UROP Research Mentor Project Submission Portal: Submission #756

Submission information
Submission Number: 756
Submission ID: 14406
Submission UUID: 2c25e436-1217-48c7-a5b2-47830dc46ae8

Created: Tue, 08/13/2024 - 10:35 AM
Completed: Tue, 08/13/2024 - 10:35 AM
Changed: Fri, 10/04/2024 - 12:20 PM

Remote IP address: 144.174.212.21
Submitted by: Anonymous
Language: English

Is draft: No
Primary Research Mentor Name Dr. Helene Tigro
Research Mentor Preferred Pronouns she her
When potential research assistants are reaching out via email, what is your preferred honorific?
Contact Email (FSU Email if affiliated) helene.tigro@med.fsu.edu
Position Title Post Doc
FSU College (if applicable) Medicine
FSU Department or Non-FSU Organization Affiliation Biomedical Sciences
Headshot (optional)
Research Assistant Supervisor (if different from above) Dr. Zucai Suo
Research Assistant Supervisor Preferred Pronouns
Research Assistant Supervisor Preferred Honorific? Dr.
Contact Email (FSU Email if affiliated) zucai.suo@med.fsu.edu
Name of Other Faculty/Collaborator(s) (if applicable)
Other Faculty/Collaborator(s) Preferred Pronouns
Other Faculty/Collaborator(s) Preferred Honorific?
Contact Email (FSU Email if affiliated)
Title of the Project Characterization of recombinant mouse Interleukin 4
Project Keywords protein purification, HPLC, gel electrophoresis, protein labeling, EV loading, mammalian cells
Are you currently looking for research assistants? No
Number of Research Assistants Needed 2
Relevant Research Assistant Major(s) Biochemistry or Biological Science
Project Location: On FSU Main Campus
If the project location is off campus, does the research assistant(s) need to provide their own transportation?
Please select the choice that most accurately describes your expectations for the research assistant(s): In-person
Approximately how many hours a week would the research assistant(s) need to work? 10 hours a week
Roughly what time frame do you expect research assistant(s) to work? During business hours
Overall Research Project Description Spinal cord injury (SCI) is a medical condition resulting in morbidity and permanent disability that can be caused by a major trauma directly to the spinal cord or vertebrae, degenerative disease or spinal infection. SCI arises from the disruption of nerve axons running through the spinal cord which leads to the loss of sensory and motor function. Following the initial trauma, a cascade of secondary events will transpire, including excessive proinflammatory response that includes the activation of CNS resident microglia to pro-inflammatory M1 phenotype, that release inflammatory cytokines, chemokines, and reactive oxygen species (ROS), contributing to further neuronal death and tissue damage. M1 phenotype is dominated by NF-kB that increases phosphodiesterase-4 (PDE4) expression, which rapidly hydrolyses cAMP. cAMP is the main regulator maintaining innate immune cell homeostasis and preventing the M1 phenotype, especially in the presence of anti-inflammatory interleukin 4 (IL4). Elevated cAMP and IL4 levels activate the anti-inflammatory M2 phenotype, that secrete factors associated with tissue repair, wound healing, and neuroprotection. Stimulating intracellular cAMP and IL4 activity remarkably promotes neuronal survival, regeneration, and enhances intrinsic axonal growth.
We propose to investigate the possibility of using engineered extracellular vesicles for therapeutic conversion that could be utilized in SCI. In vitro and in vivo experimental SCI models have shown that combining cAMP elevation with anti-inflammatory cytokines, such as IL4, phenotypically converts pro-inflammatory M1 microglia to M2, which enhances peripheral nerve recovery and regeneration, suppresses inflammation, and stimulates tissue repair and wound healing.
In the experimental set-up, mouse IL4 will be expressed as recombinant murine IL4-R9 in E. coli BL21(DE3) cells and purified by column chromatography. Purified IL4 will be analyzed by gel electrophoresis (SDS-PAGE and native PAGE) and circular dichroism (CD) spectroscopy. Purified IL4-R9 will be loaded into the MSCEVs and verified by Cys5-labelling, and immunostaining with Alexa Fluor 488-labeled anti-IL4 antibody, using TIRF and confocal microscopy.
Mouse microglia cells will be cultured in appropriate conditions and microglial classic activation will be achieved by LPS or IFN-γ induction, after which the expression of pro-inflammatory markers (iNOS, COX-2, IL-1β, TNF-α) is determined in cell lysates by Western blot or ELISA analysis.
Research Tasks Literature review, conducting laboratory experiments (recombinant protein expression in E. coli, protein purification using different methods such as HPLC, protein analysis using gel electrophoresis, protein labeling using different dyes, mammalian cell work etc), data collection, data analysis.
Skills that research assistant(s) may need: Good pipetting skills are required. Good time management skills are required. Good work ethics is required.
Mentoring Philosophy My mentoring philosophy is creating an interactive environment for learning, promoting learning through inquiry and laboratory experiments, and sharing my own experience with the mentee's.
Please provide a link to your publications, a video clip, or a website for your research project (if applicable): https://med.fsu.edu/suolab/publications
Please add any additional information here (if applicable):
Are you interested in participating in the UROP Research Mentor Roundtable? No
Roundtable times and Zoom links
Mentor Handbook, FAQs, and Communication Yes
UROP Performance Evaluation Yes
Materials Grant Yes
UROP Poster Presentation Yes
Year 2024
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