{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,19]],"date-time":"2026-08-19T18:08:57Z","timestamp":1787162937268,"version":"build-2736575974"},"reference-count":25,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,2,27]],"date-time":"2026-02-27T00:00:00Z","timestamp":1772150400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Genet."],"abstract":"<jats:p>\n                    Cytoskeletal alterations and axonal transport deficits are key factors in many neurodegenerative disorders. The neuronal kinesin family member 5A (KIF5A) is a microtubule-based motor protein critical for anterograde transport of RNA granules, organelles, and neurofilaments along axons and dendrites. Heterozygous missense and nonsense mutations in the N-terminal motor and stalk domains are associated with hereditary spastic paraplegia 10 (SPG10) and Charcot-Marie-Tooth disease type 2 (CMT2), while frameshift mutations in\n                    <jats:italic>KIF5A<\/jats:italic>\n                    \u00a0C-terminal cargo-binding domain are linked to amyotrophic lateral sclerosis (ALS). We recently reprogrammed an iPSC line from a SPG10 patient carrying the novel missense variant c.50G&amp;gt;A (p.R17Q) in the\n                    <jats:italic>KIF5A<\/jats:italic>\n                    motor domain, classified as variant of unknown significance (VUS) and predicted to affect ATP binding. Here we gene-edited this mutant iPSC line by CRISPR-Cas9 to obtain an isogenic wild-type (WT)\n                    <jats:italic>KIF5A<\/jats:italic>\n                    \u00a0cell line. We next examined functionally the impact of the p.R17Q VUS on KIF5A protein sub-cellular distribution and on axonal transport of mitochondria and lysosomes in differentiated iPSC-motoneurons (MNs). The presence of neurofilament-positive axonal swellings and an increased distribution of KIF5A protein in distal neurites was observed in the mutant p.R17Q compared to the WT\n                    <jats:italic>KIF5A<\/jats:italic>\n                    iPSC-MNs, indicating a likely defective axonal transport. The anterograde velocity and distance travelled by mitochondria and lysosomes along neurites was indeed significantly reduced in the mutant\n                    <jats:italic>KIF5A<\/jats:italic>\n                    iPSC-MNs compared to the WT ones. These findings demonstrate that the p.R17Q VUS is pathogenic, thereby extending the spectrum of\n                    <jats:italic>KIF5A<\/jats:italic>\n                    mutations causing SPG10 and support the use of patient-derived iPSC-MNs to functionally validate\n                    <jats:italic>KIF5A<\/jats:italic>\n                    -associated VUS.\n                  <\/jats:p>","DOI":"10.3389\/fgene.2026.1774170","type":"journal-article","created":{"date-parts":[[2026,2,26]],"date-time":"2026-02-26T22:10:08Z","timestamp":1772143808000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["Functional validation of the novel KIF5A p.R17Q VUS reveals defective axonal transport in iPSC-motoneurons from a SPG10 patient"],"prefix":"10.3389","volume":"17","author":[{"given":"Serena","family":"Santangelo","sequence":"first","affiliation":[{"name":"Department of Medical Biotechnology and Translational Medicine, Universit\u00e0 degli Studi di Milano","place":["Milan, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Valeria","family":"Casiraghi","sequence":"additional","affiliation":[{"name":"Department of Medical Biotechnology and Translational Medicine, Universit\u00e0 degli Studi di Milano","place":["Milan, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claudia","family":"Fallini","sequence":"additional","affiliation":[{"name":"Department of Cell and Molecular Biology, Ryan Institute for Neuroscience, University of Rhode Island","place":["Kingston, RI, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sabrina","family":"Invernizzi","sequence":"additional","affiliation":[{"name":"Department of Medical Biotechnology and Translational Medicine, Universit\u00e0 degli Studi di Milano","place":["Milan, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Silvia","family":"Peverelli","sequence":"additional","affiliation":[{"name":"Department of Neuroscience - 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