Probing regenerative heterogeneity of corticospinal neurons with scRNA-Seq - PubMed (original) (raw)

[Preprint]. 2023 Feb 21:rs.3.rs-2588274.

doi: 10.21203/rs.3.rs-2588274/v1.

Junmi Saikia 1, Katlyn Monte 1, Eunmi Ha 1, Daniel Romaus-Sanjurjo 1, Joshua Sanchez 1, Andrea Moore 1, Marc Hernaiz-Llorens 1, Carmine Chavez-Martinez 1, Chimuanya Agba 1, Haoyue Li 1, Daniel Lusk 1, Kayla Cervantes 1, Binhai Zheng 1

Affiliations

Probing regenerative heterogeneity of corticospinal neurons with scRNA-Seq

Hugo Kim et al. Res Sq. 2023.

Abstract

The corticospinal tract (CST) is clinically important for the recovery of motor functions after spinal cord injury. Despite substantial progress in understanding the biology of axon regeneration in the central nervous system (CNS), our ability to promote CST regeneration remains limited. Even with molecular interventions, only a small proportion of CST axons regenerate1. Here we investigate this heterogeneity in the regenerative ability of corticospinal neurons following PTEN and SOCS3 deletion with patch-based single cell RNA sequencing (scRNA-Seq)2,3, which enables deep sequencing of rare regenerating neurons. Bioinformatic analyses highlighted the importance of antioxidant response and mitochondrial biogenesis along with protein translation. Conditional gene deletion validated a role for NFE2L2 (or NRF2), a master regulator of antioxidant response, in CST regeneration. Applying Garnett4, a supervised classification method, to our dataset gave rise to a Regenerating Classifier (RC), which, when applied to published scRNA-Seq data, generates cell type- and developmental stage-appropriate classifications. While embryonic brain, adult dorsal root ganglion and serotonergic neurons are classified as Regenerators, most neurons from adult brain and spinal cord are classified as Non-regenerators. Adult CNS neurons partially revert to a regenerative state soon after injury, which is accelerated by molecular interventions. Our data indicate the existence of universal transcriptomic signatures underlying the regenerative abilities of vastly different neuronal populations, and further illustrate that deep sequencing of only hundreds of phenotypically identified CST neurons has the power to reveal new insights into their regenerative biology.

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Conflict of interest statement

None of the authors have significant competing financial, professional, or personal interests that might have influenced the performance or presentation of the work described in this manuscript.

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References

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References for Methods

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