PMID- 34678224 OWN - NLM STAT- MEDLINE DCOM- 20220103 LR - 20221202 IS - 1873-2968 (Electronic) IS - 0006-2952 (Print) IS - 0006-2952 (Linking) VI - 194 DP - 2021 Dec TI - Inhibition of neutral sphingomyelinase 2 reduces extracellular vesicle release from neurons, oligodendrocytes, and activated microglial cells following acute brain injury. PG - 114796 LID - S0006-2952(21)00412-3 [pii] LID - 10.1016/j.bcp.2021.114796 [doi] AB - Extracellular Vesicles (EVs) are implicated in the spread of pathogenic proteinsin a growing number of neurological diseases. Given this, there is rising interest in developing inhibitors of Neutral Sphingomyelinase 2 (nSMase2), an enzyme critical in EV biogenesis. Our group recently discovered phenyl(R)-(1-(3-(3,4-dimethoxyphenyl)-2,6-dimethylimidazo[1,2-b]pyridazin-8-yl)pyrrolidin-3-yl)carbamate (PDDC), the first potent, selective, orally-available, and brain-penetrable nSMase2 inhibitor, capable of dose-dependently reducing EVs release in vitro and in vivo. Herein, using multiplexed Surface Plasmon Resonance imaging (SPRi), we evaluated which brain cell-derived EVs were affected by PDDC following acute brain injury. Mice were fed PDDC-containing chow at doses which gave steady PDDC brain exposures exceeding its nSMase2 IC(50). Mice were then administered an intra-striatal IL-1beta injection and two hours later plasma and brain were collected. IL-1beta injection significantly increased striatal nSMase2 activity which was completely normalized by PDDC. Using SPRi, we found that IL-1beta-induced injury selectively increased plasma levels of CD171 + and PLP1 + EVs; this EV increase was normalized by PDDC. In contrast, GLAST1 + EVs were unchanged by IL-1beta or PDDC. IL-1beta injection selectively increased EVs released from activated versus non-activated microglia, indicated by the CD11b+/IB4 + ratio. The increase in EVs from CD11b + microglia was dramatically attenuated with PDDC. Taken together, our data demonstrate that following acute injury, brain nSMase2 activity is elevated. EVs released from neurons, oligodendrocytes, and activated microglial are increased in plasma and inhibition of nSMase2 with PDDC reduced these IL-1beta-induced changes implicating nSMase2 inhibition as a therapeutic target for acute brain injury. CI - Copyright (c) 2021. Published by Elsevier Inc. FAU - Tallon, Carolyn AU - Tallon C AD - Johns Hopkins Drug Discovery, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. FAU - Picciolini, Silvia AU - Picciolini S AD - IRCCS Fondazione Don Carlo Gnocchi ONLUS, Laboratory of Nanomedicine and Clinical Biophotonics (LABION), Milan, Italy. FAU - Yoo, Seung-Wan AU - Yoo SW AD - Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. FAU - Thomas, Ajit G AU - Thomas AG AD - Johns Hopkins Drug Discovery, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. FAU - Pal, Arindom AU - Pal A AD - Johns Hopkins Drug Discovery, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. FAU - Alt, Jesse AU - Alt J AD - Johns Hopkins Drug Discovery, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. FAU - Carlomagno, Cristiano AU - Carlomagno C AD - IRCCS Fondazione Don Carlo Gnocchi ONLUS, Laboratory of Nanomedicine and Clinical Biophotonics (LABION), Milan, Italy. FAU - Gualerzi, Alice AU - Gualerzi A AD - IRCCS Fondazione Don Carlo Gnocchi ONLUS, Laboratory of Nanomedicine and Clinical Biophotonics (LABION), Milan, Italy. FAU - Rais, Rana AU - Rais R AD - Johns Hopkins Drug Discovery, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. FAU - Haughey, Norman J AU - Haughey NJ AD - Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. FAU - Bedoni, Marzia AU - Bedoni M AD - IRCCS Fondazione Don Carlo Gnocchi ONLUS, Laboratory of Nanomedicine and Clinical Biophotonics (LABION), Milan, Italy. Electronic address: mbedoni@dongnocchi.it. FAU - Slusher, Barbara S AU - Slusher BS AD - Johns Hopkins Drug Discovery, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Psychiatry and Behavioral Science, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. Electronic address: bslusher@jhmi.edu. LA - eng GR - P30 MH075673/MH/NIMH NIH HHS/United States GR - R01 AG059799/AG/NIA NIH HHS/United States GR - R01 AG063831/AG/NIA NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20211020 PL - England TA - Biochem Pharmacol JT - Biochemical pharmacology JID - 0101032 RN - 0 (IL1B protein, mouse) RN - 0 (Interleukin-1beta) RN - 0 (Pyrenes) RN - 93255-34-6 (pyrenedodecanoylcarnitine) RN - EC 3.1.4.12 (Smpd3 protein, mouse) RN - EC 3.1.4.12 (Sphingomyelin Phosphodiesterase) RN - S7UI8SM58A (Carnitine) SB - IM MH - Animals MH - Brain Injuries/drug therapy/*enzymology MH - Carnitine/administration & dosage/analogs & derivatives MH - Corpus Striatum/drug effects/enzymology MH - Extracellular Vesicles/drug effects/*enzymology MH - Injections, Intraventricular MH - Interleukin-1beta/administration & dosage MH - Male MH - Mice MH - Mice, Transgenic MH - Microglia/drug effects/*enzymology MH - Neurons/drug effects/*enzymology MH - Oligodendroglia/drug effects/*enzymology MH - Pyrenes/administration & dosage MH - Sphingomyelin Phosphodiesterase/antagonists & inhibitors/*metabolism PMC - PMC8919377 MID - NIHMS1784754 OTO - NOTNLM OT - Exosomes OT - Extracellular vesicles OT - Microglia OT - Neuroinflammation OT - Neutral sphingomyelinase 2 OT - Surface plasmon resonance imaging COIS- Conflicts of interest Authors NH, RR, AGT, and BSS are inventors on patent applications filed with Johns Hopkins University which cover novel nSMase inhibitor compositions, including PDDC, and their utility in disease. EDAT- 2021/10/23 06:00 MHDA- 2022/01/04 06:00 PMCR- 2022/12/01 CRDT- 2021/10/22 20:30 PHST- 2021/07/29 00:00 [received] PHST- 2021/09/30 00:00 [revised] PHST- 2021/10/01 00:00 [accepted] PHST- 2021/10/23 06:00 [pubmed] PHST- 2022/01/04 06:00 [medline] PHST- 2021/10/22 20:30 [entrez] PHST- 2022/12/01 00:00 [pmc-release] AID - S0006-2952(21)00412-3 [pii] AID - 10.1016/j.bcp.2021.114796 [doi] PST - ppublish SO - Biochem Pharmacol. 2021 Dec;194:114796. doi: 10.1016/j.bcp.2021.114796. Epub 2021 Oct 20.