Direct reprogramming of somatic skin cells from a patient with Huntington's disease into striatal neurons to create models of pathology

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Abstract

A new in vitro model of Huntington's disease (HD) has been developed, based on the direct reprogramming of dermal fibroblasts from patients with the disease into striatal neurons. Direct reprogramming makes it possible to obtain neurons, preserving the epigenetic information inherent in the cells of a specific donor, and, consequently, the age-associated phenotype of the disease. The new model resembles the main histopathological feature of Huntington's disease – the accumulation of aggregates of the mutant huntingtin protein in striatal neurons obtained from the patient's fibroblasts. Conducting experiments on induced neurons from patients in culture, obtained using direct reprogramming, allows us to individually assess the degree of progression of neuropathology and implement a personalized approach in choosing a treatment strategy and selecting drugs for therapy. The in vitro model of Huntington's disease can be used for preclinical drug studies, for the treatment of neurodegenerative diseases and for assessing their effectiveness on neurons of specific patients.

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About the authors

N. A. Kraskovskaya

Institute of Сytology Russian Academy of Sciences

Author for correspondence.
Email: ninakraskovskaya@gmail.com
Russian Federation, Saint-Petersburg

M. G. Khotin

Institute of Сytology Russian Academy of Sciences

Email: ninakraskovskaya@gmail.com
Russian Federation, Saint-Petersburg

A. N. Tomilin

Institute of Сytology Russian Academy of Sciences

Email: ninakraskovskaya@gmail.com

Corresponding Member of the RAS

Russian Federation, Saint-Petersburg

N. A. Mikhailova

Institute of Сytology Russian Academy of Sciences

Email: ninakraskovskaya@gmail.com
Russian Federation, Saint-Petersburg

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2. Fig. 1. Micrographs of human dermal fibroblasts in vitro. (a) DF1 fibroblasts (obtained from a healthy donor) and (b) HDDF fibroblasts (obtained from a donor with Huntington's disease). Phase contrast microscopy, x10. Scale 100 microns.

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3. Fig. 2. Visualization of the diffuse distribution of huntingtin protein in the cytoplasm of dermal fibroblasts. (a) DF1 cells (healthy donor) and (b) HDDF cells (donor with Huntington's disease). Immunofluorescence staining with mEM48 huntingtin antibodies (secondary antibodies conjugated with Alexa 555) (red). The cell nuclei are visualized with DAPI dye (blue). Confocal microscopy, x60. The scale is 50 microns.

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4. Fig. 3. Visualization of huntingtin protein (green) in striatum neurons obtained by direct reprogramming from dermal fibroblasts: (a and b) DF1 cells (healthy donor) and (c, d, e, e) from cells of the line HDDF (a donor with Huntington's disease). The result of immunofluorescence staining with antibodies to the neuronal marker MAP2 (secondary antibodies conjugated with Alexa 555) and antibodies to huntingtin mEM48 (secondary antibodies Alexa 488) was shown. The co-localization of these two proteins is manifested in yellow staining. The cell nuclei are visualized with DAPI dye (blue). Confocal microscopy, x60. The scale is 30 microns. In striatum neurons obtained from dermal fibroblasts of a healthy donor, huntingtin is diffusely distributed in the cytoplasm (a and b), while in striatum neurons obtained from dermal fibroblasts of a patient with Huntington's disease, aggregates of this protein are observed (shown by arrows) (c, d, e and e).

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