A Huntingtin Knockin Pig Model Recapitulates Features of Selective Neurodegeneration in Huntington’s Disease

Abstract

Huntington's disease (HD) is characterized by preferential loss of the medium spiny neurons in the striatum. Using CRISPR/Cas9 and somatic nuclear transfer technology, we established a knockin (KI) pig model of HD that endogenously expresses full-length mutant huntingtin (HTT). By breeding this HD pig model, we have successfully obtained F1 and F2 generation KI pigs. Characterization of founder and F1 KI pigs shows consistent movement, behavioral abnormalities, and early death, which are germline transmittable. More importantly, brains of HD KI pig display striking and selective degeneration of striatal medium spiny neurons. Thus, using a large animal model of HD, we demonstrate for the first time that overt and selective neurodegeneration seen in HD patients can be recapitulated by endogenously expressed mutant proteins in large mammals, a finding that also underscores the importance of using large mammals to investigate the pathogenesis of neurodegenerative diseases and their therapeutics.

One decade ago, one decade ahead in huntington's disease

“…Improvements in viral vectors and delivery methods have already increased the brain regions that can be transfected by axonal spread from injected regions: future vectors will likely have intrinsically better ability to spread further from the injection site or may be succeeded by novel delivery methods like lipid nanoparticles, conjugation, or exosomes . A recently developed knock‐in pig model, with a high degree of genetic fidelity to the human patient thanks to the use of CRISPR in its creation, requires detailed characterization but may come to have added value for the assessment of delivery, efficacy, and safety of such therapeutics …”

Generation of transgenic‐cloned Huanjiang Xiang pigs systemically expressing enhanced green fluorescent protein

“…In addition, pigs, especially minipigs, also share a number of similarities with human, such as body size, anatomy, physiology, pathology as well as genome, providing an excellent animal model for mimicking human diseases (Nagashima et al, ; Niemann & Lucas‐Hahn, ; Prather et al, ; Whyte & Prather, ). Genetically modified pigs have also been widely used for modelling numerous human diseases, such as Alzheimer’s disease (Kragh et al, ; Lee et al, ), Huntington’s disease (Yan et al, ; Yang et al, ) and Parkinson’s disease (Wang et al, ; Zhou et al, ). Therefore, genetically modified pigs hold great promise in the fields of agriculture and biomedicine.…”

The cell re‐association‐based whole‐tooth regeneration strategies in large animal, Sus scrofa

“…Pigs serve as a promising large animal model for studying human diseases and contribute to overcome the shortage of human donor organs. [14][15][16][17] The miniature pig has proven to be a valuable animal model for diphyodont development and regeneration owing to its many similarities to human including the morphology, number and size of teeth, particularly its heterodont dentition (incisors, canines, premolars and molars) and diphyodont dentition, which are not available in rodents. 18,19 The morphology and chronology of diphyodont dentition in miniature pigs have been well characterized by our previous studies and other reports.…”