Overexpressing eukaryotic elongation factor 1 alpha (eEF1A) proteins to promote corticospinal axon repair after injury

Abstract

Although protein synthesis is hypothesized to have a pivotal role in axonal repair after central nervous system (CNS) injury, the role of core components of the protein synthesis machinery has not been examined. Notably, some elongation factors possess non-canonical functions that may further impact axonal repair. Here, we examined whether overexpressing eukaryotic elongation factor 1 alpha (eEF1A) proteins enhances the collateral sprouting of corticospinal tract (CST) neurons after unilateral pyramidotomy, along with the underlying molecular mechanisms. Compared with axonal regeneration from injured neurons, axonal sprouting from uninjured neurons occurs spontaneously after injury and may represent a more accessible form of axonal repair for clinical translation. We found that overexpressing eEF1A1, eEF1A2 or both proteins in CST neurons increased the levels of pS6, an indicator for mTOR activity, in neuronal somas. In contrast, the levels of pSTAT3 and pAKT were not increased. Strikingly, overexpressing eEF1A2 alone, but neither eEF1A1 alone nor both factors simultaneously, increased protein synthesis and actin rearrangement in CST neurons. While eEF1A1 overexpression only slightly enhanced CST sprouting across the midline into the denervated side in the cervical spinal after pyramidotomy, eEF1A2 overexpression substantially enhanced this sprouting. Surprisingly, co-overexpression of both eEF1A1 and eEF1A2 led to a sprouting phenotype similar to wild-type controls, suggesting an antagonistic effect of overexpressing both proteins. These data provide the first evidence that overexpressing a core component of the translation machinery, eEF1A2, enhances CST sprouting, likely by a combination of increased protein synthesis, mTOR signaling and actin cytoskeleton rearrangement.

Probing regenerative heterogeneity of corticospinal neurons with scRNA-Seq

“…Regenerating CST neurons differentially overexpress genes and pathways involved in protein translation including eIF2, eIF3F, eIF4 and p70S6K. This is in line with the published literature on PTEN/mTOR and other pathways demonstrating the importance of protein synthesis in CNS axon regeneration 1,45 . In our study, both regenerating and non-regenerating CST neurons underwent PTEN;SOCS3 gene deletion and hence their transcriptomic differences do not necessarily reflect differences in PTEN/mTOR or SOCS3/STAT3 signaling.…”