Metabolomic and Proteomic Composition of Human Tears using MPLEx and nanoproteomics
EMSL Project ID
60521
Abstract
BackgroundThe Aicher lab has an NEI-funded grant (R61 EY032468) to identify protein biomarkers in human tears. Specifically, we will look at patients who have LASIK surgery and compare tear proteins in patients who do or do not develop persistent pain after surgery. We recently saw a study in the literature where both proteomics and metabolomics was conducted on the same sample (Rossi C, et al. 2019 Int J Mol Sci). We are hoping to expand the scope of our current study and include metabolomic and/or lipidomic analyses of our patient tear samples.
Specific Pilot Study Plan:
The Aicher lab will submit 4 samples containing tear fluid on paper Schimer’s strips. Samples will be in individual tubes, have been stored at -80oC and will be shipped on dry ice. These samples are from 2 individuals, with 2 samples (left and right eye) from each person. The amount of fluid on each sample is variable, since fixed collection time (5 minutes) is used for each case.
We will apply a microscale implementation of the MPLEx approach developed at PNNL combined with LC-based metabolomics and nanoproteomics to provide in-depth molecular information for both omes from a single sample of human tears, as well as provide preliminary analysis of data quality and general comparisons between samples (e.g. total proteins recovered; degree of consistency between samples; data quality checks). We need sufficient data to demonstrate the approach is feasible and to convince NIH that this approach will provide further validation of prior pilot studies conducted in OHSU proteomics lab; and that we may discover additional biomarkers by looking for metabolites and lipids.
Hypothesis:
We expect within-subject variation to be less than between-subject variation. One person has had prior eye surgery and one has not; we expect potential proteomic differences due to prior surgery compared to no-surgery subject.
If pilot project is successful, we will submit a request to NIH to develop a subaward on our current grant to conduct similar analysis across groups of post-surgical patients. We will work with PNNL team to develop specific, detailed budget request prior to August 1, 2022.
The goal of this project is to leverage ultra-sensitive multi-omics capabilities developed at PNNL to provide an in-depth characterization of the proteome and metabolome from a single sample of human tears.
Project Details
Start Date
2022-06-17
End Date
N/A
Status
Active
Released Data Link
Team
Principal Investigator