C11 Transgenic Minipigs For N-terminal Part Of Human Mutated Huntingtin: Following Disease Development

Abstract

Background In order to facilitate studies of pathogenesis and therapy of Huntington’s disease (HD), we have generated transgenic minipigs using microinjection of a lentiviral vector encoding N-terminal (548 aa) part of human huntingtin (Htt) containing 126 CAG/CAA repeats under the control of the human Htt promoter. The progression of HD in human patients is predominantly linked with formation of aggregates, oligomers, and fragments of Htt. Among other factors corresponding to HD is lowered BDNF (brain-derived neurotrophic factor) expression in brain, accumulation of ACBD3 (acyl-CoA binding domain containing 3), and formation of toxic Rhes/mtHtt/ACBD3 complex. Aim To follow disease development in transgenic (TgHD) minipigs using specific biomarkers of HD. Methods At present, TgHD and WT (wild-type) minipigs of F0, F1, F2 and F3 generations are being used for invasive (24 and 36 months old animals) and non-invasive approaches (up to five years old animals). We perform biochemical assays on tissues (with main focus on brain) and cells (fibroblasts, and buccal swabs) in order to follow HD development in our model. Results To date, we have not been successful in the aggregate detection, which could be due to the young age of TgHD animals. However, we have detected Htt oligomers, several Htt fragments of lower molecular weight in samples from TgHD minipigs and not from WT siblings. In addition, we monitor altered levels of proteins previously reported as markers of HD development, namely BDNF and Rhes/mtHtt/ACBD3 complex Conclusion We provide complex information about HD phenotype development in transgenic minipig model between 24–60 months of age. Support CHDI Foundation (A-5378), TACR (TA01011466), European Regional Development FundCZ.1.05/2.1.00/03.0124