Non-electrophilic TRPA1 modulator MNP-021 protects neuronal and glial function from diabetes-associated glucotoxicity and neuroinflammation.
14 affiliations
- 1Retrovirus Center, Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, Pisa, Italy.
- 2Centre for Instrumentation Sharing, University of Pisa (CISUP), Pisa, Italy.
- 3Department of Surgical, Medical, Molecular and Critical Care Pathology, University of Pisa, Pisa, Italy.
- 4Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
- 5CNR, Institute of Neuroscience, Padua Branch, Padua, Italy.
- 6Department of Pharmacy, University of Pisa, Pisa, Italy.
- 7Department of Health Sciences, University of Milan, Milan, Italy.
- 8Unit of Pathology, Azienda Socio-Sanitaria Territoriale (ASST) dei Sette Laghi, Varese, Italy.
- 9Department of Pharmaceutical and Pharmacological Sciences, University of Padua, Padua, Italy.
- 10Diabetes Unit, Department of Medicine, Surgery and Neurosciences, University of Siena, Siena, Italy.
- 11Fondazione Umberto Di Mario ONLUS c/o Toscana Life Science, Siena, Italy.
- 12Tuscany Centre for Precision Medicine (CReMeP), Siena, Italy.
- 13Centre for Instrumentation Sharing, University of Pisa (CISUP), Pisa, Italy. [email protected].
- 14Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy. [email protected].
Abstract
Background
Diabetes-associated neurodegeneration is amplified by methylglyoxal (MGO)-driven dicarbonyl stress linking hyperglycemia to neuronal insulin resistance and maladaptive neuroinflammation. We tested the neuroprotective activity of MNP-021, a non-electrophilic transient receptor potential ankyrin 1 (TRPA1) modulator, in neurons and glial cells in vitro.Methods
SH-SY5Y neurons were pretreated with MNP-021 and challenged with MGO, then profiled by high-content imaging, RNA-seq, Seahorse Oxygen Consumption Rate (OCR) and Extracellular Acidification Rate (ECAR), glycolytic stress assays, and immunoblotting for AKT, ERK, and cAMP response element-binding protein (CREB). In an additional sustained glucotoxicity paradigm, differentiated SH-SY5Y cells were exposed to high glucose for 7 days and viability was assessed by LIVE/DEAD™ high-content imaging. HMC3 glial cells were treated with MNP-021, exposed to Lipopolysaccharide (LPS), Tumor Necrosis Factor-α (TNF-α) or Aβ(25-35), and tested for viability and inflammatory markers by Enzyme-Linked Immunosorbent Assay (ELISA) and RT-qPCR.Results
MGO increased nucleus-to-cytoplasm area ratio by 49% and dysregulated glucose handling, increasing 2-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)Amino)-2-Deoxyglucose (2-NBDG) uptake by ~ 25%, with GLUT1/GLUT4 membrane redistribution; MNP-021 normalized morphology, glucose uptake, and transporter localization without cytotoxicity up to 10 µM. RNA-seq identified 754 MGO-deregulated genes, including ISR/metabolic nodes such as glucokinase (GCK), Sestrin-2 (SESN2), phosphoglycerate dehydrogenase (PHGDH), Phosphoserine Aminotransferase 1 (PSAT1), phosphoenolpyruvate carboxykinase 2 (PCK2); MNP-021 buffered stress-induced transcription with limited baseline effects, modulated mitochondrial morphology and redox-associated fluorescence while preserving overall respiratory flux and blunting stress-evoked compensatory glycolysis. MNP-021 restored pro-survival signaling (pAKT/pERK and nuclear pCREB), including insulin responsiveness during MGO exposure. In a sustained high-glucose model, MNP-021 reduced the high-glucose-associated increase in cell death, extending its protective effects beyond the acute MGO paradigm. MNP-021 reduced Interleukine-6 (IL-6)/TNF-α release while increasing Interleukine-10 (IL-10) and Arginase 1 (ARG1) (~ 1.9-fold vs. LPS/TNF-α) in HMC3 glial cells, shifting them toward a pro-resolving IL-10/ARG1 program with reduced amyloid β-peptide ₂₅₋₃₅ (Aβ₍₂₅₋₃₅₎)-evoked cytokine release and Glucagon-Like Peptide-1 (GLP-1) remaining very low (≤ 10 pg/mL) and not significantly increased in this system.Conclusions
MNP-021 coordinates transcriptomic restraint, transporter-level glucose handling, mitochondrial resilience, and pro-survival/pro-resolving signaling in neuron-microglia cell models. These findings support further preclinical evaluation of MNP-021 in dicarbonyl-linked neuro-metabolic stress and provide pharmacological evidence consistent with a TRPA1-linked component of its activity.
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BibTeX
@article{carli2026nonelectrophilic,
title = {{Non-electrophilic TRPA1 modulator MNP-021 protects neuronal and glial function from diabetes-associated glucotoxicity and neuroinflammation.}},
author = {Alessandro De Carli and Carolina Filipponi and Beatrice Polini and Veronica Sancho-Bornez and Emy Basso and Fabio Filippini and Angela Dardano and Francesca Sardelli and Simona Daniele and Martina Contestabile and Franco Folli and Giovanna Finzi and Sara De Martin and Giuseppina Emanuela Grieco and Francesco Dotta and Guido Sebastiani and Grazia Chiellini and Mauro Pineschi and Michele Lai and Giuseppe Daniele},
journal = {Journal of translational medicine},
year = {2026},
month = oct,
doi = {10.1186/s12967-026-09028-7},
url = {https://europepmc.org/article/MED/42958409},
note = {CC BY-NC-ND},
}
RIS
TY - JOUR
TI - Non-electrophilic TRPA1 modulator MNP-021 protects neuronal and glial function from diabetes-associated glucotoxicity and neuroinflammation.
AU - Alessandro De Carli
AU - Carolina Filipponi
AU - Beatrice Polini
AU - Veronica Sancho-Bornez
AU - Emy Basso
AU - Fabio Filippini
AU - Angela Dardano
AU - Francesca Sardelli
AU - Simona Daniele
AU - Martina Contestabile
AU - Franco Folli
AU - Giovanna Finzi
AU - Sara De Martin
AU - Giuseppina Emanuela Grieco
AU - Francesco Dotta
AU - Guido Sebastiani
AU - Grazia Chiellini
AU - Mauro Pineschi
AU - Michele Lai
AU - Giuseppe Daniele
PY - 2026
DA - 2026/10/27
JO - Journal of translational medicine
DO - 10.1186/s12967-026-09028-7
UR - https://europepmc.org/article/MED/42958409
AB - Background Diabetes-associated neurodegeneration is amplified by methylglyoxal (MGO)-driven dicarbonyl stress linking hyperglycemia to neuronal insulin resistance and maladaptive neuroinflammation. We tested the neuroprotective activity of MNP-021, a non-electrophilic transient receptor potential ankyrin 1 (TRPA1) modulator, in neurons and glial cells in vitro.Methods SH-SY5Y neurons were pretreated with MNP-021 and challenged with MGO, then profiled by high-content imaging, RNA-seq, Seahorse Oxygen Consumption Rate (OCR) and Extracellular Acidification Rate (ECAR), glycolytic stress assays, and immunoblotting for AKT, ERK, and cAMP response element-binding protein (CREB). In an additional sustained glucotoxicity paradigm, differentiated SH-SY5Y cells were exposed to high glucose for 7 days and viability was assessed by LIVE/DEAD™ high-content imaging. HMC3 glial cells were treated with MNP-021, exposed to Lipopolysaccharide (LPS), Tumor Necrosis Factor-α (TNF-α) or Aβ(25-35), and tested for viability and inflammatory markers by Enzyme-Linked Immunosorbent Assay (ELISA) and RT-qPCR.Results MGO increased nucleus-to-cytoplasm area ratio by 49% and dysregulated glucose handling, increasing 2-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)Amino)-2-Deoxyglucose (2-NBDG) uptake by ~ 25%, with GLUT1/GLUT4 membrane redistribution; MNP-021 normalized morphology, glucose uptake, and transporter localization without cytotoxicity up to 10 µM. RNA-seq identified 754 MGO-deregulated genes, including ISR/metabolic nodes such as glucokinase (GCK), Sestrin-2 (SESN2), phosphoglycerate dehydrogenase (PHGDH), Phosphoserine Aminotransferase 1 (PSAT1), phosphoenolpyruvate carboxykinase 2 (PCK2); MNP-021 buffered stress-induced transcription with limited baseline effects, modulated mitochondrial morphology and redox-associated fluorescence while preserving overall respiratory flux and blunting stress-evoked compensatory glycolysis. MNP-021 restored pro-survival signaling (pAKT/pERK and nuclear pCREB), including insulin responsiveness during MGO exposure. In a sustained high-glucose model, MNP-021 reduced the high-glucose-associated increase in cell death, extending its protective effects beyond the acute MGO paradigm. MNP-021 reduced Interleukine-6 (IL-6)/TNF-α release while increasing Interleukine-10 (IL-10) and Arginase 1 (ARG1) (~ 1.9-fold vs. LPS/TNF-α) in HMC3 glial cells, shifting them toward a pro-resolving IL-10/ARG1 program with reduced amyloid β-peptide ₂₅₋₃₅ (Aβ₍₂₅₋₃₅₎)-evoked cytokine release and Glucagon-Like Peptide-1 (GLP-1) remaining very low (≤ 10 pg/mL) and not significantly increased in this system.Conclusions MNP-021 coordinates transcriptomic restraint, transporter-level glucose handling, mitochondrial resilience, and pro-survival/pro-resolving signaling in neuron-microglia cell models. These findings support further preclinical evaluation of MNP-021 in dicarbonyl-linked neuro-metabolic stress and provide pharmacological evidence consistent with a TRPA1-linked component of its activity.
SN - 1479-5876
ER -