comparative-analysis-experimental-methods-to-quantify-animal-activity

Comparative Analysis of Experimental Methods to Quantify Animal Activity in <em>Caenorhabditis elegans</em> Models of Mitochondrial Disease

Caenorhabditis elegans is widely recognized for its central utility as a translational animal model to efficiently interrogate mechanisms and therapies of ...

The Children’s Hospital of Philadelphia

University of Pennsylvania Perelman School of Medicine

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The extensive conservation of mitochondrial structure, composition, and function across evolution offers a unique opportunity to expand our understanding of human mitochondrial biology and disease. By investigating the biology of much simpler model organisms, it is often possible to answer questions that are unreachable at the clinical level. Here, we review the relative utility of four different model organisms, namely the bacterium Escherichia coli, the yeast Saccharomyces cerevisiae, the nematode Caenorhabditis elegans, and the fruit fly Drosophila melanogaster, in studying the role of mitochondrial proteins relevant to human disease. E. coli are single cell, prokaryotic bacteria that have proven to be a useful model system in which to investigate mitochondrial respiratory chain protein structure and function. S. cerevisiae is a single-celled eukaryote that can grow equally well by mitochondrial-dependent respiration or by ethanol fermentation, a property that has proven to be a veritable boon for investigating mitochondrial functionality. C. elegans is a multicellular, microscopic worm that is organized into five major tissues and has proven to be a robust model animal for in vitro and in vivo studies of primary respiratory chain dysfunction and its potential therapies in humans. Studied for over a century, D. melanogaster is a classic metazoan model system offering an abundance of genetic tools and reagents that facilitates investigations of mitochondrial biology using both forward and reverse genetics. The respective strengths and limitations of each species relative to mitochondrial studies are explored. In addition, an overview is provided of major discoveries made in mitochondrial biology in each of these four model systems.

Bacteria, yeast, worms, and flies: exploiting simple model organisms to investigate human mitochondrial diseases.

La conservation approfondie de la structure mitochondriale, composition et fonction &agrave; travers l&#39;&eacute;volution offre une occasion unique d&#39;approfondir notre connaissance de la biologie humaine mitochondriale et la maladie. En enqu&ecirc;tant sur la biologie des beaucoup plus simples organismes de mod&egrave;le, il est souvent possible de r&eacute;pondre aux questions qui sont inaccessibles au niveau clinique. Ici, nous passons en revue l&#39;utilit&eacute; relative des quatre organismes de diff&eacute;rents mod&egrave;les, &agrave; savoir la bact&eacute;rie Escherichia coli, la levure Saccharomyces cerevisiae, le n&eacute;matode Caenorhabditis elegans et la drosophile Drosophila melanogaster, en &eacute;tudiant le r&ocirc;le des prot&eacute;ines mitochondriales pertinents &agrave; la maladie humaine. E. coli sont unicellulaire, procaryotes bact&eacute;ries qui se sont av&eacute;r&eacute;s pour &ecirc;tre un syst&egrave;me mod&egrave;le utile &eacute;tudier la fonction et la structure de prot&eacute;ines de la cha&icirc;ne respiratoire mitochondriale. S. cerevisiae est un eucaryote unicellulaire qui peut pousser aussi bien par la respiration mitochondriale d&eacute;pendante ou par fermentation de l&#39;&eacute;thanol, une propri&eacute;t&eacute; qui s&#39;est av&eacute;r&eacute; pour &ecirc;tre une v&eacute;ritable aubaine pour enqu&ecirc;ter sur les fonctions mitochondriales. C. elegans est un ver multicellulaire, microscopique qui est organis&eacute; en cinq principaux tissus et s&#39;est av&eacute;r&eacute; pour &ecirc;tre un animal mod&egrave;le robuste pour des &eacute;tudes in vitro et in vivo du dysfonctionnement de la cha&icirc;ne respiratoire primaire et ses traitements possibles chez les humains. A &eacute;tudi&eacute; pendant plus d&#39;un si&egrave;cle, d. melanogaster est un syst&egrave;me de mod&egrave;le de m&eacute;tazoaires classique offrant une abondance d&#39;outils g&eacute;n&eacute;tiques et r&eacute;actifs qui facilite les recherches de biologie mitochondriale &agrave; l&#39;aide de la g&eacute;n&eacute;tique avance et arri&egrave;re. Les atouts respectifs et les limites de chaque esp&egrave;ce par rapport aux &eacute;tudes mitochondriales sont explor&eacute;es. En outre, il est donn&eacute; un aper&ccedil;u des grandes d&eacute;couvertes faites en biologie mitochondriale dans chacun de ces syst&egrave;mes de quatre mod&egrave;les.

Bact&eacute;ries, les levures, les vers et les mouches : exploiter les organismes simple mod&egrave;le pour &eacute;tudier les maladies mitochondriales humaines.

Umfangreiche Erhaltung der mitochondrialen Struktur, Zusammensetzung und Funktion &uuml;ber Evolution bietet eine einmalige Gelegenheit, unser Verst&auml;ndnis von mitochondrialen Humanbiologie und Krankheit zu erweitern. Durch die Untersuchung der Biologie viel einfacher Modellorganismen, ist es oft m&ouml;glich, Fragen zu beantworten, die auf der klinischen Ebene nicht erreichbar sind. Hier &uuml;berpr&uuml;fen wir die relative N&uuml;tzlichkeit von vier verschiedenen Modellorganismen, n&auml;mlich das Bakterium Escherichia coli, die Hefe Saccharomyces Cerevisiae, der Fadenwurm Caenorhabditis Elegans und der Fruchtfliege Drosophila Melanogaster, die Rolle der mitochondrialen Proteine f&uuml;r menschliche Krankheiten zu studieren. E. Coli sind einzelne Zelle, prokaryotischen Bakterien, die als erwiesen haben ein n&uuml;tzliches Modell-System, in dem mitochondrialen Protein-Struktur und Funktion zu untersuchen. S. Cerevisiae ist ein einzelligen Eukaryoten, die wachsen, kann ebenso gut durch mitochondriale-abh&auml;ngige Atmung oder G&auml;rung &Auml;thanol, eine Eigenschaft, die bewiesen hat, dass ein wahrer Segen f&uuml;r die Untersuchung der mitochondrialen Funktionalit&auml;t. C. Elegans ist ein Vielzeller, mikroskopisch kleinen Wurm, der gliedert sich in f&uuml;nf gro&szlig;e Gewebe und hat sich als ein stabiles Modell Tier f&uuml;r in-vitro und in vivo Studien prim&auml;re Atmungskette Dysfunktion und seine m&ouml;glichen Therapien beim Menschen. Studierte f&uuml;r &uuml;ber ein Jahrhundert, ist D. Melanogaster eine klassische indem Modell-System bietet eine F&uuml;lle von genetischen Werkzeuge und Reagenzien, die Untersuchungen der mitochondrialen Biologie mit sowohl Forward- und reverse Genetik erleichtert. Die jeweiligen St&auml;rken und Schw&auml;chen der einzelnen Arten relativ mitochondriale Studien werden untersucht. Dar&uuml;ber hinaus wird eine &Uuml;bersicht der wichtigsten Entdeckungen, die in Mitochondrien Biologie in jedem dieser vier Modellsysteme bereitgestellt.

Bakterien, Hefe, W&uuml;rmer und fliegen: Ausnutzung einfache Modellorganismen um menschliche mitochondrische Krankheiten zu untersuchen.

Il vasta conservazione della struttura mitocondriale, composizione e funzione in evoluzione offre un&#39;opportunit&agrave; unica per ampliare la nostra comprensione della biologia mitocondriale umana e malattia. Studiando la biologia dei microrganismi molto pi&ugrave; semplice, &egrave; spesso possibile a rispondere alle domande che non sono raggiungibili a livello clinico. Qui, passiamo in rassegna la relativa utilit&agrave; di quattro organismi di modello diverso, vale a dire il batterio Escherichia coli, il lievito Saccharomyces cerevisiae, il nematode Caenorhabditis elegans e il moscerino della frutta Drosophila melanogaster, studiando il ruolo delle proteine mitocondriali pertinenti alla malattia umana. E. coli sono unicellulare, procariote batteri che hanno dimostrato di essere un sistema utile modello in cui indagare la funzione e la struttura della proteina catena respiratoria mitocondriale. S. cerevisiae &egrave; un&#39;eucariote unicellulare che possono crescere altrettanto bene di respirazione mitocondriale-dipendente o dalla fermentazione di etanolo, una propriet&agrave; che ha dimostrato di essere una vera e propria manna per indagare la funzionalit&agrave; mitocondriale. C. elegans &egrave; un worm pluricellulare, microscopico che &egrave; organizzato in cinque principali tessuti e ha dimostrato di essere un animale robusto modello per gli studi in vitro e in vivo di disfunzione primaria catena respiratoria e sue terapie potenziali negli esseri umani. Ha studiato per oltre un secolo, d. melanogaster &egrave; un sistema classico modello metazoi che offre un&#39;abbondanza di reagenti e strumenti genetici che facilita le indagini di biologia mitocondriale usando genetica sia avanti e indietro. I rispettivi punti di forza e le limitazioni di ciascuna specie relativi studi mitocondriali sono esplorati. Inoltre, &egrave; fornito un quadro generale delle principali scoperte fatte in biologia mitocondriale in ciascuno di questi quattro modelli di sistemi.

Batteri, lieviti, vermi e mosche: sfruttando organismi modello semplice per indagare sulle malattie mitocondriali umane.

&#x30DF;&#x30C8;&#x30B3;&#x30F3;&#x30C9;&#x30EA;&#x30A2; dna &#x306E;&#x69CB;&#x9020;&#x3001;&#x69CB;&#x6210;&#x3001;&#x304A;&#x3088;&#x3073;&#x9032;&#x5316;&#x306E;&#x6A5F;&#x80FD;&#x306E;&#x5E83;&#x7BC4;&#x306A;&#x4FDD;&#x5168;&#x4EBA;&#x9593;&#x306E;&#x30DF;&#x30C8;&#x30B3;&#x30F3;&#x30C9;&#x30EA;&#x30A2;&#x306E;&#x751F;&#x7269;&#x5B66;&#x3068;&#x75BE;&#x60A3;&#x306E;&#x79C1;&#x9054;&#x306E;&#x7406;&#x89E3;&#x3092;&#x62E1;&#x5927;&#x3059;&#x308B;&#x30E6;&#x30CB;&#x30FC;&#x30AF;&#x306A;&#x6A5F;&#x4F1A;&#x3092;&#x63D0;&#x4F9B;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x591A;&#x304F;&#x3088;&#x308A;&#x5358;&#x7D14;&#x306A;&#x30E2;&#x30C7;&#x30EB;&#x751F;&#x7269;&#x306E;&#x751F;&#x7269;&#x5B66;&#x306E;&#x8ABF;&#x67FB;&#x306B;&#x3088;&#x3063;&#x3066;&#x3001;&#x81E8;&#x5E8A;&#x30EC;&#x30D9;&#x30EB;&#x3067;&#x306F;&#x5230;&#x9054;&#x4E0D;&#x53EF;&#x80FD;&#x306A;&#x8CEA;&#x554F;&#x306B;&#x304A;&#x7B54;&#x3048;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x53EF;&#x80FD;&#x3067;&#x3059;&#x3002;&#x3053;&#x3053;&#x3067;&#x306F;&#x3001;4 &#x3064;&#x306E;&#x7570;&#x306A;&#x308B;&#x30E2;&#x30C7;&#x30EB;&#x6709;&#x6A5F;&#x4F53;&#x3001;&#x3059;&#x306A;&#x308F;&#x3061;&#x7D30;&#x83CC;&#x5927;&#x8178;&#x83CC;&#x3001;&#x51FA;&#x82BD;&#x9175;&#x6BCD;&#x3001;&#x7DDA;&#x866B;&#x3001;&#x30B7;&#x30E7;&#x30A6;&#x30B8;&#x30E7;&#x30A6;&#x30D0;&#x30A8; &#x30AD;&#x30A4;&#x30ED;&#x30B7;&#x30E7;&#x30A6;&#x30B8;&#x30E7;&#x30A6;&#x30D0;&#x30A8;&#x4EBA;&#x9593;&#x306E;&#x75C5;&#x6C17;&#x306B;&#x95A2;&#x9023;&#x3059;&#x308B;&#x30DF;&#x30C8;&#x30B3;&#x30F3;&#x30C9;&#x30EA;&#x30A2;&#x86CB;&#x767D;&#x8CEA;&#x306E;&#x5F79;&#x5272;&#x3092;&#x52C9;&#x5F37;&#x306E;&#x76F8;&#x5BFE;&#x7684;&#x306A;&#x30E6;&#x30FC;&#x30C6;&#x30A3;&#x30EA;&#x30C6;&#x30A3;&#x3092;&#x78BA;&#x8A8D;&#x3057;&#x307E;&#x3059;&#x3002;&#x5927;&#x8178;&#x83CC;&#x306F;&#x3001;&#x5358;&#x4E00;&#x306E;&#x30BB;&#x30EB;&#x3001;&#x6709;&#x7528;&#x306A;&#x30E2;&#x30C7;&#x30EB; &#x30B7;&#x30B9;&#x30C6;&#x30E0;&#x3067;&#x3042;&#x308B;&#x30DF;&#x30C8;&#x30B3;&#x30F3;&#x30C9;&#x30EA;&#x30A2;&#x547C;&#x5438;&#x9396;&#x30BF;&#x30F3;&#x30D1;&#x30AF;&#x8CEA;&#x306E;&#x69CB;&#x9020;&#x3068;&#x6A5F;&#x80FD;&#x3092;&#x8ABF;&#x67FB;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x3042;&#x308B;&#x3068;&#x8A3C;&#x660E;&#x3057;&#x305F;&#x539F;&#x6838;&#x751F;&#x7269;&#x306E;&#x7D30;&#x83CC;&#x3067;&#x3059;&#x3002;&#x51FA;&#x82BD;&#x9175;&#x6BCD;&#x306F;&#x80B2;&#x3064;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x308B;&#x5358;&#x7D30;&#x80DE;&#x771F;&#x6838;&#x4F9D;&#x5B58;&#x30DF;&#x30C8;&#x30B3;&#x30F3;&#x30C9;&#x30EA;&#x30A2;&#x547C;&#x5438;&#x3001;&#x307E;&#x305F;&#x306F;&#x30A8;&#x30BF;&#x30CE;&#x30FC;&#x30EB;&#x767A;&#x9175;&#x3001;&#x30DF;&#x30C8;&#x30B3;&#x30F3;&#x30C9;&#x30EA;&#x30A2;&#x6A5F;&#x80FD;&#x3092;&#x8ABF;&#x67FB;&#x3059;&#x308B;&#x305F;&#x3081;&#x3001;&#x771F;&#x306E;&#x6069;&#x6075;&#x3067;&#x3042;&#x308B;&#x3053;&#x3068;&#x3092;&#x8A3C;&#x660E;&#x3057;&#x3066;&#x3044;&#x308B;&#x30D7;&#x30ED;&#x30D1;&#x30C6;&#x30A3;&#x3082;&#x540C;&#x69D8;&#x306B;&#x3002;&#x7DDA;&#x866B;&#x306F;&#x3001;5 &#x3064;&#x306E;&#x4E3B;&#x8981;&#x306A;&#x7D44;&#x7E54;&#x306B;&#x7D44;&#x7E54;&#x3055;&#x308C;&#x3001;&#x5805;&#x7262;&#x306A;&#x30E2;&#x30C7;&#x30EB;&#x52D5;&#x7269; in vitro &#x304A;&#x3088;&#x3073; in vivo &#x306E;&#x7814;&#x7A76;&#x306E;&#x4E3B;&#x306A;&#x547C;&#x5438;&#x9396;&#x4E0D;&#x5168;&#x3068;&#x305D;&#x306E;&#x6F5C;&#x5728;&#x7684;&#x306A;&#x30BB;&#x30E9;&#x30D4;&#x30FC;&#x4EBA;&#x9593;&#x3067;&#x3042;&#x308B;&#x3068;&#x8A3C;&#x660E;&#x3057;&#x305F;&#x591A;&#x7D30;&#x80DE;&#x3001;&#x9855;&#x5FAE;&#x93E1;&#x30EF;&#x30FC;&#x30E0;&#x3067;&#x3059;&#x3002;&#x4E00;&#x4E16;&#x7D00;&#x4EE5;&#x4E0A;&#x3092;&#x5B66;&#x3073;&#x3001;&#x30AD;&#x30A4;&#x30ED;&#x30B7;&#x30E7;&#x30A6;&#x30B8;&#x30E7;&#x30A6;&#x30D0;&#x30A8;&#x524D;&#x65B9;&#x304A;&#x3088;&#x3073;&#x9006;&#x907A;&#x4F1D;&#x5B66;&#x3092;&#x7528;&#x3044;&#x305F;&#x30DF;&#x30C8;&#x30B3;&#x30F3;&#x30C9;&#x30EA;&#x30A2;&#x306E;&#x751F;&#x7269;&#x5B66;&#x306E;&#x8ABF;&#x67FB;&#x3092;&#x5BB9;&#x6613;&#x306B;&#x591A;&#x91CF;&#x306E;&#x907A;&#x4F1D;&#x7684;&#x30C4;&#x30FC;&#x30EB;&#x3084;&#x8A66;&#x85AC;&#x3092;&#x63D0;&#x4F9B;&#x3059;&#x308B;&#x53E4;&#x5178;&#x7684;&#x306A;&#x5F8C;&#x751F;&#x52D5;&#x7269;&#x30E2;&#x30C7;&#x30EB; &#x30B7;&#x30B9;&#x30C6;&#x30E0;&#x3067;&#x3059;&#x3002;&#x305D;&#x308C;&#x305E;&#x308C;&#x306E;&#x9577;&#x6240;&#x3068;&#x305D;&#x308C;&#x305E;&#x308C;&#x306E;&#x7A2E;&#x306E;&#x76F8;&#x5BFE;&#x30DF;&#x30C8;&#x30B3;&#x30F3;&#x30C9;&#x30EA;&#x30A2;&#x7814;&#x7A76;&#x306E;&#x9650;&#x754C;&#x3092;&#x63A2;&#x691C;&#x3057;&#x307E;&#x3059;&#x3002;&#x307E;&#x305F;&#x3001;&#x6982;&#x8981;&#x306E;&#x5404;&#x3053;&#x308C;&#x3089; 4 &#x3064;&#x306E;&#x30E2;&#x30C7;&#x30EB; &#x30B7;&#x30B9;&#x30C6;&#x30E0;&#x306B;&#x304A;&#x3051;&#x308B;&#x30DF;&#x30C8;&#x30B3;&#x30F3;&#x30C9;&#x30EA;&#x30A2;&#x306E;&#x751F;&#x7269;&#x5B66;&#x306E;&#x4E3B;&#x8981;&#x306A;&#x767A;&#x898B;&#x306E;&#x63D0;&#x4F9B;&#x3067;&#x3059;&#x3002;

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&#xBBF8;&#xD1A0; &#xCF58; &#xB4DC;&#xB9AC; &#xC544; &#xAD6C;&#xC870; &#xAD6C;&#xC131;&#xACFC; &#xAE30;&#xB2A5; &#xC9C4;&#xD654;&#xC5D0; &#xAC78;&#xCCD0; &#xAD11;&#xBC94;&#xC704; &#xD55C; &#xBCF4;&#xC874; &#xC778;&#xAC04;&#xC758; &#xBBF8;&#xD1A0; &#xCF58; &#xB4DC; &#xB9AC;&#xC544; &#xC0DD;&#xBB3C;&#xD559; &#xBC0F; &#xC9C8;&#xBCD1;&#xC758; &#xC6B0;&#xB9AC;&#xC758; &#xC774;&#xD574;&#xB97C; &#xD655;&#xC7A5; &#xD558;&#xB294; &#xB3C5;&#xD2B9;&#xD55C; &#xAE30;&#xD68C;&#xB97C; &#xC81C;&#xACF5; &#xD569;&#xB2C8;&#xB2E4;. &#xD6E8;&#xC52C; &#xB354; &#xAC04;&#xB2E8;&#xD55C; &#xBAA8;&#xB378; &#xC0DD;&#xBB3C;&#xC758; &#xC0DD;&#xBB3C;&#xC744; &#xC870;&#xC0AC; &#xD558; &#xC5EC; &#xADF8;&#xAC83;&#xC774; &#xC885;&#xC885; &#xC784;&#xC0C1; &#xC218;&#xC900;&#xC5D0; &#xB3C4;&#xB2EC;&#xD560; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4; &#xC9C8;&#xBB38;&#xC5D0; &#xB300;&#xB2F5; &#xAC00;&#xB2A5; &#xD569;&#xB2C8;&#xB2E4;. &#xC5EC;&#xAE30;, &#xC6B0;&#xB9AC;&#xB294; 4 &#xAC1C;&#xC758; &#xB2E4;&#xB978; &#xBAA8;&#xB378; &#xC720;&#xAE30; &#xCCB4;, &#xC989; &#xBC15;&#xD14C;&#xB9AC;&#xC544; &#xB300;&#xC7A5;&#xADE0;, &#xD6A8; &#xBAA8; Saccharomyces cerevisiae, &#xC120; &#xCDA9; &#xB958; Caenorhabditis elegans&#xC640; &#xACFC;&#xC77C; &#xD30C;&#xB9AC; Drosophila melanogaster &#xC778;&#xAC04;&#xC758; &#xC9C8;&#xBCD1;&#xC5D0; &#xAD00;&#xB828; &#xB41C; &#xBBF8;&#xD1A0; &#xCF58; &#xB4DC;&#xB9AC; &#xC544; &#xB2E8;&#xBC31;&#xC9C8;&#xC758; &#xC5ED;&#xD560;&#xC744; &#xACF5;&#xBD80;&#xC5D0; &#xC0C1;&#xB300; &#xC720;&#xD2F8;&#xB9AC;&#xD2F0; &#xAC80;&#xD1A0;. &#xB300;&#xC7A5;&#xADE0;&#xC740; &#xB2E8;&#xC77C; &#xC138;&#xD3EC;, prokaryotic &#xBC15;&#xD14C;&#xB9AC;&#xC544; &#xBBF8;&#xD1A0; &#xCF58; &#xB4DC;&#xB9AC; &#xC544; &#xD638;&#xD761; &#xC0AC;&#xC2AC; &#xB2E8;&#xBC31;&#xC9C8; &#xAD6C;&#xC870;&#xC640; &#xAE30;&#xB2A5;&#xC744; &#xC870;&#xC0AC; &#xD558;&#xB294; &#xC720;&#xC6A9;&#xD55C; &#xBAA8;&#xB378; &#xC2DC;&#xC2A4;&#xD15C;&#xC744; &#xC785;&#xC99D; &#xD588;&#xB2E4;. S. cerevisiae&#xAC00; &#xC131;&#xC7A5;&#xD560; &#xC218; &#xC788;&#xB294; &#xB2E8; &#xC138;&#xD3EC; &#xC9C4;&#xD575;&#xC0DD;&#xBB3C; &#xC885;&#xC18D; &#xB41C; &#xBBF8;&#xD1A0; &#xCF58; &#xB4DC;&#xB9AC; &#xC544; &#xD638;&#xD761; &#xB610;&#xB294; &#xC5D0;&#xD0C4;&#xC62C; &#xBC1C;&#xD6A8;, &#xBBF8;&#xD1A0; &#xCF58; &#xB4DC;&#xB9AC; &#xC544; &#xAE30;&#xB2A5;&#xC744; &#xC870;&#xC0AC; &#xD558;&#xB294; &#xAC83;&#xC5D0; &#xB300; &#xD55C; &#xC9C4;&#xC815;&#xD55C; &#xBCF4;&#xD0EC;&#xC774; &#xB420; &#xC785;&#xC99D; &#xB418;&#xC5C8;&#xC2B5;&#xB2C8;&#xB2E4; &#xC18D;&#xC131;&#xC5D0; &#xC758;&#xD574; &#xB3D9;&#xB4F1; &#xD558; &#xAC8C; &#xC798;. C. elegans&#xB294; &#xB2E4;&#xC138;&#xD3EC;, &#xBBF8;&#xC138;&#xD55C; &#xBC8C;&#xB808;&#xB294; 5 &#xAC1C;&#xC758; &#xC8FC;&#xC694; &#xC870;&#xC9C1;&#xC73C;&#xB85C; &#xAD6C;&#xC131; &#xD558; &#xACE0; &#xAE30;&#xBCF8; &#xD638;&#xD761; &#xCCB4;&#xC778; &#xBD80;&#xC804; &#xBC0F; &#xC778; &#xAC04;&#xC5D0; &#xC788;&#xB294; &#xADF8;&#xAC83;&#xC758; &#xC7A0;&#xC7AC;&#xC801;&#xC778; &#xCE58;&#xB8CC;&#xC758; &#xC0DD;&#xCCB4; &#xC678; &#xBC0F; &#xC0DD;&#xCCB4; &#xC870;&#xAC74; &#xC5F0;&#xAD6C;&#xB97C; &#xC704;&#xD55C; &#xAC15;&#xB825;&#xD55C; &#xBAA8;&#xB378; &#xB3D9;&#xBB3C;&#xC744; &#xC785;&#xC99D; &#xD588;&#xB2E4;. &#xD55C; &#xC138;&#xAE30; &#xC774;&#xC0C1;&#xC5D0; &#xB300; &#xD55C; &#xACF5;&#xBD80;, &#xB514; melanogaster &#xC815;&#xBC29;&#xD5A5; &#xBC0F; &#xC5ED;&#xBC29;&#xD5A5; &#xC720;&#xC804;&#xD559;&#xC744; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; &#xBBF8;&#xD1A0; &#xCF58; &#xB4DC; &#xB9AC;&#xC544; &#xC0DD;&#xBB3C;&#xD559;&#xC758; &#xC870;&#xC0AC;&#xB97C; &#xC6A9;&#xC774; &#xD558; &#xAC8C; &#xD558;&#xB294; &#xC720;&#xC804;&#xC790; &#xB3C4;&#xAD6C; &#xBC0F; &#xC2DC; &#xC57D;&#xC758; &#xD48D;&#xBD80;&#xB97C; &#xC81C;&#xACF5; &#xD558; &#xACE0; &#xD074;&#xB798;&#xC2DD; metazoan &#xBAA8;&#xB378; &#xC2DC;&#xC2A4;&#xD15C;&#xC785;&#xB2C8;&#xB2E4;. &#xAC01;&#xAC01; &#xC7A5;&#xC810;&#xACFC; &#xBBF8;&#xD1A0; &#xCF58; &#xB4DC;&#xB9AC; &#xC544; &#xC5F0;&#xAD6C;&#xB97C; &#xAE30;&#xC900;&#xC73C;&#xB85C; &#xAC01; &#xC885;&#xC871;&#xC758; &#xD55C;&#xACC4;&#xB97C; &#xD0D0;&#xAD6C; &#xB41C;&#xB2E4;. &#xB610;&#xD55C;, &#xC774;&#xB7EC;&#xD55C; 4 &#xAC1C;&#xC758; &#xBAA8;&#xB378; &#xC2DC;&#xC2A4;&#xD15C;&#xC758; &#xAC01; &#xBBF8;&#xD1A0; &#xCF58; &#xB4DC; &#xB9AC;&#xC544; &#xC0DD;&#xBB3C;&#xD559;&#xC5D0;&#xC11C; &#xC8FC;&#xC694; &#xBC1C;&#xACAC;&#xC758; &#xAC1C;&#xC694; &#xC81C;&#xACF5; &#xB429;&#xB2C8;&#xB2E4;.

&#xBC15;&#xD14C;&#xB9AC;&#xC544;, &#xD6A8; &#xBAA8;, &#xBC8C;&#xB808;, &#xADF8;&#xB9AC;&#xACE0; &#xD30C;&#xB9AC;: &#xC778;&#xAC04;&#xC758; &#xBBF8;&#xD1A0; &#xCF58; &#xB4DC; &#xB9AC;&#xC544; &#xC9C8;&#xBCD1;&#xC744; &#xC870;&#xC0AC; &#xD558;&#xAE30; &#xC704;&#xD574; &#xAC04;&#xB2E8;&#xD55C; &#xBAA8;&#xB378; &#xC0DD;&#xBB3C;&#xC744; &#xC545;&#xC6A9;.

La amplia conservaci&oacute;n de estructura mitocondrial, composici&oacute;n y funci&oacute;n a trav&eacute;s de la evoluci&oacute;n ofrece una oportunidad &uacute;nica para ampliar nuestra comprensi&oacute;n de la biolog&iacute;a humana mitocondrial y la enfermedad. Al investigar la biolog&iacute;a del mucho m&aacute;s simples organismos modelo, a menudo es posible responder a preguntas que son inalcanzables en el nivel cl&iacute;nico. Aqu&iacute;, repasamos la utilidad relativa de cuatro organismos modelo diferente, es decir, la bacteria Escherichia coli, la levadura Saccharomyces cerevisiae, el nematodo Caenorhabditis elegans y la mosca de la fruta Drosophila melanogaster, estudiar el papel de las prote&iacute;nas mitocondriales relevantes para la enfermedad humana. E. coli son unicelulares, procariotas bacterias que han demostrado para ser un sistema modelo &uacute;til para investigar la funci&oacute;n y estructura de la prote&iacute;na de la cadena respiratoria mitocondrial. S. cerevisiae es un eucariotas unicelulares que pueden crecer tanto por la respiraci&oacute;n mitocondrial dependiente o por fermentaci&oacute;n de etanol, una propiedad que ha demostrado para ser una verdadera bendici&oacute;n para investigar la funcionalidad mitocondrial. C. elegans es un gusano microsc&oacute;pico, multicelular que se organiza en cinco grandes tejidos y ha demostrado para ser un animal robusto modelo para estudios in vitro e in vivo de disfunci&oacute;n primaria de cadena respiratoria y sus posibles tratamientos en los seres humanos. Estudi&oacute; durante m&aacute;s de un siglo, D. melanogaster es un sistema de modelo metazoarios cl&aacute;sico que ofrece una abundancia de herramientas gen&eacute;ticas y reactivos que facilita las investigaciones de Biolog&iacute;a mitocondrial gen&eacute;tica tanto en avance y retroceso. Se exploran las respectivas fortalezas y limitaciones de cada especie en relaci&oacute;n con estudios mitocondriales. Adem&aacute;s, se proporciona un Resumen de importantes descubrimientos realizados en biolog&iacute;a mitocondrial en cada uno de estos sistemas de modelo de cuatro.

Bacterias, levaduras, gusanos y moscas: explotaci&oacute;n de organismos modelo simple para investigar enfermedades mitocondriales humanas.

&#x7EBF;&#x7C92;&#x4F53;&#x547C;&#x5438;&#x94FE;&#x75BE;&#x75C5;&#x6CBB;&#x7597;&#x88AB;&#x590D;&#x6742;&#x5BFC;&#x81F4;&#x5E7F;&#x6CDB;&#x53D8;&#x91CF;&#x7684;&#x4E34;&#x5E8A;&#x8C03;&#x67E5;&#x7ED3;&#x679C;&#x7684;&#x7EC6;&#x80DE;&#x673A;&#x5236;&#x7684;&#x4E86;&#x89E3;&#x6709;&#x9650;&#x3002;&#x5728;&#x8FD9;&#x91CC;&#xFF0C;&#x6211;&#x4EEC;&#x5728; Pdss2 &#x7A81;&#x53D8;&#x4F53;&#x52A8;&#x7269;&#x4E3B;&#x8981;&#x8F85;&#x9176; Q (CoQ) &#x865A;&#x660E;&#x663E;&#x5730;&#x6539;&#x5584;&#x7531;&#x666E;&#x7F57;&#x5E03;&#x8003; (1 %w / w) &#x53E3;&#x670D;&#x6CBB;&#x7597;&#x4E2D;&#x663E;&#x793A;&#x8BE5;&#x5C40;&#x7076;&#x6027;&#x8282;&#x6BB5;&#x6027;&#x80BE;&#x5C0F;&#x7403;&#x75BE;&#x75C5;&#x7C7B;&#x4F3C;&#x80BE;&#x810F;&#x75BE;&#x75C5;&#x3002;&#x9519;&#x4E49;&#x7A81;&#x53D8;&#x5C0F;&#x9F20;&#x7684;&#x9884;&#x9632;&#x6548;&#x679C;&#x662F;&#x7C7B;&#x4F3C;&#x7F8E;&#x8054;&#x50A8;&#x662F;&#x5426;&#x666E;&#x7F57;&#x5E03;&#x8003;&#x4ECE;&#x65AD;&#x5976;&#x6216;&#x5178;&#x578B;&#x80BE;&#x708E;&#x53D1;&#x75C5;&#x524D; 3 &#x5468;&#x3002;&#x6B64;&#x5916;&#xFF0C;&#x4E0E;&#x666E;&#x7F57;&#x5E03;&#x8003;&#x7684; 2 &#x5468;&#x5185;&#x663E;&#x8457;&#x6CBB;&#x7597;&#x75C7;&#x72B6;&#x6027;&#x52A8;&#x7269;&#x51CF;&#x5C11;&#x86CB;&#x767D;&#x5C3F;&#x3002;&#x666E;&#x7F57;&#x5E03;&#x8003;&#x4E86;&#x4E00;&#x4E2A;&#x66F4;&#x660E;&#x663E;&#x7684;&#x5065;&#x5EB7;&#x798F;&#x5229;&#x6BD4;&#x9AD8;&#x5242;&#x91CF; CoQ(10) &#x8865;&#x5145;&#x5E76;&#x552F;&#x4E00;&#x8FD8;&#x539F; CoQ(9) &#x7A81;&#x53D8;&#x4F53;&#x80BE;&#x810F;&#x4E2D;&#x7684;&#x5185;&#x5BB9;&#x3002;&#x666E;&#x7F57;&#x5E03;&#x8003;&#x5927;&#x5927;&#x7F13;&#x89E3;&#x8DE8;&#x8BB8;&#x591A;&#x4E2D;&#x95F4;&#x4EE3;&#x8C22;&#x57DF;&#xFF0C;&#x5305;&#x62EC;&#x8FC7;&#x6C27;&#x5316;&#x7269;&#x9176;&#x4F53;&#x589E;&#x6B96;&#x7269;&#x6FC0;&#x6D3B;&#x53D7;&#x4F53; &#xFF08;PPAR&#xFF09; &#x901A;&#x8DEF;&#x4FE1;&#x53F7;&#x8F6C;&#x5F55;&#x6539;&#x5EFA;&#x3002;&#x666E;&#x7F57;&#x5E03;&#x8003;&#x7684;&#x6709;&#x76CA;&#x5F71;&#x54CD;&#x80BE;&#x529F;&#x80FD;&#x548C;&#x4EE3;&#x8C22;&#x75C5;&#x8868;&#x73B0;&#x7684; Pdss2 &#x867D;&#x7136;&#x6E29;&#x548C;&#x8BF1;&#x5BFC;&#x6C27;&#x5316;&#x5E94;&#x6FC0;&#x7684;&#x53D1;&#x751F;&#x548C;&#x51FA;&#x73B0;&#x72EC;&#x7ACB;&#x4E8E;&#x5176;&#x964D;&#x8102;&#x4F5C;&#x7528;&#x3002;&#x76F8;&#x53CD;&#xFF0C;&#x4E0B;&#x964D; CoQ(9) &#x5185;&#x5BB9;&#x548C;&#x6539;&#x53D8; PPAR &#x901A;&#x8DEF;&#x4FE1;&#x53F7;&#xFF0C;&#x5206;&#x522B;&#x663E;&#x793A;&#xFF0C;&#x4EE5;&#x534F;&#x8C03;&#x80BE;&#x5C0F;&#x7403;&#x548C;&#x5168;&#x7403;&#x4EE3;&#x8C22;&#x7684;&#x540E;&#x679C;&#x4E3B;&#x8981; CoQ &#x4E0D;&#x8DB3;&#x4E4B;&#x5904;&#xFF0C;&#x8FD9;&#x662F;&#x53EF;&#x4EE5;&#x9884;&#x9632;&#x548C;&#x6CBB;&#x6108;&#x6CBB;&#x7597;&#x53E3;&#x8154;&#x666E;&#x7F57;&#x5E03;&#x8003;&#x3002;

&#x666E;&#x7F57;&#x5E03;&#x8003;&#x6539;&#x5584;&#x80BE;&#x529F;&#x80FD;&#x548C;&#x4EE3;&#x8C22; Pdss2 &#x57FA;&#x56E0;&#x7A81;&#x53D8;&#x5C0F;&#x9F20;&#x521D;&#x7EA7; CoQ &#x7F3A;&#x5931;&#x7684;&#x540E;&#x9057;&#x75C7;&#x3002;

Therapy of mitochondrial respiratory chain diseases is complicated by limited understanding of cellular mechanisms that cause the widely variable clinical findings. Here, we show that focal segmental glomerulopathy-like kidney disease in Pdss2 mutant animals with primary coenzyme Q (CoQ) deficiency is significantly ameliorated by oral treatment with probucol (1%â€‰w/w). Preventative effects in missense mutant mice are similar whether fed probucol from weaning or for 3 weeks prior to typical nephritis onset. Furthermore, treating symptomatic animals for 2 weeks with probucol significantly reduces albuminuria. Probucol has a more pronounced health benefit than high-dose CoQ(10) supplementation and uniquely restores CoQ(9) content in mutant kidney. Probucol substantially mitigates transcriptional alterations across many intermediary metabolic domains, including peroxisome proliferator-activated receptor (PPAR) pathway signaling. Probucol's beneficial effects on the renal and metabolic manifestations of Pdss2 disease occur despite modest induction of oxidant stress and appear independent of its hypolipidemic effects. Rather, decreased CoQ(9) content and altered PPAR pathway signaling appear, respectively, to orchestrate the glomerular and global metabolic consequences of primary CoQ deficiency, which are both preventable and treatable with oral probucol therapy.

Probucol ameliorates renal and metabolic sequelae of primary CoQ deficiency in Pdss2 mutant mice.

Th&eacute;rapie des maladies de la cha&icirc;ne respiratoire mitochondriale est compliqu&eacute;e par une compr&eacute;hension limit&eacute;e des m&eacute;canismes cellulaires qui provoquent des r&eacute;sultats cliniques extr&ecirc;mement variables. Nous d&eacute;montrons ici que la maladie focale segmentaire ressemblant immunotacto&iuml;de rein en Pdss2 animaux mutants pr&eacute;sentant un d&eacute;ficit en primaire coenzyme Q (CoQ) est significativement am&eacute;lior&eacute;e par un traitement oral avec probucol (1 % w/w). Effet pr&eacute;ventif chez des souris mutantes faux-sens est semblables si nourris probucol du sevrage ou 3 semaines pr&eacute;c&eacute;dant l&#39;apparition de la n&eacute;phrite typique. En outre, traiter les animaux symptomatique pendant 2 semaines avec probucol significativement r&eacute;duit albuminurie. Probucol a une prestation de sant&eacute; plus prononc&eacute;e que la suppl&eacute;mentation de haut-dose CoQ(10) et restaure uniquement CoQ(9) contenu dans le rein mutant. Probucol att&eacute;nue sensiblement les alt&eacute;rations transcriptionnelles dans de nombreux domaines m&eacute;taboliques interm&eacute;diaires, y compris peroxisome proliferator-activated receptor (PPAR) voie de signalisation. Les effets b&eacute;n&eacute;fiques du probucol sur les manifestations r&eacute;nales et m&eacute;taboliques de la maladie de Pdss2 se produisent malgr&eacute; la modeste induction du stress oxydant et apparaissent ind&eacute;pendamment de ses effets hypolip&eacute;miants. Au contraire, a diminu&eacute; CoQ(9) contenu et alt&eacute;r&eacute;e PPAR voie de signalisation figurent, respectivement, d&#39;orchestrer les cons&eacute;quences m&eacute;taboliques glom&eacute;rulaires et globales du d&eacute;ficit primaire de CoQ, soit &agrave; la fois &eacute;vitable et traitable avec la th&eacute;rapie orale probucol.

Probucol am&eacute;liore les s&eacute;quelles du d&eacute;ficit primaire de CoQ dans les souris mutantes Pdss2 r&eacute;nales et m&eacute;taboliques.

Therapie der mitochondrialen Krankheiten wird durch begrenztes Verst&auml;ndnis der zellul&auml;ren Mechanismen erschwert, die sehr unterschiedlich klinische Ergebnisse verursachen. Hier zeigen wir die fokale segmentale Glomerulopathie-wie Nierenkrankheit in Pdss2 mutierte Tiere mit prim&auml;ren Coenzym Q (CoQ) Mangel ist durch orale Behandlung mit induzierender (1 % w/w) deutlich verbessert. Pr&auml;ventive Wirkung Missense Mutanten M&auml;usen sind &auml;hnlich, ob gef&uuml;ttert induzierender Entw&ouml;hnung oder f&uuml;r 3 Wochen vor Beginn der typischen Lupusnephritis. Dar&uuml;ber hinaus reduziert die Behandlung von symptomatische Tieren f&uuml;r 2 Wochen mit induzierender deutlich Albuminurie. Induzierender hat einen deutlichen Nutzen f&uuml;r die Gesundheit als Hochdosis-CoQ(10) Erg&auml;nzung und eindeutig CoQ(9) Inhalt in mutant Niere wiederhergestellt. Induzierender verringert erheblich transkriptionelle &Auml;nderungen dom&auml;nen&uuml;bergreifend viele zwischengeschaltete metabolische, einschlie&szlig;lich Peroxisom-Proliferator-aktivierter Rezeptor (PPAR)-Weg-Signalisierung. Induzierender die wohltuende Wirkung auf die renale und metabolischen Erscheinungsformen von Pdss2 Krankheit auftreten, trotz bescheidener Induktion von oxidativen Stress und erscheinen unabh&auml;ngig von seiner lipidsenkende Wirkung. Vielmehr verringert CoQ(9) Inhalt und ver&auml;nderte PPAR Stoffwechselweg Signalisierung angezeigt, bzw. die glomerul&auml;ren und globalen metabolischen Folgen der prim&auml;ren CoQ-Mangel, zu orchestrieren, die vermeidbare und mit m&uuml;ndlichen induzierender Therapie behandelbar sind.

Induzierender aerztlichen Nieren- und Stoffwechselerkrankungen Folgeerscheinungen von prim&auml;ren CoQ Mangel Pdss2 Mutanten M&auml;usen.

La terapia delle malattie della catena respiratoria mitocondriale &egrave; complicata dalla limitata comprensione dei meccanismi cellulari che causano i rilievi clinici ampiamente variabili. Qui, vi mostriamo quella malattia focale segmentale Rene immunotattoide-come in Pdss2 animali mutanti con deficit primario coenzima Q (CoQ) &egrave; notevolmente migliorata dal trattamento orale con probucol (1% w/w). Effetti di prevenzione nei topi mutanti missenso sono simili se alimentato probucol dallo svezzamento o per 3 settimane prima dell&#39;esordio tipico di nefrite. Inoltre, nel trattamento sintomatici degli animali per 2 settimane con probucol significativamente riduce albuminuria. Probucol ha un beneficio di salute pi&ugrave; pronunciato rispetto a un&#39;integrazione di alte dosi CoQ(10) e ripristina in modo univoco CoQ(9) contenuto nel rene mutante. Probucol riduce sostanzialmente transcriptional alterazioni in molti domini metaboliche intermediari, tra cui peroxisome proliferator-activated receptor (PPAR) signaling pathway. Gli effetti benefici del Probucol sulle manifestazioni renali e metabolici della malattia Pdss2 verificano nonostante la modesta induzione dello stress ossidante e appaiono indipendenti dei suoi effetti ipolipemizzanti. Piuttosto, &egrave; diminuito CoQ(9) contenuto e alterata PPAR pathway di segnalazione appaiono, rispettivamente, a orchestrare le conseguenze metaboliche glomerulare e globale di deficit primario di CoQ, che sono prevenibili e curabili con terapia orale probucol.

Probucol migliora sequele renali e metaboliche di primario CoQ deficit nel topo mutante Pdss2.

&#x30DF;&#x30C8;&#x30B3;&#x30F3;&#x30C9;&#x30EA;&#x30A2;&#x547C;&#x5438;&#x9396;&#x75BE;&#x60A3;&#x306E;&#x6CBB;&#x7642;&#x306E;&#x5E83;&#x304F;&#x53EF;&#x5909;&#x306E;&#x81E8;&#x5E8A;&#x6240;&#x898B;&#x3092;&#x5F15;&#x304D;&#x8D77;&#x3053;&#x3059;&#x7D30;&#x80DE;&#x306E;&#x30E1;&#x30AB;&#x30CB;&#x30BA;&#x30E0;&#x306E;&#x9650;&#x3089;&#x308C;&#x305F;&#x7406;&#x89E3;&#x306B;&#x3088;&#x3063;&#x3066;&#x8907;&#x96D1;&#x306B;&#x306A;&#x308A;&#x307E;&#x3059;&#x3002;&#x3053;&#x3053;&#x3067;&#x306F;&#x3001;&#x30D7;&#x30E9;&#x30A4;&#x30DE;&#x30EA; &#x30B3;&#x30A8;&#x30F3;&#x30B6;&#x30A4;&#x30E0; Q (CoQ) &#x6B20;&#x640D;&#x75C7;&#x306E;&#x5909;&#x7570;&#x52D5;&#x7269;&#x304C;&#x5927;&#x5E45;&#x306B;&#x6539;&#x5584;&#x3055;&#x308C;&#x308B;&#x30D7;&#x30ED;&#x30D6;&#x30B3;&#x30FC;&#x30EB; (1 &#xFF05; w/w&#xFF09; &#x7D4C;&#x53E3;&#x6295;&#x4E0E;&#x306B;&#x3088;&#x308B; Pdss2 &#x3067;&#x305D;&#x306E;&#x7126;&#x70B9;&#x306E;&#x5206;&#x7BC0;&#x306E;&#x7403;&#x4F53;&#x306E;&#x3088;&#x3046;&#x306A;&#x814E;&#x81D3;&#x75C5;&#x3092;&#x8868;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;&#x30DF;&#x30B9;&#x30BB;&#x30F3;&#x30B9;&#x5909;&#x7570;&#x30DE;&#x30A6;&#x30B9;&#x306B;&#x304A;&#x3051;&#x308B;&#x4E88;&#x9632;&#x52B9;&#x679C;&#x304C;&#x4F9B;&#x7D66;&#x3059;&#x308B;&#x304B;&#x3069;&#x3046;&#x304B;&#x4F3C;&#x3066;&#x30D7;&#x30ED;&#x30D6;&#x30B3;&#x30FC;&#x30EB;&#x96E2;&#x4E73;&#x304B;&#x3089;&#x307E;&#x305F;&#x306F;&#x5178;&#x578B;&#x7684;&#x306A;&#x814E;&#x708E;&#x767A;&#x75C7;&#x307E;&#x3067; 3 &#x9031;&#x9593;&#x3002;&#x3055;&#x3089;&#x306B;&#x3001;&#x75C7;&#x5019;&#x6027;&#x52D5;&#x7269;&#x30D7;&#x30ED;&#x30D6;&#x30B3;&#x30FC;&#x30EB;&#x3067; 2 &#x9031;&#x9593;&#x5927;&#x5E45;&#x306B;&#x6CBB;&#x7642;&#x86CB;&#x767D;&#x5C3F;&#x3092;&#x6E1B;&#x5C11;&#x3057;&#x307E;&#x3059;&#x3002;&#x30D7;&#x30ED;&#x30D6;&#x30B3;&#x30FC;&#x30EB;&#x306F;&#x3088;&#x308A;&#x9855;&#x8457;&#x306A;&#x5065;&#x5EB7;&#x52B9;&#x679C;&#x306E;&#x3088;&#x308A;&#x9AD8;&#x7528;&#x91CF; CoQ(10) &#x6442;&#x53D6;&#x304C;&#x3042;&#x308A;&#x3001;CoQ(9) &#x7A81;&#x7136;&#x5909;&#x7570;&#x4F53;&#x306E;&#x814E;&#x81D3;&#x306E;&#x30B3;&#x30F3;&#x30C6;&#x30F3;&#x30C4;&#x3092;&#x4E00;&#x610F;&#x306B;&#x5FA9;&#x5143;&#x3057;&#x307E;&#x3059;&#x3002;&#x30D7;&#x30ED;&#x30D6;&#x30B3;&#x30FC;&#x30EB;&#x306F;&#x5B9F;&#x8CEA;&#x7684;&#x306B;&#x4EF2;&#x4ECB;&#x3055;&#x308C;&#x305F;&#x4EE3;&#x8B1D;&#x30DA;&#x30EB;&#x30AA;&#x30AD;&#x30B7;&#x30BD;&#x30FC;&#x30E0;&#x5897;&#x6B96;&#x56E0;&#x5B50;&#x6D3B;&#x6027;&#x5316;&#x53D7;&#x5BB9;&#x4F53; (PPAR) &#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x4F1D;&#x9054;&#x7D4C;&#x8DEF;&#x3092;&#x542B;&#x3080;&#x30C9;&#x30E1;&#x30A4;&#x30F3;&#x9593;&#x3067;&#x306F;&#x591A;&#x304F;&#x3001;&#x8EE2;&#x5199;&#x306E;&#x5909;&#x5316;&#x3092;&#x7DE9;&#x548C;&#x3057;&#x307E;&#x3059;&#x3002;Pdss2 &#x75C5;&#x306E;&#x814E;&#x30FB;&#x4EE3;&#x8B1D;&#x306E;&#x75C7;&#x72B6;&#x306B;&#x30D7;&#x30ED;&#x30D6;&#x30B3;&#x30FC;&#x30EB;&#x306E;&#x6709;&#x76CA;&#x306A;&#x52B9;&#x679C;&#x306F;&#x9178;&#x5316;&#x30B9;&#x30C8;&#x30EC;&#x30B9;&#x306E;&#x3055;&#x3055;&#x3084;&#x304B;&#x306A;&#x8A98;&#x5C0E;&#x306B;&#x3082;&#x304B;&#x304B;&#x308F;&#x3089;&#x305A;&#x306B;&#x767A;&#x751F;&#x3059;&#x308B;&#x3001;&#x305D;&#x306E;&#x8102;&#x8CEA;&#x4EE3;&#x8B1D;&#x306B;&#x53CA;&#x307C;&#x3059;&#x72EC;&#x7ACB;&#x8868;&#x793A;&#x3055;&#x308C;&#x307E;&#x3059;&#x3002;&#x3080;&#x3057;&#x308D;&#x3001;CoQ(9) &#x30B3;&#x30F3;&#x30C6;&#x30F3;&#x30C4;&#x3092;&#x6E1B;&#x5C11;&#x3057;&#x3001;&#x5909;&#x8CEA; PPAR &#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x4F1D;&#x9054;&#x7D4C;&#x8DEF;&#x8868;&#x793A;&#x3001;&#x305D;&#x308C;&#x305E;&#x308C;&#x3001;&#x4E21;&#x65B9;&#x306E;&#x4E88;&#x9632;&#x3068;&#x6CBB;&#x7642;&#x7D4C;&#x53E3;&#x30D7;&#x30ED;&#x30D6;&#x30B3;&#x30FC;&#x30EB;&#x7642;&#x6CD5;&#x3067;&#x3042;&#x308B;&#x7CF8;&#x7403;&#x4F53;&#x304A;&#x3088;&#x3073;&#x30B0;&#x30ED;&#x30FC;&#x30D0;&#x30EB;&#x4EE3;&#x8B1D;&#x7D50;&#x679C;&#x30D7;&#x30E9;&#x30A4;&#x30DE;&#x30EA; CoQ &#x6B20;&#x4E4F;&#x3092;&#x8ABF;&#x6574;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x3002;

&#x30D7;&#x30ED;&#x30D6;&#x30B3;&#x30FC;&#x30EB; &#x30D7;&#x30E9;&#x30A4;&#x30DE;&#x30EA; CoQ &#x6B20;&#x4E4F;&#x3067; Pdss2 &#x907A;&#x4F1D;&#x5B50;&#x5909;&#x7570;&#x30DE;&#x30A6;&#x30B9;&#x306E;&#x814E;&#x30FB;&#x4EE3;&#x8B1D;&#x306E;&#x5F8C;&#x907A;&#x75C7;&#x3092;&#x6539;&#x5584;&#x3057;&#x307E;&#x3059;&#x3002;

&#xBBF8;&#xD1A0; &#xCF58; &#xB4DC;&#xB9AC; &#xC544; &#xD638;&#xD761; &#xC0AC;&#xC2AC; &#xC9C8;&#xBCD1;&#xC758; &#xCE58;&#xB8CC; &#xC6D0;&#xC778; &#xB110;&#xB9AC; &#xBCC0;&#xC218; &#xC784;&#xC0C1; &#xC5F0;&#xAD6C; &#xACB0;&#xACFC; &#xC138;&#xD3EC; &#xBA54;&#xCEE4;&#xB2C8;&#xC998;&#xC758; &#xD55C;&#xC815; &#xB41C; &#xC774;&#xD574;&#xC5D0; &#xBCF5;&#xC7A1; &#xD569;&#xB2C8;&#xB2E4;. &#xC5EC;&#xAE30;, &#xC6B0;&#xB9AC;&#xB294; &#xAE30;&#xBCF8; &#xCF54;&#xC5D4;&#xC790;&#xC784; Q (CoQ) &#xACB0;&#xD54D;&#xACFC; &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xB3D9;&#xBB3C; probucol (1 %w / w)&#xC640; &#xD568;&#xAED8; &#xAD6C;&#xAC15; &#xCE58;&#xB8CC;&#xC5D0; &#xC758;&#xD574; ameliorated &#xD06C;&#xAC8C;&#xB294; pdss2&#xC5D0; &#xADF8; &#xCD08;&#xC810; segmental glomerulopathy &#xAC19;&#xC740; &#xC2E0;&#xC7A5; &#xC9C8;&#xD658;&#xC744; &#xBCF4;&#xC5EC;&#xC90D;&#xB2C8;&#xB2E4;. Missense &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xC0DD;&#xC950;&#xC5D0; &#xC608;&#xBC29; &#xD6A8;&#xACFC; &#xC720;&#xC0AC; &#xBA39;&#xC774; &#xC5EC;&#xBD80; probucol weaning &#xB610;&#xB294; &#xC77C;&#xBC18;&#xC801;&#xC778; &#xC2E0;&#xC7A5; &#xC5FC; &#xBC1C;&#xBCD1; &#xC804;&#xC5D0; 3 &#xC8FC; &#xB3D9;&#xC548;. &#xB610;&#xD55C;, probucol&#xC640; &#xD568;&#xAED8; 2 &#xC8FC; &#xB3D9;&#xC548; &#xD06C;&#xAC8C; &#xC99D;&#xC0C1; &#xB3D9;&#xBB3C; &#xCE58;&#xB8CC; &#xB2E8;&#xBC31;&#xB1E8;&#xB97C; &#xAC10;&#xC18C; &#xC2DC;&#xD0A8;&#xB2E4;. Probucol &#xB192;&#xC740; &#xBCF5;&#xC6A9;&#xB7C9; CoQ(10) &#xBCF4;&#xCDA9; &#xBCF4;&#xB2E4; &#xB354; &#xB69C;&#xB837;&#xD55C; &#xAC74;&#xAC15; &#xC774;&#xC810;&#xC774; &#xACE0; &#xC720;&#xC77C; &#xD558; &#xAC8C; &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xC2E0;&#xC7A5;&#xC5D0;&#xC11C; CoQ(9) &#xCF58;&#xD150;&#xCE20;&#xB97C; &#xBCF5;&#xC6D0; &#xD569;&#xB2C8;&#xB2E4;. Probucol&#xB294; &#xC2E4;&#xC9C8;&#xC801;&#xC73C;&#xB85C; peroxisome proliferator &#xD65C;&#xC131;&#xD654; &#xC218;&#xC6A9; &#xCCB4; (PPAR) &#xD1B5;&#xB85C; &#xC2E0;&#xD638;&#xB97C; &#xD3EC;&#xD568; &#xD558; &#xC5EC; &#xB9CE;&#xC740; &#xC911;&#xAC04; &#xB300;&#xC0AC; &#xB3C4;&#xBA54;&#xC778;&#xC5D0; &#xAC78;&#xCCD0; transcriptional &#xBCC0;&#xACBD;&#xC744; &#xC644;&#xD654; &#xD569;&#xB2C8;&#xB2E4;. Pdss2 &#xC9C8;&#xBCD1;&#xC758; &#xC2E0;&#xC7A5; &#xBC0F; &#xB300;&#xC0AC; &#xBC1C;&#xD604;&#xC5D0; Probucol&#xC758; &#xC720;&#xC775;&#xD55C; &#xD6A8;&#xACFC; &#xC81C; &#xC2A4;&#xD2B8;&#xB808;&#xC2A4;&#xC758; &#xACB8;&#xC190; &#xD55C; &#xC720;&#xB3C4;&#xB3C4; &#xBD88;&#xAD6C; &#xD558; &#xACE0; &#xBC1C;&#xC0DD; &#xD558; &#xACE0; &#xB0AE;&#xCD94;&#xB294; &#xD6A8;&#xACFC; &#xBB34;&#xAD00; &#xB098;&#xD0C0;&#xB0A9;&#xB2C8;&#xB2E4;. &#xC624;&#xD788;&#xB824;, CoQ(9) &#xCF58;&#xD150;&#xCE20;&#xB97C; &#xAC10;&#xC18C; &#xBC0F; &#xBCC0;&#xACBD; &#xB41C; PPAR &#xD1B5;&#xB85C; &#xC2E0;&#xD638; &#xD45C;&#xC2DC;, &#xAC01;&#xAC01; &#xC0AC; &#xBC0F; &#xAE00;&#xB85C;&#xBC8C; &#xB300;&#xC0AC; &#xACB0;&#xACFC; &#xAE30;&#xBCF8; CoQ &#xACB0;&#xD54D;&#xC99D;&#xC758; &#xC608;&#xBC29; &#xBC0F; &#xAD6C;&#xAC15; probucol &#xC694;&#xBC95;&#xC73C;&#xB85C; &#xCE58;&#xB8CC;&#xD560; &#xC218; &#xC788;&#xB3C4;&#xB85D; &#xC870;&#xC728;&#xD558;.

Probucol ameliorates Pdss2 &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xC0DD;&#xC950;&#xC5D0;&#xC11C; &#xAE30;&#xBCF8; CoQ &#xACB0;&#xD54D;&#xC758; &#xC2E0;&#xC7A5; &#xBC0F; &#xB300;&#xC0AC; sequelae.

Terapia de enfermedades de la cadena respiratoria mitocondrial se complica por la comprensi&oacute;n limitada de los mecanismos celulares que causan los resultados cl&iacute;nicos ampliamente variables. Aqu&iacute;, demostramos focal segmentaria glomerulopat&iacute;a como nefropat&iacute;a en Pdss2 animales mutantes con deficiencia primaria coenzima Q (CoQ) se mejor&oacute; significativamente por el tratamiento oral con probucol (1% w/w). Efectos preventivos en ratones mutantes sin sentido son similares si alimenta probucol desde el destete o durante 3 semanas antes del inicio de la nefritis t&iacute;pico. Adem&aacute;s, tratar a los animales sintom&aacute;ticos durante 2 semanas con probucol significativamente reduce la albuminuria. Probucol tiene un beneficio de salud m&aacute;s pronunciado que los suplementos de dosis alta CoQ(10) y restaura &uacute;nicamente CoQ(9) contenido en ri&ntilde;&oacute;n mutante. Probucol mitiga sustancialmente transcripcionales alteraciones en muchos dominios metab&oacute;licos intermediarios, incluyendo peroxisome proliferator-activado receptores (PPAR) v&iacute;a se&ntilde;alizaci&oacute;n. Efectos beneficiosos del probucol sobre las manifestaciones renales y metab&oacute;licas de Pdss2 enfermedad ocurren a pesar de la modesta inducci&oacute;n de estr&eacute;s oxidante y aparecen independientes de sus efectos hipolipid&eacute;micos. Por el contrario, disminuy&oacute; CoQ(9) contenido y alterado PPAR v&iacute;a se&ntilde;alizaci&oacute;n aparecen, respectivamente, para orquestar las consecuencias metab&oacute;licas glomerulares y globales de la deficiencia primaria de CoQ, que son prevenibles y tratables con terapia oral probucol.

Probucol mejora las secuelas renales y metab&oacute;licas de la deficiencia primaria de CoQ en ratones mutantes Pdss2.

Leber congenital amaurosis (LCA) is an infantile-onset form of inherited retinal degeneration characterized by severe vision loss(1,2). Two-thirds of LCA cases are caused by mutations in 17 known disease-associated genes(3) (Retinal Information Network (RetNet)). Using exome sequencing we identified a homozygous missense mutation (c.25G>A, p.Val9Met) in NMNAT1 that is likely to be disease causing in two siblings of a consanguineous Pakistani kindred affected by LCA. This mutation segregated with disease in the kindred, including in three other children with LCA. NMNAT1 resides in the previously identified LCA9 locus and encodes the nuclear isoform of nicotinamide mononucleotide adenylyltransferase, a rate-limiting enzyme in nicotinamide adenine dinucleotide (NAD(+)) biosynthesis(4,5). Functional studies showed that the p.Val9Met alteration decreased NMNAT1 enzyme activity. Sequencing NMNAT1 in 284 unrelated families with LCA identified 14 rare mutations in 13 additional affected individuals. These results are the first to link an NMNAT isoform to disease in humans and indicate that NMNAT1 mutations cause LCA.

NMNAT1 mutations cause Leber congenital amaurosis.

Primary mitochondrial respiratory chain (RC) diseases are heterogeneous in etiology and manifestations but collectively impair cellular energy metabolism. Mechanism(s) by which RC dysfunction causes global cellular sequelae are poorly understood. To identify a common cellular response to RC disease, integrated gene, pathway, and systems biology analyses were performed in human primary RC disease skeletal muscle and fibroblast transcriptomes. Significant changes were evident in muscle across diverse RC complex and genetic etiologies that were consistent with prior reports in other primary RC disease models and involved dysregulation of genes involved in RNA processing, protein translation, transport, and degradation, and muscle structure. Global transcriptional and post-transcriptional dysregulation was also found to occur in a highly tissue-specific fashion. In particular, RC disease muscle had decreased transcription of cytosolic ribosomal proteins suggestive of reduced anabolic processes, increased transcription of mitochondrial ribosomal proteins, shorter 5'-UTRs that likely improve translational efficiency, and stabilization of 3'-UTRs containing AU-rich elements. RC disease fibroblasts showed a strikingly similar pattern of global transcriptome dysregulation in a reverse direction. In parallel with these transcriptional effects, RC disease dysregulated the integrated nutrient-sensing signaling network involving FOXO, PPAR, sirtuins, AMPK, and mTORC1, which collectively sense nutrient availability and regulate cellular growth. Altered activities of central nodes in the nutrient-sensing signaling network were validated by phosphokinase immunoblot analysis in RC inhibited cells. Remarkably, treating RC mutant fibroblasts with nicotinic acid to enhance sirtuin and PPAR activity also normalized mTORC1 and AMPK signaling, restored NADH/NAD(+) redox balance, and improved cellular respiratory capacity. These data specifically highlight a common pathogenesis extending across different molecular and biochemical etiologies of individual RC disorders that involves global transcriptome modifications. We further identify the integrated nutrient-sensing signaling network as a common cellular response that mediates, and may be amenable to targeted therapies for, tissue-specific sequelae of primary mitochondrial RC disease. 

Primary respiratory chain disease causes tissue-specific dysregulation of the global transcriptome and nutrient-sensing signaling network.

Mitochondrial DNA (mtDNA) sequence variation can influence the penetrance of complex diseases and climatic adaptation. While studies in geographically defined human populations suggest that mtDNA mutations become fixed when they have conferred metabolic capabilities optimally suited for a specific environment, it has been challenging to definitively assign adaptive functions to specific mtDNA sequence variants in mammals. We investigated whether mtDNA genome variation functionally influences Caenorhabditis elegans wild isolates of distinct mtDNA lineages and geographic origins. We found that, relative to N2 (England) wild-type nematodes, CB4856 wild isolates from a warmer native climate (Hawaii) had a unique p.A12S amino acid substitution in the mtDNA-encoded COX1 core catalytic subunit of mitochondrial complex IV (CIV). Relative to N2, CB4856 worms grown at 20°C had significantly increased CIV enzyme activity, mitochondrial matrix oxidant burden, and sensitivity to oxidative stress but had significantly reduced lifespan and mitochondrial membrane potential. Interestingly, mitochondrial membrane potential was significantly increased in CB4856 grown at its native temperature of 25°C. A transmitochondrial cybrid worm strain, chpIR (M, CB4856>N2), was bred as homoplasmic for the CB4856 mtDNA genome in the N2 nuclear background. The cybrid strain also displayed significantly increased CIV activity, demonstrating that this difference results from the mtDNA-encoded p.A12S variant. However, chpIR (M, CB4856>N2) worms had significantly reduced median and maximal lifespan relative to CB4856, which may relate to their nuclear-mtDNA genome mismatch. Overall, these data suggest that C. elegans wild isolates of varying geographic origins may adapt to environmental challenges through mtDNA variation to modulate critical aspects of mitochondrial energy metabolism. 

Mitochondrial DNA variant in COX1 subunit significantly alters energy metabolism of geographically divergent wild isolates in Caenorhabditis elegans.

Mitochondrial respiratory chain (RC) diseases are highly morbid multi-systemic conditions for which few effective therapies exist. Given the essential role of sirtuin and PPAR signaling in mediating both mitochondrial physiology and the cellular response to metabolic stress in RC complex I (CI) disease, we postulated that drugs that alter these signaling pathways either directly (resveratrol for sirtuin, rosiglitazone for PPARγ, fenofibrate for PPARα), or indirectly by increasing NAD(+) availability (nicotinic acid), might offer effective treatment strategies for primary RC disease. Integrated effects of targeting these cellular signaling pathways on animal lifespan and multi-dimensional in vivo parameters were studied in gas-1(fc21) relative to wild-type (N2 Bristol) worms. Specifically, animal lifespan, transcriptome profiles, mitochondrial oxidant burden, mitochondrial membrane potential, mitochondrial content, amino acid profiles, stable isotope-based intermediary metabolic flux, and total nematode NADH and NAD(+) concentrations were compared. Shortened gas-1(fc21) mutant lifespan was rescued with either resveratrol or nicotinic acid, regardless of whether treatments were begun at the early larval stage or in young adulthood. Rosiglitazone administration beginning in young adult stage animals also rescued lifespan. All drug treatments reversed the most significant transcriptome alterations at the biochemical pathway level relative to untreated gas-1(fc21) animals. Interestingly, increased mitochondrial oxidant burden in gas-1(fc21) was reduced with nicotinic acid but exacerbated significantly by resveratrol and modestly by fenofibrate, with little change by rosiglitazone treatment. In contrast, the reduced mitochondrial membrane potential of mutant worms was further decreased by nicotinic acid but restored by either resveratrol, rosiglitazone, or fenofibrate. Using a novel HPLC assay, we discovered that gas-1(fc21) worms have significant deficiencies of NAD(+) and NADH. Whereas resveratrol restored concentrations of both metabolites, nicotinic acid only restored NADH. Characteristic branched chain amino acid elevations in gas-1(fc21) animals were normalized completely by nicotinic acid and largely by resveratrol, but not by either rosiglitazone or fenofibrate. We developed a visualization system to enable objective integration of these multi-faceted physiologic endpoints, an approach that will likely be useful to apply in future drug treatment studies in human patients with mitochondrial disease. Overall, these data demonstrate that direct or indirect pharmacologic restoration of altered sirtuin and PPAR signaling can yield significant health and longevity benefits, although by divergent bioenergetic mechanism(s), in a nematode model of mitochondrial RC complex I disease. Thus, these animal model studies introduce important, integrated insights that may ultimately yield rational treatment strategies for human RC disease. 

Pharmacologic targeting of sirtuin and PPAR signaling improves longevity and mitochondrial physiology in respiratory chain complex I mutant Caenorhabditis elegans.

Mitochondrial respiratory chain (RC) disease is a heterogeneous and highly morbid group of energy deficiency disorders for which no proven effective therapies exist. Robust vertebrate animal models of primary RC dysfunction are needed to explore the effects of variation in RC disease subtypes, tissue-specific manifestations, and major pathogenic factors contributing to each disorder, as well as their pre-clinical response to therapeutic candidates. We have developed a series of zebrafish (Danio rerio) models that inhibit, to variable degrees, distinct aspects of RC function, and enable quantification of animal development, survival, behaviors, and organ-level treatment effects as well as effects on mitochondrial biochemistry and physiology. Here, we characterize four pharmacologic inhibitor models of mitochondrial RC dysfunction in early larval zebrafish, including rotenone (complex I inhibitor), azide (complex IV inhibitor), oligomycin (complex V inhibitor), and chloramphenicol (mitochondrial translation inhibitor that leads to multiple RC complex dysfunction). A range of concentrations and exposure times of each RC inhibitor were systematically evaluated on early larval development, animal survival, integrated behaviors (touch and startle responses), organ physiology (brain death, neurologic tone, heart rate), and fluorescence-based analyses of mitochondrial physiology in zebrafish skeletal muscle. Pharmacologic RC inhibitor effects were validated by spectrophotometric analysis of Complex I, II and IV enzyme activities, or relative quantitation of ATP levels in larvae. Outcomes were prioritized that utilize in vivo animal imaging and quantitative behavioral assessments, as may optimally inform the translational potential of pre-clinical drug screens for future clinical study in human mitochondrial disease subjects. The RC complex inhibitors each delayed early embryo development, with short-term exposures of these three agents or chloramphenicol from 5 to 7 days post fertilization also causing reduced larval survival and organ-specific defects ranging from brain death, behavioral and neurologic alterations, reduced mitochondrial membrane potential in skeletal muscle (rotenone), and/or cardiac edema with visible blood pooling (oligomycin). Remarkably, we demonstrate that treating animals with probucol, a nutrient-sensing signaling network modulating drug that has been shown to yield therapeutic effects in a range of other RC disease cellular and animal models, both prevented acute rotenone-induced brain death in zebrafish larvae, and significantly rescued early embryo developmental delay from either rotenone or oligomycin exposure. Overall, these zebrafish pharmacologic RC function inhibition models offer a unique opportunity to gain novel insights into diverse developmental, survival, organ-level, and behavioral defects of varying severity, as well as their individual response to candidate therapies, in a highly tractable and cost-effective vertebrate animal model system.

Pharmacologic modeling of primary mitochondrial respiratory chain dysfunction in zebrafish.

Cysteamine bitartrate is a US Food and Drug Administration-approved therapy for nephropathic cystinosis also postulated to enhance glutathione biosynthesis. We hypothesized this antioxidant effect may reduce oxidative stress in primary mitochondrial respiratory chain (RC) disease, improving cellular viability and organismal health. Here, we systematically evaluated the therapeutic potential of cysteamine bitartrate in RC disease models spanning three evolutionarily distinct species. These pre-clinical studies demonstrated the narrow therapeutic window of cysteamine bitartrate, with toxicity at millimolar levels directly correlating with marked induction of hydrogen peroxide production. Micromolar range cysteamine bitartrate treatment in Caenorhabditis elegans gas-1(fc21) RC complex I (NDUFS2-/-) disease invertebrate worms significantly improved mitochondrial membrane potential and oxidative stress, with corresponding modest improvement in fecundity but not lifespan. At 10 to 100 μm concentrations, cysteamine bitartrate improved multiple RC complex disease FBXL4 human fibroblast survival, and protected both complex I (rotenone) and complex IV (azide) Danio rerio vertebrate zebrafish disease models from brain death. Mechanistic profiling of cysteamine bitartrate effects showed it increases aspartate levels and flux, without increasing total glutathione levels. Transcriptional normalization of broadly dysregulated intermediary metabolic, glutathione, cell defense, DNA, and immune pathways was greater in RC disease human cells than in C. elegans, with similar rescue in both models of downregulated ribosomal and proteasomal pathway expression. Overall, these data suggest cysteamine bitartrate may hold therapeutic potential in RC disease, although not through obvious modulation of total glutathione levels. Careful consideration is required to determine safe and effective cysteamine bitartrate concentrations to further evaluate in clinical trials of human subjects with primary mitochondrial RC disease.

Pre-clinical evaluation of cysteamine bitartrate as a therapeutic agent for mitochondrial respiratory chain disease.

Diseases associated with mitochondrial DNA (mtDNA) mutations are highly variable in phenotype, in large part because of differences in the percentage of normal and mutant mtDNAs (heteroplasmy) present within the cell. For example, increasing heteroplasmy levels of the mtDNA tRNA nucleotide (nt) 3243A > G mutation result successively in diabetes, neuromuscular degenerative disease, and perinatal lethality. These phenotypes are associated with differences in mitochondrial function and nuclear DNA (nDNA) gene expression, which are recapitulated in cybrid cell lines with different percentages of m.3243G mutant mtDNAs. Using metabolic tracing, histone mass spectrometry, and NADH fluorescence lifetime imaging microscopy in these cells, we now show that increasing levels of this single mtDNA mutation cause profound changes in the nuclear epigenome. At high heteroplasmy, mitochondrially derived acetyl-CoA levels decrease causing decreased histone H4 acetylation, with glutamine-derived acetyl-CoA compensating when glucose-derived acetyl-CoA is limiting. In contrast, α-ketoglutarate levels increase at midlevel heteroplasmy and are inversely correlated with histone H3 methylation. Inhibition of mitochondrial protein synthesis induces acetylation and methylation changes, and restoration of mitochondrial function reverses these effects. mtDNA heteroplasmy also affects mitochondrial NAD/NADH ratio, which correlates with nuclear histone acetylation, whereas nuclear NAD/NADH ratio correlates with changes in nDNA and mtDNA transcription. Thus, mutations in the mtDNA cause distinct metabolic and epigenomic changes at different heteroplasmy levels, potentially explaining transcriptional and phenotypic variability of mitochondrial disease.

Regulation of nuclear epigenome by mitochondrial DNA heteroplasmy.

Mitochondrial respiratory chain disorders are empirically managed with variable antioxidant, cofactor, and vitamin 'cocktails'. However, clinical trial validated and approved compounds, or doses, do not exist for any single or combinatorial mitochondrial disease therapy. Here, we sought to pre-clinically evaluate whether rationally-designed mitochondrial medicine combinatorial regimens might synergistically improve survival, health, and physiology in translational animal models of respiratory chain complex I disease. Having previously demonstrated that gas-1(fc21) complex I subunit NDUFS2-/-  C. elegans have short lifespan that can be significantly rescued with 17 different metabolic modifiers, signaling modifiers, or antioxidants, here we evaluated 11 random combinations of these 3 treatment classes on gas-1(fc21) lifespan. Synergistic rescue occurred only with glucose, nicotinic acid, and N-acetylcysteine (Glu + NA + NAC), yielding improved mitochondrial membrane potential that reflects integrated respiratory chain function, without exacerbating oxidative stress and while reducing mitochondrial stress (UPRmt) and improving intermediary metabolic disruptions at the levels of the transcriptome, steady-state metabolites, and intermediary metabolic flux. Equimolar Glu + NA + NAC dosing in a zebrafish vertebrate model of rotenone-based complex I inhibition synergistically rescued larval activity, brain death, lactate, ATP, and glutathione levels. Overall, these data provide objective preclinical evidence in two evolutionary-divergent animal models of mitochondrial complex I disease to demonstrate that combinatorial Glu + NA + NAC therapy significantly improved animal resiliency in the face of stressors that exacerbate their underlying metabolic deficiency, thereby preventing acute neurologic and biochemical decompensation. Clinical trials are warranted to evaluate the efficacy of this lead combinatorial therapy regimen to improve resiliency and health outcomes in human subjects with mitochondrial disease.

Combinatorial glucose, nicotinic acid, and N-acetylcysteine therapy has synergistic effect in preclinical C. elegans and zebrafish models of mitochondrial complex I disease.

Article	Year
Bacteria, yeast, worms, and flies: exploiting simple model organisms to investigate human mitochondrial diseases. Developmental disabilities research reviews\| PubMed ID: 20818735	2010
Probucol ameliorates renal and metabolic sequelae of primary CoQ deficiency in Pdss2 mutant mice. EMBO molecular medicine\| PubMed ID: 21567994	2011
NMNAT1 mutations cause Leber congenital amaurosis. Nature genetics\| PubMed ID: 22842227	2012
Primary respiratory chain disease causes tissue-specific dysregulation of the global transcriptome and nutrient-sensing signaling network. PloS one\| PubMed ID: 23894440	2013
Mitochondrial DNA variant in COX1 subunit significantly alters energy metabolism of geographically divergent wild isolates in Caenorhabditis elegans. Journal of molecular biology\| PubMed ID: 24534730	2014
Pharmacologic targeting of sirtuin and PPAR signaling improves longevity and mitochondrial physiology in respiratory chain complex I mutant Caenorhabditis elegans. Mitochondrion\| PubMed ID: 25744875	2015
Pharmacologic modeling of primary mitochondrial respiratory chain dysfunction in zebrafish. Neurochemistry international\| PubMed ID: 28732770	2018
Pre-clinical evaluation of cysteamine bitartrate as a therapeutic agent for mitochondrial respiratory chain disease. Human molecular genetics\| PubMed ID: 30668749	2019
Regulation of nuclear epigenome by mitochondrial DNA heteroplasmy. Proceedings of the National Academy of Sciences of the United States of America\| PubMed ID: 31253706	2019
Combinatorial glucose, nicotinic acid, and N-acetylcysteine therapy has synergistic effect in preclinical C. elegans and zebrafish models of mitochondrial complex I disease. Human molecular genetics\| PubMed ID: 33640978	2021

Eiko Nakamaru-Ogiso

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