Inflammation and immunological mechanisms are at the epicenter of all disease mechanisms. High-parameter flow cytometry is a powerful analytical tool that will enable the Birmingham VA Medical Center (BVAMC) researchers to identify and analyze distinct phenotypes in the very heterogeneous populations of cells derived from various sources of animal models and human specimens. There is no high dimensional flow cytometer at the BVAMC or the affiliate, the University of Alabama at Birmingham (UAB), yet there is an acute need for BVAMC investigators to have access to such an instrument. The BD FACSymphony features an ultra-quiet VPX (also known as VITA 46), which is the next generation of ruggedized compact embedded electronic systems that supports up to 50 high-performance photomultiplier tubes (PMTs) and improves detection sensitivity to enable the user to identify and analyze rare cell subtypes in advance. In order to understand the causes of diseases such as cancer, kidney disease, heart failure, diabetes, or degenerative diseases of the nervous system, it is important to define the cell population not at just one point in time but at multiple times in a disease process. Multiparameter analysis improves the efficiency by requiring smaller sample volume and by increasing sample throughput. This is critical for human samples available at limited amounts from human tissue and body fluids. Increased number of parameters facilitates improved simultaneous functional characterization of cells by intracellular staining of phosphorylated targets, cytokine and growth factor production, analysis of cell cycle, apoptosis, mitochondria membrane polarization, etc. These capacities are critical for understanding a broad spectrum of single cells or fluids derived from VA patients in active studies by the BVAMC investigators. For example, in Dr. Mountz's work in Systemic Lupus Erythematosis (SLE), the BD FACSymphony will be used to specifically and very individually determine the pathogenic phenotype of autoreactive B cells and other cells in SLE patients and enable tailored precision medical therapy approaches. The important precision medicine approaches can be best utilized and targeted when the abnormal cell subpopulation in a specific individual and disease can be identified, and specifically suppressed. This is of considerable importance to facilitate the development of effective therapies for VA patients who have developed inflammatory and autoimmune diseases presented in this proposal. In another novel approach, Drs. Dell'Italia and Gaggar will utilize the BD Symphony to track the secretion of exosomes in acute and chronic lung and heart injury. Exosomes are membrane-bound structures secreted by a wide range of mammalian cell types carrying and transporting cellular cargo in normal and pathologic states. The ability to utilize flow cytometry to directly interrogate individual exosomes and their content is tremendously valuable and offers insight to the pathogenic features of a given exosome population. In each of the projects presented in this proposal, there is a similar targeted cellular approach for cardiac, pulmonary, vascular kidney disease and cancer, as well as immunological disease of the gastrointestinal tract and nervous system. The application of this exciting new technology will enhance funding capabilities and most importantly our research goals and mission of Veteran-centric medical care.
The Birmingham VA Medical Center (BVAMC) has always embodied an interdisciplinary research program that is Veteran centered, results-driven, and disease oriented. The current proposal is the first step in our forward thinking delivery of health care in the burgeoning field of precision medicine. We believe that these approaches can be best utilized when the abnormal subpopulation in a specific individual can be identified and specifically targeted with rational therapy. By understanding these diseases at the cellular level at various time points from patient blood, body fluids or tissue biopsies, we are working for a future in which these diseases can be prevented or treated. The current request is for a new cutting-edge technology that can specifically and very individually determine the pathogenic phenotype of various cell types, including inflammatory, malignant, stem cells, and microvesicles in patients that enable potentially novel tailored precision medical therapy approaches.
