Establishment of cellular and rodent models for amyotrophic lateral sclerosis using recombinant viral vectors

2015 Fiscal Year Final Research Report

• Project/Area Number: 24500428
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Nerve anatomy/Neuropathology
• Research Institution: Tokyo Metropolitan Institute of Medical Science
• Principal Investigator: WATABE Kazuhiko 公益財団法人東京都医学総合研究所, 運動・感覚システム研究分野, 副参事研究員 (30240477)
• Project Period (FY): 2012-04-01 – 2016-03-31
• Keywords: 運動ニューロン / TDP-43 / FUS / 筋萎縮性側索硬化症 / 凝集体 / 組換えウイルス / プロテアソーム / オートファジー

Outline of Final Research Achievements

Formation of TDP-43- or FUS-positive cytoplasmic aggregates in neuronal and glial cells is one of the pathological hallmarks of amyotrophic lateral sclerosis (ALS). We have demonstrated that inhibition of protein degradation pathways enhanced adenovirus-induced neuronal cytoplasmic aggregate formation of TDP-43 and FUS in vitro and in vivo. We then produced recombinant adeno-associated virus type 9 (AAV9) and lentivirus vectors encoding wild type and mutant TDP-43 or FUS, and those encoding shRNAs for protein degradation machineries and demonstrated their long-term retrograde transduction of the foreign genes and formation of cytoplasmic aggregates in adult mouse facial and spinal motoneurons. We also performed time-lapse imaging analysis of neuronal and glial cells infected with adenoviruses encoding TDP-43 and FUS cDNAs under conditions of proteasome inhibition to examine cytoplasmic aggregate formation, cell death, and cell to cell spreading of these aggregates.

Free Research Field

神経科学・神経解剖学・神経病理学