Javier Riancho1,2,3, David Castanedo-Vázquez4, Francisco Gil-Bea5,6, Olga Tapia5,7, Jana Arozamena7, Carlos Durán-Vían8, María José Sedano5,9, Maria Teresa Berciano5,7, Adolfo Lopez de Munain5,6,10,11, Miguel Lafarga5,7. 1. Service of Neurology, Hospital Sierrallana-IDIVAL, Barrio Ganzo s/n, 39300, Torrelavega, Spain. javier.riancho86@gmail.com. 2. Department of Medicine and Psychiatry, University of Cantabria, Santander, Spain. javier.riancho86@gmail.com. 3. Centro de Investigación en Red de Enfermedades Neurodegenerativas, CIBERNED. Instituto Carlos III, Madrid, Spain. javier.riancho86@gmail.com. 4. Service of Radiology, Hospital, Universitario Marqués de Valdecilla, Santander, Spain. 5. Centro de Investigación en Red de Enfermedades Neurodegenerativas, CIBERNED. Instituto Carlos III, Madrid, Spain. 6. Neurosciences Area. Biodonostia Research Institute, San Sebastián, Spain. 7. Department of Anatomy and Cell Biology, University of Cantabria-IDIVAL, Santander, Spain. 8. Service of Dermatology, Hospital Universitario Marqués de Valdecilla, Santander, Spain. 9. Service of Neurology, Hospital Universitario Marqués de Valdecilla-IDIVAL, Santander, Spain. 10. Service of Neurology, Hospital Universitario Donostia, San Sebastián, Spain. 11. Department of Neurosciences. School of Medicine and Nursery, University of the Basque Country, San Sebastián, Spain.
Abstract
BACKGROUND: Dermic fibroblasts have been proposed as a potential genetic-ALS cellular model. This study aimed to explore whether dermic fibroblasts from patients with sporadic-ALS (sALS) recapitulate alterations typical of ALS motor neurons and exhibit abnormal DNA-damage response. METHODS: Dermic fibroblasts were obtained from eight sALS patients and four control subjects. Cellular characterization included proliferation rate analysis, cytoarchitecture studies and confocal immunofluorescence assessment for TDP-43. Additionally, basal and irradiation-induced DNA damage was evaluated by confocal immunofluorescence and biochemical techniques. RESULTS: sALS-fibroblasts showed decreased proliferation rates compared to controls. Additionally, whereas control fibroblasts exhibited the expected normal spindle-shaped morphology, ALS fibroblasts were thinner, with reduced cell size and enlarged nucleoli, with frequent cytoplasmic TDP-43aggregates. Also, baseline signs of DNA damage were evidenced more frequently in ALS-derived fibroblasts (11 versus 4% in control-fibroblasts). Assays for evaluating the irradiation-induced DNA damage demonstrated that DNA repair was defective in ALS-fibroblasts, accumulating more than double of γH2AX-positive DNA damage foci than controls. Very intriguingly, the proportion of fibroblasts particularly vulnerable to irradiation (with more than 15 DNA damage foci per nucleus) was seven times higher in ALS-derived fibroblasts than in controls. CONCLUSIONS: Dermic-derived ALS fibroblasts recapitulate relevant cellular features of sALS and show a higher susceptibility to DNA damage and defective DNA repair responses. Altogether, these results support that dermic fibroblasts may represent a convenient and accessible ALS cellular model to study pathogenetic mechanisms, particularly those related to DNA damage response, as well as the eventual response to disease-modifying therapies.
BACKGROUND: Dermic fibroblasts have been proposed as a potential genetic-ALS cellular model. This study aimed to explore whether dermic fibroblasts from patients with sporadic-ALS (sALS) recapitulate alterations typical of ALS motor neurons and exhibit abnormal DNA-damage response. METHODS: Dermic fibroblasts were obtained from eight sALS patients and four control subjects. Cellular characterization included proliferation rate analysis, cytoarchitecture studies and confocal immunofluorescence assessment for TDP-43. Additionally, basal and irradiation-induced DNA damage was evaluated by confocal immunofluorescence and biochemical techniques. RESULTS: sALS-fibroblasts showed decreased proliferation rates compared to controls. Additionally, whereas control fibroblasts exhibited the expected normal spindle-shaped morphology, ALS fibroblasts were thinner, with reduced cell size and enlarged nucleoli, with frequent cytoplasmic TDP-43aggregates. Also, baseline signs of DNA damage were evidenced more frequently in ALS-derived fibroblasts (11 versus 4% in control-fibroblasts). Assays for evaluating the irradiation-induced DNA damage demonstrated that DNA repair was defective in ALS-fibroblasts, accumulating more than double of γH2AX-positive DNA damage foci than controls. Very intriguingly, the proportion of fibroblasts particularly vulnerable to irradiation (with more than 15 DNA damage foci per nucleus) was seven times higher in ALS-derived fibroblasts than in controls. CONCLUSIONS: Dermic-derived ALS fibroblasts recapitulate relevant cellular features of sALS and show a higher susceptibility to DNA damage and defective DNA repair responses. Altogether, these results support that dermic fibroblasts may represent a convenient and accessible ALS cellular model to study pathogenetic mechanisms, particularly those related to DNA damage response, as well as the eventual response to disease-modifying therapies.
Authors: Hazel Quek; Carla Cuní-López; Romal Stewart; Tiziana Colletti; Antonietta Notaro; Tam Hong Nguyen; Yifan Sun; Christine C Guo; Michelle K Lupton; Tara L Roberts; Yi Chieh Lim; Lotta E Oikari; Vincenzo La Bella; Anthony R White Journal: J Neuroinflammation Date: 2022-02-28 Impact factor: 9.587