Huntingtin polyglutamine expansions misdirect axonal transport by perturbing motor and adaptor recruitment

Prowse, E. N. P.; Turkalj, B. A.; Sebastien, M.; Beaudet, D.; McBride, H. M.; Brouhard, G. J.; Pouladi, M. A.; Hendricks, A. G.

doi:10.1101/2024.04.12.589210

bioRxiv · 10.1101/2024.04.12.589210

Huntingtin polyglutamine expansions misdirect axonal transport by perturbing motor and adaptor recruitment

Prowse, E. N. P.·Turkalj, B. A.·Sebastien, M.·Beaudet, D.·McBride, H. M.·Brouhard, G. J.·Pouladi, M. A.·Hendricks, A. G.

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Abstract

Huntingtons disease (HD) is caused by polyglutamine (polyQ) expansions in huntingtin (HTT). Polyglutamine repeat lengths >35Q lead to neurodegeneration and longer repeats correspond to earlier symptom onset. HTT scaffolds kinesin-1 and dynein to a variety of vesicles and organelles directly and through adaptors. To characterize the effects of HTT polyQ expansions on axonal transport, we tracked BDNF vesicles, mitochondria, and lysosomes in neurons induced from an isogenic set of human stem cell lines with repeat lengths of 30, 45, 65, and 81Q. Mild and intermediate pathogenic polyQ expansions caused increased BDNF motility, while HTT-81Q misdirected BDNF towards the distal tip. In comparison, mitochondria and lysosome transport showed mild defects with polyQHTT. We next examined the effect of polyQHTT in combination with neuroinflammatory stress. Under stress, BDNF cargoes in HTT-30Q neurons were more processive. Stress in HTT-81Q resulted in a stark decrease in the number of BDNF cargoes. However, the few remaining BDNF cargoes displayed more frequent long-range motility in both directions. Under neuroinflammatory stress, lysosomes were more abundant in HTT-81Q neurons, and motile lysosomes moved less processively and had an anterograde bias while lysosomes in HTT-30Q where not strongly affected. To examine how HTT-polyQ expansions altered the motors and adaptors on vesicular cargoes, we isolated BDNF cargoes from neurons and quantified the proteins associated with them. BDNF-endosomes isolated from HTT-81Q neurons associated with 2.5 kinesin-1 and 3.9 HAP1 molecules on average, compared to 1.0 kinesin-1 and 1.0 HAP1 molecule for HTT-30Q neurons. Together, these results show that polyQ expansions in HTT cause aberrant motor and adaptor recruitment to cargoes, resulting in dysregulated transport and responses to neuroinflammatory stress.

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Prowse, E. N. P., Turkalj, B. A., Sebastien, M., Beaudet, D., McBride, H. M., Brouhard, G. J., Pouladi, M. A., Hendricks, A. G.. 2024-04-12. Huntingtin polyglutamine expansions misdirect axonal transport by perturbing motor and adaptor recruitment. https://doi.org/10.1101/2024.04.12.589210

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