Supramolecular Nanostructure Activates TrkB Receptor Signaling of Neuronal Cells by Mimicking Brain-Derived Neurotrophic Factor

Alexandra N. Edelbrock; Zaida Àlvarez; Dina Simkin; Timmy Fyrner; Stacey M. Chin; Kohei Sato; Evangelos Kiskinis; Samuel I. Stupp

doi:10.1021/acs.nanolett.8b02317

Supramolecular Nanostructure Activates TrkB Receptor Signaling of Neuronal Cells by Mimicking Brain-Derived Neurotrophic Factor

Alexandra N. Edelbrock, Zaida Àlvarez, Dina Simkin, Timmy Fyrner, Stacey M. Chin, Kohei Sato, Evangelos Kiskinis, Samuel I. Stupp^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

75 Scopus citations

Abstract

Brain-derived neurotrophic factor (BDNF), a neurotrophin that binds specifically to the tyrosine kinase B (TrkB) receptor, has been shown to promote neuronal differentiation, maturation, and synaptic plasticity in the central nervous system (CNS) during development or after injury and onset of disease. Unfortunately, native BDNF protein-based therapies have had little clinical success due to their suboptimal pharmacological properties. In the past 20 years, BDNF mimetic peptides have been designed with the purpose of activating certain cell pathways that mimic the functional activity of native BDNF, but the interaction of mimetic peptides with cells can be limited due to the conformational specificity required for receptor activation. We report here on the incorporation of a BDNF mimetic sequence into a supramolecular peptide amphiphile filamentous nanostructure capable of activating the BDNF receptor TrkB and downstream signaling in primary cortical neurons in vitro. Interestingly, we found that this BDNF mimetic peptide is only active when displayed on a peptide amphiphile supramolecular nanostructure. We confirmed that increased neuronal maturation is linked to TrkB signaling pathways by analyzing the phosphorylation of downstream signaling effectors and tracking electrical activity over time. Furthermore, three-dimensional gels containing the BDNF peptide amphiphile (PA) nanostructures encourage cell infiltration while increasing functional maturation. Our findings suggest that the BDNF mimetic PA nanostructure creates a highly bioactive matrix that could serve as a biomaterial therapy in injured regions of the CNS. This new strategy has the potential to induce endogenous cell infiltration and promote functional neuronal maturation through the presentation of the BDNF mimetic signal.

Original language	English (US)
Pages (from-to)	6237-6247
Number of pages	11
Journal	Nano letters
Volume	18
Issue number	10
DOIs	https://doi.org/10.1021/acs.nanolett.8b02317
State	Published - Oct 10 2018

Funding

*S.I. Stupp. E-mail: [email protected]. ORCID Timmy Fyrner: 0000-0003-2119-9883 Kohei Sato: 0000-0002-8948-8537 Samuel I. Stupp: 0000-0002-5491-7442 Present Address ∇Department of Life Science and Technology, Tokyo Institute of Technology, Yokohama 226−8501, Japan Author Contributions ◆A.N.E. and Z.A. contributed equally to this work. A.N.E. synthesized materials, A.N.E. and Z.A. designed and performed experiments, analyzed data, and wrote the manuscript. D.S. performed all electrophysiology experiments and analyzed data. S.M.C. performed analysis experiments and analyzed data. T.F. and K.S. carried out synthetic work, material characterization, and took part in discussions. E.K. supervised the research. S.I.S. wrote the manuscript and supervised the research. The manuscript was written through contributions of all authors. All authors have given approval to the final version of the manuscript. Funding This work was supported by National Institutes of Health/ National Institute of Biomedical Imaging and Bioengineering Award Number 5R01EB003806-07 and by the Center for Regenerative Nanomedicine at the Simpson Querrey Institute at Northwestern. The E.K. lab is supported by research grants from the Les Turner ALS Foundation, the Muscular Dystrophy Association, the Dravet Foundation and NIH/NINDS R01NS104219. E.K. is a Les Turner ALS Center Investigator. A.N.E. and S.M.C. received graduate research fellowships through the National Science Foundation. Z.A. received postdoctoral support from the Beatriu de Pinoś Fellowship 2014 BP-A 00007 (Ageǹcia de Gestió d’Ajust Universitaris i de Recerca, AGAUR) and the PVA Grant # PVA17_RF_0008 from the Paralyzed Veterans of America (PVA) Research Foundation. Notes The authors declare no competing financial interest. We are grateful to the following core facilities at Northwestern University: the Peptide Synthesis Core and the Analytical Bionanotechnology Equipment Core both at the Simpson Querrey Institute for BioNanotechnology. The U.S. Army Research Office, the U.S. Army Medical Research Materiel Command, and Northwestern University provided funding to develop these facilities and ongoing support is being received from the Soft and Hybrid Nanotechnology Experimental (SHyNE) Resource (NSF NNCI-1542205). The Biological Imaging Facility (supported by the Northwestern University Office for Research), the Center for Advanced Microscopy (NCI CCSG P30 CA060553), Flow Cytometry Core (supported by Cancer Center Support Grant NCI CA060553), the Electron Probe Instrumentation Center (EPIC) facility of Northwestern University’s NUANCE Center, which has received support from the Soft and Hybrid Nanotechnology Experimental (SHyNE) Resource (NSF ECCS-1542205), and this work made use of the Integrated Molecular Structure Education and Research Center (IM-SERC) at Northwestern University, which has received support from the Soft and Hybrid Nanotechnology Experimental (SHyNE) Resource (NSF ECCS-1542205); the State of Illinois and International Institute for Nanotechnology (IIN). Portions of this work were performed at the DuPont-Northwestern-Dow Collaborative Access Team (DND-CAT) located at Sector 5 of the Advanced Photon Source (APS). DND-CAT is supported by Northwestern University, E.I. DuPont de Nemours & Co., and The Dow Chemical Company. This research used resources of the Advanced Photon Source, a U.S. Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under Contract No. DE-AC02-06CH11357. The authors also thank Prof. Alfred George for the use of his facilities for all patch clamp experiments, Armando Hernandez Garcia for assistance with flow cytometry experiments, Prof. Liam Palmer for helpful discussions, as well as Jack Edelbrock and Mark Seniw for their design of schematic drawings in the manuscript. The authors would also like to thank Dr. Ivań Sasselli for his assistance with all FT-IR experiments.

Keywords

BDNF mimetic
TrkB receptor
maturation
nanostructure
neurons
peptide amphiphile

ASJC Scopus subject areas

General Chemistry
Condensed Matter Physics
Mechanical Engineering
Bioengineering
General Materials Science

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10.1021/acs.nanolett.8b02317

Cite this

@article{c3d0474cd64a4855ac5b3c508484135a,

title = "Supramolecular Nanostructure Activates TrkB Receptor Signaling of Neuronal Cells by Mimicking Brain-Derived Neurotrophic Factor",

abstract = "Brain-derived neurotrophic factor (BDNF), a neurotrophin that binds specifically to the tyrosine kinase B (TrkB) receptor, has been shown to promote neuronal differentiation, maturation, and synaptic plasticity in the central nervous system (CNS) during development or after injury and onset of disease. Unfortunately, native BDNF protein-based therapies have had little clinical success due to their suboptimal pharmacological properties. In the past 20 years, BDNF mimetic peptides have been designed with the purpose of activating certain cell pathways that mimic the functional activity of native BDNF, but the interaction of mimetic peptides with cells can be limited due to the conformational specificity required for receptor activation. We report here on the incorporation of a BDNF mimetic sequence into a supramolecular peptide amphiphile filamentous nanostructure capable of activating the BDNF receptor TrkB and downstream signaling in primary cortical neurons in vitro. Interestingly, we found that this BDNF mimetic peptide is only active when displayed on a peptide amphiphile supramolecular nanostructure. We confirmed that increased neuronal maturation is linked to TrkB signaling pathways by analyzing the phosphorylation of downstream signaling effectors and tracking electrical activity over time. Furthermore, three-dimensional gels containing the BDNF peptide amphiphile (PA) nanostructures encourage cell infiltration while increasing functional maturation. Our findings suggest that the BDNF mimetic PA nanostructure creates a highly bioactive matrix that could serve as a biomaterial therapy in injured regions of the CNS. This new strategy has the potential to induce endogenous cell infiltration and promote functional neuronal maturation through the presentation of the BDNF mimetic signal.",

keywords = "BDNF mimetic, TrkB receptor, maturation, nanostructure, neurons, peptide amphiphile",

author = "Edelbrock, {Alexandra N.} and Zaida {\`A}lvarez and Dina Simkin and Timmy Fyrner and Chin, {Stacey M.} and Kohei Sato and Evangelos Kiskinis and Stupp, {Samuel I.}",

note = "Funding Information: *S.I. Stupp. E-mail: [email protected]. ORCID Timmy Fyrner: 0000-0003-2119-9883 Kohei Sato: 0000-0002-8948-8537 Samuel I. Stupp: 0000-0002-5491-7442 Present Address ∇Department of Life Science and Technology, Tokyo Institute of Technology, Yokohama 226−8501, Japan Author Contributions ◆A.N.E. and Z.A. contributed equally to this work. A.N.E. synthesized materials, A.N.E. and Z.A. designed and performed experiments, analyzed data, and wrote the manuscript. D.S. performed all electrophysiology experiments and analyzed data. S.M.C. performed analysis experiments and analyzed data. T.F. and K.S. carried out synthetic work, material characterization, and took part in discussions. E.K. supervised the research. S.I.S. wrote the manuscript and supervised the research. The manuscript was written through contributions of all authors. All authors have given approval to the final version of the manuscript. Funding This work was supported by National Institutes of Health/ National Institute of Biomedical Imaging and Bioengineering Award Number 5R01EB003806-07 and by the Center for Regenerative Nanomedicine at the Simpson Querrey Institute at Northwestern. The E.K. lab is supported by research grants from the Les Turner ALS Foundation, the Muscular Dystrophy Association, the Dravet Foundation and NIH/NINDS R01NS104219. E.K. is a Les Turner ALS Center Investigator. A.N.E. and S.M.C. received graduate research fellowships through the National Science Foundation. Z.A. received postdoctoral support from the Beatriu de Pino{\'s} Fellowship 2014 BP-A 00007 (Ageǹcia de Gesti{\'o} d{\textquoteright}Ajust Universitaris i de Recerca, AGAUR) and the PVA Grant # PVA17_RF_0008 from the Paralyzed Veterans of America (PVA) Research Foundation. Notes The authors declare no competing financial interest. Funding Information: We are grateful to the following core facilities at Northwestern University: the Peptide Synthesis Core and the Analytical Bionanotechnology Equipment Core both at the Simpson Querrey Institute for BioNanotechnology. The U.S. Army Research Office, the U.S. Army Medical Research Materiel Command, and Northwestern University provided funding to develop these facilities and ongoing support is being received from the Soft and Hybrid Nanotechnology Experimental (SHyNE) Resource (NSF NNCI-1542205). The Biological Imaging Facility (supported by the Northwestern University Office for Research), the Center for Advanced Microscopy (NCI CCSG P30 CA060553), Flow Cytometry Core (supported by Cancer Center Support Grant NCI CA060553), the Electron Probe Instrumentation Center (EPIC) facility of Northwestern University{\textquoteright}s NUANCE Center, which has received support from the Soft and Hybrid Nanotechnology Experimental (SHyNE) Resource (NSF ECCS-1542205), and this work made use of the Integrated Molecular Structure Education and Research Center (IM-SERC) at Northwestern University, which has received support from the Soft and Hybrid Nanotechnology Experimental (SHyNE) Resource (NSF ECCS-1542205); the State of Illinois and International Institute for Nanotechnology (IIN). Portions of this work were performed at the DuPont-Northwestern-Dow Collaborative Access Team (DND-CAT) located at Sector 5 of the Advanced Photon Source (APS). DND-CAT is supported by Northwestern University, E.I. DuPont de Nemours & Co., and The Dow Chemical Company. This research used resources of the Advanced Photon Source, a U.S. Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under Contract No. DE-AC02-06CH11357. The authors also thank Prof. Alfred George for the use of his facilities for all patch clamp experiments, Armando Hernandez Garcia for assistance with flow cytometry experiments, Prof. Liam Palmer for helpful discussions, as well as Jack Edelbrock and Mark Seniw for their design of schematic drawings in the manuscript. The authors would also like to thank Dr. Iva{\'n} Sasselli for his assistance with all FT-IR experiments. Publisher Copyright: {\textcopyright} 2018 American Chemical Society.",

year = "2018",

month = oct,

day = "10",

doi = "10.1021/acs.nanolett.8b02317",

language = "English (US)",

volume = "18",

pages = "6237--6247",

journal = "Nano letters",

issn = "1530-6984",

publisher = "American Chemical Society",

number = "10",

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TY - JOUR

T1 - Supramolecular Nanostructure Activates TrkB Receptor Signaling of Neuronal Cells by Mimicking Brain-Derived Neurotrophic Factor

AU - Edelbrock, Alexandra N.

AU - Àlvarez, Zaida

AU - Simkin, Dina

AU - Fyrner, Timmy

AU - Chin, Stacey M.

AU - Sato, Kohei

AU - Kiskinis, Evangelos

AU - Stupp, Samuel I.

N1 - Funding Information: *S.I. Stupp. E-mail: [email protected]. ORCID Timmy Fyrner: 0000-0003-2119-9883 Kohei Sato: 0000-0002-8948-8537 Samuel I. Stupp: 0000-0002-5491-7442 Present Address ∇Department of Life Science and Technology, Tokyo Institute of Technology, Yokohama 226−8501, Japan Author Contributions ◆A.N.E. and Z.A. contributed equally to this work. A.N.E. synthesized materials, A.N.E. and Z.A. designed and performed experiments, analyzed data, and wrote the manuscript. D.S. performed all electrophysiology experiments and analyzed data. S.M.C. performed analysis experiments and analyzed data. T.F. and K.S. carried out synthetic work, material characterization, and took part in discussions. E.K. supervised the research. S.I.S. wrote the manuscript and supervised the research. The manuscript was written through contributions of all authors. All authors have given approval to the final version of the manuscript. Funding This work was supported by National Institutes of Health/ National Institute of Biomedical Imaging and Bioengineering Award Number 5R01EB003806-07 and by the Center for Regenerative Nanomedicine at the Simpson Querrey Institute at Northwestern. The E.K. lab is supported by research grants from the Les Turner ALS Foundation, the Muscular Dystrophy Association, the Dravet Foundation and NIH/NINDS R01NS104219. E.K. is a Les Turner ALS Center Investigator. A.N.E. and S.M.C. received graduate research fellowships through the National Science Foundation. Z.A. received postdoctoral support from the Beatriu de Pinoś Fellowship 2014 BP-A 00007 (Ageǹcia de Gestió d’Ajust Universitaris i de Recerca, AGAUR) and the PVA Grant # PVA17_RF_0008 from the Paralyzed Veterans of America (PVA) Research Foundation. Notes The authors declare no competing financial interest. Funding Information: We are grateful to the following core facilities at Northwestern University: the Peptide Synthesis Core and the Analytical Bionanotechnology Equipment Core both at the Simpson Querrey Institute for BioNanotechnology. The U.S. Army Research Office, the U.S. Army Medical Research Materiel Command, and Northwestern University provided funding to develop these facilities and ongoing support is being received from the Soft and Hybrid Nanotechnology Experimental (SHyNE) Resource (NSF NNCI-1542205). The Biological Imaging Facility (supported by the Northwestern University Office for Research), the Center for Advanced Microscopy (NCI CCSG P30 CA060553), Flow Cytometry Core (supported by Cancer Center Support Grant NCI CA060553), the Electron Probe Instrumentation Center (EPIC) facility of Northwestern University’s NUANCE Center, which has received support from the Soft and Hybrid Nanotechnology Experimental (SHyNE) Resource (NSF ECCS-1542205), and this work made use of the Integrated Molecular Structure Education and Research Center (IM-SERC) at Northwestern University, which has received support from the Soft and Hybrid Nanotechnology Experimental (SHyNE) Resource (NSF ECCS-1542205); the State of Illinois and International Institute for Nanotechnology (IIN). Portions of this work were performed at the DuPont-Northwestern-Dow Collaborative Access Team (DND-CAT) located at Sector 5 of the Advanced Photon Source (APS). DND-CAT is supported by Northwestern University, E.I. DuPont de Nemours & Co., and The Dow Chemical Company. This research used resources of the Advanced Photon Source, a U.S. Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under Contract No. DE-AC02-06CH11357. The authors also thank Prof. Alfred George for the use of his facilities for all patch clamp experiments, Armando Hernandez Garcia for assistance with flow cytometry experiments, Prof. Liam Palmer for helpful discussions, as well as Jack Edelbrock and Mark Seniw for their design of schematic drawings in the manuscript. The authors would also like to thank Dr. Ivań Sasselli for his assistance with all FT-IR experiments. Publisher Copyright: © 2018 American Chemical Society.

PY - 2018/10/10

Y1 - 2018/10/10

N2 - Brain-derived neurotrophic factor (BDNF), a neurotrophin that binds specifically to the tyrosine kinase B (TrkB) receptor, has been shown to promote neuronal differentiation, maturation, and synaptic plasticity in the central nervous system (CNS) during development or after injury and onset of disease. Unfortunately, native BDNF protein-based therapies have had little clinical success due to their suboptimal pharmacological properties. In the past 20 years, BDNF mimetic peptides have been designed with the purpose of activating certain cell pathways that mimic the functional activity of native BDNF, but the interaction of mimetic peptides with cells can be limited due to the conformational specificity required for receptor activation. We report here on the incorporation of a BDNF mimetic sequence into a supramolecular peptide amphiphile filamentous nanostructure capable of activating the BDNF receptor TrkB and downstream signaling in primary cortical neurons in vitro. Interestingly, we found that this BDNF mimetic peptide is only active when displayed on a peptide amphiphile supramolecular nanostructure. We confirmed that increased neuronal maturation is linked to TrkB signaling pathways by analyzing the phosphorylation of downstream signaling effectors and tracking electrical activity over time. Furthermore, three-dimensional gels containing the BDNF peptide amphiphile (PA) nanostructures encourage cell infiltration while increasing functional maturation. Our findings suggest that the BDNF mimetic PA nanostructure creates a highly bioactive matrix that could serve as a biomaterial therapy in injured regions of the CNS. This new strategy has the potential to induce endogenous cell infiltration and promote functional neuronal maturation through the presentation of the BDNF mimetic signal.

AB - Brain-derived neurotrophic factor (BDNF), a neurotrophin that binds specifically to the tyrosine kinase B (TrkB) receptor, has been shown to promote neuronal differentiation, maturation, and synaptic plasticity in the central nervous system (CNS) during development or after injury and onset of disease. Unfortunately, native BDNF protein-based therapies have had little clinical success due to their suboptimal pharmacological properties. In the past 20 years, BDNF mimetic peptides have been designed with the purpose of activating certain cell pathways that mimic the functional activity of native BDNF, but the interaction of mimetic peptides with cells can be limited due to the conformational specificity required for receptor activation. We report here on the incorporation of a BDNF mimetic sequence into a supramolecular peptide amphiphile filamentous nanostructure capable of activating the BDNF receptor TrkB and downstream signaling in primary cortical neurons in vitro. Interestingly, we found that this BDNF mimetic peptide is only active when displayed on a peptide amphiphile supramolecular nanostructure. We confirmed that increased neuronal maturation is linked to TrkB signaling pathways by analyzing the phosphorylation of downstream signaling effectors and tracking electrical activity over time. Furthermore, three-dimensional gels containing the BDNF peptide amphiphile (PA) nanostructures encourage cell infiltration while increasing functional maturation. Our findings suggest that the BDNF mimetic PA nanostructure creates a highly bioactive matrix that could serve as a biomaterial therapy in injured regions of the CNS. This new strategy has the potential to induce endogenous cell infiltration and promote functional neuronal maturation through the presentation of the BDNF mimetic signal.

KW - BDNF mimetic

KW - TrkB receptor

KW - maturation

KW - nanostructure

KW - neurons

KW - peptide amphiphile

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Supramolecular Nanostructure Activates TrkB Receptor Signaling of Neuronal Cells by Mimicking Brain-Derived Neurotrophic Factor

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