17urn:lsid:arphahub.com:pub:8E638694-B4E0-570A-856A-746FF325BF6BResearch Ideas and OutcomesRIO2367-7163Pensoft Publishers10.3897/rio.2.e926892685485PhD Project PlanBiochemistryDiagnostic medicineMedicine & Health sciencesMolecular biologySurgery & Invasive treatmentBi-directional immuno-modulation by Matrix Metalloproteinase-7 (MMP-7) and A Disintegrin And Metalloproteinase-17 (ADAM-17) as transplantation rejection-tolerance spectrumChenRobert Jeenchenrjcc@ntu.edu.twhttps://orcid.org/0000-0001-6892-0602598Cardiovascular Surgery, Taipei Tzuchi Hospital, New Taipei City, TaiwanCardiovascular Surgery, Taipei Tzuchi HospitalNew Taipei CityTaiwan
Corresponding author: Robert Jeenchen Chen (rjcc@ntu.edu.tw).
Academic editor:
2016260520162e9268E8359944-B06B-5099-AC8B-40FED3DDF4AC34418518052016Robert ChenThis is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Theoretically, Matrix Metalloproteinase-7 (MMP-7) leads to allograft rejection, and A Disintegrin and Metalloproteinase-17 (ADAM-17) results in allograft tolerance. The research proposal utilizes the animal model of knock-out mice to perform transplant surgery and then detect or measure allograft rejection by selected serum biomarker and tissue typing. Comparisons will be made for knock-out, wild-type, and wild-type treated with proteinase inhibitors. Methodological and theoretical details will be elucidated and revised as the research goes on.
Matrix Metalloproteinase-7 (MMP-7)A Disintegrin And Metalloproteinase-17 (ADAM-17)transplantationrejectionallograftPending.Funding programe
Pending.
Hosting institution
Biochemistry & Molecular Biology, National Taiwan University College of Medicine, Taipei, Taiwan;
Cardiovascular Surgery, Taipei Tzuchi Hospital, New Taipei City, Taiwan;
Tzuchi University College of Medicine, Hualian, Taiwan.
Ethics and security
Institutional Animal Care and Utilization Committee (IACUC) will review and approve the proposal. The whole research will comply with the guidelines with IACUC approval.
Conflicts of interest
None.
Objectives, Concept and Approach
Objectives:
Transplant immunity spectrum measures from none (tolerance) to full (rejection);
Immuno-modulator drugs that does not only suppress but also enhance immunity in transplant.
Concept
Currently, we only have immuno-suppressants (Neoral; Tacrolimus; Cellcept, etc.) “Sew-saw” roles of MMP-7 and ADAM-17 can up- (Gill et al. 2016, Li et al. 2002, Ra et al. 2009, Yu et al. 2002, Yu and Woessner 2001) and down-regulate immunity (Esteso et al. 2014, Isernhagen et al. 2016, Platt et al. 2015, Ra et al. 2009) more timely, to better overcome infection events in post-transplant patients.
We can quantify post-transplant patients’ immunity status and adjust bi-directionally the immunity levels whenever rejection or infection occurs.
Current clinical practice only has immuno-suppressant, such as Neoral, Tacrolimus, Cellcept, but no immuno-enhancers. Infection becomes worrisome whenever immune is over-suppressed.
Prior studies have shown that MMP-7 promotes rejection (immuno-enhancer) and ADAM-17 promotes tolerance (immuno-suppressant) (Yu et al. 2002, van Zandbergen et al. 1999).
Our lab have established animal model of knockout (KO) mice (MMP-7; ADAM-17), and qualified proteomics/degradomics facilities.
The immuno-modulatory mechanism of MMP-7 and ADAM-17 in transplant is still a hypothesis (Yu and Woessner 2000, Yu et al. 2012)
Approach
Animal model of transplant surgery; protease epitopes quantification and analysis; proteomics, degradomics; immune synapses networks.
Project description
Animal model: protease gene-knockout (KO) and wild-type (WT) mice;
Surgery: transplantation of s specified organ, such as kidney, heart, aorta, lungs, etc.
Measurement of rejection (cellular, humoral, acute, subacute, chronic?) by serum biomarker and tissue typing;
Data collection and analysis: proteomics, degradomics, statistical modeling;
Drafting of immune synapse networks from MMP-7 and ADAM-17.
ImplementationMethodology
Transplant surgery on KO- and WT-mice;
Measurement of rejection by serum marker and tissue typing;
Degradomics analysis;
Repeat the above steps 1 to 3 on mice treated with protease inhibitors;
Repeat the experiment on MMP-7 and ADAM-17 mice;
Repeat the whole experiment for replicability and reproducibility;
Results interpretatiom and report writing.
Work plan
The research will be carried out with the mentorship of Dr. Wei-Hsuan Yu, PhD, Laboratory of Matrix Biology, Institute of Biochemistry and Molecular Biology, College of Medicine, National Taiwan University, Taipei, Taiwan.
Decision of which organ to transplant on mice;
Decision of which serum marker and tissue typing to use;
Repeat the experiment for MMP-7 and ADAM-17;
Repeat the whole experiment for replicability and reproducibility;
Analysis and interpretation;
Report writing and publication.
Details for replicability and reproducibility
To achieve replicability and reproducibility, we will repeat the same surgery, sample collection, lab analysis, and statistical analyses on at least two individual mice. If time and funding permit, we will repeat the experiment on more mice.
Timeline
Systematic review of knowledge: 2 months;
Animal model for transplant and rejection: 4 months;
Experiment: 6 months;
Troubleshooting (buffer): 2 months;
Interpretation: 4 months;
Thesis writing: 4 months.
Data resources
Matrix biology database (Launay et al. 2015)
Experimental data obtained from our animal model and degradomics will be compared and explored with the use of MatrixDB (Launay et al. 2015)
Expected results and impact
Expected Results
MMP-7 shedding products are detected in transplant rejection, or MMP7-wild type (WT) mice;
ADAM-17 shedding products are detected in transplant rejection, or ADAM17-KO mice;
MMP7-KO mice have no rejection, similar to MMP7-WT mice treated with MMP7-inhibitor.
ADAM17-WT mice treated with ADAM17-inhibitor have minimal rejection.
Impact
MMP-7 inhibitor will suppress rejection and ADAM-17 inhibitor will enhance immunity. Whenever there is suspicious or confirmed rejection event, we can use MMP-7 inhibitor to rescue allograft. If there are signs of infection, we can use ADAM-17 inhibitor to conquest pathogens by upgraded immunity.
Acknowledgements
Special thanks to animal and lab technicians for their efforts.
Funding program
Pending.
Project
Shedding of MMP-7 and ADAM-17 will be analyzed and reported. Effect of proteases and their inhibitors on KO-and WT-mice on detected ejection will be assessed.
Hosting institution
Biochemistry & Molecular Biology, National Taiwan University College of Medicine, Taipei, Taiwan;
Cardiovascular Surgery, Taipei Tzuchi Hospital, New Taipei City, Taiwan;
Tzuchi University College of Medicine, Hualian, Taiwan.
Ethics and security
Institutional Animal Care and Utilization Committee (IACUC) will review and approve the proposal. The whole research will comply with the guidelines with IACUC approval.
Author contributions
Chen RJ: systematic review, study design, methodology, proposal writing; transplant surgery on mice, data sampling, lab analysis, statistical analysis.
Conflicts of interest
None.
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