In vitro Characterization of Gut Microbiota-Derived Bacterial Strains With Neuroprotective Properties

Suaad Ahmed, Alessandro Busetti, Parthena Fotiadou, Nisha Vincy Jose, Sarah Reid, Marieta Georgieva, Samantha Brown, Hayley Dunbar, Gloria Beurket-Ascencio, Margaret I Delday, Anna Ettorre*, Imke E Mulder

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

Neurodegenerative diseases are disabling, incurable, and progressive conditions characterized by neuronal loss and decreased cognitive function. Changes in gut microbiome composition have been linked to a number of neurodegenerative diseases, indicating a role for the gut-brain axis. Here, we show how specific gut-derived bacterial strains can modulate neuroinflammatory and neurodegenerative processes in vitro through the production of specific metabolites and discuss the potential therapeutic implications for neurodegenerative disorders. A panel of fifty gut bacterial strains was screened for their ability to reduce pro-inflammatory IL-6 secretion in U373 glioblastoma astrocytoma cells. Parabacteroides distasonis MRx0005 and Megasphaera massiliensis MRx0029 had the strongest capacity to reduce IL-6 secretion in vitro. Oxidative stress plays a crucial role in neuroinflammation and neurodegeneration, and both bacterial strains displayed intrinsic antioxidant capacity. While MRx0005 showed a general antioxidant activity on different brain cell lines, MRx0029 only protected differentiated SH-SY5Y neuroblastoma cells from chemically induced oxidative stress. MRx0029 also induced a mature phenotype in undifferentiated neuroblastoma cells through upregulation of microtubule-associated protein 2. Interestingly, short-chain fatty acid analysis revealed that MRx0005 mainly produced C1-C3 fatty acids, while MRx0029 produced C4-C6 fatty acids, specifically butyric, valeric and hexanoic acid. None of the short-chain fatty acids tested protected neuroblastoma cells from chemically induced oxidative stress. However, butyrate was able to reduce neuroinflammation in vitro, and the combination of butyrate and valerate induced neuronal maturation, albeit not to the same degree as the complex cell-free supernatant of MRx0029. This observation was confirmed by solvent extraction of cell-free supernatants, where only MRx0029 methanolic fractions containing butyrate and valerate showed an anti-inflammatory activity in U373 cells and retained the ability to differentiate neuroblastoma cells. In summary, our results suggest that the pleiotropic nature of live biotherapeutics, as opposed to isolated metabolites, could be a promising novel drug class in drug discovery for neurodegenerative disorders.

Original languageEnglish
Article number402
JournalFrontiers in cellular neuroscience
Volume13
DOIs
Publication statusPublished - 20 Sep 2019

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Neuroblastoma
Neurodegenerative Diseases
Butyrates
Valerates
Oxidative Stress
Volatile Fatty Acids
Interleukin-6
Megasphaera
Fatty Acids
Antioxidants
Aptitude
Microtubule-Associated Proteins
Gastrointestinal Microbiome
In Vitro Techniques
Astrocytoma
Brain
Drug Discovery
Glioblastoma
Cognition
Anti-Inflammatory Agents

Keywords

  • gut microbiota-derived bacterial strains
  • microbiome
  • neurodegenerative diseases
  • neuroinflammation
  • neuroprotection
  • oxidative stress
  • gut-brain axis
  • short-chain fatty acids
  • cells
  • brain axis
  • involvement
  • disorders
  • model
  • gas-chromatography
  • distasonis
  • chain fatty-acids
  • stress
  • Parkinson's disease

Cite this

Ahmed, S., Busetti, A., Fotiadou, P., Vincy Jose, N., Reid, S., Georgieva, M., ... Mulder, I. E. (2019). In vitro Characterization of Gut Microbiota-Derived Bacterial Strains With Neuroprotective Properties. Frontiers in cellular neuroscience, 13, [402]. https://doi.org/10.3389/fncel.2019.00402

In vitro Characterization of Gut Microbiota-Derived Bacterial Strains With Neuroprotective Properties. / Ahmed, Suaad; Busetti, Alessandro; Fotiadou, Parthena; Vincy Jose, Nisha; Reid, Sarah; Georgieva, Marieta; Brown, Samantha; Dunbar, Hayley; Beurket-Ascencio, Gloria; Delday, Margaret I; Ettorre, Anna; Mulder, Imke E.

In: Frontiers in cellular neuroscience, Vol. 13, 402, 20.09.2019.

Research output: Contribution to journalArticle

Ahmed, S, Busetti, A, Fotiadou, P, Vincy Jose, N, Reid, S, Georgieva, M, Brown, S, Dunbar, H, Beurket-Ascencio, G, Delday, MI, Ettorre, A & Mulder, IE 2019, 'In vitro Characterization of Gut Microbiota-Derived Bacterial Strains With Neuroprotective Properties', Frontiers in cellular neuroscience, vol. 13, 402. https://doi.org/10.3389/fncel.2019.00402
Ahmed, Suaad ; Busetti, Alessandro ; Fotiadou, Parthena ; Vincy Jose, Nisha ; Reid, Sarah ; Georgieva, Marieta ; Brown, Samantha ; Dunbar, Hayley ; Beurket-Ascencio, Gloria ; Delday, Margaret I ; Ettorre, Anna ; Mulder, Imke E. / In vitro Characterization of Gut Microbiota-Derived Bacterial Strains With Neuroprotective Properties. In: Frontiers in cellular neuroscience. 2019 ; Vol. 13.
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abstract = "Neurodegenerative diseases are disabling, incurable, and progressive conditions characterized by neuronal loss and decreased cognitive function. Changes in gut microbiome composition have been linked to a number of neurodegenerative diseases, indicating a role for the gut-brain axis. Here, we show how specific gut-derived bacterial strains can modulate neuroinflammatory and neurodegenerative processes in vitro through the production of specific metabolites and discuss the potential therapeutic implications for neurodegenerative disorders. A panel of fifty gut bacterial strains was screened for their ability to reduce pro-inflammatory IL-6 secretion in U373 glioblastoma astrocytoma cells. Parabacteroides distasonis MRx0005 and Megasphaera massiliensis MRx0029 had the strongest capacity to reduce IL-6 secretion in vitro. Oxidative stress plays a crucial role in neuroinflammation and neurodegeneration, and both bacterial strains displayed intrinsic antioxidant capacity. While MRx0005 showed a general antioxidant activity on different brain cell lines, MRx0029 only protected differentiated SH-SY5Y neuroblastoma cells from chemically induced oxidative stress. MRx0029 also induced a mature phenotype in undifferentiated neuroblastoma cells through upregulation of microtubule-associated protein 2. Interestingly, short-chain fatty acid analysis revealed that MRx0005 mainly produced C1-C3 fatty acids, while MRx0029 produced C4-C6 fatty acids, specifically butyric, valeric and hexanoic acid. None of the short-chain fatty acids tested protected neuroblastoma cells from chemically induced oxidative stress. However, butyrate was able to reduce neuroinflammation in vitro, and the combination of butyrate and valerate induced neuronal maturation, albeit not to the same degree as the complex cell-free supernatant of MRx0029. This observation was confirmed by solvent extraction of cell-free supernatants, where only MRx0029 methanolic fractions containing butyrate and valerate showed an anti-inflammatory activity in U373 cells and retained the ability to differentiate neuroblastoma cells. In summary, our results suggest that the pleiotropic nature of live biotherapeutics, as opposed to isolated metabolites, could be a promising novel drug class in drug discovery for neurodegenerative disorders.",
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author = "Suaad Ahmed and Alessandro Busetti and Parthena Fotiadou and {Vincy Jose}, Nisha and Sarah Reid and Marieta Georgieva and Samantha Brown and Hayley Dunbar and Gloria Beurket-Ascencio and Delday, {Margaret I} and Anna Ettorre and Mulder, {Imke E}",
note = "Acknowledgments The authors are grateful to Dr. Nicole Reichardt and the 4D Pharma Isolation Team for their technical assistance with bacterial culture. Funding This work was privately funded by 4D Pharma PLC. The authors of the study, who are employees of (or in the case of MID, are funded by) 4D Pharma Research Ltd., a wholly owned subsidiary of the funder, were responsible for the design and execution of the study, as well as the analysis of the results obtained. Author Contributions In vitro experiments: AE, SA, and AB designed the experiments; SA, PF, NV, MG, SB, GB-A, and MD performed the majority of the in vitro experiments; SR, HD, and AB validated, performed, and analyzed the microbiological-related data output; AE coordinated and managed the research project; AE, SA, PF, and AB analyzed the data. IM oversaw the overall research plan. AE wrote the manuscript with the assistance of SA and AB. All authors have read and commented on the manuscript and have approved the final version of the manuscript. Data Availability: 16S gene sequences for MRx0005 and MRx0029 are disclosed in International Patent Publication Nos. WO2018/229189 and WO2018/229216, respectively, filed by 4D Pharma Research Ltd. The data supporting the findings in this paper are available within the article and its Supplementary Information Files.",
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T1 - In vitro Characterization of Gut Microbiota-Derived Bacterial Strains With Neuroprotective Properties

AU - Ahmed, Suaad

AU - Busetti, Alessandro

AU - Fotiadou, Parthena

AU - Vincy Jose, Nisha

AU - Reid, Sarah

AU - Georgieva, Marieta

AU - Brown, Samantha

AU - Dunbar, Hayley

AU - Beurket-Ascencio, Gloria

AU - Delday, Margaret I

AU - Ettorre, Anna

AU - Mulder, Imke E

N1 - Acknowledgments The authors are grateful to Dr. Nicole Reichardt and the 4D Pharma Isolation Team for their technical assistance with bacterial culture. Funding This work was privately funded by 4D Pharma PLC. The authors of the study, who are employees of (or in the case of MID, are funded by) 4D Pharma Research Ltd., a wholly owned subsidiary of the funder, were responsible for the design and execution of the study, as well as the analysis of the results obtained. Author Contributions In vitro experiments: AE, SA, and AB designed the experiments; SA, PF, NV, MG, SB, GB-A, and MD performed the majority of the in vitro experiments; SR, HD, and AB validated, performed, and analyzed the microbiological-related data output; AE coordinated and managed the research project; AE, SA, PF, and AB analyzed the data. IM oversaw the overall research plan. AE wrote the manuscript with the assistance of SA and AB. All authors have read and commented on the manuscript and have approved the final version of the manuscript. Data Availability: 16S gene sequences for MRx0005 and MRx0029 are disclosed in International Patent Publication Nos. WO2018/229189 and WO2018/229216, respectively, filed by 4D Pharma Research Ltd. The data supporting the findings in this paper are available within the article and its Supplementary Information Files.

PY - 2019/9/20

Y1 - 2019/9/20

N2 - Neurodegenerative diseases are disabling, incurable, and progressive conditions characterized by neuronal loss and decreased cognitive function. Changes in gut microbiome composition have been linked to a number of neurodegenerative diseases, indicating a role for the gut-brain axis. Here, we show how specific gut-derived bacterial strains can modulate neuroinflammatory and neurodegenerative processes in vitro through the production of specific metabolites and discuss the potential therapeutic implications for neurodegenerative disorders. A panel of fifty gut bacterial strains was screened for their ability to reduce pro-inflammatory IL-6 secretion in U373 glioblastoma astrocytoma cells. Parabacteroides distasonis MRx0005 and Megasphaera massiliensis MRx0029 had the strongest capacity to reduce IL-6 secretion in vitro. Oxidative stress plays a crucial role in neuroinflammation and neurodegeneration, and both bacterial strains displayed intrinsic antioxidant capacity. While MRx0005 showed a general antioxidant activity on different brain cell lines, MRx0029 only protected differentiated SH-SY5Y neuroblastoma cells from chemically induced oxidative stress. MRx0029 also induced a mature phenotype in undifferentiated neuroblastoma cells through upregulation of microtubule-associated protein 2. Interestingly, short-chain fatty acid analysis revealed that MRx0005 mainly produced C1-C3 fatty acids, while MRx0029 produced C4-C6 fatty acids, specifically butyric, valeric and hexanoic acid. None of the short-chain fatty acids tested protected neuroblastoma cells from chemically induced oxidative stress. However, butyrate was able to reduce neuroinflammation in vitro, and the combination of butyrate and valerate induced neuronal maturation, albeit not to the same degree as the complex cell-free supernatant of MRx0029. This observation was confirmed by solvent extraction of cell-free supernatants, where only MRx0029 methanolic fractions containing butyrate and valerate showed an anti-inflammatory activity in U373 cells and retained the ability to differentiate neuroblastoma cells. In summary, our results suggest that the pleiotropic nature of live biotherapeutics, as opposed to isolated metabolites, could be a promising novel drug class in drug discovery for neurodegenerative disorders.

AB - Neurodegenerative diseases are disabling, incurable, and progressive conditions characterized by neuronal loss and decreased cognitive function. Changes in gut microbiome composition have been linked to a number of neurodegenerative diseases, indicating a role for the gut-brain axis. Here, we show how specific gut-derived bacterial strains can modulate neuroinflammatory and neurodegenerative processes in vitro through the production of specific metabolites and discuss the potential therapeutic implications for neurodegenerative disorders. A panel of fifty gut bacterial strains was screened for their ability to reduce pro-inflammatory IL-6 secretion in U373 glioblastoma astrocytoma cells. Parabacteroides distasonis MRx0005 and Megasphaera massiliensis MRx0029 had the strongest capacity to reduce IL-6 secretion in vitro. Oxidative stress plays a crucial role in neuroinflammation and neurodegeneration, and both bacterial strains displayed intrinsic antioxidant capacity. While MRx0005 showed a general antioxidant activity on different brain cell lines, MRx0029 only protected differentiated SH-SY5Y neuroblastoma cells from chemically induced oxidative stress. MRx0029 also induced a mature phenotype in undifferentiated neuroblastoma cells through upregulation of microtubule-associated protein 2. Interestingly, short-chain fatty acid analysis revealed that MRx0005 mainly produced C1-C3 fatty acids, while MRx0029 produced C4-C6 fatty acids, specifically butyric, valeric and hexanoic acid. None of the short-chain fatty acids tested protected neuroblastoma cells from chemically induced oxidative stress. However, butyrate was able to reduce neuroinflammation in vitro, and the combination of butyrate and valerate induced neuronal maturation, albeit not to the same degree as the complex cell-free supernatant of MRx0029. This observation was confirmed by solvent extraction of cell-free supernatants, where only MRx0029 methanolic fractions containing butyrate and valerate showed an anti-inflammatory activity in U373 cells and retained the ability to differentiate neuroblastoma cells. In summary, our results suggest that the pleiotropic nature of live biotherapeutics, as opposed to isolated metabolites, could be a promising novel drug class in drug discovery for neurodegenerative disorders.

KW - gut microbiota-derived bacterial strains

KW - microbiome

KW - neurodegenerative diseases

KW - neuroinflammation

KW - neuroprotection

KW - oxidative stress

KW - gut-brain axis

KW - short-chain fatty acids

KW - cells

KW - brain axis

KW - involvement

KW - disorders

KW - model

KW - gas-chromatography

KW - distasonis

KW - chain fatty-acids

KW - stress

KW - Parkinson's disease

U2 - 10.3389/fncel.2019.00402

DO - 10.3389/fncel.2019.00402

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VL - 13

JO - Frontiers in cellular neuroscience

JF - Frontiers in cellular neuroscience

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