• Author
    Stuart Harper
  • Discovery PI

    Kunal Patel, MD

  • Project Co-Author

  • Abstract Title

    SLC4A10 as a Novel Molecular Target to Modulate Cerebrospinal Fluid Production in Human Choroid Plexus

  • Discovery AOC Petal or Dual Degree Program

    Basic, Clinical, & Translational Research

  • Abstract

    Background: Modulation of cerebrospinal fluid (CSF) production has significant implications in the management of many neurosurgical disorders, particularly hydrocephalus. However, the molecular targets within the human choroid plexus (CP) that regulate CSF production remain poorly defined. This study utilized single-cell sequencing to identify novel molecular targets involved in CSF secretion, followed by development of a functional, fluid-secreting model of human CP to evaluate the impact of target modulation on CSF production.

    Methods: CP was acquired intraoperatively in patients undergoing intraventricular surgery. Specimens underwent single-cell RNA sequencing (n=9) and immunohistochemistry (n=15). CP epithelial cells (CPEC) were cultured in a two-chamber Transwell system. Lentiviral knockdown of SLC4A10 was performed, followed by Western blot and immunofluorescence to validate and characterize the knockdown. Fluorescent dextran was used to measure fluid production in wildtype and SLC4A10 knockdown conditions (n=10).

    Results: CP sampling was safe and feasible in all patients. Specimens had classic morphology on H&E with positive transthyretin (TTR) staining. Single-cell analysis of CPEC demonstrated strong expression of sodium transporters and aquaporins including ATP1A1, NKCC1, AQP1 and SLC4A10. Immunohistochemistry showed strong expression of Na+/K+ ATPase, NKCC1 and AQP1 on the apical surface. The Na+/HCO3- co-transporter SLC4A10 was the primary basolateral sodium transporter and is hypothesized to be responsible for replenishing the intracellular sodium gradient which drives CSF production. CPEC were cultured on the Transwell model to confluence with positive TTR and SLC4A10 immunofluorescence. As no selective SLC4A10 small molecule inhibitor currently exists, lentiviral knockdown of SLC4A10 reduced fluid secretion by 40% compared to wildtype CPEC cells in the transwell model (p<0.0001).

    Conclusions: To the best of our knowledge, we have compiled one of the largest single-cell databases of human CP and developed a functional, in vitro model of human CP capable of secreting fluid. We identify the Na+/HCO3- co-transporter SLC4A10 as a key basolateral sodium transporter responsible for replenishing the intracellular Na+ gradient that drives CSF secretion and find that its knockdown significantly reduces CSF secretion. Development of a novel SLC4A10 inhibitor may reduce CSF secretion, potentially offering the first pharmacologic therapy for hydrocephalus and making invasive ventricular shunting procedures obsolete.

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