was cardiopulmonary arrest (55.0%, n = 22). The most prevalent condition was developmental delay (67.9%). Around 56.9% were diagnosed using Whole Exome Sequencing (WES), 10.3% by Whole Genome Sequencing due to negative WES, 23.3% through a single-gene approach, 7.8% were analyzed through a mitochondrial panel, and 1.7% via a gene panel. The distributions of current age and age at diagnosis were significantly different between the nuclear and mitochondrial gene types. Notably, the diagnostic delay time (time taken from symptom onset to genetic diagnosis) averaged 1.5 years for nDNA variants compared to an average of 10 years for mDNA variants.
    CONCLUSIONS: This study shows that gene type affects clinical characteristics, highlighting the importance of genetic studies in disease manifestation.
    Keywords:  Saudi Arabia; disorders; genetic disorders; mitochondria; mitochondrial diseases; mitochondrial dysfunction
    DOI:  https://doi.org/10.3390/children12081102
  14. J Adv Res. 2025 Aug 21. pii: S2090-1232(25)00644-7. [Epub ahead of print]
       BACKGROUND: Mitochondrial DNA (mtDNA), a circular genome essential for cellular energy production, is increasingly recognized to exhibit aberrant methylation under pathological conditions. Dysregulated methylation in regulatory regions can impair mtDNA replication, transcription, and metabolic homeostasis, thereby promoting disease progression, including neurodegenerative diseases, cardiovascular diseases, metabolic disorders, as well as aging. Despite challenges posed by nuclear pseudogene interference, advanced detection technologies have significantly improved the resolution of mtDNA methylation analysis.
    AIM OF REVIEW: This review focuses on three key mtDNA methylation patterns, 5-methylcytosine (5mC), 5-hydroxymethylcytosine (5hmC), and N6-methyladenine (6mA), summarizing the evidence for their existence as well as their molecular mechanisms in diseases and offering insights into recent advances in mtDNA detection techniques. Key Scientific Concepts of Review: Under pathological conditions, the dysregulation of mtDNA methylation highlights its emerging promise as both a biomarker and therapeutic target. Therefore, this epigenetic aberration provides a foundational framework for elucidating the molecular mechanisms underlying mitochondrial dysfunction across diverse diseases and advancing precision medicine strategies.
    Keywords:  5-hydroxymethylcytosine; 5-methylcytosine; MtDNA methylation; N6-methyladenine
    DOI:  https://doi.org/10.1016/j.jare.2025.08.029
  15. Biol Chem. 2025 Jun 24.
      The mitochondrial solute carrier family, also called SLC25 family, comprises a group of structurally and evolutionary related transporters that are embedded in the mitochondrial inner membrane. About 35 and 53 mitochondrial carrier proteins are known in yeast and human cells, respectively, which transport nucleotides, metabolites, amino acids, fatty acids, inorganic ions and cofactors across the inner membrane. They are proposed to function by a common rocker-switch mechanism, alternating between conformations that expose substrate-binding pockets to the intermembrane space (cytoplasmic state) and to the matrix (matrix state). The substrate specificities of both states differ so that carriers can operate as antiporters, symporters or uniporters. Carrier proteins share a characteristic structure comprising six transmembrane domains and expose both termini to the intermembrane space. Most carriers lack N-terminal presequences but use carrier-specific internal targeting signals that direct them into mitochondria via a specific import route, known as the 'carrier pathway'. Owing to their hydrophobicity and aggregation-prone nature, the mistargeting of carriers can lead to severe proteotoxic stress and diseases. In this review article, we provide an overview about the structure, biogenesis and physiology of carrier proteins, focusing on baker's yeast where their biology is particularly well characterized.
    Keywords:  TIM22 complex; membrane transport; metabolism; mitochondria; protein translocation
    DOI:  https://doi.org/10.1515/hsz-2025-0152
  16. STAR Protoc. 2025 Aug 23. pii: S2666-1667(25)00451-4. [Epub ahead of print]6(3): 104045
      Here, we present a protocol for differentiating 3T3-L1 preadipocytes and stromal vascular fraction (SVF)-derived preadipocytes from mice into mature adipocytes, followed by the isolation of crude mitochondrial fractions. This cost-effective and reproducible protocol is optimized for small-plate formats, compatible with standard reagents, and suitable for metabolic studies such as insulin resistance and mitochondrial function.
    Keywords:  Cell culture; Metabolism; Molecular Biology
    DOI:  https://doi.org/10.1016/j.xpro.2025.104045
  17. Ann Intern Med Clin Cases. 2025 Aug;pii: e250080. [Epub ahead of print]4(8):
      Mitochondrial 3-hydroxy-3-methylglutaryl-coenzyme A synthase (mHS) deficiency is an ultra-rare inborn error of ketone body synthesis that is caused by biallelic mutations in HMGCS2. The manifestations of mHS deficiency can include hypoketotic hypoglycemia, metabolic acidosis, lethargy, encephalopathy, hyperammonemia, and hepatomegaly. Here, we report a case of movement disorder following hypoglycemic encephalopathy involving the basal ganglia in a patient with mHS deficiency. Exome sequencing showed novel compound heterozygous variants in HMGCS2, a partial gene deletion (classified as pathogenic) and c.704T>A (p.M235K) variant that was deemed to be likely pathogenic. Our findings suggest that mHS deficiency can result in basal ganglia injury and movement disorder.
    Keywords:  Basal ganglia; Brain; Coagulopathy; Exome; Hypoglycemia; Hypoglycemics; Ischemic stroke; Medical risk factors; Mitochondria; Movement disorders; Pathogenesis; Stroke
    DOI:  https://doi.org/10.7326/aimcc.2025.0080
  18. EMBO Mol Med. 2025 Aug 26.
    Undiagnosed Diseases Network
      ATP5F1A encodes the α-subunit of complex V of the respiratory chain, which is responsible for mitochondrial ATP synthesis. We describe 6 probands with heterozygous de novo missense ATP5F1A variants that presented with developmental delay, intellectual disability, and movement disorders. All variants were located at the contact points between the α- and β-subunits. Functional studies in C. elegans revealed that the variants were damaging via a dominant negative genetic mechanism. Biochemical and proteomics studies of proband-derived cells showed a marked reduction in complex V abundance and activity. Mitochondrial physiology studies revealed increased oxygen consumption, yet decreased mitochondrial membrane potential and ATP levels indicative of uncoupled oxidative phosphorylation as a pathophysiologic mechanism. Our findings contrast with the previously reported ATP5F1A variant, p.Arg207His, indicating a different pathological mechanism. This study expands the phenotypic and genotypic spectrum of ATP5F1A-associated conditions and highlights how functional studies can provide an understanding of the genetic, molecular, and cellular mechanisms of ATP5F1A variants of uncertain significance. With 12 heterozygous individuals now reported, ATP5F1A is the most frequent nuclear genome cause of complex V deficiency.
    Keywords:  ATP Synthase; ATP5F1A; Complex V; Mitochondriopathy; Oxidative Phosphorylation
    DOI:  https://doi.org/10.1038/s44321-025-00290-8
  19. Nat Commun. 2025 Aug 23. 16(1): 7863
      Protein AMPylation, the covalent addition of adenosine monophosphate (AMP) to protein substrates, has been known as a post translational modification for over 50 years. Research in this field is largely underdeveloped due to the lack of tools that enable the systematic identification of AMPylated substrates. Here, we address this gap by developing an enrichment technique to isolate and study AMPylated proteins using a nucleotide-binding protein, hinT. Cryo-EM reconstruction of an AMPylated protein bound to hinT provides a structural basis for AMP selectivity. Using structure guided mutagenesis, we optimize enrichment to identify novel substrates of the evolutionarily conserved AMPylase, Selenoprotein O. We show that mammalian Selenoprotein O regulates metabolic flux through AMPylation of key mitochondrial proteins including glutamate dehydrogenase and pyruvate dehydrogenase. Our findings highlight the broader significance of AMPylation, an emerging post translational modification with critical roles in signal transduction and disease pathology. Furthermore, we establish a powerful enrichment platform for the discovery of novel AMPylated proteins to study the mechanisms and significance of protein AMPylation in cellular function.
    DOI:  https://doi.org/10.1038/s41467-025-63014-z
  20. Muscles. 2025 Jul 31. pii: 26. [Epub ahead of print]4(3):
      Skeletal muscle aging and related diseases are characterized by progressive loss of muscle mass, strength, and metabolic function. Central to these processes is mitochondrial dysfunction, which impairs energy metabolism, redox homeostasis, and proteostasis. In addition, non-mitochondrial factors such as muscle stem cell exhaustion, neuromuscular junction remodeling, and chronic inflammation also contribute significantly to muscle degeneration. This review integrates recent advances in understanding mitochondrial and non-mitochondrial mechanisms underlying muscle aging and disease. Additionally, we discuss emerging therapeutic approaches targeting these pathways to preserve muscle health and promote healthy aging.
    Keywords:  inflammation; mitochondrial dysfunction; muscle aging; muscle stem cells; neuromuscular junction; therapy
    DOI:  https://doi.org/10.3390/muscles4030026
  21. Nat Commun. 2025 Aug 25. 16(1): 7671
      Mitochondrial dynamics enable cellular adaptation to fluctuations in energy demand, such as those imposed on skeletal muscle by exercise, metabolic disorders, or aging. Here, we report a novel pathway that modulates mitochondria dynamics in skeletal muscle involving the scaffolding protein ankyrin-B. Rare variants in ankyrin-B, encoded by ANK2, increase risk for cardio-metabolic syndrome in humans and mice. We show that mice selectively lacking skeletal muscle ankyrin-B have reduced endurance exercise capacity without alterations in muscle strength or systemic glucose regulation. Muscle fibers in these mice have increased oxidative stress, reduced fatty acid oxidation, and enlarged and hyperconnected mitochondria. We found that ankyrin-B interacts with and is required for efficient mitochondria recruitment of fission modulators and sarcoplasmic reticulum-mitochondria coupling. Thus, we conclude that ankyrin-B enables substrate adaptability and bioenergetic homeostasis under energetic stress, and exercise capacity by promoting efficient mitochondrial fission in skeletal muscle.
    DOI:  https://doi.org/10.1038/s41467-025-62977-3
  22. Curr Pharm Des. 2025 Aug 21.
      Mitochondria are known as the powerhouse of eukaryotic cells. They play a crucial role in several biological processes and maintain cellular health. The ideal condition of mitochondria depends not only on their morphology but also on various micro-environmental factors, including pH, polarity, and temperature. Changes in these factors or malfunctions of mitochondrial species, such as Reactive Oxygen Species (ROS), active nitrogen species, metal cations, anions, and protons, can lead to several diseases in humans, including heart failure, kidney disorders, diabetes, Alzheimer's disease, and Parkinson's disease. Therefore, monitoring Reactive Small Molecules (RSMs), maintaining micro-environmental factors, and estimating ROS levels in mitochondria are essential for understanding physiological behaviour and the pathogenesis of related diseases. Irregularities in mitochondrial function are closely linked to a range of clinical conditions, highlighting the importance of targeting mitochondria for therapeutic benefits. Over the last decade, numerous studies have focused on the development of small organic conjugated systems for mitochondrial imaging, utilizing optical signal transduction pathways. In this review, the design and synthetic strategies for small organic fluorophores conjugated with a pyridinium acceptor, their applications in mitochondrial imaging, and the detection of RSMs in mitochondria have been discussed. Studies have revealed that small-molecule fluorescent probes are being widely used for the detection and imaging of RSMs located in mitochondria. Moreover, this review covers the mechanistic insights, photophysical properties, biological characteristics of fluorophores, and therapeutic strategies targeting the mitochondria of human cells.
    Keywords:  Fluorophores; fluorescence; mitochondrial targeting; organic molecules.; pyridinium moiety
    DOI:  https://doi.org/10.2174/0113816128395084250708080830
  23. Autophagy. 2025 Sep 01. 1-18
      The neuromuscular junction (NMJ) is essential for transmitting neural stimulus to muscles, triggering muscle contraction. Mitochondria are enriched in NMJ to support the energy needs required for neuromuscular function and stability. Thus, maintaining mitochondrial homeostasis through the clearance of damaged mitochondria, a process known as mitophagy, is vital for preserving neuromuscular health. Here, we highlight the crucial role of muscle PRMT1 in maintaining NMJ and mitochondrial homeostasis via mitophagy regulation. PRMT1 is distinctively expressed in myofibers, accumulating in the postsynaptic area, with its levels upregulated in denervated muscles. PRMT1-ablated muscles displayed disrupted NMJs and an accumulation of abnormal mitochondria, accompanied by increased mitochondrial oxidative stress. Additionally, prmt1 depletion in muscles specifically impaired TBK1 (TANK binding kinase 1)-OPTN (optineurin)-mediated mitophagy. Overall, our findings suggest that PRMT1 plays a critical role in maintaining NMJ and mitochondrial health by regulating selective mitophagy through TBK1-OPTN.Abbreviations: ADMA: asymmetric arginine dimethylation; BTX: α-bungarotoxin; EDL: extensor digitorum longus; FDB: flexor digitorum brevis; GAS: gastrocnemius; NMJ: Neuromuscular junction; Mko: mice with muscle-specific prmt1 ablation; MTOR: mechanistic target of rapamycin kinase; OPTN: optineurin; PRMT1: protein arginine methyltransferase 1; SA: sodium arsenate; SNI: sciatic nerve crush injury; Sol: soleus; SQSTM1/p62: sequestosome 1; TBK1: TANK binding kinase 1; TOMM20: translocase of outer mitochondrial membrane 20; TA: tibialis anterior; VDAC1: voltage dependent anion channel 1.
    Keywords:  Mitophagy; PRMT1; TBK1; neuromuscular junction; skeletal muscle
    DOI:  https://doi.org/10.1080/15548627.2025.2551477
  24. Adv Mater. 2025 Aug 25. e04509
      Gene therapy offers a promising treatment for Leber's hereditary optic neuropathy (LHON), a disease of retinal ganglion cell (RGC) degeneration with severe vision loss caused by mitochondria-NADH dehydrogenase 4 (MT-ND4) mutations. However, optimizing mitochondria-targeted gene delivery to promote RGC regeneration and visual-photoreception recovery remains challenging in LHON. Here, mitochondria-targeted wireless charging gold nanoparticles (WCGs), doubling as a wireless charging-mediated gene-delivery platform and electric stimulus-restored phototransduction, are developed for LHON treatment. Upon high-frequency magnetic field (HFMF) irradiation, WCGs enhanced MT-ND4 transfection efficiency, restored complex I activity and mitochondrial homeostasis in vitro, and further promoted RGC neurite outgrowth in LHON patient-derived iPSC-differentiated retinal organoids. Wireless charging combined with electric stimulation also facilitated in vivo gene delivery, effectively promoting neuronal recovery, preventing RGC degeneration, suppressing inflammation, and enhancing retinal electrophysiological function in the damaged retinas of LHON mice. Furthermore, single-cell RNA sequencing and spatiotemporal transcriptomic analysis revealed that HFMF-treatment reprograms Müller glia to enhance dendritic development, restore mitochondrial function, and express phototransduction genes to support photoreceptor function in vivo. Finally, functional retinal-optic-electrophysiological findings with spatiotemporal transcriptomic analysis at the single-cell level support that HFMF synergized with WCG/hND4 therapy, promoting retinal repairment with RGC neurite regeneration and recovering visual phototransduction in LHON mouse models.
    Keywords:  Leber's hereditary optic neuropathy (LHON); high‐frequency magnetic field (HFMF); induced pluripotent stem cells (iPSCs); phototransduction; retinal organoid; wireless charging gold nanoparticles (WCGs)
    DOI:  https://doi.org/10.1002/adma.202504509
  25. Nat Commun. 2025 Aug 23. 16(1): 7864
      Mitochondrial fatty acid β-oxidation (FAO) is essential for energy production and cellular homeostasis, yet its role in sperm function has remained unclear. Through whole-exome sequencing (WES) of 800 patients with asthenozoospermia, we identified biallelic Testis-Expressed Protein 44 (TEX44) variants in six individuals, all of whom exhibited defective mitochondrial sheath assembly and impaired sperm motility. Using Tex44 knockout mice, we show that TEX44 interacts with carnitine palmitoyltransferase 1B (CPT1B) to form a mitochondrial glue, anchoring adjacent mitochondria and facilitating the assembly of the sperm-specific mitochondrial sheath. In vitro, we show that purified TEX44 protein can modulate CPT1B enzymatic activity, limiting the conversion of long-chain fatty acids such as palmitic acid and myristic acid into acyl-carnitines, thereby reducing reactive oxygen species (ROS) production. Loss of TEX44 disrupts this regulatory mechanism, leading to unregulated FAO, excessive ROS generation, and severe oxidative damage to sperm DNA and flagellar structure. Additionally, germ cell-specific Cpt1b knockout mice exhibit phenotypes similar to TEX44 deficiency, including mitochondrial sheath defects and reduced sperm motility. These findings reveal a sperm-specific mechanism by which TEX44 regulates CPT1B activity to balance FAO and ROS generation, providing critical insights into energy metabolism, mitochondrial integrity, and male infertility.
    DOI:  https://doi.org/10.1038/s41467-025-63280-x
  26. Genes (Basel). 2025 Aug 20. pii: 982. [Epub ahead of print]16(8):
       BACKGROUND: L-2-hydroxyglutaric aciduria (L2HGA) is a rare autosomal recessive neurometabolic disorder caused by biallelic loss-of-function variants in the L-2-hydroxyglutarate dehydrogenase (L2HGDH) gene, leading to accumulation of L-2-hydroxyglutarate in the brain and other tissues. While various variants have been reported, the pathogenic mechanism of specific variants remains unclear. In this study, we aimed to investigate the molecular consequences of the c.905C>T p.(Pro302Leu) variant, identified in two siblings with typical symptoms of L2HGA, by analyzing its effects on protein localization and enzymatic activity in a cell model.
    METHODS: HA-tagged wild-type and p.(Pro302Leu) mutant L2HGDH constructs were overexpressed in HEK293T cells. We assessed protein expression, subcellular localization, and enzymatic activity using Western blot analysis, immunofluorescence microscopy, and a specific enzyme assay measuring 2,6-dichloroindophenol (DCIP) reduction to assess L2HGDH enzymatic activity.
    RESULTS: Western blotting showed that wild-type L2HGDH exists primarily in the processed, mature mitochondrial form, whereas the p.(Pro302Leu) mutant remained largely in the unprocessed precursor form. Immunofluorescence and differential centrifugation revealed that wild-type protein localized to mitochondria, while the mutant protein accumulated in the cytoplasm in a diffuse or punctate pattern. Enzyme activity assays demonstrated that the mutant retained <30% of wild-type activity.
    CONCLUSIONS: These findings indicate that the p.(Pro302Leu) variant leads to aggregation of mislocalized protein, thereby impairing L2HGDH function rather than decreasing enzymatic function. This study provides clinical and molecular evidence supporting the pathogenicity of this previously reported mutation and highlights the importance of mitochondrial import for enzyme functionality in L2HGA.
    Keywords:  L-2-hydroxyglutarate dehydrogenase 3; L-2-hydroxyglutaric aciduria 2; metabolic disorder disease
    DOI:  https://doi.org/10.3390/genes16080982
  27. Crit Rev Anal Chem. 2025 Aug 20. 1-23
      Mitochondria are key cellular organelles that perform essential functions, including energy generation, calcium regulation, and cell signaling. Disruptions in mitochondrial function are linked to various conditions, such as neurodegenerative disorders, metabolic diseases, and cardiovascular issues. This review critically analyzes the intricate relationship between mitochondrial health and disease, emphasizing the role of proteomics and lipidomics in enhancing the understanding of these processes. Powerful analytical strategies including various mass spectrometric techniques employed in these omic studies have been evaluated for their suitability and effectiveness. Additionally, it highlights the broader field of omics sciences and their integration into mitochondrial research. The aim is to illustrate how omics approaches play a crucial role in advancing biological knowledge related to mitochondria, surpassing the limitations of traditional analytical methods.
    Keywords:  HRMS-based proteomics; mitochondrial health and dysfunction
    DOI:  https://doi.org/10.1080/10408347.2025.2545295
  28. Genes (Basel). 2025 Aug 13. pii: 957. [Epub ahead of print]16(8):
       BACKGROUND: The mitochondrial integrated stress response (ISR) represents a fundamental cellular adaptation mechanism with dual protective and pathological roles. We critically analyzed current literature on ISR mechanisms, focusing on recent paradigm shifts including the 2020 discovery of the OMA1-DELE1-HRI axis, emerging controversies over context-dependent activation patterns, and the January 2025 clinical trial failures that have reshaped the therapeutic landscape.
    METHODS: We reviewed recent literature (2020-2025) examining ISR mechanisms, clinical trials, and therapeutic developments through comprehensive database searches.
    RESULTS: The field has evolved from simple linear pathway models to recognition of complex, context-dependent networks. Recent findings reveal that ISR activation mechanisms vary dramatically based on cellular metabolic state, with distinct pathways operating in proliferating versus differentiated cells. The "dark microglia" phenotype in neurodegeneration and DR5-mediated apoptotic switches exemplify pathological ISR manifestations, while adaptive responses include metabolic reprogramming and quality control enhancement.
    CONCLUSIONS: The 2025 failures of DNL343 and ABBV-CLS-7262 in ALS trials underscore the need for precision medicine approaches that account for context-dependent ISR functions, temporal dynamics, and disease-specific mechanisms.
    Keywords:  cellular adaptation; eIF2α phosphorylation; integrated stress response; mitochondrial dysfunction; neurodegeneration; precision medicine
    DOI:  https://doi.org/10.3390/genes16080957
  29. J Neurol. 2025 Aug 22. 272(9): 587
       BACKGROUND: Primary Lateral Sclerosis (PLS) is a rare, adult-onset neurodegenerative disease that predominantly affects upper motor neurons. Despite being considered mostly sporadic, familial cases and rare genetic variants in genes associated with amyotrophic lateral sclerosis, hereditary spastic paraplegia and other neurological disorders have been reported in some PLS cases. Due to its rare prevalence among general population, large genetic studies of PLS are lacking.
    METHODS: Fifty patients diagnosed with PLS based on consensus criteria were enrolled between 2013 and 2022 for comprehensive phenotypic and genotypic analysis using whole genome sequencing. We analyzed rare single nucleotide variants (SNVs), deemed pathogenic, likely pathogenic or of uncertain significance (VUS) based on the American College of Medical Genetics and Genomics criteria, and repeat expansions (REs) exceeding the pathogenic threshold, in 290 genes involved in neurological disorders.
    RESULTS: We identified mutations in 7 patients (13.7%), specifically SNVs in CAPN1 (Spastic paraplegia 76), TBK1 (amyotrophic lateral sclerosis/frontotemporal dementia, ALS4/FTD), LITAF (Charcot-Marie-Tooth disease 1C), POLG (chronic progressive external ophthalmoplegia), APP (Alzheimer's disease) and OPTN (ALS12 ± FTD), and one RE in ATXN8OS (spinocerebellar ataxia 8). Additionally, two VUS were found in ANTXR2, a candidate gene for PLS recently identified via truncating variant collapsing analysis, but none of them was loss-of-function (one synonymous and one in-frame insertion).
    CONCLUSIONS: Our study demonstrates a notable genetic intersection between PLS and various neurological disorders, including motor neuron diseases, neuropathies, mitochondrial disorders, ataxias, and dementias. These findings underscore the relevance of further investigation in larger cohorts to fully elucidate PLS genetic architecture and highlight the need to reconsider the role of genetic testing in its diagnostic criteria.
    Keywords:  Ataxia; Hereditary; Mitochondrial diseases; Motor neuron disease; Neurodegenerative diseases; Peripheral nervous system diseases; Primary lateral sclerosis; Sequence analysis; Spastic paraplegia; Tandem repeat sequences; Whole genome sequencing
    DOI:  https://doi.org/10.1007/s00415-025-13328-1
  30. Eur J Hum Genet. 2025 Aug 22.
      Genomic sequencing technologies, which include both exome and genome sequencing, as well as panels or targeted analyses using genome-wide approaches, are being implemented across healthcare. Implementation, however, varies greatly by application and jurisdiction, with a diversity of approaches being employed around the world. This review aims to summarise the current state of implementation of genomic testing in mainstream healthcare for the detection of rare disease throughout the lifespan. Through a discussion of evidence gathered to date, highlighting exemplar studies, the following applications of genomic testing will be covered: (1) routine diagnostic genomic testing in the clinic; (2) rapid diagnostic genomic testing in the intensive care unit; (3) genomic newborn screening; and, (4) reproductive genetic carrier screening. Mainstream implementation necessarily extends beyond the clinical genetics service, where genomic testing has historically been offered. Given that the involvement of non-genetics clinicians in the delivery of these technologies has important implications for models of care and education, related areas of growing evidence are also discussed: (5) genetic counsellors working outside clinical genetics services; and, (6) workforce development considerations for mainstream genomics. The diversity of approaches and examples illustrates that integration of genomic technologies into mainstream healthcare is complex and requires significant health system transformation. Efforts to evaluate services, guided by implementation science, will be essential to ensure lessons are shared across jurisdictions and benefit is delivered to patients and the system at-large.
    DOI:  https://doi.org/10.1038/s41431-025-01925-7
  31. Brain Sci. 2025 Jul 22. pii: 777. [Epub ahead of print]15(8):
      Mitochondrial Membrane Protein-Associated Neurodegeneration is a rare monogenic form of neurodegeneration characterized by iron accumulation in the brain. It is due to variants in the orphan gene C19orf12. Since its definition in 2011, many scientific groups have investigated the clinical features and molecular underpinnings of the disorder. In this review, we summarize the main points of progress in this field, trying to highlight the issues that need further attention and efforts to speed up the diagnostic path, improve the existing treatment options, and define targeted therapies.
    Keywords:  C19orf12; Mitochondrial Membrane Protein-Associated Neurodegeneration (MPAN); Neurodegeneration with Brain Iron Accumulation (NBIA); movement disorders; rare hereditary diseases
    DOI:  https://doi.org/10.3390/brainsci15080777
  32. Biomater Sci. 2025 Aug 26.
      Astrocytes, the abundant glial cells, maintain cerebral homeostasis and cognitive functions through calcium signalling - a regulatory pathway that is frequently altered in brain disease. Mitochondria serve as thermal hubs in living systems, generating metabolic heat during respiratory substrate oxidation and ATP synthesis. Crucially, mitochondrial temperature variations directly reflect metabolic status, as impaired ATP production induces thermodynamic shifts. Here, we utilized a fluorescent thermometer probe MTY for in vitro determination and visualization of intracellular mitochondrial temperatures at the single-cell level. Through precisely controlled thermal modulation of fixed, living, and laser-stimulated astrocytes, we established a platform extendable to MCF-7 and Panc02 cell lines. The methodology enabled real-time tracking of near-infrared-induced thermal perturbations and FCCP-mediated uncoupling effects. Spinning-disk confocal microscopy revealed synchronized mitochondrial thermogenesis and calcium transients, with thermal/laser stimulation inducing 2-4-fold greater calcium spiking versus controls. Mechanistic analysis suggested this response was likely mediated through TRPV4 channel-mediated extracellular Ca2+ influx and/or intracellular calcium release from mitochondrial and endoplasmic reticulum stores.
    DOI:  https://doi.org/10.1039/d5bm00691k
  33. Life (Basel). 2025 Aug 11. pii: 1273. [Epub ahead of print]15(8):
      Migraine is a complex neurological disorder characterized by recurrent headaches and sensory disturbances. Emerging evidence highlights a critical role for mitochondrial dysfunction in migraine pathophysiology, including impairments in oxidative phosphorylation, disruptions in mitochondrial dynamics, and altered biogenesis. Experimental migraine models-ranging from nitroglycerin-induced attacks to inflammatory stimuli-consistently demonstrate mitochondrial swelling, cristae disruption, decreased ATP production, and increased oxidative stress. These findings are accompanied by the altered expression of key mitochondrial regulators such as PGC-1α, Drp1, and Mfn1. Recent studies have further identified distinct metabolic subtypes of mitochondria, including P5CS-containing subsets, which exhibit unique structural and functional profiles, including cristae loss and reduced ATP synthase expression. Notably, the mitochondrial alterations observed in migraine models show remarkable parallels to those described in P5CS-related mitochondrial subsets. These similarities suggest a potential mechanistic link between metabolic reprogramming within mitochondria and migraine pathogenesis. Understanding the contribution of these newly defined mitochondrial populations could offer novel insights into migraine biology and open new avenues for targeted therapeutic strategies.
    Keywords:  migraine; mitochondrial dynamics; mitochondrial dysfunction; mitochondrial subtypes
    DOI:  https://doi.org/10.3390/life15081273
  34. Front Cardiovasc Med. 2025 ;12 1572559
      Mitochondrial Antiviral Signaling Protein (MAVS) is a pivotal adaptor protein in the innate immune response, mediating the activation of NF-κB and type I interferon signaling pathways during viral infections. As an integral component of the mitochondrial outer membrane, MAVS also plays critical roles in the regulation of apoptosis, cellular metabolism, and the activation of inflammasomes, including NLRP3 and caspase family members. Emerging evidence indicates that MAVS is not only essential in antiviral defense but also contributes significantly to the pathogenesis of various diseases, notably cardiovascular diseases. In this review, we provide a comprehensive overview of the molecular structure of MAVS and the regulatory mechanisms modulating its activity. We further highlight the involvement of MAVS in the development of cardiovascular diseases through its participation in innate immune signaling and mitochondrial dynamics. Particular attention is given to the regulation of MAVS by post-translational modifications-such as ubiquitination, methylation, and acetylation-as well as by microRNAs and other mitochondria-associated proteins. These insights aim to deepen the understanding of MAVS as a potential biomarker and therapeutic target, offering novel perspectives for the prevention, diagnosis, and immunotherapeutic intervention of cardiovascular diseases.
    Keywords:  cardiovascular diseases; inflammation; innate immunity; mitochondrial antiviral signaling protein (MAVS); mitochondrial homeostasis
    DOI:  https://doi.org/10.3389/fcvm.2025.1572559
  35. Vascul Pharmacol. 2025 Aug 21. pii: S1537-1891(25)00070-9. [Epub ahead of print]160 107531
      The heart is an organ that depends significantly on mitochondria to operate, since it requires a lot of energy, which mitochondria create, making them essential for the efficient functioning of the heart. The term "mitochondrial dynamics" refers to extremely dynamic organelles known as mitochondria that undergo cycles of fusion and fission to modify their appearance, distribution, and function. Drp1 or Dynamin-related protein 1, a primary fission protein, strictly regulates the elimination of damaged mitochondria by mitophagy. This ensures that the complex processes of organ and cellular dynamics in the heart are strictly managed. Phosphorylation, SUMOylation, palmitoylation, ubiquitination, S-nitrosylation, and O-GlcNAcylation are some of the posttranslational modifications (PTMs) of Drp1 that contribute to the regulation of mitochondrial dynamics. While abnormalities in mitochondrial dynamics are a crucial component of the pathophysiology of a number of cardiovascular diseases (CVDs), the heart requires an effective mitochondrial balance to sustain cardiomyocyte metabolism along with contractile activity. This review summarizes the current knowledge of the crucial function of Drp1 inhibitors in the pathophysiology of cardiovascular diseases, including myocardial ischemia-reperfusion, dysfunction of endothelial cells, smooth muscle remodelling, hypertrophy of the heart, high blood pressure, and myocardial infarction. We further highlighted the possible advantages of treating CVDs by specifically targeting Drp1.
    Keywords:  CVDs; Drp1; Heart; Mitochondrial dynamics; SUMOylation
    DOI:  https://doi.org/10.1016/j.vph.2025.107531
  36. Int J Mol Sci. 2025 Aug 11. pii: 7740. [Epub ahead of print]26(16):
      Ectopic lipid accumulation is a core contributor to insulin resistance and metabolic diseases, including type 2 diabetes, dyslipidemia, and non-alcoholic fatty liver disease. Conventional therapies have primarily focused on redistributing lipid burden across tissues or modulating specific pathways. However, this often causes compensatory responses that merely shift the burden rather than resolve the underlying lipid excess. In this review, we introduce the concept of the ballooning effect, wherein single-target interventions inadvertently exacerbate lipid accumulation in non-target tissues. We then explore fundamental strategies for true lipid disposal, which aim either to prevent lipid influx or to promote complete lipid oxidation. Among these, mild mitochondrial uncoupling emerges as a promising solution. By dissipating substrate energy as heat, mitochondrial uncoupling reduces ectopic lipid burden without relying on redistribution. Recent advances have yielded safer chemical uncouplers and novel endogenous protein-based mechanisms that enable controlled uncoupling with minimal toxicity. Together, these provide a new framework for next-generation metabolic therapies that move beyond lipid redistribution and aim for a true lipid disposal, potentially offering a safe and effective strategy.
    Keywords:  ballooning effects; ectopic lipids; metabolic diseases; mitochondrial uncoupling
    DOI:  https://doi.org/10.3390/ijms26167740
  37. Mol Biol Rep. 2025 Aug 26. 52(1): 846
      Mitochondria serve as an important cellular organelle for maintaining neurotransmission and synaptic plasticity in neuronal cells by playing a key role in ATP generation, maintaining calcium homeostasis, and regulating the levels of reactive oxygen species (ROS), etc. The regulation of the dynamic nature of mitochondria, including their fission, fusion, and removal of damaged mitochondria by mitophagy, is also very important for neuronal health. Abnormalities in mitochondrial processes, including but not limited to fission, fusion, and mitophagy, are known to be associated with numerous neurodegenerative diseases (NDDs), such as Parkinson's disease (PD), Alzheimer's disease (AD), Amyotrophic lateral sclerosis (ALS), and Huntington's disease (HD). In the recent past, the Rho kinase (ROCK) isoforms, particularly ROCK1 and ROCK2, have gained a lot of attention in NDDs, mainly for their role in regulating the dynamics of the mitochondria, mitophagy, and other cell signalling pathways. By adding phosphate groups to Drp1, ROCK1 is crucial in supporting excessive mitochondrial fission, causing the death of neuronal cells. On the other hand, ROCK2 inhibits Parkin-dependent mitophagy by inhibiting the PTEN protein, the activator of Parkin-dependent mitophagy. This impaired mitochondrial quality control via reduced mitophagic flux leads to oxidative stress and neuronal degeneration, the central pathological feature of NDDs. The inhibition of ROCK isoforms has shown great promise in neuroprotective effects, controlling the dynamics of mitochondria in neuronal cells, lowering inflammation, and improving motor and cognitive functions in preclinical models of different NDDs, indicating ROCK isoforms as an attractive therapeutic target in different NDDs. This review aims to highlight the therapeutic significance of targeting ROCK isoforms in different NDDs.
    Keywords:  Mitochondrial dynamics; Mitophagy; Neurodegenerative diseases; Neuroprotective; ROCK isoforms
    DOI:  https://doi.org/10.1007/s11033-025-10947-9
  38. Trends Cell Biol. 2025 Aug 26. pii: S0962-8924(25)00175-8. [Epub ahead of print]
      Defects in ribosomal machinery cause ribosomopathies such as Diamond Blackfan anemia, classically linked to impaired protein synthesis. However, emerging evidence places mitochondrial dysfunction as a critical downstream consequence of ribosomal insufficiency. Thus, is impaired energy metabolism, rather than translation alone, a key driver of ribosomopathies such as Diamond Blackfan anemia? This insight could reframe our understanding of disease mechanisms and could identify metabolic pathways as promising therapeutic targets.
    Keywords:  mitochondrial function; mitochondriopathies; ribosome biogenesis; ribosomopathies
    DOI:  https://doi.org/10.1016/j.tcb.2025.07.007
  39. Life (Basel). 2025 Jul 23. pii: 1168. [Epub ahead of print]15(8):
      An explorative study was conducted to evaluate the efficacy and safety of 5-aminolevulinic acid hydrochloride combined with sodium ferrous citrate (SPP-004) in 10 pediatric patients with Leigh syndrome (LS) aged 3-24 months in 10 institutions between December 2014 and July 2019. The patients were randomized and allocated to the SPP-004 or placebo group for a 12-week double-blind period, followed by a 12-week open-label period with SPP-004 and then a long-term study of up to 180 weeks. The efficacy and safety were evaluated using the Newcastle Pediatric Mitochondrial Disease Scale (NPMDS) and adverse events (AEs), respectively. No significant differences were found between groups in NPMDS scores, but prolonged SPP-004 treatment stabilized or improved scores. During the initial double-blind phase, the serum lactate levels increased in the placebo group but not in the SPP-004 group. Over the period of prolonged treatment with SPP-004, the average serum lactate level gradually decreased to a normal level. One patient died due to heart failure, presumably due to an underlying disease. Overall, 7 out of 10 patients received SPP-004 without developing severe AEs until the termination of the long-term study. Given the severe symptoms and poor prognosis of pediatric LS, NPMDS scores were indicative of stabilization in pediatric LS patients treated with SPP-004.
    Keywords:  5-aminolevulinic acid; Leigh syndrome; mitochondrial disease; mitochondrial respiratory chain disorder; sodium ferrous citrate
    DOI:  https://doi.org/10.3390/life15081168
  40. Bioorg Med Chem. 2025 Aug 11. pii: S0968-0896(25)00291-3. [Epub ahead of print]130 118350
      Mitochondrial proteostasis is essential for tumorigenesis, and mitochondrial inner membrane proteins have emerged as meaningful targets due to their crucial functions in regulating apoptosis, maintaining oxidative phosphorylation, and influencing tumor initiation and progression. Targeted protein degradation (TPD) has garnered significant attention as a promising therapeutic approach. However, conventional TPD platforms relying on the ubiquitin-proteasome system or lysosomal pathways encounter inherent obstacles in targeting proteins sequestered within the mitochondrial compartment and cannot degrade mitochondrial inner membrane proteins. Utilizing our previously established MtPTAC system, we selected dihydroorotate dehydrogenase (DHODH), the rate-limiting enzyme in de novo pyrimidine biosynthesis, as a model substrate. We designed and synthesized a series of degraders, with 3D-2 achieving over 50 % degradation efficiency of DHODH via the ClpP protease. This degrader can form a stable ternary complex with DHODH and ClpP, and it exhibits significant inhibitory effects across various tumor cell lines. This technological innovation is the first to successfully degrade endogenous mitochondrial inner membrane proteins. It provides a diverse toolkit for investigating mitochondrial protein functions and paving the way for novel anticancer therapies.
    Keywords:  DHODH degrader; Inner mitochondrial membrane; MtPTAC; Targeting protein degradation
    DOI:  https://doi.org/10.1016/j.bmc.2025.118350
  41. J Genet Genomics. 2025 Aug 22. pii: S1673-8527(25)00229-2. [Epub ahead of print]
      The mitochondrial 3243A>G mutation (m.3243A>G) is associated with diverse clinical phenotypes. To elucidate the underlying mechanisms and explore intervention strategies in m.3243A>G patients, urine-derived stem cells (USCs) and a mitochondrial leucyl-tRNA synthetase (lars-2) deficient Caenorhabditis elegans (C. elegans) model are used to assess mitochondrial homeostasis and neuromuscular dysfunction. Patient-derived USCs with high levels of m.3243A>G heteroplasmy exhibit impaired mitochondrial function, disrupted mitochondrial dynamics, and inhibited mitophagy, which are reversed by MitoQ through suppression of OMA1 zinc metallopeptidase (OMA1)-induced mitochondrial phosphatase and tensin (PTEN) induced kinase 1 (PINK1) degradation. Furthermore, lars-2 knockdown in C. elegans induces mitochondrial stress and mimics the loss of neural and muscle functions observed in patients with the m.3243A>G mutation. MitoQ treatment partially improves neurobehavioral function by promoting the PINK1 pathway. These findings suggest that MitoQ has therapeutic potential in the context of the m.3243A>G mutation.
    Keywords:  C. elegans; MitoQ; Mitochondrial quality control; USCs; m.3243A>G
    DOI:  https://doi.org/10.1016/j.jgg.2025.08.007
  42. Methods Protoc. 2025 Aug 02. pii: 87. [Epub ahead of print]8(4):
      Mitochondria play a crucial role in adapting to fluctuating energy demands, particularly in various heart diseases. In addition to functional analyses such as the measurement of ROS or ATP, analysis of mitochondrial ultrastructure can be used to draw further conclusions about their functions and effects in tissue. In this protocol, we introduce a set of measurements to compare the ultrastructural and functional characteristics of human left ventricular mitochondria, using transmission electron microscopy (TEM). Measured parameters included mean size in µm2, elongation, count, percental mitochondrial area in the measuring frame, and a conglomeration score. We also introduce a novel method of defining hydropic mitochondria as a comparable evaluation standard. With this cluster of measurement parameters, we aim to contribute a protocol for studying human mitochondrial morphology, distribution, and functionality.
    Keywords:  cardiomyocytes; electron microscopy; mitochondria; quantitative image analysis; ultrastructure
    DOI:  https://doi.org/10.3390/mps8040087
  43. Mol Metab. 2025 Aug 25. pii: S2212-8778(25)00146-2. [Epub ahead of print] 102239
      Diets influence metabolism and disease susceptibility, with lysine acetyltransferases (KATs) serving as key regulators through acetyl-CoA. We have previously demonstrated that a ketogenic diet alleviates cardiac pathology, though the underlying mechanisms remain largely unknown. Here we show that KAT6A acetylation is crucial for mitochondrial function and cell growth. Proteomic analysis revealed that KAT6A is acetylated at lysine (K)816 in the hearts of mice fed a ketogenic diet under hypertension, which enhances its interaction with AMPK regulatory subunits. RNA-sequencing analysis demonstrated that the KAT6A acetylation-mimetic mutant stimulates AMPK signaling in cardiomyocytes. Moreover, the acetylation-mimetic mutant mitigated phenylephrine-induced mitochondrial dysfunction and cardiomyocyte hypertrophy via AMPK activation. However, KAT6A-K816R acetylation-resistant knock-in mice unexpectedly exhibited smaller hearts with enhanced AMPK activity, conferring protection against neurohumoral stress-induced cardiac hypertrophy and remodeling. These findings indicate that KAT6A regulates metabolism and cellular growth by interacting with and modulating AMPK activity through K816-acetylation in a cell type-specific manner.
    Keywords:  AMPK; KAT6A; acetylation; acetyltransferase; cardiac hypertrophy; heart failure; hypertensive cardiomyopathy; ketogenic diet; ketone body; lysine acetyltransferase 6A
    DOI:  https://doi.org/10.1016/j.molmet.2025.102239
  44. Nucleic Acids Res. 2025 Aug 27. pii: gkaf803. [Epub ahead of print]53(16):
      The mitochondrial DNA helicase TWINKLE, a hexameric ring-shaped helicase, plays a crucial role in maintaining mitochondrial DNA integrity. TWINKLE translocates along one DNA strand, unwinding the duplex by excluding the complementary strand through coordinated ATP hydrolysis. However, the precise mechanisms underlying this process remain incompletely understood. In this study, we utilized single-molecule Förster Resonance Energy Transfer (smFRET) to investigate the mechanisms of TWINKLE-mediated DNA unwinding. Our results reveal that TWINKLE occasionally pauses during unwinding, with the rate of unwinding and the duration of pausing strongly influenced by ATP concentration, but not by the presence of DNA mismatches or mitochondrial single-stranded DNA-binding protein (mtSSB). These findings suggest that the pausing events primarily arise from stochastic ATP hydrolysis within the helicase subunits. DNA mismatches exacerbate TWINKLE's pausing and dissociation from DNA, thereby impairing DNA unwinding. In contrast, mtSSB significantly mitigates helicase dissociation by stabilizing TWINKLE-DNA interactions. This study provides novel insights into the functional dynamics of TWINKLE, highlighting the role of ATP hydrolysis in orchestrating single-stranded DNA translocation, the detrimental effects of DNA mismatches on DNA unwinding, and the critical role of mtSSB in supporting helicase function.
    DOI:  https://doi.org/10.1093/nar/gkaf803
  45. JACC Case Rep. 2025 Aug 20. pii: S2666-0849(25)01456-1. [Epub ahead of print]30(24): 104676
       BACKGROUND: Nonischemic cardiomyopathy (NICM) can be caused by single-gene mutations, including genes such as inorganic pyrophosphatase 2 (PPA2) with multisystem effects.
    CASE SUMMARY: A 28-year-old woman presenting with respiratory symptoms was discharged with a diagnosis of decompensated idiopathic NICM. Her NICM progressively worsened, and the patient underwent a heart transplant at the age of 38 and again at the age of 42. At age 47, genetic testing confirmed 2 mutations in the PPA2 gene that had caused her NICM.
    DISCUSSION: This patient is to our knowledge the oldest published to date (48 years) presenting with cardiac symptoms who has PPA2 deficiency, a mitochondrial disease characterized by sudden cardiac death in infancy.
    TAKE-HOME MESSAGE: This case exemplifies the utility of employing genetic testing early in the diagnostic workup of NICM before applying the designation "idiopathic."
    Keywords:  PPA2; cardiomyopathy; mitochondrial disease; monogenic
    DOI:  https://doi.org/10.1016/j.jaccas.2025.104676
  46. Aging (Albany NY). 2025 Aug 25. 17
      Research in the field of mitochondrial biomarkers plays an important role in understanding the processes of cellular aging. Mitochondria are not only the energy centers of the cell, but also key regulators of signaling within the cell. They significantly affect the life and function of the cell. The aging process of cells is associated with various factors, including DNA damage, disruption of the cell cycle, changes in mitochondria, and problems with signal transmission. Mitochondrial dysfunction is a major contributor to cellular and organismal aging. As we age, there is an accumulation of dysfunctional mitochondria, leading to decreased efficiency of oxidative phosphorylation and increased production of reactive oxygen species. This review focuses on the main mitochondrial markers involved in the mechanisms of cell aging: DRP1, Prohibitin, Parkin, PINK1, MFF, VDAC, TOM. These signaling molecules are involved in mitochondrial fission and the mechanisms of mitochondria-dependent apoptosis, in the regulation of mitochondrial respiratory activity, ensuring the stability of the organization and copying of mitochondrial DNA, protecting cells from oxidative stress, in the process of autophagy of damaged mitochondria, in protective mechanisms during stress-induced mitochondrial dysfunction. Analysis of mitochondrial markers can provide valuable information about the state of cells and their functional significance at various stages of aging, which could promote our understanding of cellular aging mechanisms and developing corrective methods. These insights highlight mitochondrial proteins as potential therapeutic targets to combat age-related diseases.
    Keywords:  age-associated diseases; biomarkers; cellular senescence; mitochondria; mitochondrial proteins
    DOI:  https://doi.org/10.18632/aging.206305
  47. Bioessays. 2025 Aug 22. e70058
      Recent advances in genomics uncovered a large number of microproteins, which are peptides of less than 100 amino-acids encoded by small open reading frames. In contrast to their identification, the validation of the functions of microproteins remains challenging. Especially, what are their biological functions in the cell and how this relates to disease conditions are still largely unknown. Although microproteins ensure a plethora of cellular functions, recent evidence demonstrate that they may disproportionately affect cellular metabolism. In this review, we will address the roles of mitochondrial-targeted microproteins, and especially how this class of protein regulates neuronal metabolism in neurodevelopment and neurodegeneration, and may contribute to axonal and dendritic metabolic disorders.
    Keywords:  metabolism; microproteins; mitochondria; neurodegeneration; neurodevelopment; neuron
    DOI:  https://doi.org/10.1002/bies.70058
  48. Cells. 2025 Aug 18. pii: 1278. [Epub ahead of print]14(16):
      Aging, a state of progressive decline in physiological function, is an important risk factor for chronic diseases, ranging from cancer and musculoskeletal frailty to cardiovascular and neurodegenerative diseases. Understanding its cellular basis is critical for developing interventions to extend human health span. This review highlights the crucial role of in vitro models, discussing foundational discoveries like the Hayflick limit and the senescence-associated secretory phenotype (SASP), the utility of immortalized cell lines, and transformative human induced pluripotent stem cells (iPSCs) for aging and disease modeling and rejuvenation studies. We also examine methods to induce senescence and discuss the distinction between chronological time and biological clock, with examples of applying cells from progeroid syndromes and mitochondrial diseases to recapitulate some signaling mechanisms in aging. Although no in vitro model can perfectly recapitulate organismal aging, well-chosen models are invaluable for addressing specific mechanistic questions. We focus on experimental strategies to manipulate cellular aging: from "steering" cells toward resilience to "reversing" age-related phenotypes via senolytics, partial epigenetic reprogramming, and targeted modulation of proteostasis and mitochondrial health. This review ultimately underscores the value of in vitro systems for discovery and therapeutic testing while acknowledging the challenge of translating insights from cell studies into effective, organism-wide strategies to promote healthy aging.
    Keywords:  cardiovascular aging; cellular aging; epigenetic reprogramming; in vitro models; induced pluripotent stem cells (iPSCs); mitochondrial dysfunction; neurodegeneration; progeroid syndromes; senescence; senolytics
    DOI:  https://doi.org/10.3390/cells14161278
  49. Endocrine. 2025 Aug 23.
      
    Keywords:   ACADVL ; Compound heterozygous variants; Next-generation sequencing; Very long-chain acyl-CoA dehydrogenase deficiency
    DOI:  https://doi.org/10.1007/s12020-025-04399-1
  50. Commun Biol. 2025 Aug 26. 8(1): 1280
      Protein kinase A (PKA) plays a crucial and conserved role in various biological processes across species. Despite its significance, tissue-level regulatory networks controlling PKA activity remain incompletely characterized. In this study, we develop a live animal PKA sensor that can faithfully indicate changes in PKA activity in the intestines of Caenorhabditis elegans (C. elegans). Using complementary genome-wide and intestine-specific RNAi screens, we reveal both intestine-autonomous and non-autonomous regulators of intestinal PKA. Notably, we show that inhibiting mitochondrial oxidative phosphorylation, either through RNAi or chemical treatment, leads to a marked increase in intestinal PKA activity. Additionally, we demonstrate that the pyrimidine synthesis pathway serves as a critical autonomous regulator of PKA in the intestines of C. elegans, operating mechanistically in a cAMP-independent manner. Furthermore, this pathway holds a critical and conserved role in regulating PKA in cultured human cells. Overall, our study uncovers the first set of intestinal PKA regulators in C. elegans, with potential implications for PKA modulation under physiological or pathological conditions across species.
    DOI:  https://doi.org/10.1038/s42003-025-08718-0
  51. Am J Physiol Endocrinol Metab. 2025 Aug 26.
      The mitochondrial tRNA-derived fragment mt-tRF-LeuTAA couples mitochondrial metabolism to insulin secretion. While its role in pancreatic β-cell function is well established, its broader impact on multi-organ glucose homeostasis remains unclear. In insulin target tissues, the presence, regulation, and mechanism of action of mt-tRF-LeuTAA are entirely unexplored. This study addresses this gap by investigating the impact of diet, nutritional status and diabetes on mt-tRF-LeuTAA regulation and by assessing its role in insulin sensitivity. We examined mt-tRF-LeuTAA levels in different insulin target tissues, including skeletal muscle, liver, and epididymal white adipose tissue, of rodents under physiological and pathological conditions. In skeletal muscle myotubes, we combined subcellular fractionation, antisense oligonucleotide-mediated knockdown and glucose uptake assays to determine mt-tRF-LeuTAA's mitochondrial localization and its influence on insulin sensitivity. mt-tRF-LeuTAA levels in mouse skeletal muscle decreased twofold in response to fasting. In myotubes, this tRNA fragment was enriched in mitochondria, and its downregulation enhanced glucose uptake. While the levels of mt-tRF-LeuTAA remained unchanged in insulin target tissues of diabetic mice, we observed a skeletal muscle-specific downregulation of mt-tRF-LeuTAA in young adult rats exhibiting insulin hypersensitivity. This study identifies mt-tRF-LeuTAA as a candidate regulator of skeletal muscle insulin response. By modulating both insulin secretion and action, mt-tRF-LeuTAA appears to play a notable role in systemic metabolic control, and may represent a promising target for diabetes treatment.
    Keywords:  Diabetes susceptibility; Mitochondrially-encoded tRNAs; Muscle glucose uptake; Nutritional status; mt-tRNA-derived fragments
    DOI:  https://doi.org/10.1152/ajpendo.00284.2025
  52. Antioxidants (Basel). 2025 Jul 29. pii: 923. [Epub ahead of print]14(8):
      Platelets have long been known to be critically involved in hemostasis and thrombosis. However, platelets are also recognized as metabolically active cells that require well-regulated mitochondrial function to support their multiple functions in hemostasis, thrombosis, and inflammation. Mitochondrial activity has also recently been shown to play a crucial role in determining platelet activation, survival, and pro-inflammatory potential. A key nexus in these processes is the mitochondrial permeability transition pore (mPTP), a high-conductance channel in the inner mitochondrial membrane. Sustained mPTP opening triggers mitochondrial depolarization, the cessation of ATP synthesis, osmotic swelling, and, finally, platelet dysfunction or clearance. However, its transient opening might play physiological signaling roles. This review summarizes the current understanding of the molecular components and regulatory factors governing the platelet mPTP, explores its physiological and pathological relevance, and evaluates its potential as a therapeutic target in cardiovascular disease, inflammation, cancer, and potentially neurodegenerative diseases. We also highlight the ongoing challenges and crucial future directions in deciphering the complexities of platelet mitochondrial dynamics and mPTP functions.
    Keywords:  mitochondrial permeability transition pore; platelets; therapeutic targets
    DOI:  https://doi.org/10.3390/antiox14080923
  53. Mol Genet Metab. 2025 Aug 14. pii: S1096-7192(25)00212-4. [Epub ahead of print]146(1-2): 109221
      Citrin deficiency is an autosomal recessive disorder caused by mutations in SLC25A13, encoding the inner mitochondrial membrane protein citrin. This disease manifests in age-dependent forms: neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD), failure to thrive and dyslipidemia caused by citrin deficiency, and adolescent and adult citrin deficiency (AACD). While NICCD often resolves with early treatment, AACD symptoms tend to persist or worsen over time. However, the long-term outcome in adult patients with citrin deficiency, particularly those affected by the symptoms of AACD, is not well understood. To address this gap, we conducted a new study from 2022 to 2024 that focused specifically on adult patients with citrin deficiency. This retrospective study investigated the long-term outcomes in a total of 128 adult patients, categorized depending on their onset of disease into NICCD, Post-NICCD, and AACD groups. Significant differences in height were observed: NICCD patients had taller median heights than AACD patients (Males: 170.6 cm vs. 168.0 cm, P = 0.016; Females: 156.0 cm vs. 153.0 cm, P = 0.007). Former NICCD patients generally achieved favorable long-term outcomes with early intervention, while AACD patients often experienced persistent or worsening symptoms, including irreversible liver damage with impaired urea cycle function. In conclusion, this study suggests that early intervention during infancy or childhood may improve long-term prognosis in citrin deficiency, particularly for NICCD patients. Conversely, outcome for AACD patients remains a concern, highlighting the need for improved management strategies in adulthood. This study emphasizes the importance of timely diagnosis and treatment to mitigate the progressive nature of citrin deficiency.
    Keywords:  Adolescent and adult citrin deficiency; End stage liver disease; Hepatic encephalopathy; Intellectual disability; Liver transplantation
    DOI:  https://doi.org/10.1016/j.ymgme.2025.109221
  54. Nat Commun. 2025 Aug 22. 16(1): 7811
      Mitochondria-lysosome interactions are critical for maintaining cellular homeostasis. Although genetically encoded protein based optogenetic technique is developed to regulate such interactions, it still suffers from shortcomings including complicated operation and potential interference to organelle functions. Here, we present a fast, simple, biocompatible and programmable platform via activable DNA regulators to achieve spatiotemporal regulation of mitochondria-lysosome interactions in living cells. In our system, two locked DNA regulators, OK-MLIR and DK-MLIR, that can be respectively activated with UV light (One Key) as well as UV light and endogenous glutathione (Dual Keys), are modularly designed for modulating mitochondria-lysosome contacts. We show that these DNA regulators can be used for facilitating mitochondrial fission and autophagy. Moreover, the DK-MLIR enables selective and efficient manipulation of target cell migration and proliferation with highly temporal and spatial controllability. This programmable and modular design principle provides a platform for organelle interaction study, cellular regulation and precision therapy.
    DOI:  https://doi.org/10.1038/s41467-025-63040-x
  55. Cell. 2025 Aug 13. pii: S0092-8674(25)00863-3. [Epub ahead of print]
      RNA contains diverse post-transcriptional modifications, and its catabolic breakdown yields numerous modified nucleosides requiring correct processing, but the mechanisms remain unknown. Here, we demonstrate that three RNA-derived modified adenosines, N6-methyladenosine (m6A), N6,N6-dimethyladenosine (m6,6A), and N6-isopentenyladenosine (i6A), are sequentially metabolized into inosine monophosphate (IMP) to mitigate their intrinsic cytotoxicity. After phosphorylation by adenosine kinase (ADK), they undergo deamination by adenosine deaminase-like (ADAL). In Adal knockout mice, N6-modified adenosine monophosphates (AMPs) accumulate and allosterically inhibit AMP-activated protein kinase (AMPK), dysregulating glucose metabolism. Furthermore, ADK deficiency, linked to human inherited disorders of purine metabolism, elevates levels of the three modified adenosines, resulting in early lethality in mice. Mechanistically, excessive m6A, m6,6A, and i6A impair lysosomal function by interfering with lysosomal membrane proteins, thereby disrupting lipid metabolism and causing cellular toxicity. Through this nucleotide metabolism pathway and mechanism, cells detoxify modified adenosines, linking modified RNA metabolism to human disease.
    Keywords:  ADAL; ADK; AMP-activated protein kinase; AMPK; Adenosine deaminase-like; Adenosine kinase; Lipid metabolism; Lysosome; Modified RNA metabolism; Purine metabolism; RNA modification; m(6)A
    DOI:  https://doi.org/10.1016/j.cell.2025.07.041
  56. Nature. 2025 Aug 27.
      
    Keywords:  Cell biology; Metabolism; Structural biology
    DOI:  https://doi.org/10.1038/d41586-025-02694-5