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Spinal cord injury regulates circular RNA expression in axons.
Siddiq, Mustafa M; Toro, Carlos A; Johnson, Nicholas P; Hansen, Jens; Xiong, Yuguang; Mellado, Wilfredo; Tolentino, Rosa E; Johnson, Kaitlin; Jayaraman, Gomathi; Suhail, Zaara; Harlow, Lauren; Dai, Jinye; Beaumont, Kristin G; Sebra, Robert; Willis, Dianna E; Cardozo, Christopher P; Iyengar, Ravi.
Afiliação
  • Siddiq MM; Pharmacological Sciences and Institute for Systems Biomedicine, Icahn School of Medicine at Mount Sinai, New York, NY, United States.
  • Toro CA; Spinal Cord Damage Research Center, James J. Peters VA Medical Center, Bronx, NY, United States.
  • Johnson NP; Department of Medicine, Icahn School of Medicine at Mount Sinai, New York, NY, United States.
  • Hansen J; Pharmacological Sciences and Institute for Systems Biomedicine, Icahn School of Medicine at Mount Sinai, New York, NY, United States.
  • Xiong Y; Pharmacological Sciences and Institute for Systems Biomedicine, Icahn School of Medicine at Mount Sinai, New York, NY, United States.
  • Mellado W; Pharmacological Sciences and Institute for Systems Biomedicine, Icahn School of Medicine at Mount Sinai, New York, NY, United States.
  • Tolentino RE; Burke Neurological Institute, White Plains, NY, United States.
  • Johnson K; Pharmacological Sciences and Institute for Systems Biomedicine, Icahn School of Medicine at Mount Sinai, New York, NY, United States.
  • Jayaraman G; Spinal Cord Damage Research Center, James J. Peters VA Medical Center, Bronx, NY, United States.
  • Suhail Z; Pharmacological Sciences and Institute for Systems Biomedicine, Icahn School of Medicine at Mount Sinai, New York, NY, United States.
  • Harlow L; Pharmacological Sciences and Institute for Systems Biomedicine, Icahn School of Medicine at Mount Sinai, New York, NY, United States.
  • Dai J; Spinal Cord Damage Research Center, James J. Peters VA Medical Center, Bronx, NY, United States.
  • Beaumont KG; Department of Medicine, Icahn School of Medicine at Mount Sinai, New York, NY, United States.
  • Sebra R; Pharmacological Sciences and Institute for Systems Biomedicine, Icahn School of Medicine at Mount Sinai, New York, NY, United States.
  • Willis DE; Nash Family Department of Neuroscience, Icahn School of Medicine at Mount Sinai, New York, NY, United States.
  • Cardozo CP; Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY, United States.
  • Iyengar R; Department of Genetics and Genomic Studies, Icahn School of Medicine at Mount Sinai, New York, NY, United States.
Front Mol Neurosci ; 16: 1183315, 2023.
Article em En | MEDLINE | ID: mdl-37692100
ABSTRACT

Introduction:

Neurons transport mRNA and translational machinery to axons for local translation. After spinal cord injury (SCI), de novo translation is assumed to enable neurorepair. Knowledge of the identity of axonal mRNAs that participate in neurorepair after SCI is limited. We sought to identify and understand how axonal RNAs play a role in axonal regeneration.

Methods:

We obtained preparations enriched in axonal mRNAs from control and SCI rats by digesting spinal cord tissue with cold-active protease (CAP). The digested samples were then centrifuged to obtain a supernatant that was used to identify mRNA expression. We identified differentially expressed genes (DEGS) after SCI and mapped them to various biological processes. We validated the DEGs by RT-qPCR and RNA-scope.

Results:

The supernatant fraction was highly enriched for mRNA from axons. Using Gene Ontology, the second most significant pathway for all DEGs was axonogenesis. Among the DEGs was Rims2, which is predominately a circular RNA (circRNA) in the CNS. We show that Rims2 RNA within spinal cord axons is circular. We found an additional 200 putative circRNAs in the axonal-enriched fraction. Knockdown in primary rat cortical neurons of the RNA editing enzyme ADAR1, which inhibits formation of circRNAs, significantly increased axonal outgrowth and increased the expression of circRims2. Using Rims2 as a prototype we used Circular RNA Interactome to predict miRNAs that bind to circRims2 also bind to the 3'UTR of GAP-43, PTEN or CREB1, all known regulators of axonal outgrowth. Axonally-translated GAP-43 supports axonal elongation and we detect GAP-43 mRNA in the rat axons by RNAscope.

Discussion:

By enriching for axonal RNA, we detect SCI induced DEGs, including circRNA such as Rims2. Ablation of ADAR1, the enzyme that regulates circRNA formation, promotes axonal outgrowth of cortical neurons. We developed a pathway model using Circular RNA Interactome that indicates that Rims2 through miRNAs can regulate the axonal translation GAP-43 to regulate axonal regeneration. We conclude that axonal regulatory pathways will play a role in neurorepair.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Tipo de estudo: Prognostic_studies Idioma: En Ano de publicação: 2023 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Tipo de estudo: Prognostic_studies Idioma: En Ano de publicação: 2023 Tipo de documento: Article