bims-mitmed 2022-04-17 papers

bims-mitmed

Biomed News

on Mitochondrial medicine

Issue of 2022–04–17
34 papers selected by
Dario Brunetti, Fondazione IRCCS Istituto Neurologico

SnapShot: Regulation and biology of PGC-1α.
Mitochondrial DNA Transport in Drosophila Neurons.
L-carnitine rescue for neonatal intractable mitochondrial cardiomyopathy.
Mitochondrial-to-nuclear communication in aging: an epigenetic perspective.
High-Resolution Imaging of Mitochondria and Mitochondrial Nucleoids in Differentiated SH-SY5Y Cells.
Cloning and Organelle Expression of Bamboo Mitochondrial Complex I Subunits Nad1, Nad2, Nad4, and Nad5 in the Yeast Saccharomyces cerevisiae.
The Role of Impaired Mitochondrial Dynamics in MFN2-Mediated Pathology.
Mitochondrial Lipids: From Membrane Organization to Apoptotic Facilitation.
Mitochondrial supplementation of Sus scrofa metaphase II oocytes alters DNA methylation and gene expression profiles of blastocysts.
Clinical and genetic features of a cohort of patients with MFN2-related neuropathy.
L-arginine in Mitochondrial Encephalopathy, Lactic Acidosis, and Stroke-like Episodes: A Systematic Review.
Cerebellar Pathology in an Inducible Mouse Model of Friedreich Ataxia.
Mitochondrial Dysfunction and Acute Fatty Liver of Pregnancy.
Mitochondrial shape alteration by metabolites.
Pharmacologic enrichment of exosome yields and mitochondrial cargo.
Assessing Energy Substrate Oxidation In Vitro with 14CO2 Trapping.
Detailed Imaging of Mitochondrial Transport and Precise Manipulation of Mitochondrial Function with Genetically Encoded Photosensitizers in Adult Drosophila Neurons.
The great astroglial metabolic revolution: Mitochondria fuel astrocyte homeostatic support and neuroprotection.
The smoothened agonist SAG reduces mitochondrial dysfunction and neurotoxicity of frataxin-deficient astrocytes.
Mitochondrial function in spinal cord injury and regeneration.
In Vitro Reconstitution of Molecular Motor-Driven Mitochondrial Transport.
Erodible thermogelling hydrogels for localized mitochondrial transplantation to the spinal cord.
Induction of Mitochondrial Fragmentation and Mitophagy after Neonatal Hypoxia-Ischemia.
The molecular basis of spinocerebellar ataxia type 48 caused by a de novo mutation in the ubiquitin ligase CHIP.
Mitolysosome exocytosis, a mitophagy-independent mitochondrial quality control in flunarizine-induced parkinsonism-like symptoms.
Do compromised mitochondria aggravate severity and fatality by SARS-CoV-2?
DGAT1 activity synchronises with mitophagy to protect cells from metabolic rewiring by iron depletion.
Activation of mitophagy by rapamycin eliminated the accumulation of TDP-43 on mitochondrial and promoted the resolution of carbon tetrachloride-induced liver fibrosis in mice.
Proteolysis of adaptor protein Mmr1 during budding is necessary for mitochondrial homeostasis in Saccharomyces cerevisiae.
Mitochondrial Respiratory Complexes as Targets of Drugs: The PPAR Agonist Example.
Age-related disruption of the proteome and acetylome in mouse hearts is associated with loss of function and attenuated by elamipretide (SS-31) and nicotinamide mononucleotide (NMN) treatment.
Clinical trials for cerebellar ataxia.
Mitochondrial protease ClpP supplementation ameliorates diet-induced NASH in mice.
Metabolic Syndrome and β-Oxidation of Long-Chain Fatty Acids in the Brain, Heart, and Kidney Mitochondria.

Cell. 2022 Apr 14. pii: S0092-8674(22)00337-3. [Epub ahead of print]185(8): 1444-1444.e1

SnapShot: Regulation and biology of PGC-1α.

Paulo R Jannig, Phillip A Dumesic, Bruce M Spiegelman, Jorge L Ruas.

The peroxisome proliferator-activated receptor γ coactivator-1α (Ppargc1a) gene encodes several PGC-1α isoforms that regulate mitochondrial bioenergetics and cellular adaptive processes. Expressing specific PGC-1α isoforms in mice can confer protection in different disease models. This SnapShot summarizes how regulation of Ppargc1a transcription, splicing, translation, protein stability, and activity underlies its multifaceted functions. To view this SnapShot, open or download the PDF.

DOI: https://doi.org/10.1016/j.cell.2022.03.027
Methods Mol Biol. 2022 ;2431 409-416

Mitochondrial DNA Transport in Drosophila Neurons.

Joseph M Bateman.

  Mitochondria are essential organelles that generate energy and play vital roles in cellular metabolism. The small circular mitochondrial genome encodes key components of the mitochondrial respiratory apparatus. Depletion of, or mutations in mitochondrial DNA (mtDNA) cause mitochondrial dysfunction and disease. mtDNA is packaged into nucleoids, which are transported throughout the cell within mitochondria. Efficient transport of nucleoids is essential in neurons, where mitochondrial function is required locally at synapses. Here I describe methods for visualization of nucleoids in Drosophila neurons using a GFP fusion of the mitochondrial transcription factor TFAM. TFAM-GFP, together with mCherry-labeled mitochondria, was used to visualize nucleoids in fixed larval segmental nerves. I also describe how these tools can be used for live imaging of nucleoid dynamics. Using Drosophila as a model system, these methods will enable further characterization and analysis of nucleoid dynamics in neurons.

Keywords:  Drosophila; Live imaging; Mitochondrial DNA; Nucleoid; TFAM

DOI:  https://doi.org/10.1007/978-1-0716-1990-2_21
Pediatr Int. 2022 Jan;64(1): e15143

L-carnitine rescue for neonatal intractable mitochondrial cardiomyopathy.

Yuka Toyama, Yoshimitsu Fujii, Shin-Ichiro Hori, Ken Yoshimura, Kazunari Kaneko.



Keywords:   SLC25A26 ; L-carnitine; mitochondrial cardiomyopathy; mitochondrial diseases; neonates

DOI:  https://doi.org/10.1111/ped.15143
Trends Biochem Sci. 2022 Apr 06. pii: S0968-0004(22)00067-6. [Epub ahead of print]

Mitochondrial-to-nuclear communication in aging: an epigenetic perspective.

Di Zhu, Xinyu Li, Ye Tian.

  Age-associated changes in mitochondria are closely involved in aging. Apart from the established roles in bioenergetics and biosynthesis, mitochondria are signaling organelles that communicate their fitness to the nucleus, triggering transcriptional programs to adapt homeostasis stress that is essential for organismal health and aging. Emerging studies revealed that mitochondrial-to-nuclear (mito-nuclear) communication via altered levels of mitochondrial metabolites or stress signals causes various epigenetic changes, facilitating efforts to maintain homeostasis and affect aging. Here, we summarize recent studies on the mechanisms by which mito-nuclear communication modulates epigenomes and their effects on regulating the aging process. Insights into understanding how mitochondrial metabolites serve as prolongevity signals and how aging affects this communication will help us develop interventions to promote longevity and health.

Keywords:  UPR(mt); aging; epigenetic regulation; longevity; mitochondrial metabolites; mitochondrial–nuclear communication

DOI:  https://doi.org/10.1016/j.tibs.2022.03.008
Methods Mol Biol. 2022 ;2431 291-310

High-Resolution Imaging of Mitochondria and Mitochondrial Nucleoids in Differentiated SH-SY5Y Cells.

Emily Annuario, Kristal Ng, Alessio Vagnoni.

  Mitochondria are highly dynamic organelles which form intricate networks with complex dynamics. Mitochondrial transport and distribution are essential to ensure proper cell function, especially in cells with an extremely polarised morphology such as neurons. A layer of complexity is added when considering mitochondria have their own genome, packaged into nucleoids. Major mitochondrial morphological transitions, for example mitochondrial division, often occur in conjunction with mitochondrial DNA (mtDNA) replication and changes in the dynamic behaviour of the nucleoids. However, the relationship between mtDNA dynamics and mitochondrial motility in the processes of neurons has been largely overlooked. In this chapter, we describe a method for live imaging of mitochondria and nucleoids in differentiated SH-SY5Y cells by instant structured illumination microscopy (iSIM). We also include a detailed protocol for the differentiation of SH-SY5Y cells into cells with a pronounced neuronal-like morphology and show examples of coordinated mitochondrial and nucleoid motility in the long processes of these cells.

Keywords:  Axonal transport; Instant structured illumination microscopy (iSIM); Mitochondria; Mitochondrial DNA; Mitochondrial fission; Neuronal differentiation; Nucleoids; SH-SY5Y cells; Superresolution

DOI:  https://doi.org/10.1007/978-1-0716-1990-2_15
Int J Mol Sci. 2022 Apr 06. pii: 4054. [Epub ahead of print]23(7):

Cloning and Organelle Expression of Bamboo Mitochondrial Complex I Subunits Nad1, Nad2, Nad4, and Nad5 in the Yeast Saccharomyces cerevisiae.

Hsieh-Chin Tsai, Cheng-Hung Hsieh, Ching-Wen Hsu, Yau-Heiu Hsu, Lee-Feng Chien.

  Mitochondrial respiratory complex I catalyzes electron transfer from NADH to ubiquinone and pumps protons from the matrix into the intermembrane space. In particular, the complex I subunits Nad1, Nad2, Nad4, and Nad5, which are encoded by the nad1, nad2, nad4, and nad5 genes, reside at the mitochondrial inner membrane and possibly function as proton (H+) and ion translocators. To understand the individual functional roles of the Nad1, Nad2, Nad4, and Nad5 subunits in bamboo, each cDNA of these four genes was cloned into the pYES2 vector and expressed in the mitochondria of the yeast Saccharomyces cerevisiae. The mitochondrial targeting peptide mt gene (encoding MT) and the egfp marker gene (encoding enhanced green fluorescent protein, EGFP) were fused at the 5'-terminal and 3'-terminal ends, respectively. The constructed plasmids were then transformed into yeast. RNA transcripts and fusion protein expression were observed in the yeast transformants. Mitochondrial localizations of the MT-Nad1-EGFP, MT-Nad2-EGFP, MT-Nad4-EGFP, and MT-Nad5-EGFP fusion proteins were confirmed by fluorescence microscopy. The ectopically expressed bamboo subunits Nad1, Nad2, Nad4, and Nad5 may function in ion translocation, which was confirmed by growth phenotype assays with the addition of different concentrations of K+, Na+, or H+.

Keywords:  bamboo; mitochondrial complex I; organelle expression; proton pumping subunits

DOI:  https://doi.org/10.3390/ijms23074054
Front Cell Dev Biol. 2022 ;10 858286

The Role of Impaired Mitochondrial Dynamics in MFN2-Mediated Pathology.

Mashiat Zaman, Timothy E Shutt.

  The Mitofusin 2 protein (MFN2), encoded by the MFN2 gene, was first described for its role in mediating mitochondrial fusion. However, MFN2 is now recognized to play additional roles in mitochondrial autophagy (mitophagy), mitochondrial motility, lipid transfer, and as a tether to other organelles including the endoplasmic reticulum (ER) and lipid droplets. The tethering role of MFN2 is an important mediator of mitochondrial-ER contact sites (MERCs), which themselves have many important functions that regulate mitochondria, including calcium homeostasis and lipid metabolism. Exemplifying the importance of MFN2, pathogenic variants in MFN2 are established to cause the peripheral neuropathy Charcot-Marie-Tooth Disease Subtype 2A (CMT2A). However, the mechanistic basis for disease is not clear. Moreover, additional pathogenic phenotypes such as lipomatosis, distal myopathy, optic atrophy, and hearing loss, can also sometimes be present in patients with CMT2A. Given these variable patient phenotypes, and the many cellular roles played by MFN2, the mechanistic underpinnings of the cellular impairments by which MFN2 dysfunction leads to disease are likely to be complex. Here, we will review what is known about the various functions of MFN2 that are impaired by pathogenic variants causing CMT2A, with a specific emphasis on the ties between MFN2 variants and MERCs.

Keywords:  CMT2A; MFN2; lipid homeostasis; mitochondria; mitochondrial dynamics; mitochondrial endoplasmic reticulum contact sites; mitophagy; mtDNA

DOI:  https://doi.org/10.3389/fcell.2022.858286
Int J Mol Sci. 2022 Mar 29. pii: 3738. [Epub ahead of print]23(7):

Mitochondrial Lipids: From Membrane Organization to Apoptotic Facilitation.

Aikaterini Poulaki, Stavroula Giannouli.

  Mitochondria are the most complex intracellular organelles, their function combining energy production for survival and apoptosis facilitation for death. Such a multivariate physiology is structurally and functionally reflected upon their membrane configuration and lipid composition. Mitochondrial double membrane lipids, with cardiolipin as the protagonist, show an impressive level of complexity that is mandatory for maintenance of mitochondrial health and protection from apoptosis. Given that lipidomics is an emerging field in cancer research and that mitochondria are the organelles with the most important role in malignant maintenance knowledge of the mitochondrial membrane, lipid physiology in health is mandatory. In this review, we will thus describe the delicate nature of the healthy mitochondrial double membrane and its role in apoptosis. Emphasis will be given on mitochondrial membrane lipids and the changes that they undergo during apoptosis induction and progression.

Keywords:  apoptosis; cardiolipin; ceramide; cytochrome C; double membrane; mitochondria; mitochondrial lipids

DOI:  https://doi.org/10.3390/ijms23073738
Epigenetics Chromatin. 2022 Apr 15. 15(1): 12

Mitochondrial supplementation of Sus scrofa metaphase II oocytes alters DNA methylation and gene expression profiles of blastocysts.

Takashi Okada, Stephen McIlfatrick, Nhi Hin, Nader Aryamanesh, James Breen, Justin C St John.

   BACKGROUND: Mitochondrial DNA (mtDNA) copy number in oocytes correlates with oocyte quality and fertilisation outcome. The introduction of additional copies of mtDNA through mitochondrial supplementation of mtDNA-deficient Sus scrofa oocytes resulted in: (1) improved rates of fertilisation; (2) increased mtDNA copy number in the 2-cell stage embryo; and (3) improved development of the embryo to the blastocyst stage. Furthermore, a subset of genes showed changes in gene expression. However, it is still unknown if mitochondrial supplementation alters global and local DNA methylation patterns during early development.
RESULTS: We generated a series of embryos in a model animal, Sus scrofa, by intracytoplasmic sperm injection (ICSI) and mitochondrial supplementation in combination with ICSI (mICSI). The DNA methylation status of ICSI- and mICSI-derived blastocysts was analysed by whole genome bisulfite sequencing. At a global level, the additional copies of mtDNA did not affect nuclear DNA methylation profiles of blastocysts, though over 2000 local genomic regions exhibited differential levels of DNA methylation. In terms of the imprinted genes, DNA methylation patterns were conserved in putative imprint control regions; and the gene expression profile of these genes and genes involved in embryonic genome activation were not affected by mitochondrial supplementation. However, 52 genes showed significant differences in expression as demonstrated by RNAseq analysis. The affected gene networks involved haematological system development and function, tissue morphology and cell cycle. Furthermore, seven mtDNA-encoded t-RNAs were downregulated in mICSI-derived blastocysts suggesting that extra copies of mtDNA affected tRNA processing and/or turnover, hence protein synthesis in blastocysts. We also showed a potential association between differentially methylated regions and changes in expression for 55 genes due to mitochondrial supplementation.
CONCLUSIONS: The addition of just an extra ~ 800 copies of mtDNA into oocytes can have a significant impact on both gene expression and DNA methylation profiles in Sus scrofa blastocysts by altering the epigenetic programming established during oogenesis. Some of these changes may affect specific tissue-types later in life. Consequently, it is important to determine the longitudinal effect of these molecular changes on growth and development before considering human clinical practice.

Keywords:  Assisted reproductive technology; Blastocyst; DNA methylation; Mitochondrial DNA; Mitochondrial supplementation; Oocyte; Sus scrofa; Transcriptome analysis; Whole genome bisulfite sequencing

DOI:  https://doi.org/10.1186/s13072-022-00442-x
Sci Rep. 2022 Apr 13. 12(1): 6181

Clinical and genetic features of a cohort of patients with MFN2-related neuropathy.

Elena Abati, Arianna Manini, Daniele Velardo, Roberto Del Bo, Laura Napoli, Federica Rizzo, Maurizio Moggio, Nereo Bresolin, Emilia Bellone, Maria Teresa Bassi, Maria Grazia D'Angelo, Giacomo Pietro Comi, Stefania Corti.

Charcot-Marie-Tooth disease type 2A (CMT2A) is a rare inherited axonal neuropathy caused by mutations in MFN2 gene, which encodes Mitofusin 2, a transmembrane protein of the outer mitochondrial membrane. We performed a cross-sectional analysis on thirteen patients carrying mutations in MFN2, from ten families, describing their clinical and genetic characteristics. Evaluated patients presented a variable age of onset and a wide phenotypic spectrum, with most patients presenting a severe phenotype. A novel heterozygous missense variant was detected, p.K357E. It is located at a highly conserved position and predicted as pathogenic by in silico tools. At a clinical level, the p.K357E carrier shows a severe sensorimotor axonal neuropathy. In conclusion, our work expands the genetic spectrum of CMT2A, disclosing a novel mutation and its related clinical effect, and provides a detailed description of the clinical features of a cohort of patients with MFN2 mutations. Obtaining a precise genetic diagnosis in affected families is crucial both for family planning and prenatal diagnosis, and in a therapeutic perspective, as we are entering the era of personalized therapy for genetic diseases.

DOI: https://doi.org/10.1038/s41598-022-10220-0
Neurology. 2022 Apr 15. pii: 10.1212/WNL.0000000000200299. [Epub ahead of print]

L-arginine in Mitochondrial Encephalopathy, Lactic Acidosis, and Stroke-like Episodes: A Systematic Review.

Renae Stefanetti, Yi Ng, Linda Errington, Alasdair Blain, Robert McFarland, Gráinne Siobhan Gorman.

OBJECTIVE: Stroke management in the context of primary mitochondrial disease is clinically challenging and the best treatment options for patients with stroke-like episodes remain uncertain. We sought to perform a systematic review on the safety and efficacy of L-arginine use in the acute and prophylactic management of stroke-like episodes in patients with mitochondrial disease.
METHODS: The systematic review was registered in PROSPERO (CRD42020181230). We searched six databases from inception - 15/01/2021: MEDLINE, Embase, Scopus, Web of Science, CINAHL and ClinicalTrials.gov. Original articles and registered trials available, in English, reporting L-arginine use in the acute or prophylactic management of stroke-like episodes in patients with genetically confirmed mitochondrial disease were eligible for inclusion.Data on safety and treatment response were extracted and summarized by multiple observers. Risk of bias was assessed by the methodological quality of case reports, case series and a risk-of-bias checklist for non-randomized studies. Quality of evidence was synthesized using the Oxford Centre for Evidence-Based Medicine Levels of Evidence and Grade of Recommendations. The predetermined main outcome measures were clinical response to L-arginine treatment, adverse events, withdrawals, and deaths (on treatment and/or during follow up), as defined by the author.
RESULTS: Thirty-seven articles met inclusion [0=randomised controlled trials (RCTs); 3 open-label; 1 retrospective cohort; 33 case reports/case series] (N = 91 patients; 86% m.3243A>G). In the case reports, 54% of patients reported a positive clinical response to acute L-arginine, of which 40% were concomitantly treated with AEDs. Improved headache at 24-hours was the greatest reported benefit in response to IV L-arginine in the open-label trials (31/39, 79%). Of 15/48 patients (31%) who positively responded to prophylactic L-arginine, AEDs were either used (7/15) or unreported (8/15). Moderate adverse events were reported in the follow-up of both IV and oral L-arginine treatment and 11 patients (12%) died during follow up or while on prophylactic treatment.
CONCLUSION: The available evidence is of poor methodological quality and classified as Level 5. IV and/or oral L-arginine confers no demonstrable clinical benefit in either the acute or prophylactic treatment of MELAS, with more robust controlled trials required to assess its efficacy and safety profile.

DOI: https://doi.org/10.1212/WNL.0000000000200299
Front Neurosci. 2022 ;16 819569

Cerebellar Pathology in an Inducible Mouse Model of Friedreich Ataxia.

Elizabeth Mercado-Ayón, Nathan Warren, Sarah Halawani, Layne N Rodden, Lucie Ngaba, Yi Na Dong, Joshua C Chang, Carlos Fonck, Fulvio Mavilio, David R Lynch, Hong Lin.

  Friedreich ataxia (FRDA) is an autosomal recessive neurodegenerative disorder caused by deficiency of the mitochondrial protein frataxin. Lack of frataxin causes neuronal loss in various areas of the CNS and PNS. In particular, cerebellar neuropathology in FRDA patients includes loss of large principal neurons and synaptic terminals in the dentate nucleus (DN), and previous studies have demonstrated early synaptic deficits in the Knockin-Knockout mouse model of FRDA. However, the exact correlation of frataxin deficiency with cerebellar neuropathology remains unclear. Here we report that doxycycline-induced frataxin knockdown in a mouse model of FRDA (FRDAkd) leads to synaptic cerebellar degeneration that can be partially reversed by AAV8-mediated frataxin restoration. Loss of cerebellar Purkinje neurons and large DN principal neurons are observed in the FRDAkd mouse cerebellum. Levels of the climbing fiber-specific glutamatergic synaptic marker VGLUT2 decline starting at 4 weeks after dox induction, whereas levels of the parallel fiber-specific synaptic marker VGLUT1 are reduced by 18-weeks. These findings suggest initial selective degeneration of climbing fiber synapses followed by loss of parallel fiber synapses. The GABAergic synaptic marker GAD65 progressively declined during dox induction in FRDAkd mice, while GAD67 levels remained unaltered, suggesting specific roles for frataxin in maintaining cerebellar synaptic integrity and function during adulthood. Expression of frataxin following AAV8-mediated gene transfer partially restored VGLUT1/2 levels. Taken together, our findings show that frataxin knockdown leads to cerebellar degeneration in the FRDAkd mouse model, suggesting that frataxin helps maintain cerebellar structure and function.

Keywords:  cerebellum; frataxin; gene therapy; glutamatergic and GABAergic synapses; mitochondria dynamics

DOI:  https://doi.org/10.3389/fnins.2022.819569
Int J Mol Sci. 2022 Mar 25. pii: 3595. [Epub ahead of print]23(7):

Mitochondrial Dysfunction and Acute Fatty Liver of Pregnancy.

Raghu Ramanathan, Jamal A Ibdah.

  The liver is one of the richest organs in mitochondria, serving as a hub for key metabolic pathways such as β-oxidation, the tricarboxylic acid (TCA) cycle, ketogenesis, respiratory activity, and adenosine triphosphate (ATP) synthesis, all of which provide metabolic energy for the entire body. Mitochondrial dysfunction has been linked to subcellular organelle dysfunction in liver diseases, particularly fatty liver disease. Acute fatty liver of pregnancy (AFLP) is a life-threatening liver disorder unique to pregnancy, which can result in serious maternal and fetal complications, including death. Pregnant mothers with this disease require early detection, prompt delivery, and supportive maternal care. AFLP was considered a mysterious illness and though its pathogenesis has not been fully elucidated, molecular research over the past two decades has linked AFLP to mitochondrial dysfunction and defects in fetal fatty-acid oxidation (FAO). Due to deficient placental and fetal FAO, harmful 3-hydroxy fatty acid metabolites accumulate in the maternal circulation, causing oxidative stress and microvesicular fatty infiltration of the liver, resulting in AFLP. In this review, we provide an overview of AFLP and mitochondrial FAO followed by discussion of how altered mitochondrial function plays an important role in the pathogenesis of AFLP.

Keywords:  acute fatty liver of pregnancy; liver; long chain 3-hydroxyacyl Co-A; mitochondrial dysfunction; mitochondrial trifunctional protein; β-oxidation

DOI:  https://doi.org/10.3390/ijms23073595
Nat Cell Biol. 2022 Apr 13.

Mitochondrial shape alteration by metabolites.

Till Klecker, Benedikt Westermann.

DOI: https://doi.org/10.1038/s41556-022-00889-w
Mitochondrion. 2022 Apr 06. pii: S1567-7249(22)00027-7. [Epub ahead of print]64 136-144

Pharmacologic enrichment of exosome yields and mitochondrial cargo.

Xiaowan Wang, Alexandra Berkowicz, Kirsten King, Blaise Menta, Alexander P Gabrielli, Lesya Novikova, Benjamin Troutwine, Joseph Pleen, Heather M Wilkins, Russell H Swerdlow.

  In studies with human participants, exosome-based biospecimens can facilitate unique biomarker assessments. As exosome cargos can include mitochondrial components, there is interest in using exosomes to inform the status of an individual's mitochondria. Here, we evaluated whether targeted pharmacologic manipulations could influence the quantity of exosomes shed by cells, and whether these manipulations could impact their mitochondrial cargos. We treated human SH-SY5Y cells with bafilomycin A1, which interferes with general autophagy and mitophagy by inhibiting lysosome acidification and lysosome-autophagosome fusion; deferiprone (DFP), which enhances receptor-mediated mitophagy; or both. Exosome fractions from treated cells were harvested from the cell medium and analyzed for content including mitochondria-derived components. We found bafilomycin increased particle yields, and a combination of bafilomycin plus DFP consistently increased particle yields and mitochondria-associated content. Specifically, the exosome fractions from the bafilomycin plus DFP-treated cells contained more mitochondrial DNA (mtDNA), mtDNA-derived mRNA transcripts, and citrate synthase protein. Our data suggest pharmacologic manipulations that enhance mitophagy initiation, while inhibiting the lysosomal digestion of autophagosomes and multivesicular bodies, could potentially enhance the sensitivity of exosome-based biomarker assays intended to inform the status of an individual's mitochondria.

Keywords:  Biomarker; Exosome; Mitochondria; Mitochondrial DNA; Mitophagy

DOI:  https://doi.org/10.1016/j.mito.2022.04.001
J Vis Exp. 2022 Mar 23.

Assessing Energy Substrate Oxidation In Vitro with 14CO2 Trapping.

Chao Song, Arianna Valeri, Fanxin Long.

Mitochondria host the machinery for the tricarboxylic acid (TCA) cycle and electron transport chain (ETC), which generate adenosine triphosphate (ATP) to maintain energy homeostasis. Glucose, fatty acids, and amino acids are the major energy substrates fueling mitochondrial respiration in most somatic cells. Evidence shows that different cell types may have a distinct preference for certain substrates. However, substrate utilization by various cells in the skeleton has not been studied in detail. Moreover, as cellular metabolism is attuned to physiological and pathophysiological changes, direct assessments of substrate dependence in skeletal cells may provide important insights into the pathogenesis of bone diseases. The following protocol is based on the principle of carbon dioxide release from substrate molecules following oxidative phosphorylation. By using substrates containing radioactively labeled carbon atoms (14C), the method provides a sensitive and easy-to-use assay for the rate of substrate oxidation in cell culture. A case study with primary calvarial preosteoblasts versus bone marrow-derived macrophages (BMMs) demonstrates different utilization of the main substrates between the two cell types.

DOI: https://doi.org/10.3791/63568
Methods Mol Biol. 2022 ;2431 385-407

Detailed Imaging of Mitochondrial Transport and Precise Manipulation of Mitochondrial Function with Genetically Encoded Photosensitizers in Adult Drosophila Neurons.

Francesca Mattedi, George Chennell, Alessio Vagnoni.

  Precise distribution of mitochondria is essential for maintaining neuronal homeostasis. Although detailed mechanisms governing the transport of mitochondria have emerged, it is still poorly understood how the regulation of transport is coordinated in space and time within the physiological context of an organism. How alteration in mitochondrial functionality may trigger changes in organellar dynamics also remains unclear in this context. Therefore, the use of genetically encoded tools to perturb mitochondrial functionality in real time would be desirable. Here we describe methods to interfere with mitochondrial function with high spatiotemporal precision with the use of photosensitizers in vivo in the intact wing nerve of adult Drosophila. We also provide details on how to visualize the transport of mitochondria and to improve the quality of the imaging to attain super-resolution in this tissue.

Keywords:  Axonal transport; Drosophila; Intravital imaging; KillerRed; Mitochondria; Neurons; Reactive oxygen species (ROS); Super-resolution radial fluctuations (SRRF); SuperNova

DOI:  https://doi.org/10.1007/978-1-0716-1990-2_20
Cell Calcium. 2022 Apr 01. pii: S0143-4160(22)00058-6. [Epub ahead of print]104 102583

The great astroglial metabolic revolution: Mitochondria fuel astrocyte homeostatic support and neuroprotection.

Alexei Verkhratsky, Alexey Semyanov.



Keywords:  ATP; Astrocyte; Metabolism; Mitochondrial N(a+) C(a2+) exchanger; Neurodegeneration; Neuroptotection

DOI:  https://doi.org/10.1016/j.ceca.2022.102583
J Neuroinflammation. 2022 Apr 12. 19(1): 93

The smoothened agonist SAG reduces mitochondrial dysfunction and neurotoxicity of frataxin-deficient astrocytes.

Andrés Vicente-Acosta, Alfredo Giménez-Cassina, Javier Díaz-Nido, Frida Loria.

   BACKGROUND: Friedreich's ataxia is a rare hereditary neurodegenerative disease caused by decreased levels of the mitochondrial protein frataxin. Similar to other neurodegenerative pathologies, previous studies suggested that astrocytes might contribute to the progression of the disease. To fully understand the mechanisms underlying neurodegeneration in Friedreich's ataxia, we investigated the reactivity status and functioning of cultured human astrocytes after frataxin depletion using an RNA interference-based approach and tested the effect of pharmacologically modulating the SHH pathway as a novel neuroprotective strategy.
RESULTS: We observed loss of cell viability, mitochondrial alterations, increased autophagy and lipid accumulation in cultured astrocytes upon frataxin depletion. Besides, frataxin-deficient cells show higher expression of several A1-reactivity markers and release of pro-inflammatory cytokines. Interestingly, most of these defects were prevented by chronically treating the cells with the smoothened agonist SAG. Furthermore, in vitro culture of neurons with conditioned medium from frataxin-deficient astrocytes results in a reduction of neuronal survival, neurite length and synapse formation. However, when frataxin-deficient astrocytes were chronically treated with SAG, we did not observe these alterations in neurons.
CONCLUSIONS: Our results demonstrate that the pharmacological activation of the SHH pathway could be used as a target to modulate astrocyte reactivity and neuron-glia interactions to prevent neurodegeneration in Friedreich's ataxia.

Keywords:  Frataxin; Mitochondrial dysfunction; Neurotoxicity; Reactive astrocytes; Smoothened agonist; Sonic hedgehog

DOI:  https://doi.org/10.1186/s12974-022-02442-w
Cell Mol Life Sci. 2022 Apr 13. 79(5): 239

Mitochondrial function in spinal cord injury and regeneration.

Paula G Slater, Miguel E Domínguez-Romero, Maximiliano Villarreal, Verónica Eisner, Juan Larraín.

  Many people around the world suffer from some form of paralysis caused by spinal cord injury (SCI), which has an impact on quality and life expectancy. The spinal cord is part of the central nervous system (CNS), which in mammals is unable to regenerate, and to date, there is a lack of full functional recovery therapies for SCI. These injuries start with a rapid and mechanical insult, followed by a secondary phase leading progressively to greater damage. This secondary phase can be potentially modifiable through targeted therapies. The growing literature, derived from mammalian and regenerative model studies, supports a leading role for mitochondria in every cellular response after SCI: mitochondrial dysfunction is the common event of different triggers leading to cell death, cellular metabolism regulates the immune response, mitochondrial number and localization correlate with axon regenerative capacity, while mitochondrial abundance and substrate utilization regulate neural stem progenitor cells self-renewal and differentiation. Herein, we present a comprehensive review of the cellular responses during the secondary phase of SCI, the mitochondrial contribution to each of them, as well as evidence of mitochondrial involvement in spinal cord regeneration, suggesting that a more in-depth study of mitochondrial function and regulation is needed to identify potential targets for SCI therapeutic intervention.

Keywords:  Axon regeneration; Cell death; Cell metabolism; Immune response; Mitochondria; Neurogenesis; Spinal cord

DOI:  https://doi.org/10.1007/s00018-022-04261-x
Methods Mol Biol. 2022 ;2431 533-546

In Vitro Reconstitution of Molecular Motor-Driven Mitochondrial Transport.

Verena Puttrich, Jakub Rohlena, Marcus Braun, Zdenek Lansky.

  Intracellular trafficking of organelles driven by molecular motors underlies essential cellular processes. Mitochondria, the powerhouses of the cell, are one of the major cargoes of molecular motors. Efficient distribution of mitochondria ensures cellular fitness while defects in this process contribute to severe pathologies, such as neurodegenerative diseases. Reconstitution of the mitochondrial microtubule-based transport in vitro in a bottom-up approach provides a powerful tool to investigate the mitochondrial trafficking machinery in a controlled environment in the absence of complex intracellular interactions. In this chapter, we describe the procedures for achieving such reconstitution of mitochondrial transport.

Keywords:  Adaptor proteins; Interference reflection microscopy; Kinesin-1; Mitochondria; Molecular motors; Motility assay; TIRF microscopy; TRAK

DOI:  https://doi.org/10.1007/978-1-0716-1990-2_28
Mitochondrion. 2022 Apr 06. pii: S1567-7249(22)00028-9. [Epub ahead of print]64 145-155

Erodible thermogelling hydrogels for localized mitochondrial transplantation to the spinal cord.

Samir P Patel, Felicia M Michael, M Arif Khan, Brian Duggan, Sam Wyse, Daniel R Darby, Krishnaroop Chaudhuri, Jonathan T Pham, Jenna Gollihue, Jason E DeRouchey, Patrick G Sullivan, Tom D Dziubla, Alexander G Rabchevsky.

  We developed a thermal-gelling, erodible hydrogel system for localized delivery of viable mitochondria in vivo, as well as labeled transplanted mitochondria with specific dyes and/or genetically modified mitochondria tagged with red fluorescence protein (RFP). We also employed cell lines to optimize a hydrogel composed of methylcellulose and hyaluronic acid designed to preserve bioenergetics while facilitating mitochondrial release. We further investigated how transplantation of allogeneic or xenogeneic mitochondria into respective cell lines affects host cellular metabolism, as measured by MTS assay. We found that 70% of mitochondria are released from the hydrogel within 20 min at 37 °C, that the respiratory capacity of hydrogel-released mitochondria over 60 min was greater than those without gel, and that MTR-labeling of mitochondria is not indelible. RFP-tagged transgenic mitochondria isolated from modified SH-SY5Y human neuroblastoma cells showed effective uptake into both naïve SH-SY5Y cells and rat PC-12 cells, notably when released from hydrogel. The hydrogel both protected the mitochondria at physiological conditions in vitro while solidifying and diffusing within 60 min locally in situ. To assess metabolic effects, both cell lines were transplanted with different concentrations of SH-SY5Y or PC-12 cell line-derived mitochondria and all resulted in significant increases in metabolism at 6- and 24-hour after transplantation. Alternatively, transplanted mitochondria at highest concentration from rat brain and spinal cord tissues reduced metabolic activities after 24-hour. Along with hydrogel refinements, we are further investigating whether such metabolic changes are due to alterations in cell proliferation or the number of exogenous mitochondria incorporated into individual host cells.

Keywords:  Hydrogel; MTR; MTS; Metabolism; Mitochondrial transplantation; Spinal cord

DOI:  https://doi.org/10.1016/j.mito.2022.04.002
Cells. 2022 Apr 01. pii: 1193. [Epub ahead of print]11(7):

Induction of Mitochondrial Fragmentation and Mitophagy after Neonatal Hypoxia-Ischemia.

Syam Nair, Anna-Lena Leverin, Eridan Rocha-Ferreira, Kristina S Sobotka, Claire Thornton, Carina Mallard, Henrik Hagberg.

  Hypoxia-ischemia (HI) leads to immature brain injury mediated by mitochondrial stress. If damaged mitochondria cannot be repaired, mitochondrial permeabilization ensues, leading to cell death. Non-optimal turnover of mitochondria is critical as it affects short and long term structural and functional recovery and brain development. Therefore, disposal of deficient mitochondria via mitophagy and their replacement through biogenesis is needed. We utilized mt-Keima reporter mice to quantify mitochondrial morphology (fission, fusion) and mitophagy and their mechanisms in primary neurons after Oxygen Glucose Deprivation (OGD) and in brain sections after neonatal HI. Molecular mechanisms of PARK2-dependent and -independent pathways of mitophagy were investigated in vivo by PCR and Western blotting. Mitochondrial morphology and mitophagy were investigated using live cell microscopy. In primary neurons, we found a primary fission wave immediately after OGD with a significant increase in mitophagy followed by a secondary phase of fission at 24 h following recovery. Following HI, mitophagy was upregulated immediately after HI followed by a second wave at 7 days. Western blotting suggests that both PINK1/Parkin-dependent and -independent mechanisms, including NIX and FUNDC1, were upregulated immediately after HI, whereas a PINK1/Parkin mechanism predominated 7 days after HI. We hypothesize that excessive mitophagy in the early phase is a pathologic response which may contribute to secondary energy depletion, whereas secondary mitophagy may be involved in post-HI regeneration and repair.

Keywords:  metabolism; mitochondria; mitochondrial fission; mitophagy; neonatal brain injury; neonatal hypoxia–ischemia; reactive oxygen species

DOI:  https://doi.org/10.3390/cells11071193
J Biol Chem. 2022 Apr 07. pii: S0021-9258(22)00339-8. [Epub ahead of print] 101899

The molecular basis of spinocerebellar ataxia type 48 caused by a de novo mutation in the ubiquitin ligase CHIP.

A Umano, K Fang, Z Qu, J B Scaglione, S Altinok, C J Treadway, E T Wick, E Paulakonis, C Karunanayake, S Chou, T M Bardakjian, P Gonzalez-Alegre, R C Page, J C Schisler, N G Brown, D Yan, K M Scaglione.

  The spinocerebellar ataxias (SCAs) are a class of incurable diseases characterized by degeneration of the cerebellum that results in movement disorder. Recently, a new heritable form of SCA, SCA48, was attributed to dominant mutations in STUB1; however, little is known about how these mutations cause SCA48. STUB1 encodes for the protein C-terminus of Hsc70 Interacting Protein (CHIP), an E3 ubiquitin ligase. CHIP is known to regulate proteostasis by recruiting chaperones via a N-terminal tetratricopeptide repeat (TPR) domain and recruiting E2 ubiquitin-conjugating enzymes via a C-terminal U-box domain. These interactions allow CHIP to mediate the ubiquitination of chaperone-bound, misfolded proteins to promote their degradation via the proteasome. Here we have identified a novel, de novo mutation in STUB1 in a patient with spinocerebellar ataxia type 48 (SCA48) encoding for an A52G point mutation in the TPR domain of CHIP. Utilizing an array of biophysical, biochemical, and cellular assays, we demonstrate that the CHIPA52G point mutant retains E3-ligase activity but has decreased affinity for chaperones. We further show that this mutant decreases cellular fitness in response to certain cellular stressors and induces neurodegeneration in a transgenic Caenorhabditis elegans model of SCA48. Together, our data identify the A52G mutant as a cause of SCA48 and provide molecular insight into how mutations in STUB1 cause SCA48.

Keywords:  E3 ubiquitin-ligase; ataxia; chaperone; neurodegeneration; neurodegenerative disease

DOI:  https://doi.org/10.1016/j.jbc.2022.101899
Sci Adv. 2022 Apr 15. 8(15): eabk2376

Mitolysosome exocytosis, a mitophagy-independent mitochondrial quality control in flunarizine-induced parkinsonism-like symptoms.

Feixiang Bao, Lingyan Zhou, Rui Zhou, Qiaoying Huang, Junguo Chen, Sheng Zeng, Yi Wu, Liang Yang, Shufang Qian, Mengfei Wang, Xueying He, Shan Liang, Juntao Qi, Ge Xiang, Qi Long, Jingyi Guo, Zhongfu Ying, Yanshuang Zhou, Qiuge Zhao, Jiwei Zhang, Di Zhang, Wei Sun, Mi Gao, Hao Wu, Yifan Zhao, Jinfu Nie, Min Li, Quan Chen, Jiekai Chen, Xiao Zhang, Guangjin Pan, Hong Zhang, Mingtao Li, Mei Tian, Xingguo Liu.

Mitochondrial quality control plays an important role in maintaining mitochondrial homeostasis and function. Disruption of mitochondrial quality control degrades brain function. We found that flunarizine (FNZ), a drug whose chronic use causes parkinsonism, led to a parkinsonism-like motor dysfunction in mice. FNZ induced mitochondrial dysfunction and decreased mitochondrial mass specifically in the brain. FNZ decreased mitochondrial content in both neurons and astrocytes, without affecting the number of nigral dopaminergic neurons. In human neural progenitor cells, FNZ also induced mitochondrial depletion. Mechanistically, independent of ATG5- or RAB9-mediated mitophagy, mitochondria were engulfed by lysosomes, followed by a vesicle-associated membrane protein 2- and syntaxin-4-dependent extracellular secretion. A genome-wide CRISPR knockout screen identified genes required for FNZ-induced mitochondrial elimination. These results reveal not only a previously unidentified lysosome-associated exocytosis process of mitochondrial quality control that may participate in the FNZ-induced parkinsonism but also a drug-based method for generating mitochondria-depleted mammal cells.

DOI: https://doi.org/10.1126/sciadv.abk2376
Curr Med Res Opin. 2022 Apr 11. 1-10

Do compromised mitochondria aggravate severity and fatality by SARS-CoV-2?

Nilesh Kumar Sharma, Sachin C Sarode.

  At global level, pandemic coronavirus disease 2019 (COVID-19) is known to be caused by an etiologic agent severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Numerous evidence and propositions have emerged on the molecular and cellular attributes that cause COVID-19. Notwithstanding, still several key questions with reference to molecular aspects of severity of infection by SARS-CoV-2 need to be answered. In the same line, the role of healthy mitochondria to maintain intracellular temperature and their association with the severity of SARS-CoV-2 is completely missing. In this direction, preclinical and clinical data on the comorbidities in case of mitochondrial defective disease and COVID-19 are not available. The authors propose that patients harboring primary mitochondrial Disease and secondary mitochondrial dysfunction will display higher severity and death rate compared to healthy mitochondria harboring patients.

Keywords:  ACE-2; COVID-19; Intracellular temperature; Mitochondria; SPIKE protein

DOI:  https://doi.org/10.1080/03007995.2022.2065140
EMBO J. 2022 Apr 12. e109390

DGAT1 activity synchronises with mitophagy to protect cells from metabolic rewiring by iron depletion.

Maeve Long, Alvaro Sanchez-Martinez, Marianna Longo, Fumi Suomi, Hans Stenlund, Annika I Johansson, Homa Ehsan, Veijo T Salo, Lambert Montava-Garriga, Seyedehshima Naddafi, Elina Ikonen, Ian G Ganley, Alexander J Whitworth, Thomas G McWilliams.

  Mitophagy removes defective mitochondria via lysosomal elimination. Increased mitophagy coincides with metabolic reprogramming, yet it remains unknown whether mitophagy is a cause or consequence of such state changes. The signalling pathways that integrate with mitophagy to sustain cell and tissue integrity also remain poorly defined. We performed temporal metabolomics on mammalian cells treated with deferiprone, a therapeutic iron chelator that stimulates PINK1/PARKIN-independent mitophagy. Iron depletion profoundly rewired the metabolome, hallmarked by remodelling of lipid metabolism within minutes of treatment. DGAT1-dependent lipid droplet biosynthesis occurred several hours before mitochondrial clearance, with lipid droplets bordering mitochondria upon iron chelation. We demonstrate that DGAT1 inhibition restricts mitophagy in vitro, with impaired lysosomal homeostasis and cell viability. Importantly, genetic depletion of DGAT1 in vivo significantly impaired neuronal mitophagy and locomotor function in Drosophila. Our data define iron depletion as a potent signal that rapidly reshapes metabolism and establishes an unexpected synergy between lipid homeostasis and mitophagy that safeguards cell and tissue integrity.

Keywords:  DGAT1; iron; lipid droplet; metabolism; mitophagy

DOI:  https://doi.org/10.15252/embj.2021109390
Toxicology. 2022 Apr 09. pii: S0300-483X(22)00088-9. [Epub ahead of print]471 153176

Activation of mitophagy by rapamycin eliminated the accumulation of TDP-43 on mitochondrial and promoted the resolution of carbon tetrachloride-induced liver fibrosis in mice.

Hui Yong, Shulin Shan, Shuai Wang, Zhidan Liu, Zhaoxiong Liu, Cuiqin Zhang, Yiyu Yang, Zhengcheng Huang, Fuyong Song.

  Liver fibrosis can lead to liver cirrhosis and hepatocellular carcinoma, and no effective treatment is available in clinical practice. Mitochondrial dysfunction is thought to be closely related to the development of liver fibrosis. Recent studies have reported that abnormal accumulation of TDP-43 on mitochondria may interfere with mitochondrial function in neurodegenerative disorders. However, whether aberrant TDP-43 aggregation is also involved in liver fibrosis has not been investigated. In this study, C57/BL6 mice were treated with CCl4 (escalating doses, three times a week) for 8 weeks to establish a model of liver fibrosis. Furthermore, mitophagy intervention experiment was achieved by the activator rapamycin (RAPA). The results demonstrated that chronic CCl4 exposure resulted in severe mitochondrial damage, inflammatory response and hepatic fibrogenesis. Interestingly, abnormal aggregation of TDP-43 on mitochondria was observed. By contrast, RAPA administration could promote the regression of liver fibrosis. Mechanistically, RAPA could eliminate the accumulation of TDP-43 on mitochondrial through enhancing mitophagy, thereby improving mitochondrial function. Taken together, our study revealed that mitochondrial damage induced by abnormal accumulation of TDP-43 has been implicated in the progression of liver fibrosis. Targeted clearance of mitochondrial TDP-43 may lead to the development of some anti-fibrotic therapies.

Keywords:  Carbon tetrachloride; Liver fibrosis; Mitophagy; TDP-43

DOI:  https://doi.org/10.1016/j.tox.2022.153176
Nat Commun. 2022 Apr 14. 13(1): 2005

Proteolysis of adaptor protein Mmr1 during budding is necessary for mitochondrial homeostasis in Saccharomyces cerevisiae.

Keisuke Obara, Taku Yoshikawa, Ryu Yamaguchi, Keiko Kuwata, Kunio Nakatsukasa, Kohei Nishimura, Takumi Kamura.

In yeast, mitochondria are passed on to daughter cells via the actin cable, motor protein Myo2, and adaptor protein Mmr1. They are released from the actin-myosin machinery after reaching the daughter cells. We report that Mmr1 is rapidly degraded by the ubiquitin-proteasome system in Saccharomyces cerevisiae. Redundant ubiquitin ligases Dma1 and Dma2 are responsible for Mmr1 ubiquitination. Dma1/2-mediated Mmr1 ubiquitination requires phosphorylation, most likely at S414 residue by Ste20 and Cla4. These kinases are mostly localized to the growing bud and nearly absent from mother cells, ensuring phosphorylation and ubiquitination of Mmr1 after the mitochondria enter the growing bud. In dma1Δ dma2Δ cells, transported mitochondria are first stacked at the bud-tip and then pulled back to the bud-neck. Stacked mitochondria in dma1Δ dma2Δ cells exhibit abnormal morphology, elevated respiratory activity, and increased level of reactive oxygen species, along with hypersensitivity to oxidative stresses. Collectively, spatiotemporally-regulated Mmr1 turnover guarantees mitochondrial homeostasis.

DOI: https://doi.org/10.1038/s41467-022-29704-8
Cells. 2022 Mar 30. pii: 1169. [Epub ahead of print]11(7):

Mitochondrial Respiratory Complexes as Targets of Drugs: The PPAR Agonist Example.

Patrizia Bottoni, Alessandro Pontoglio, Salvatore Scarà, Luisa Pieroni, Andrea Urbani, Roberto Scatena.

  Mitochondrial bioenergetics are progressively acquiring significant pathophysiological roles. Specifically, mitochondria in general and Electron Respiratory Chain in particular are gaining importance as unintentional targets of different drugs. The so-called PPAR ligands are a class of drugs which not only link and activate Peroxisome Proliferator-Activated Receptors but also show a myriad of extrareceptorial activities as well. In particular, they were shown to inhibit NADH coenzyme Q reductase. However, the molecular picture of this intriguing bioenergetic derangement has not yet been well defined. Using high resolution respirometry, both in permeabilized and intact HepG2 cells, and a proteomic approach, the mitochondrial bioenergetic damage induced by various PPAR ligands was evaluated. Results show a derangement of mitochondrial oxidative metabolism more complex than one related to a simple perturbation of complex I. In fact, a partial inhibition of mitochondrial NADH oxidation seems to be associated not only with hampered ATP synthesis but also with a significant reduction in respiratory control ratio, spare respiratory capacity, coupling efficiency and, last but not least, serious oxidative stress and structural damage to mitochondria.

Keywords:  cancer; complex I (NADH: ubiquinone oxidoreductase); drug toxicity; mitochondria; reactive oxygen species (ROS); therapeutic drug monitoring

DOI:  https://doi.org/10.3390/cells11071169
Geroscience. 2022 Apr 13.

Age-related disruption of the proteome and acetylome in mouse hearts is associated with loss of function and attenuated by elamipretide (SS-31) and nicotinamide mononucleotide (NMN) treatment.

Jeremy A Whitson, Richard Johnson, Lu Wang, Theo K Bammler, Shin-Ichiro Imai, Huiliang Zhang, Jeanne Fredrickson, Elena Latorre-Esteves, Alessandro Bitto, Michael J MacCoss, Peter S Rabinovitch.

  We analyzed the effects of aging on protein abundance and acetylation, as well as the ability of the mitochondrial-targeted drugs elamipretide (SS-31) and nicotinamide mononucleotide (NMN) to reverse aging-associated changes in mouse hearts. Both drugs had a modest effect on restoring the abundance and acetylation of proteins that are altered with age, while also inducing additional changes. Age-related increases in protein acetylation were predominantly in mitochondrial pathways such as mitochondrial dysfunction, oxidative phosphorylation, and TCA cycle signaling. We further assessed how these age-related changes associated with diastolic function (Ea/Aa) and systolic function (fractional shortening under higher workload) measurements from echocardiography. These results identify a subset of protein abundance and acetylation changes in muscle, mitochondrial, and structural proteins that appear to be essential in regulating diastolic function in old hearts.

Keywords:  Acetylomics; Aging; Elamipretide; Heart; Mitochondria; NMN; Proteomics; SS-31

DOI:  https://doi.org/10.1007/s11357-022-00564-w
J Neurol. 2022 Apr 15.

Clinical trials for cerebellar ataxia.

Benjamin E Schroeder, Neil P Robertson.

DOI: https://doi.org/10.1007/s00415-022-11088-w
J Hepatol. 2022 Apr 11. pii: S0168-8278(22)00226-4. [Epub ahead of print]

Mitochondrial protease ClpP supplementation ameliorates diet-induced NASH in mice.

Sung-E Choi, Yoonjung Hwang, Soo-Jin Lee, Hyunkyung Jung, Tae Hwan Shin, Youngho Son, Seokho Park, Seung Jin Han, Hae Jin Kim, Kwan Woo Lee, Gwang Lee, Jongsook Kim Kemper, Hyun Kyu Song, Yup Kang.

   BACKGROUND & AIMS: Mitochondrial dysfunction is considered a pathogenic linker in the development of non-alcoholic steatohepatitis (NASH). Inappropriate mitochondrial protein-quality control, possibly induced due to insufficiency of the mitochondrial matrix caseinolytic protease P (ClpP), can potentially cause mitochondrial dysfunction. Herein, we aimed to investigate hepatic ClpP levels in a diet-induced model of NASH and determine whether supplementation of ClpP can ameliorate the diet-induced NASH.
METHODS: NASH was induced by a high fat/high fructose (HF/HFr) diet in C57BL/6J mice. Stress/inflammatory signals were induced in mouse primary hepatocytes (MPHs) by treatment with palmitate/oleate (PA/OA). ClpP levels in hepatocytes were reduced using the RNAi-mediated gene knockdown technique but increased through the viral transduction of ClpP. ClpP activation was induced by administering a chemical activator of ClpP.
RESULTS: Hepatic ClpP protein levels in C57BL/6J mice fed a HF/HFr diet were lower than the levels in those fed a normal chow diet. PA/OA treatment also decreased the ClpP protein levels in MPHs. Overexpression or activation of ClpP reversed the PA/OA-induced mitochondrial dysfunction and stress/inflammatory signal activation in MPHs, whereas ClpP knockdown induced mitochondrial dysfunction and stress/inflammatory signals in these cells. On the other hand, ClpP overexpression or activation improved HF/HFr-induced NASH characteristics such as hepatic steatosis, inflammation, fibrosis, and injury in the C57BL/6J mice, whereas ClpP knockdown further augmented steatohepatitis in mice fed a HF/HFr diet.
CONCLUSIONS: Reduced ClpP expression and subsequent mitochondrial dysfunction are key to the development of diet-induced NASH. ClpP supplementation through viral transduction or chemical activation represents potential therapeutic strategies for preventing diet-induced NASH.

Keywords:  inflammation; mitochondrial dysfunction; proteostasis; steatohepatitis

DOI:  https://doi.org/10.1016/j.jhep.2022.03.034
Int J Mol Sci. 2022 Apr 06. pii: 4047. [Epub ahead of print]23(7):

Metabolic Syndrome and β-Oxidation of Long-Chain Fatty Acids in the Brain, Heart, and Kidney Mitochondria.

Alexander Panov, Vladimir I Mayorov, Sergey Dikalov.

  We present evidence that metabolic syndrome (MetS) represents the postreproductive stage of the human postembryonic ontogenesis. Accordingly, the genes governing this stage experience relatively weak evolutionary selection pressure, thus representing the metabolic phenotype of distant ancestors with β-oxidation of long-chain fatty acids (FAs) as the primary energy source. Mitochondria oxidize at high-rate FAs only when succinate, glutamate, or pyruvate are present. The heart and brain mitochondria work at a wide range of functional loads and possess an intrinsic inhibition of complex II to prevent oxidative stress at periods of low functional activity. Kidney mitochondria constantly work at a high rate and lack inhibition of complex II. We suggest that in people with MetS, oxidative stress is the central mechanism of the heart and brain pathologies. Oxidative stress is a secondary pathogenetic mechanism in the kidney, while the primary mechanisms are kidney hypoxia caused by persistent hyperglycemia and hypertension. Current evidence suggests that most of the nongenetic pathologies associated with MetS originate from the inconsistencies between the metabolic phenotype acquired after the transition to the postreproductive stage and excessive consumption of food rich in carbohydrates and a sedentary lifestyle.

Keywords:  brain mitochondria; heart; human postembryonic ontogenesis; kidney; long-chain fatty acids; metabolic syndrome; oxidative stress; β-oxidation

DOI:  https://doi.org/10.3390/ijms23074047