Study Rationale:
This project will standardize common biological assays to provide reproducible results related to immune cell function for patients with Parkinson’s Disease (PD). This involves analysis of peripheral blood mononuclear cells (PBMCs) from healthy controls in several biological assays, which will, in the future, be extended to analysis of PBMCs from PD patients. The goal will be to generate protocols for labs not familiar with such assays so they may conduct analysis based on standardized protocols. This work is being done with three other investigators: Dr. Malu Tansey, Dr. Bradlee Heckman and Dr. Elizabeth Bradshaw.
Hypothesis:
Making recommendations for critical assays to obtain a comprehensive picture of immune system function/dysfunction in PD will allow for reproducible results and confidence in results obtained by multiple labs.
Study Design:
This coordinated study plan being conducted by four separate labs consists of validation and cross-correlation of immunophenotypic and effector function assays in PBMCs, enriched subsets of immune cells (monocytes, B-cells, T-cells), and monocyte-derived macrophages (MDMi), as well as transcriptional profile comparison of primary monocytes and MDMi cells. Four major categoriesof experiments will be conducted: 1. Immune cell identification and characterization; 2. Immune effector functional assays; 3. Transcriptional assays on monocytes and MDMis; and 4. Cross-correlational analyses of transcriptional profiles on monocytes with MDMi cells.
Impact on Diagnosis/Treatment of Parkinson’s disease:
First, establishes the reproducibility (operator- and site-independence) and robustness of key immune assays to investigators in the Parkinson’s field wanting to conduct deep immunophenotyping of human peripheral blood for cellular biomarker studies, patient stratification for trials, and response monitoring during clinical trials. Second, it utilizes the same subset of assays in conjunction with single-cell transcriptional profiling of paired primary monocyte and MDMi samples for cross-correlational analyses of phenotypes, which will reveal the extent to which MDMi respond similarly or in a distinct and more graded fashion relative to their parent monocytes.
Next Steps for Development:
Successful completion of the proposed studies lays the foundation for future studies focused on other important functional immune cell comparisons, biomarker validation, patient stratification for clinical trials, and response monitoring during clinical trials.