Huntington's disease (HD) is a monogenic, progressive, neurodegenerative disorder with currently no available treatment. The Libechov transgenic minipig model for HD (TgHD) displays neuroanatomical similarities to humans and exhibits slow disease progression, and is therefore more powerful than available mouse models for the development of therapy. The phenotypic characterization of this model is still ongoing, and it is essential to validate biomarkers to monitor disease progression and intervention. In this study, the behavioral phenotype (cognitive, motor and behavior) of the TgHD model was assessed, along with biomarkers for mitochondrial capacity, oxidative stress, DNA integrity and DNA repair at different ages (24, 36 and 48 months), and compared with age-matched controls. The TgHD minipigs showed progressive accumulation of the mutant huntingtin (mHTT) fragment in brain tissue and exhibited locomotor functional decline at 48 months. Interestingly, this neuropathology progressed without any significant age-dependent changes in any of the other biomarkers assessed. Rather, we observed genotype-specific effects on mitochondrial DNA (mtDNA) damage, mtDNA copy number, 8-oxoguanine DNA glycosylase activity and global level of the epigenetic marker 5-methylcytosine that we believe is indicative of a metabolic alteration that manifests in progressive neuropathology. Peripheral blood mononuclear cells (PBMCs) were relatively spared in the TgHD minipig, probably due to the lack of detectable mHTT. Our data demonstrate that neuropathology in the TgHD model has an age of onset of 48 months, and that oxidative damage and electron transport chain impairment represent later states of the disease that are not optimal for assessing interventions.This article has an associated First Person interview with the first author of the paper.

Department of Cell Biology Faculty of Science Charles University Prague Prague 12843 Czech Republic

Department of Medical Biochemistry Institute of Clinical Medicine University of Oslo 0372 Oslo Norway

Department of Microbiology Oslo University Hospital 0372 Oslo Norway

Department of Neurology and Centre of Clinical Neuroscience 1st Faculty of Medicine Charles University Prague and General University Hospital Prague Prague 12821 Czech Republic

Department of Pediatrics and Adolescent Medicine 1st Faculty of Medicine Charles University Prague and General University Hospital Prague Prague 12808 Czech Republic

Laboratory of Cell Regeneration and Plasticity Institute of Animal Physiology and Genetics Czech Academy of Science Libechov 27721 Czech Republic

Proteomics and Metabolomics Core Facility PROMEC Department of Clinical and Molecular Medicine Norwegian University of Science and Technology 7491 Trondheim Norway

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Askeland G., Dosoudilova Z., Rodinova M., Klempir J., Liskova I., Kuśnierczyk A., Bjørås M., Nesse G., Klungland A., Hansikova H. et al. (2018). Increased nuclear DNA damage precedes mitochondrial dysfunction in peripheral blood mononuclear cells from Huntington's disease patients. PubMed DOI PMC

Baxa M., Hruska-Plochan M., Juhas S., Vodicka P., Pavlok A., Juhasova J., Miyanohara A., Nejime T., Klima J., Macakova M. et al. (2013). A transgenic minipig model of Huntington's Disease. PubMed

Benn C. L., Sun T., Sadri-Vakili G., McFarland K. N., DiRocco D. P., Yohrling G. J., Clark T. W., Bouzou B. and Cha J.-H. J. (2008). Huntingtin modulates transcription, occupies gene promoters in vivo, and binds directly to DNA in a polyglutamine-dependent manner. PubMed DOI PMC

Björkqvist M., Wild E. J., Thiele J., Silvestroni A., Andre R., Lahiri N., Raibon E., Lee R. V., Benn C. L., Soulet D. et al. (2008). A novel pathogenic pathway of immune activation detectable before clinical onset in Huntington's disease. PubMed DOI PMC

Bogdanov M. B., Andreassen O. A., Dedeoglu A., Ferrante R. J. and Beal M. F. (2001). Increased oxidative damage to DNA in a transgenic mouse model of Huntington's disease. PubMed DOI

Borowsky B., Warner J., Leavitt B. R., Tabrizi S. J., Roos R. A. C., Durr A., Becker C., Sampaio C., Tobin A. J. and Schulman H. (2013). 8OHdG is not a biomarker for Huntington disease state or progression. PubMed DOI PMC

Browne S. E., Bowling A. C., MacGarvey U., Baik M. J., Berger S. C., Muquit M. M. K., Bird E. D. and Beal M. F. (1997). Oxidative damage and metabolic dysfunction in Huntington's disease: selective vulnerability of the basal ganglia. PubMed DOI

Budworth H., Harris F. R., Williams P., Lee D. Y., Holt A., Pahnke J., Szczesny B., Acevedo-Torres K., Ayala-Peña S. and McMurray C. T. (2015). Suppression of somatic expansion delays the onset of pathophysiology in a mouse model of Huntington's disease. PubMed DOI PMC

Cahova M., Chrastina P., Hansikova H., Drahota Z., Trnovska J., Skop V., Spacilova J., Malinska H., Oliyarnyk O., Papackova Z. et al. (2015). Carnitine supplementation alleviates lipid metabolism derangements and protects against oxidative stress in non-obese hereditary hypertriglyceridemic rats. PubMed DOI

Choudhry S., Mukerji M., Srivastava A. K., Jain S. and Brahmachari S. K. (2001). CAG repeat instability at SCA2 locus: anchoring CAA interruptions and linked single nucleotide polymorphisms. PubMed DOI

Davies S. W., Turmaine M., Cozens B. A., DiFiglia M., Sharp A. H., Ross C. A., Scherzinger E., Wanker E. E., Mangiarini L. and Bates G. P. (1997). Formation of neuronal intranuclear inclusions underlies the neurological dysfunction in mice transgenic for the HD mutation. PubMed DOI

Dedeoglu A., Kubilus J. K., Yang L., Ferrante K. L., Hersch S. M., Beal M. F. and Ferrante R. J. (2003). Creatine therapy provides neuroprotection after onset of clinical symptoms in Huntington's disease transgenic mice. PubMed DOI PMC

DiFiglia M., Sapp E., Chase K. O., Davies S. W., Bates G. P., Vonsattel J. P. and Aronin N. (1997). Aggregation of huntingtin in neuronal intranuclear inclusions and dystrophic neurites in brain. PubMed DOI

Ferrante R. J., Andreassen O. A., Jenkins B. G., Dedeoglu A., Kuemmerle S., Kubilus J. K., Kaddurah-Daouk R., Hersch S. M. and Beal M. F. (2000). Neuroprotective effects of creatine in a transgenic mouse model of Huntington's disease. PubMed DOI PMC

Ferrante R. J., Andreassen O. A., Dedeoglu A., Ferrante K. L., Jenkins B. G., Hersch S. M. and Beal M. F. (2002). Therapeutic effects of coenzyme Q10 and remacemide in transgenic mouse models of Huntington's disease. PubMed DOI PMC

Ferrante R. J., Kubilus J. K., Lee J., Ryu H., Beesen A., Zucker B., Smith K., Kowall N. W., Ratan R. R., Luthi-Carter R. et al. (2003). Histone deacetylase inhibition by sodium butyrate chemotherapy ameliorates the neurodegenerative phenotype in Huntington's disease mice. PubMed DOI PMC

Gray M., Shirasaki D. I., Cepeda C., Andre V. M., Wilburn B., Lu X.-H., Tao J., Yamazaki I., Li S.-H., Sun Y. E. et al. (2008). Full-length human mutant huntingtin with a stable polyglutamine repeat can elicit progressive and selective neuropathogenesis in BACHD mice. PubMed DOI PMC

Gu M., Gash M. T., Mann V. M., Javoy-Agid F., Cooper J. M. and Schapira A. H. V. (1996). Mitochondrial defect in Huntington's disease caudate nucleus. PubMed DOI

Gustafson E. L., Ehrlich M. E., Trivedi P. and Greengard P. (1992). Developmental regulation of phosphoprotein gene expression in the caudate-putamen of rat: an in situ hybridization study. PubMed DOI

Hands S., Sajjad M. U., Newton M. J. and Wyttenbach A. (2011). In vitro and in vivo aggregation of a fragment of huntingtin protein directly causes free radical production. PubMed DOI PMC

Hersch S. M., Schifitto G., Oakes D., Bredlau A.-L., Meyers C. M., Nahin R., Rosas H. D. and Huntington Study Group CREST-E Investigators and Coordinators (2017). The CREST-E study of creatine for Huntington disease: a randomized controlled trial. PubMed DOI PMC

Huntington Study Group. (2001). A randomized, placebo-controlled trial of coenzyme Q10 and remacemide in Huntington's disease. PubMed DOI

Jonson I., Ougland R., Klungland A. and Larsen E. (2013). Oxidative stress causes DNA triplet expansion in Huntington's disease mouse embryonic stem cells. PubMed DOI

Jozefovicova M., Herynek V., Jiru F., Dezortova M., Juhasova J., Juhas S., Motlik J. and Hajek M. (2016). Minipig model of Huntington's disease: (1)H magnetic resonance spectroscopy of the brain. PubMed

Klungland A., Rosewell I., Hollenbach S., Larsen E., Daly G., Epe B., Seeberg E., Lindahl T. and Barnes D. E. (1999). Accumulation of premutagenic DNA lesions in mice defective in removal of oxidative base damage. PubMed DOI PMC

Kovtun I. V., Liu Y., Bjoras M., Klungland A., Wilson S. H. and McMurray C. T. (2007). OGG1 initiates age-dependent CAG trinucleotide expansion in somatic cells. PubMed DOI PMC

Krizova J., Stufkova H., Rodinova M., Macakova M., Bohuslavova B., Vidinska D., Klima J., Ellederova Z., Pavlok A., Howland D. S. et al. (2017). Mitochondrial metabolism in a large-animal model of Huntington disease: the hunt for biomarkers in the spermatozoa of Presymptomatic Minipigs. PubMed DOI

Lai Y., Budworth H., Beaver J. M., Chan N. L. S., Zhang Z., McMurray C. T. and Liu Y. (2016). Crosstalk between MSH2-MSH3 and polβ promotes trinucleotide repeat expansion during base excision repair. PubMed DOI PMC

Lee J., Hwang Y. J., Kim K. Y., Kowall N. W. and Ryu H. (2013). Epigenetic mechanisms of neurodegeneration in Huntington's disease. PubMed DOI PMC

Logan A., Shabalina I. G., Prime T. A., Rogatti S., Kalinovich A. V., Hartley R. C., Budd R. C., Cannon B. and Murphy M. P. (2014). In vivo levels of mitochondrial hydrogen peroxide increase with age in mtDNA mutator mice. PubMed DOI PMC

Long J. D., Matson W. R., Juhl A. R., Leavitt B. R., Paulsen J. S., PREDICT-HD Investigators and Coordinators of the Huntington Study Group (2012). 8OHdG as a marker for Huntington disease progression. PubMed DOI PMC

Lowry O. H., Rosebrough N. J., Farr A. L. and Randall R. J. (1951). Protein measurement with the Folin phenol reagent. PubMed

Macakova M., Bohuslavova B., Vochozkova P., Pavlok A., Sedlackova M., Vidinska D., Vochyanova K., Liskova I., Valekova I., Baxa M. et al. (2016). Mutated Huntingtin causes testicular pathology in transgenic minipig boars. PubMed DOI

McGarry A., McDermott M., Kieburtz K., de Blieck E. A., Beal F., Marder K., Ross C., Shoulson I., Gilbert P., Mallonee W. M. et al. (2017). A randomized, double-blind, placebo-controlled trial of coenzyme Q10 in Huntington disease. PubMed DOI PMC

Mende-Mueller L. M., Toneff T., Hwang S.-R., Chesselet M.-F. and Hook V. Y. H. (2001). Tissue-specific proteolysis of Huntingtin (htt) in human brain: evidence of enhanced levels of N- and C-terminal htt fragments in Huntington's disease striatum. PubMed DOI PMC

Miller J. P., Holcomb J., Al-Ramahi I., de Haro M., Gafni J., Zhang N., Kim E., Sanhueza M., Torcassi C., Kwak S. et al. (2010). Matrix metalloproteinases are modifiers of huntingtin proteolysis and toxicity in Huntington's disease. PubMed DOI PMC

Mollersen L., Rowe A. D., Illuzzi J. L., Hildrestrand G. A., Gerhold K. J., Tveteras L., Bjolgerud A., Wilson D. M. III, Bjoras M. and Klungland A. (2012). Neil1 is a genetic modifier of somatic and germline CAG trinucleotide repeat instability in R6/1 mice. PubMed DOI PMC

Ng C. W., Yildirim F., Yap Y. S., Dalin S., Matthews B. J., Velez P. J., Labadorf A., Housman D. E. and Fraenkel E. (2013). Extensive changes in DNA methylation are associated with expression of mutant huntingtin. PubMed DOI PMC

Pearson C. E., Eichler E. E., Lorenzetti D., Kramer S. F., Zoghbi H. Y., Nelson D. L. and Sinden R. R. (1998). Interruptions in the triplet repeats of SCA1 and FRAXA reduce the propensity and complexity of slipped strand DNA (S-DNA) formation. PubMed DOI

Pinto R. M., Dragileva E., Kirby A., Lloret A., Lopez E., St Claire J., Panigrahi G. B., Hou C., Holloway K., Gillis T. et al. (2013). Mismatch repair genes Mlh1 and Mlh3 modify CAG instability in Huntington's disease mice: genome-wide and candidate approaches. PubMed DOI PMC

Polidori M. C., Mecocci P., Browne S. E., Senin U. and Beal M. F. (1999). Oxidative damage to mitochondrial DNA in Huntington's disease parietal cortex. PubMed DOI

Rustin P., Chretien D., Bourgeron T., Gérard B., Rötig A., Saudubray J. M. and Munnich A. (1994). Biochemical and molecular investigations in respiratory chain deficiencies. PubMed DOI

Sathasivam K., Neueder A., Gipson T. A., Landles C., Benjamin A. C., Bondulich M. K., Smith D. L., Faull R. L. M., Roos R. A. C., Howland D. et al. (2013). Aberrant splicing of HTT generates the pathogenic exon 1 protein in Huntington disease. PubMed DOI PMC

Schramke S., Schuldenzucker V., Schubert R., Frank F., Wirsig M., Ott S., Motlik J., Fels M., Kemper N., Hölzner E. et al. (2016). Behavioral phenotyping of minipigs transgenic for the Huntington gene. PubMed DOI

Schuldenzucker V., Schubert R., Muratori L. M., Freisfeld F., Rieke L., Matheis T., Schramke S., Motlik J., Kemper N., Radespiel U. et al. (2017). Behavioral testing of minipigs transgenic for the Huntington gene-A three-year observational study. PubMed DOI PMC

Siddiqui A., Rivera-Sánchez S., Castro M. R., Acevedo-Torres K., Rane A., Torres-Ramos C. A., Nicholls D. G., Andersen J. K. and Ayala-Torres S. (2012). Mitochondrial DNA damage is associated with reduced mitochondrial bioenergetics in Huntington's disease. PubMed DOI PMC

Srere P. A. (1969). Citrate synthase: [EC 4.1.3.7. Citrate oxaloacetate-lyase (CoA-acetylating)].

Tabrizi S. J., Workman J., Hart P. E., Mangiarini L., Mahal A., Bates G., Cooper J. M. and Schapira A. H. V. (2000). Mitochondrial dysfunction and free radical damage in the Huntington R6/2 transgenic mouse. PubMed DOI

Träger U., Andre R., Magnusson-Lind A., Miller J. R. C., Connolly C., Weiss A., Grueninger S., Silajdžić E., Smith D. L., Leavitt B. R. et al. (2015). Characterisation of immune cell function in fragment and full-length Huntington's disease mouse models. PubMed DOI PMC

Trifunovic A., Hansson A., Wredenberg A., Rovio A. T., Dufour E., Khvorostov I., Spelbrink J. N., Wibom R., Jacobs H. T. and Larsson N.-G. (2005). Somatic mtDNA mutations cause aging phenotypes without affecting reactive oxygen species production. PubMed DOI PMC

Underwood B. R., Broadhurst D., Dunn W. B., Ellis D. I., Michell A. W., Vacher C., Mosedale D. E., Kell D. B., Barker R. A., Grainger D. J. et al. (2006). Huntington disease patients and transgenic mice have similar pro-catabolic serum metabolite profiles. PubMed DOI

Villar-Menéndez I., Blanch M., Tyebji S., Pereira-Veiga T., Albasanz J. L., Martín M., Ferrer I., Pérez-Navarro E. and Barrachina M. (2013). Increased 5-methylcytosine and decreased 5-hydroxymethylcytosine levels are associated with reduced striatal A2AR levels in Huntington's disease. PubMed DOI

Wang W., Esbensen Y., Scheffler K. and Eide L. (2015). Analysis of mitochondrial DNA and RNA integrity by a real-time qPCR-based method. PubMed DOI

Wang G., Liu X., Gaertig M. A., Li S. and Li X.-J. (2016a). Ablation of huntingtin in adult neurons is nondeleterious but its depletion in young mice causes acute pancreatitis. PubMed DOI PMC

Wang W., Scheffler K., Esbensen Y. and Eide L. (2016b). Quantification of DNA damage by real-time qPCR. PubMed DOI

Weiss A., Träger U., Wild E. J., Grueninger S., Farmer R., Landles C., Scahill R. I., Lahiri N., Haider S., Macdonald D. et al. (2012). Mutant huntingtin fragmentation in immune cells tracks Huntington's disease progression. PubMed DOI PMC

Yuzefovych L. V., Schuler A. M., Chen J., Alvarez D. F., Eide L., Ledoux S. P., Wilson G. L. and Rachek L. I. (2013). Alteration of mitochondrial function and insulin sensitivity in primary mouse skeletal muscle cells isolated from transgenic and knockout mice: role of ogg1. PubMed DOI PMC

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