Abstract
Disclosure: L. Kolatorova: None. T. Skodova: None. J. Vitku: None. M. Hill: None. P. Skalicky: None. O. Bradac: None. Background: Normal pressure hydrocephalus (NPH) is neurodegenerative disease manifested by balance impairment, urinary incontinence and dementia development, which can easily be mistaken for Alzheimer's or Parkinson's disease. The commonly used treatment is implantation of a ventriculoperitoneal shunt leading to a clinical improvement in 70–90% of patients, in contrast to other poorly treatable neurodegenerative disorder. Nervous system is an important target and source of steroids. In neurodegenerative disorders, the steroid metabolic pathways and transport channels in the nervous system may be disrupted, these changes are most likely to be reflected in the cerebrospinal fluid (CSF). All major steroid classes were considered to be active within the nervous system and those are termed neuroactive steroids (NAS). Several NAS (dehydroepiandrosterone (DHEA), pregnenolone, progesterone and their metabolites) are known to be synthesized directly in the brain, while others (androgens, corticoids etc.) are of peripheral origin. The aim of our study is to find CSF steroids that could be helpful in differential diagnosis of NPH. Methods: Three newly developed, highly sensitive LC-MS/MS methods for quantifying 17 NAS including androgens, gestagens, corticoids and estrogens were developed and validated. The methods can quantify NAS within order of pg/mL units. Results: First, we compared CSF samples from men (n=20) and woman (n=18) obtained within the NPH diagnostics. We found significantly elevated levels of testosterone and 17-hydroxyprogesterone in men. Second, we make a pilot study observing NPH patients (n=11) before, 3 and 12 months after the shunt implantation. We found significantly increased CSF levels of 7-ketoDHEA after 3 months, after 1year, the levels dropped back. The opposite results were found for estrone. The CSF 17-hydroxypregnenolone levels were significantly increased after 3 months and remained stable after 1 year. CSF 11-hydroxytestosterone increased after 3 months and its increase was also significant after 1 year. Surprising results were found for corticoids. Cortisol, cortisone, corticosterone and 11-deoxycortisol increased significantly 3 and 12 months after surgery. Conclusion: In accordance with our previous research, we found increased levels of neuroprotective 7-ketoDHEA. To the best of our knowledge, we have quantified for the first time the CSF levels of 17-hydroxypregnenolone, which were significantly increased in the 3- and 12-months follow-up. The increase was found also in 11-hydroxytestosterone, which metabolism within the nervous system is actually not fully understood. Until 2026, we plan to quantify the spectrum of NAS in CSF and plasma in much larger cohort of NPH patients as well as controls, and contribute to advancing the diagnosis and pathophysiology of NPH.Acknowledgement: Supported by NU23-04-00551 of Czech Ministry of Health. Presentation: Sunday, July 13, 2025