TY - JOUR
T1 - Intranasally administered human MSC-derived extracellular vesicles inhibit NLRP3-p38/MAPK signaling after TBI and prevent chronic brain dysfunction
AU - Kodali, Maheedhar
AU - Madhu, Leelavathi N.
AU - Reger, Roxanne L.
AU - Milutinovic, Bojana
AU - Upadhya, Raghavendra
AU - Gonzalez, Jenny J.
AU - Attaluri, Sahithi
AU - Shuai, Bing
AU - Gitai, Daniel L.G.
AU - Rao, Shama
AU - Choi, Jong M.
AU - Jung, Sung Y.
AU - Shetty, Ashok K.
N1 - Funding Information:
This work was primarily supported by a grant from the National Institute of Neurological Disorders and Stroke (1R01NS106907 to A.K.S.). The authors thank Dr. Darwin Prockop for insightful suggestions on culturing human mesenchymal stem cells and isolating and characterizing the therapeutic properties of human mesenchymal cell-derived extracellular vesicles. The Small RNA sequencing was performed at the Institute for Genome Sciences and Society, Texas A&M University. The mass spectrometry analyses were performed at the Mass Spectrometry based proteomics core at the Baylor College of Medicine, supported by the N.I.H. Grant P30CA125123. The TEM images of hMSC-EVs were performed in the Image Analysis Laboratory, Texas A&M Veterinary Medicine & Biomedical Sciences (RRIS: SCR_022479). Concept: AKS. Research design: AKS, MK, LNM, RR, BM, RU, SA, DLGG, and SYJ. Data collection, analysis, and interpretation: MK, LNM, RLR, BM, RU, JJG, SA, BS, SR, DLGG, JMC, SYJ, and AKS. Preparation of figure composites: MK, SYJ, and AKS. Manuscript writing: MK, SYJ, and AKS. All authors provided feedback, edits, and additions to the manuscript text and approved the final version of the manuscript.
Funding Information:
This work was primarily supported by a grant from the National Institute of Neurological Disorders and Stroke (1R01NS106907 to A.K.S.). The authors thank Dr. Darwin Prockop for insightful suggestions on culturing human mesenchymal stem cells and isolating and characterizing the therapeutic properties of human mesenchymal cell-derived extracellular vesicles. The Small RNA sequencing was performed at the Institute for Genome Sciences and Society, Texas A&M University. The mass spectrometry analyses were performed at the Mass Spectrometry based proteomics core at the Baylor College of Medicine, supported by the N.I.H. Grant P30CA125123. The TEM images of hMSC-EVs were performed in the Image Analysis Laboratory, Texas A&M Veterinary Medicine & Biomedical Sciences (RRIS: SCR_022479).
Publisher Copyright:
© 2022 The Author(s)
PY - 2023/2
Y1 - 2023/2
N2 - Traumatic brain injury (TBI) leads to lasting brain dysfunction with chronic neuroinflammation typified by nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 (NLRP3) inflammasome activation in microglia. This study probed whether a single intranasal (IN) administration of human mesenchymal stem cell-derived extracellular vesicles (hMSC-EVs) naturally enriched with activated microglia-modulating miRNAs can avert chronic adverse outcomes of TBI. Small RNA sequencing confirmed the enrichment of miRNAs capable of modulating activated microglia in hMSC-EV cargo. IN administration of hMSC-EVs into adult mice ninety minutes after the induction of a unilateral controlled cortical impact injury resulted in their incorporation into neurons and microglia in both injured and contralateral hemispheres. A single higher dose hMSC-EV treatment also inhibited NLRP3 inflammasome activation after TBI, evidenced by reduced NLRP3, apoptosis-associated speck-like protein containing a CARD, activated caspase-1, interleukin-1 beta, and IL-18 levels in the injured brain. Such inhibition in the acute phase of TBI endured in the chronic phase, which could also be gleaned from diminished NLRP3 inflammasome activation in microglia of TBI mice receiving hMSC-EVs. Proteomic analysis and validation revealed that higher dose hMSC-EV treatment thwarted the chronic activation of the p38 mitogen-activated protein kinase (MAPK) signaling pathway by IL-18, which decreased the release of proinflammatory cytokines. Inhibition of the chronic activation of NLRP3-p38/MAPK signaling after TBI also prevented long-term cognitive and mood impairments. Notably, the animals receiving higher doses of hMSC-EVs after TBI displayed better cognitive and mood function in all behavioral tests than animals receiving the vehicle after TBI. A lower dose of hMSC-EV treatment also partially improved cognitive and mood function. Thus, an optimal IN dose of hMSC-EVs naturally enriched with activated microglia-modulating miRNAs can inhibit the chronic activation of NLRP3-p38/MAPK signaling after TBI and prevent lasting brain dysfunction.
AB - Traumatic brain injury (TBI) leads to lasting brain dysfunction with chronic neuroinflammation typified by nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 (NLRP3) inflammasome activation in microglia. This study probed whether a single intranasal (IN) administration of human mesenchymal stem cell-derived extracellular vesicles (hMSC-EVs) naturally enriched with activated microglia-modulating miRNAs can avert chronic adverse outcomes of TBI. Small RNA sequencing confirmed the enrichment of miRNAs capable of modulating activated microglia in hMSC-EV cargo. IN administration of hMSC-EVs into adult mice ninety minutes after the induction of a unilateral controlled cortical impact injury resulted in their incorporation into neurons and microglia in both injured and contralateral hemispheres. A single higher dose hMSC-EV treatment also inhibited NLRP3 inflammasome activation after TBI, evidenced by reduced NLRP3, apoptosis-associated speck-like protein containing a CARD, activated caspase-1, interleukin-1 beta, and IL-18 levels in the injured brain. Such inhibition in the acute phase of TBI endured in the chronic phase, which could also be gleaned from diminished NLRP3 inflammasome activation in microglia of TBI mice receiving hMSC-EVs. Proteomic analysis and validation revealed that higher dose hMSC-EV treatment thwarted the chronic activation of the p38 mitogen-activated protein kinase (MAPK) signaling pathway by IL-18, which decreased the release of proinflammatory cytokines. Inhibition of the chronic activation of NLRP3-p38/MAPK signaling after TBI also prevented long-term cognitive and mood impairments. Notably, the animals receiving higher doses of hMSC-EVs after TBI displayed better cognitive and mood function in all behavioral tests than animals receiving the vehicle after TBI. A lower dose of hMSC-EV treatment also partially improved cognitive and mood function. Thus, an optimal IN dose of hMSC-EVs naturally enriched with activated microglia-modulating miRNAs can inhibit the chronic activation of NLRP3-p38/MAPK signaling after TBI and prevent lasting brain dysfunction.
KW - Human mesenchymal stem cell-derived extracellular vesicles
KW - Interleukin-18
KW - Mitogen-activated protein kinase signaling
KW - NLRP3 inflammasomes
KW - Proinflammatory microglia
KW - Traumatic brain injury
UR - http://www.scopus.com/inward/record.url?scp=85143633996&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85143633996&partnerID=8YFLogxK
U2 - 10.1016/j.bbi.2022.11.014
DO - 10.1016/j.bbi.2022.11.014
M3 - Article
C2 - 36427808
AN - SCOPUS:85143633996
SN - 0889-1591
VL - 108
SP - 118
EP - 134
JO - Brain, behavior, and immunity
JF - Brain, behavior, and immunity
ER -