Premise:
Adequate glucose uptake and release is one of the key mechanisms to fuel cells with energy and thereby maintain energy balance, metabolic health and body functions. High glucose levels in the bloodstream is directly linked with insufficient insulin release and/or responses, which ultimately lead to cell starvation and detrimental health conditions. Diabetes is known for insufficient insulin production due to beta-cell destruction during autoimmune attacks (Type 1 diabetes), or inability for cells to respond to insulin, often caused by severe obesity-induced insulin resistance (Type 2 diabetes). Despite the increased biomedical research efforts to overcome diabetes, and the increased availability of effective drugs for chronic treatment of disease state, there is no definitive cure or fully established molecular mechanism that can help identify specific targets for development of long-term sustained therapeutic approaches. The Elizondo Lab is working to delineate the immunometabolic mechanisms implicated in the development of diabetes and metabolic dysfunction, which concomitantly serve as a training tool for both undergraduate and graduate students at UMBC.
Undergraduate Students’ Project:
Title: Identify Candidate Genes Implicated in Deficient Immunometabolic Responses.
This project is designed for undergraduate students with the goal of granting them with wide hands-on experience on genetic, proteomic and cellular techniques. We aim to reinforce coursework knowledge foundations through implemented wet lab experience, while equipping students with required skillsets for successful transition into competitive PhD programs. Briefly, students will have the opportunity to explore available scRNAseq data derived from non-obese type 1 diabetic (T1D) or diet-induced obese type 2 diabetic (T2D) mice and select deregulated genes with potential roles in diabetes, chronic inflammation and/or metabolic dysfunction. The laboratory will train students to study selected candidate genes in either T1D or T2D, via corroborative expression assays such as qPCR, western blot, fluorescence and bright field microscopy, coupled with cell culture, RNAi, flow cytometry and ELISA techniques for gene characterization.
Outcomes of this project will be presented at internal and national scientific conferences, generate co-authorship manuscripts and will serve as potential foundations for long-term projects in the laboratory.
Graduate Students’ Projects:
Title: Delineate Molecular Immunometabolic Mechanisms Implicated in Diet-Induced Obesity Metabolic Dysfunction.
Obesity has increased in prevalence steadily over the last decade. Importantly, it can be characterized as metabolically healthy or unhealthy, with the unhealthy state associated with adipose tissue hypertrophy, restrained normal adipocyte proliferation, infiltration of macrophages, skewed inflammatory responses, and excess collagen deposition coupled with fibrosis. Invariably, this leads to adipose tissue dysfunction and ectopic lipid deposition, along with increased risk for type 2 diabetes (T2D), cardiovascular dysfunction, certain types of cancers and fatty liver disease. Although the interplay between infiltrating myeloid cells, altered inflammatory states and adipose tissue dysfunction during unhealthy obese states has been well described, the underlying mechanisms of these immunometabolic activities are not well understood. This project is designed for incoming graduate students interested in delineating the signaling pathways associated with diet-induced obesity metabolic dysfunction/T2D. We will study selected genes of interest in conditionally knockout (cKO) mice, generated via in vivo lentivirus-mediated gene silencing approach, or by transgenic mice breeding approaches. Assigned projects will involve physiological assessments of cKO mice under diet-induced obesity treatment by continuous body weight monitoring, glucose/insulin tolerance testing, energy expenditure evaluation and DEXA body scanning. Further assessments, include ability to monitor mice’s changes in peripheral tissue health and inflammatory responses through a combination of adipocyte or myeloid/lymphoid cell culture assays. These assays will be complemented with multiplex readout indices, such as Luminex, flow cytometry, qPCR, fluorescence microscopy and western blot techniques.
Title: Study of Myeloid Cell Mechanisms Implicated in Type 1 Diabetes Autoimmunity.
Approximately 1 million people nationwide have type 1 Diabetes (T1D), which is a chronic condition well characterized by the body’s immune system mistakenly destroying pancreatic insulin-producing beta cells, thereby leading to insufficient insulin production; current therapies include exogenous insulin administration. Unfortunately, this approach does not prevent ongoing autoimmune destruction and requires a life-long dependency on insulin treatment. This project is designed for incoming graduate students interested in delineating the molecular mechanisms involved in the development of autoreactive myeloid cell populations in T1D, which can further serve as key targets to alleviate beta cell autoreactivity. To do so, we will study selected candidate immunomodulatory genes in non-obese diabetic conditional knockout (NOD-cKO) mice generated via in vivo lentivirus-mediated gene silencing approach, or by transgenic mice breeding approaches. Assigned projects will evaluate reduced T1D pathology by measuring reduced diabetes onset, restoration of normal insulin production, restrained infiltration of autoreactive T cells into the pancreas and induction of T regulatory cell subsets. Success of the studies will generate tolerogenic dendritic cells to restrain autoimmune events, thereby providing a potential immunotherapeutic approach addressing the challenges faced with current exogenous insulin therapies.
Our goal is to train graduate students with required knowledge and technical foundations required for a successful career path in immunometabolism. Outcomes of this project will be presented at internal and national scientific conferences, first author manuscripts and will serve as a premise for successful attaining of graduate students’ fellowship applications.