Profiling of metalloprotease activities in cerebrospinal fluids of patients with neoplastic meningitis
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Author(s)
Conrad, Catharina
Dorzweiler, Kristina
Strik, Herwig
Bartsch, Jörg W.
Miller, Miles Aaron
Lauffenburger, Douglas A
Date Issued
August 2017
Journal
Fluids and Barriers of the CNS
Publisher
Biomed Central Ltd
Citation
Conrad, Catharina et al. “Profiling of Metalloprotease Activities in Cerebrospinal Fluids of Patients with Neoplastic Meningitis.” Fluids and Barriers of the CNS 14, 1 (August 2017): 22 © 2017 The Author(s)
Version
Final published version
Abstract
Background: Neoplastic invasion into leptomeninges and subarachnoid space, resulting in neoplastic meningitis (NM) is a fatal complication of advanced solid and hematological neoplasms. Identification of malignant involvement of the cerebrospinal fluid (CSF) early in the disease course has crucial prognostic and therapeutic implications, but remains challenging. As indicators of extracellular matrix (ECM) degradation and breakdown of the blood-brain-barrier, Matrix Metalloproteases (MMPs) and A Disintegrin and Metalloproteases (ADAMs) are potential analytes for cerebral pathophysiology and metastatic dissemination of tumor cells into the CSF. Methods: We compared protease activities in CSF samples from patients with NM and control individuals using FRET-based metalloprotease substrates with distinct enzyme selectivity profiles in a real-time, multiplex approach termed "proteolytic activity matrix assay" (PrAMA). Protease activity dynamics can be tracked by fluorescence changes over time. By simultaneously monitoring a panel of 5 FRET-substrate cleavages, a proteolytic signature can be identified and analyzed to infer the activities of multiple specific proteases. Distinct patterns of substrate cleavage comparing disease vs. control samples allow rapid, reproducible and sensitive discrimination even in small volumes of CSF. Results: Individual substrate cleavage rates were linked to distinct proteases, and PrAMA computational inference implied increased activities of MMP-9, ADAM8 and ADAM17 (4-5-fold on average) in CSF samples from NM patients that were inhibitable by the metalloprotease inhibitor batimastat (BB-94). The activities of these proteases correlated with blood-brain barrier impairment. Notably, CSF cell counts were not found to directly reflect the protease activities observed in CSF samples from NM patients; this may explain the frequent clinical observation of negative cytology in NM patients. Conclusion: PrAMA analysis of CSF samples is a potential diagnostic method for sensitive detection of NM and may be suitable for the clinical routine. Keywords: Neoplastic meningitis; Metalloproteases; CSF; Real-time protease activities; FRET-substrates
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
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Creative Commons Attribution 4.0 International License
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DOI of Published Version
https://doi.org/10.1186/S12987-017-0070-5
