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This study created a brain endothelial cell atlas to identify targets for receptor-mediated transcytosis (RMT) and designed novel constructs to enhance drug delivery across the blood-brain barrier (BBB). Findings inform target selection for improved trans-BBB therapies.

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Area of Science:

  • Proteomics and transcriptomics
  • Drug delivery
  • Neuroscience

Background:

  • Receptor-mediated transcytosis (RMT) is a key strategy for brain drug delivery.
  • Developing multi-organ atlases aids in identifying RMT targets by comparing brain capillary endothelial cells (BCEC) with peripheral cells.
  • Protein turnover analysis is crucial for optimizing RMT targets, especially for those hijacking cellular recycling pathways.

Purpose of the Study:

  • To create a comprehensive proteomics atlas of BCEC for RMT target identification.
  • To analyze protein turnover and identify targets suitable for triggered endocytosis.
  • To design novel constructs for enhanced cargo transport across the blood-brain barrier (BBB).

Main Methods:

  • Utilized cell type-resolved proteomics and transcriptomics to build the BCEC atlas.
  • Analyzed protein isoforms, turnover, and trafficking using peptide-level proteomics and various transcriptomics data.
  • Employed in vivo proteomics, protein-protein interaction, and organelle fractionation data to identify intracellular targets and design endocytosis-triggering constructs.

Main Results:

  • The proteomics atlas enables RMT target identification, with some discrepancies compared to transcriptomics.
  • Protein turnover data and isoform analysis provide insights into target specificity and utility.
  • Designed constructs for triggered endocytosis show promise, and endosomal targets for cargo interception were predicted.

Conclusions:

  • The developed atlas and identified targets can inform the selection of optimal candidates for RMT.
  • Refined constructs show potential for improving drug delivery across the BBB.
  • This work lays the foundation for more effective targeted therapies for brain disorders.