The development of a combined cell therapy for huntington's disease

• Autores: Cristina Salado Manzano
• Directores de la Tesis: Josep Maria Canals Coll (dir. tes.), Phil Sanders (codir. tes.)
• Lectura: En la Universitat de Barcelona ( España ) en 2021
• Idioma: español
• Tribunal Calificador de la Tesis: Jordi Alberch Vié (presid.), Joaquim Vives Armengol (secret.), Francisco Javier Rodríguez Muñoz (voc.)
• Programa de doctorado: Programa de Doctorado en Medicina e Investigación Traslacional por la Universidad de Barcelona
• Materias:
  • Ciencias de la vida
    • Biología celular
    • Genética
      • Ingeniería genética
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• Resumen

  • Huntington’s disease (HD) is a neurodegenerative disease caused by a mutation in the huntingtin gene (HTT). The imbalances triggered by mutant HTT, including reduced expression and transport of the neurotrophic factor BDNF, increase striatal medium spiny neuron (MSN) vulnerability which leads to MSN dysfunction and death. Massive MSN death causes striatal atrophy which is the main neuropathological sign of HD and starts before the onset of symptoms. Current therapies are based on the administration of drugs to treat symptoms, although no disease-modifying treatment is available to cure HD. Through this thesis, a combined cell-based therapy for HD has been developed, which carries the potential to slow down the neurodegeneration that occurs in HD and to balance the loss of BDNF in the striatum. We have designed and produced the necessary tools for the manufacturing of lentiviral vectors (LVs) carrying BDNF as a transgene. After this step, the procedure was established at a research scale and adapted for large-scale production in a good manufacture practices (GMP)-like environment. To adapt to a GMP-compatible clinical scale, a system to concentrate LVs through tangential flow filtration (TFF) and diafiltration (DF) was implemented. The LVs were used to transduce human bone marrow derived mesenchymal stem cells (hBM-MSC). After obtaining hBM-MSCsBDNF, the released BDNF was quantified by ELISA. The cells were co-transplanted with striatal neuron precursors (NPCs) into the striatum of adult knock-in HD and wild-type mice. The effects of the combined cell therapy were studied one week after transplantation. Our results demonstrated the feasibility of the production of LV_BDNF particles in a GMP-like environment, and the ability of TFF system to successfully concentrate those particles. Immunohistochemistry results showed an activation of the immune reaction upon co-transplantation and an increase in the amount of vessel-like structures, indicating the enhancement in the angiogenesis in the co-transplanted hemisphere.