Characterizing CSF inflammatory proteomics in pediatric post-hemorrhagic hydrocephalus and Anti-NMDAR encephalitis

Authors

Taylor Broudy, Center for Neuroscience Research, Children's National Hospital, Washington, DC, 20010, USA.
Ankush Bansal, Center for Neuroscience Research, Children's National Hospital, Washington, DC, 20010, USA.
Akilah Pascall, Department of Critical Care, Children's National Hospital, Washington, DC, 20010, USA.
William Suslovic, Department of Laboratory Medicine, Children's National Hospital, Washington, DC, 20010, USA.
Nhu To Chau, Department of Laboratory Medicine, Children's National Hospital, Washington, DC, 20010, USA.
Leigh Sepeta, Center for Neuroscience Research, Children's National Hospital, Washington, DC, 20010, USA.
Courtney Lowe, Center for Neuroscience Research, Children's National Hospital, Washington, DC, 20010, USA.
Shani Israel, Center for Neuroscience Research, Children's National Hospital, Washington, DC, 20010, USA.
Alexandra B. Kornbluh, Department of Neurology, Children's National Hospital, Washington, DC, 20010, USA.
Claire Marie Har, Department of Neurology, Children's National Hospital, Washington, DC, 20010, USA.
Hayley Roper, Department of Neurology, Children's National Hospital, Washington, DC, 20010, USA.
Ilana Kahn, Department of Neurology, Children's National Hospital, Washington, DC, 20010, USA.
Hasan Syed, Department of Neurosurgery, Children's National Hospital, Washington, DC, 20010, USA.
Chima Oluigbo, Department of Neurosurgery, Children's National Hospital, Washington, DC, 20010, USA.
John Myseros, Department of Neurosurgery, Children's National Hospital, Washington, DC, 20010, USA.
Robert Keating, Department of Neurosurgery, Children's National Hospital, Washington, DC, 20010, USA.
Elizabeth Wells, Department of Neurology, Children's National Hospital, Washington, DC, 20010, USA.
Meghan Delaney, Department of Laboratory Medicine, Children's National Hospital, Washington, DC, 20010, USA.
Daniel Donoho, Department of Neurosurgery, Children's National Hospital, Washington, DC, 20010, USA.
Kazue Hashimoto-Torii, Center for Neuroscience Research, Children's National Hospital, Washington, DC, 20010, USA.
Terry Dean, Center for Neuroscience Research, Children's National Hospital, Washington, DC, 20010, USA. tdean@childrensnational.org.

Document Type

Journal Article

Publication Date

12-10-2025

Journal

Journal of neuroinflammation

DOI

10.1186/s12974-025-03642-w

Keywords

Brain tumor; CSF; NMDA receptor encephalitis; Post-hemorrhagic hydrocephalus; Proteomics

Abstract

BACKGROUND: Neuroinflammation is a key contributor to pathology in many central nervous system (CNS) diseases. While cerebrospinal fluid (CSF) proteomic studies in adult neurologic conditions have identified insightful inflammatory signatures, fewer studies have been conducted in pediatric diseases. Moreover, past studies primarily employed proteomic approaches that are less suited to detect novel, low-abundance inflammatory mediators that may be critical in pediatric CNS pathophysiology. To address this gap, we applied high-sensitivity, multi-targeted proteomic profiling to characterize the neuroinflammatory signatures across three distinct pediatric neurologic diseases: post-hemorrhagic hydrocephalus (PHH), N-methyl-D-aspartate receptor encephalitis (NMDARE), and brain tumor-associated hydrocephalus. METHODS: CSF samples from controls (n = 5) and patients with PHH (n = 9), NMDARE (n = 5), and brain tumor-associated hydrocephalus (n = 10) were obtained from a pediatric CSF biorepository. After proteomic profiling using the Olink Explore platform, 641 inflammation-related proteins were retained for analysis. Differentially abundant proteins (DAPs) were identified using limma with false discovery rate (FDR) correction (FDR < 0.05, |log₂FC| >1). Pathway enrichment of DAPs was performed with Reactome via Enrichr, and protein-protein interaction networks were constructed using STRING to identify functional modules and key hub proteins. RESULTS: Principal component and hierarchical clustering analyses revealed separation of PHH and brain tumor samples from controls, while NMDARE partially overlapped. CSF in PHH contained 532 DAPs, with pathway enrichment analysis identifying alternative complement activation, coagulation, and platelet degranulation pathways as top hits. Conversely, CSF in NMDARE showed only 65 DAPs, with the top pathways involving IL-10 and IL-18 signaling, and the top 3 DAPs involving humoral immunity (IGLC2, MZB1, CD79B). DAPs did not meet statistical significance in brain tumor patients. Longitudinal analysis of serial collections from PHH patients suggested a persistence of coagulation- and complement-related neuroinflammation over time. NELL2 emerged as a consistently downregulated protein in PHH for weeks after the initial hemorrhage. CONCLUSIONS: PHH and NMDARE revealed distinct neuroinflammatory proteomic signatures compared to our control samples. PHH was marked by a broad increase in detection of the majority of inflammation-related proteins, with highest representation among the alternative complement and coagulation-related pathways. The persistent detection of these proteins for weeks after the initial hemorrhage may be indicative of chronic neuroinflammation, even at the time of permanent CSF diversion. Conversely, NMDARE induced a narrower lymphocyte-driven profile, more consistent with an antibody-mediated autoimmune disease. Furthermore, suppression of NELL2 and up-regulation of immunoglobulin-related markers (IGLC2, MZB1, CD79B) are potential candidates for biomarkers in PHH and NMDARE, respectively.

Department

Pediatrics

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