Recently developed therapeutic approaches for the treatment of Huntington's disease (HD) require preclinical testing in large animal models. The minipig is a suitable experimental animal because of its large gyrencephalic brain, body weight of 70-100 kg, long lifespan, and anatomical, physiological and metabolic resemblance to humans. The Libechov transgenic minipig model for HD (TgHD) has proven useful for proof of concept of developing new therapies. However, to evaluate the efficacy of different therapies on disease progression, a broader phenotypic characterization of the TgHD minipig is needed. In this study, we analyzed the brain tissues of TgHD minipigs at the age of 48 and 60-70 months, and compared them to wild-type animals. We were able to demonstrate not only an accumulation of different forms of mutant huntingtin (mHTT) in TgHD brain, but also pathological changes associated with cellular damage caused by mHTT. At 48 months, we detected pathological changes that included the demyelination of brain white matter, loss of function of striatal neurons in the putamen and activation of microglia. At 60-70 months, we found a clear marker of neurodegeneration: significant cell loss detected in the caudate nucleus, putamen and cortex. This was accompanied by clusters of structures accumulating in the neurites of some neurons, a sign of their degeneration that is also seen in Alzheimer's disease, and a significant activation of astrocytes. In summary, our data demonstrate age-dependent neuropathology with later onset of neurodegeneration in TgHD minipigs.

Department of Histology and Embryology Masaryk University in Brno Faculty of Medicine 62500 Brno Czech Republic

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

Zobrazit více v PubMed

Askeland G., Rodinova M., Štufková H., Dosoudilova Z., Baxa M., Smatlikova P., Bohuslavova B., Klempir J., Nguyen T. D., Kuśnierczyk A. et al. (2018). A transgenic minipig model of Huntington's disease shows early signs of behavioral and molecular pathologies. PubMed DOI PMC

Aziz N. A., van der Burg J. M. M., Tabrizi S. J. and Landwehrmeyer G. B. (2018). Overlap between age-at-onset and disease-progression determinants in Huntington disease. PubMed DOI PMC

Bartzokis G., Lu P. H., Tishler T. A., Fong S. M., Oluwadara B., Finn J. P., Huang D., Bordelon Y., Mintz J. and Perlman S. (2007). Myelin breakdown and iron changes in Huntington's disease: pathogenesis and treatment implications. PubMed DOI

Bates G. P., Dorsey R., Gusella J. F., Hayden M. R., Kay C., Leavitt B. R., Nance M., Ross C. A., Scahill R. I., Wetzel R. et al. (2015). Huntington disease. PubMed DOI

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 DOI

Benn C. L., Landles C., Li H., Strand A. D., Woodman B., Sathasivam K., Li S.-H., Ghazi-Noori S., Hockly E., Faruque S. M. N. N. et al. (2005). Contribution of nuclear and extranuclear polyQ to neurological phenotypes in mouse models of Huntington's disease. PubMed DOI

Chen Y. and Swanson R. A. (2003). Astrocytes and brain injury. PubMed DOI

Davies S. W. and Scherzinger E. (1997). Nuclear inclusions in Huntington's disease. 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

Evers M. M., Miniarikova J., Juhas S., Vallès A., Bohuslavova B., Juhasova J., Skalnikova H. K., Vodicka P., Valekova I., Brouwers C. et al. (2018). AAV5-miHTT gene therapy demonstrates broad distribution and strong human mutant huntingtin lowering in a Huntington's disease minipig model. PubMed DOI PMC

Genetic Modifiers of Huntington's Disease (GeM-HD) Consortium (2015). Identification of genetic factors that modify clinical onset of Huntington's disease. PubMed DOI PMC

Graham R. K., Slow E. J., Deng Y., Bissada N., Lu G., Pearson J., Shehadeh J., Leavitt B. R., Raymond L. A. and Hayden M. R. (2006). Levels of mutant huntingtin influence the phenotypic severity of Huntington disease in YAC128 mouse models. PubMed DOI

Gusella J. F., Macdonald M. E. and Lee J. M. (2014). Genetic modifiers of Huntington's disease. PubMed DOI

Gutiérrez M. L., Guevara J. and Barrera L. A. (2012). Semi-automatic grading system in histologic and immunohistochemistry analysis to evaluate in vitro chondrogenesis. DOI

Harjes P. and Wanker E. E. (2003). The hunt for huntingtin function: interaction partners tell many different stories. PubMed DOI

Heng M. Y., Tallaksen-Greene S. J., Detloff P. J. and Albin R. L. (2007). Longitudinal evaluation of the Hdh(CAG)150 knock-in murine model of Huntington's disease. 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

Hinkle J. T., Dawson V. L. and Dawson T. M. (2019). The A1 astrocyte paradigm: new avenues for pharmacological intervention in neurodegeneration. PubMed DOI PMC

Hoffner G., Soues S. and Djian P. (2007). Aggregation of expanded huntingtin in the brains of patients with Huntington disease. PubMed DOI PMC

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

Jacobsen J. C., Bawden C. S., Rudiger S. R., McLaughlan C. J., Reid S. J., Waldvogel H. J., MacDonald M. E., Gusella J. F., Walker S. K., Kelly J. M. et al. (2010). An ovine transgenic Huntington's disease model. PubMed DOI PMC

Jansen A. H. P., van Hal M., Op, den Kelder I. C., Meier R. T., de Ruiter A.-A., Schut M. H., Smith D. L., Grit C., Brouwer N.,  et al. (2017). Frequency of nuclear mutant huntingtin inclusion formation in neurons and glia is cell-type-specific. PubMed DOI PMC

Jortner B. S. (2006). The return of the dark neuron. a histological artifact complicating contemporary neurotoxicologic evaluation. PubMed DOI

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

Lajoie P. and Snapp E. L. (2010). Formation and toxicity of soluble polyglutamine oligomers in living cells. PubMed DOI PMC

Li H., Li S.-H., Yu Z.-X., Shelbourne P. and Li X.-J. (2001). Huntingtin aggregate-associated axonal degeneration is an early pathological event in Huntington's disease mice. PubMed DOI PMC

Liddelow S. A., Guttenplan K. A., Clarke L. E., Bennett F. C., Bohlen C. J., Schirmer L., Bennett M. L., Münch A. E., Chung W.-S., Peterson T. C. et al. (2017). Neurotoxic reactive astrocytes are induced by activated microglia. PubMed DOI PMC

Liedtke W., Edelmann W., Bieri P. L., Chiu F.-C., Cowan N. J., Kucherlapati R. and Raine C. S. (1996). GFAP is necessary for the integrity of CNS white matter architecture and long-term maintenance of myelination. PubMed DOI

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

Menalled L., El-Khodor B. F., Patry M., Suárez-Fariñas M., Orenstein S. J., Zahasky B., Leahy C., Wheeler V., Yang X. W., MacDonald M. et al. (2009). Systematic behavioral evaluation of Huntington's disease transgenic and knock-in mouse models. 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

Nixon R. A., Wegiel J., Kumar A., Yu W. H., Peterhoff C., Cataldo A. and Cuervo A. M. (2005). Extensive involvement of autophagy in Alzheimer disease: an immuno-electron microscopy study. PubMed DOI

Paulsen J. S. (2010). Early detection of Huntington's disease. PubMed DOI PMC

Peferoen L., Kipp M., van der Valk P., van Noort J. M. and Amor S. (2014). Oligodendrocyte-microglia cross-talk in the central nervous system. PubMed DOI PMC

Politis M., Lahiri N., Niccolini F., Su P., Wu K., Giannetti P., Scahill R. I., Turkheimer F. E., Tabrizi S. J. and Piccini P. (2015). Increased central microglial activation associated with peripheral cytokine levels in premanifest Huntington's disease gene carriers. PubMed DOI

Rodinova M., Krizova J., Stufkova H., Bohuslavova B., Askeland G., Dosoudilova Z., Juhas S., Juhasova J., Ellederova Z., Zeman J. et al. (2019). Deterioration of mitochondrial bioenergetics and ultrastructure impairment in skeletal muscle of a transgenic minipig model in the early stages of Huntington's disease. PubMed DOI PMC

Rosas H. D., Salat D. H., Lee S. Y., Zaleta A. K., Hevelone N. and Hersch S. M. (2008). Complexity and heterogeneity: what drives the ever-changing brain in Huntington's disease? PubMed DOI PMC

Sathasivam K., Lane A., Legleiter J., Warley A., Woodman B., Finkbeiner S., Paganetti P., Muchowski P. J., Wilson S. and Bates G. P. (2010). Identical oligomeric and fibrillar structures captured from the brains of R6/2 and knock-in mouse models of Huntington's disease. PubMed DOI PMC

Saudou F., Finkbeiner S., Devys D. and Greenberg M. E. (1998). Huntingtin acts in the nucleus to induce apoptosis but death does not correlate with the formation of intranuclear inclusions. PubMed DOI

Swami M., Hendricks A. E., Gillis T., Massood T., Mysore J., Myers R. H. and Wheeler V. C. (2009). Somatic expansion of the Huntington's disease CAG repeat in the brain is associated with an earlier age of disease onset. PubMed DOI PMC

Teo R. T. Y., Hong X., Yu-Taeger L., Huang Y., Tan L. J., Xie Y., To X. V., Guo L., Rajendran R., Novati A. et al. (2016). Structural and molecular myelination deficits occur prior to neuronal loss in the YAC128 and BACHD models of Huntington disease. PubMed DOI PMC

Truant R., Atwal R. S., Desmond C., Munsie L. and Tran T. (2008). Huntington's disease: revisiting the aggregation hypothesis in polyglutamine neurodegenerative diseases. PubMed DOI

Turmaine M., Raza A., Mahal A., Mangiarini L., Bates G. P. and Davies S. W. (2000). Nonapoptotic neurodegeneration in a transgenic mouse model of Huntington's disease. PubMed DOI PMC

Uchida M., Shimatsu Y., Onoe K., Matsuyama N., Niki R., Ikeda J.-E. and Imai H. (2001). Production of transgenic miniature pigs by pronuclear microinjection. PubMed DOI

Usdin M. T., Shelbourne P. F., Myers R. M. and Madison D. V. (1999). Impaired synaptic plasticity in mice carrying the Huntington's disease mutation. PubMed DOI

Valekova I., Jarkovska K., Kotrcova E., Bucci J., Ellederova Z., Juhas S., Motlik J., Gadher S. J. and Kovarova H. (2016). Revelation of the IFNalpha, IL-10, IL-8 and IL-1beta as promising biomarkers reflecting immuno-pathological mechanisms in porcine Huntington's disease model. PubMed DOI

Vidinská D., Vochozková P., Šmatlíková P., Ardan T., Klíma J., Juhás ., Juhásová J., Bohuslavová B., Baxa M., Valeková I. et al. (2018). Gradual phenotype development in Huntington disease transgenic minipig model at 24 months of age. PubMed DOI

Vodička P., Smetana K. Jr., Dvořánková B., Emerick T., Xu Y. Z., Ourednik J., Ourednik V. and Motlík J. (2005). The miniature pig as an animal model in biomedical research. PubMed DOI

Walz W. (2000). Role of astrocytes in the clearance of excess extracellular potassium. PubMed DOI

Woodman B., Butler R., Landles C., Lupton M. K., Tse J., Hockly E., Moffitt H., Sathasivam K. and Bates G. P. (2007). The Hdh(Q150/Q150) knock-in mouse model of HD and the R6/2 exon 1 model develop comparable and widespread molecular phenotypes. PubMed DOI

Yan S., Tu Z., Liu Z., Fan N., Yang H., Yang S., Yang W., Zhao Y., Ouyang Z., Lai C. et al. (2018). A huntingtin knockin pig model recapitulates features of selective neurodegeneration in Huntington's Disease. PubMed DOI PMC

Yang D., Wang C.-E., Zhao B., Li W., Ouyang Z., Liu Z., Yang H., Fan P., O'Neill A., Gu W. et al. (2010). Expression of Huntington's disease protein results in apoptotic neurons in the brains of cloned transgenic pigs. PubMed DOI PMC

Zuccato C., Valenza M. and Cattaneo E. (2010). Molecular mechanisms and potential therapeutical targets in Huntington's disease. PubMed DOI

Huntingtin Co-Isolates with Small Extracellular Vesicles from Blood Plasma of TgHD and KI-HD Pig Models of Huntington's Disease and Human Blood Plasma

Longitudinal study revealing motor, cognitive and behavioral decline in a transgenic minipig model of Huntington's disease