Team:Nanjing high school/Description
Project Description
1. Why do we want to do this project?
Inflammatory neurodegeneration is
neuronal death/loss
caused by inflammation and is a process (like energy depletion, protein aggregation, and excitotoxicity)
contributing to neuronal loss or dysfunction in many different neurological diseases. Many severe
degenerative diseases are connected to the inflammatory reaction in our brain. For example, Alzheimer's
Disease, Amyotrophic Laternal Sclerosis, and stroke. These diseases are very terrible, they can only be
prevented and delayed, but not cured. there is evidence that blocking inflammation can either delay onset or
reduce symptoms. Through this project, our goal is to find a compound that may inhibit the inflammatory
response in the brain. Meanwhile, we hope that our project will raise more people’s awareness to these
diseases.
2. The theory of the project
Microglia is a type of resident
immune cell that balances
inflammatory function parts of the brain. But once they become overly inflammatory, they would differentiate
into dendritic cells, which in turn may affect many other cells in the brain. This would cause apoptosis
injury of other cells and interfere with the balance of microenvironment in the brain.
Once the microglia are activated,
they secrete
pro-inflammatory cytokines, including TNFα, IL-1β, and
IL-6. Most of the expression changes are a result of activation of the transcription factor NF-κB via
phosphorylation-induced activation of IκB kinase. Notably, PPM1A activation could inhibit inflammatory
response in peripheral macrophages, and directly dephosphorylate RelA subunit at residues S536 and S276,
which is necessary for transcriptional competence of NF-κB.
3. What do we want to get from the project
Based on the above, we expressed
and purified the
recombinant human PPM1A protein, and constructed a PPM1A enzyme activity screening platform for screening
out PPM1A agonists from FDA-approved drugs library. In this experiment, starting with culturing PPM1A
protein, the purified protein was subjected to enzyme activity experiment to evaluate the activation
efficiency of the compound, then BV-2 cells (mouse microglia cell line) were cultured, and finally
quantitative PCR (Q-PCR) experiment was performed to detect the transcription level of the inflammatory
factor, which was calculated to determine the effectiveness of the lead compound, and finally an effective
lead compound that can be used for follow-up research is obtained.
Reference:
1) Cohen, P.T.W. Overview of protein serine/threonine phosphatases. Protein
Phosphatases 2004.
2) Smith SR, Schaaf K, Rajabalee N, et al. The phosphatase PPM1A controls
monocyte-to-macrophage differentiation. Sci Rep. 2018; 8(1):902.
3) Hansen DV, Hanson J E, Sheng M. Microglia in alzheimer's disease. J Cell Biol
2017; 217:459-472.
4) Paolicelli, R. C., Jawaid, A., Henstridge, C. M., Valeri, A., Merlini, M.,
Robinson, J. L., Lee, E. B., Rose, J., Appel, S., Lee, V. M. Y., Trojanowski, J. Q., Spires-Jones, T.,
Schulz, P. E. & Rajendran, L. 2017. Tdp-43 Depletion In Microglia Promotes Amyloid Clearance But Also
Induces Synapse Loss. Neuron, 95, 297-308.E6. doi: 10.1016/j.neuron.2017.05.037
5) Wilson, R. S., Yu L Fau – Trojanowski, J. Q., Trojanowski Jq Fau – Chen, E.-Y.,
Chen Ey Fau – Boyle, P. A., Boyle Pa Fau – Bennett, D. A., Bennett Da Fau – Schneider, J. A. & Schneider, J.
A. 2013. Tdp-43 Pathology, Cognitive Decline, And Dementia In Old Age. JAMA Neurol, 70, 1418-1424. doi:
10.1001/jamaneurol.2013.3961.