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[[Image:TAT.jpg|thumb|400px|right|Cell penetrating peptides, like the TAT peptide, can deliver cargos to different cells and tissues.  We have been working to evolve new specific CPPs for targeted therapeutic delivery]]


'''Protein-based Hydrogels for Bioelectrocatalysis'''


Self-assembly is an essential process for all forms of life.  For example, proteins spontaneously fold into well-defined 3-dimensional structures, and cellular organelles form that spatially segregate diverse cellular processes.  As engineers aim to create  new devices and systems at ever decreasing size scales, self-assembly processes become increaseingly attactive techniques.
'''Cell Penetrating Peptides for Targeted Drug Delivery'''


We are collaborating with Plamen Atanassov at University of New Mexico, Scott Calabrese-Barton at Michigan State University and Shelley Minteer at Saint Louis University to make improved electrodes for biofuel cells.  In a biofuel cell, redox enzymes are immobilized on electrode surfaces.  Enzymes located at the anode are able to oxidize substrates and the electrons move through an external circuit to create power.  On the cathode, other redox enzymes are able to use the electrons to reduce oxygen to water.  The net result of this process is the generation of electricity from a variety of readily available biofuel sources, with oxygen as the terminal electron acceptor.
The plasma membrane protects cells by regulating the access of molecules to the cellular cytoplasm.  Only compounds within a narrow range of size, charge, and polarity are able to passively diffuse across the membrane.  Recent advances in the developmental, molecular, and cellular biology of many devastating human diseases has led to the discovery of “mechanism-based” therapies, such as gene therapy and monoclonal antibodies, which show great promise in vitro.  However, these successes have been difficult to translate in vivo due to difficulties in targeting and delivering large exogenous therapeutics specifically to the appropriate cells.


The architecture of the electrodes is crucial for biofuel cell performance.  The enzymes on the electrodes must be positioned so that electrons can easily move between the enzymatic active site and the electorde surface (Direct Electron Transfer (DET)).  Alternatively, the enzymes can be immoblilized with redox mediators, such as osmium, that facilitate the transport of electrons from the electrodes to the enzymes (Fig. 1).  In this Mediated Electron Transport (MET) configuration, the enzyme and mediators are immobilized in a polymer matrix on the electrode surface.  While this system has been used to demonstrate impressive biofuel cell performances, it is potentially hampered by poor dispersion of the enzyme and mediator within the polymer matrix, and complex manufacturing requirments.
Cell penetrating peptides (CPPs) are short, mostly basic peptides can traverse biological membranes and enter cells.  Since their original discovery, CPPs have been used to deliver a wide variety of cargos to different cell types, including fluorochromes, enzymes, antibodies, DNA, phage particles, and nanoparticles.  It has been suggested that CPPs, such as the TAT CPP, could be valuable vectors for the delivery of therapeutic agents to different cell and tissue types in vivo.  Unfortunately, the use of CPPs in the therapeutic arena has been problematic due to their general lack of cellular specificity.


We are using biological self-assembly to improve the biofuel cell electrode construction and performance (Fig. 2).  Instead of combing enzymes and mediators in a polymer matrix, we are creating self-assembling protein-based hydrogels that intrinsically include the redox enzymes and the mediators.  In this configuration the loading of the enzyme and the mediators into the hydrogel can be finely controlled, and the hydrogel assembly process will be well-defined and repeatable.  These new bioelectrocatalytic hydrogels will have the potential to significantly improve biofuel cell performance.
An alternative approach used to ensure cell type specificity is the development of peptides that interact with specific cell surface receptors.  It has been proposed that all tissue types have specific “zip code” molecules on their vasculature.  Phage display has been used to map this system leading to the identification of homing peptides (HPs) that localize to different cell subtypes.  This represents a valuable new approach to targeting material to different cell and tissue types, but applications using HPs are limited by the fact that the HPs generally do not enter the targeted cells.
 
We are using Directed Evolution to create Specific Cell Penetrating Peptides (SCPPs).  The SCPPs will exhibit combined CPP and HP behavior.    These peptides will target specific cell and tissue types in vivo, and they will enable the delivery of therapeutic cargos, such as DNA, proteins, or other exogenous materials, to targeted cellular cytoplasms.  We are collaborating with Prof. Barclay Morrison's laboratory in the Department of Biomedical Engineering in order to create SCPPs that are specific for different brain cell types.  We are especially interested in engineering SCPPs that can target cells that are damaged following traumatic brain injury.  There is a narrow window of time following a brain injury where the targeted delivery of neurotrophic agents to injured cells could provide a significant benefit to the head injured patient.




'''Related Publications'''
'''Related Publications'''


<biblio>
<biblio>
#Paper4 pmid=19061242
#Paper7 pmid=21291271
#Paper3 pmid=18824691
#Paper6 pmid=21510821
#Paper2 pmid=18096378
#Paper5 pmid=20938973
#Paper1 pmid=17887795
#Paper4 pmid=20851169
#Paper3 pmid=20737289
#Paper2 pmid=19449355
#Paper1 pmid=19402206
</biblio>
</biblio>

Latest revision as of 12:21, 17 July 2014

Banta Lab

Protein and Metabolic Engineering

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Cell penetrating peptides, like the TAT peptide, can deliver cargos to different cells and tissues. We have been working to evolve new specific CPPs for targeted therapeutic delivery


Cell Penetrating Peptides for Targeted Drug Delivery

The plasma membrane protects cells by regulating the access of molecules to the cellular cytoplasm. Only compounds within a narrow range of size, charge, and polarity are able to passively diffuse across the membrane. Recent advances in the developmental, molecular, and cellular biology of many devastating human diseases has led to the discovery of “mechanism-based” therapies, such as gene therapy and monoclonal antibodies, which show great promise in vitro. However, these successes have been difficult to translate in vivo due to difficulties in targeting and delivering large exogenous therapeutics specifically to the appropriate cells.

Cell penetrating peptides (CPPs) are short, mostly basic peptides can traverse biological membranes and enter cells. Since their original discovery, CPPs have been used to deliver a wide variety of cargos to different cell types, including fluorochromes, enzymes, antibodies, DNA, phage particles, and nanoparticles. It has been suggested that CPPs, such as the TAT CPP, could be valuable vectors for the delivery of therapeutic agents to different cell and tissue types in vivo. Unfortunately, the use of CPPs in the therapeutic arena has been problematic due to their general lack of cellular specificity.

An alternative approach used to ensure cell type specificity is the development of peptides that interact with specific cell surface receptors. It has been proposed that all tissue types have specific “zip code” molecules on their vasculature. Phage display has been used to map this system leading to the identification of homing peptides (HPs) that localize to different cell subtypes. This represents a valuable new approach to targeting material to different cell and tissue types, but applications using HPs are limited by the fact that the HPs generally do not enter the targeted cells.

We are using Directed Evolution to create Specific Cell Penetrating Peptides (SCPPs). The SCPPs will exhibit combined CPP and HP behavior. These peptides will target specific cell and tissue types in vivo, and they will enable the delivery of therapeutic cargos, such as DNA, proteins, or other exogenous materials, to targeted cellular cytoplasms. We are collaborating with Prof. Barclay Morrison's laboratory in the Department of Biomedical Engineering in order to create SCPPs that are specific for different brain cell types. We are especially interested in engineering SCPPs that can target cells that are damaged following traumatic brain injury. There is a narrow window of time following a brain injury where the targeted delivery of neurotrophic agents to injured cells could provide a significant benefit to the head injured patient.


Related Publications

  1. Gao S, Simon MJ, Hue CD, Morrison B 3rd, and Banta S. An unusual cell penetrating peptide identified using a plasmid display-based functional selection platform. ACS Chem Biol. 2011 May 20;6(5):484-91. DOI:10.1021/cb100423u | PubMed ID:21291271 | HubMed [Paper7]
  2. Kang WH, Simon MJ, Gao S, Banta S, and Morrison B 3rd. Attenuation of astrocyte activation by TAT-mediated delivery of a peptide JNK inhibitor. J Neurotrauma. 2011 Jul;28(7):1219-28. DOI:10.1089/neu.2011.1879 | PubMed ID:21510821 | HubMed [Paper6]
  3. Gao S, Simon MJ, Morrison B 3rd, and Banta S. A plasmid display platform for the selection of peptides exhibiting a functional cell-penetrating phenotype. Biotechnol Prog. 2010 Nov-Dec;26(6):1796-800. DOI:10.1002/btpr.490 | PubMed ID:20938973 | HubMed [Paper5]
  4. Simon MJ, Kang WH, Gao S, Banta S, and Morrison B 3rd. Increased delivery of TAT across an endothelial monolayer following ischemic injury. Neurosci Lett. 2010 Dec 3;486(1):1-4. DOI:10.1016/j.neulet.2010.09.029 | PubMed ID:20851169 | HubMed [Paper4]
  5. Simon MJ, Kang WH, Gao S, Banta S, and Morrison B 3rd. TAT is not capable of transcellular delivery across an intact endothelial monolayer in vitro. Ann Biomed Eng. 2011 Jan;39(1):394-401. DOI:10.1007/s10439-010-0144-x | PubMed ID:20737289 | HubMed [Paper3]
  6. Simon MJ, Gao S, Kang WH, Banta S, and Morrison B 3rd. TAT-mediated intracellular protein delivery to primary brain cells is dependent on glycosaminoglycan expression. Biotechnol Bioeng. 2009 Sep 1;104(1):10-9. DOI:10.1002/bit.22377 | PubMed ID:19449355 | HubMed [Paper2]
  7. Gao S, Simon MJ, Morrison B 3rd, and Banta S. Bifunctional chimeric fusion proteins engineered for DNA delivery: optimization of the protein to DNA ratio. Biochim Biophys Acta. 2009 Mar;1790(3):198-207. DOI:10.1016/j.bbagen.2009.01.001 | PubMed ID:19402206 | HubMed [Paper1]

All Medline abstracts: PubMed | HubMed

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