Juvenile amyotrophic lateral sclerosis (jALS) is a rare condition affecting motor neurons, marked by their gradual deterioration. Recent findings have linked mutations in the human SPTLC1 gene to jALS. This gene encodes Serine Palmitoyltransferase, a vital enzyme in sphingolipid production. Studies have revealed that missense and deletion mutations in SPTLC1 are linked to ALS onset in childhood. Notably, these mutations occur within the gene’s transmembrane domain, leading to an overproduction of sphingolipids. However, the precise mechanism by which changes in sphingolipid levels contribute to the disease remains unclear. We analyzed SPTLC1-jALS mutation using both in vitro and in vivo models. We identified alterations in sphingolipid profiles in the SPTLC1-jALS mutant through lipidomics studies. These findings suggest that mutations in SPTLC1 disrupt sphingolipid metabolism, causing cellular abnormalities that might result in neuronal degeneration in jALS. Our goal is to delve deeper into the role and regulation of sphingolipid signaling and biology in ALS.