$56,788 Awarded to Date; $86,662: Total Intended Award; NSF Cellular and Biochemical Engineering (CBE)

Investigating the role and interplay of microenvironment, manufacturing, and metabolism on MSC production of extracellular vesicles

Principal Investigator: Sudhir Khetan, associate professor of biomedical engineering

Mesenchymal stromal cells (MSCs) are similar to stem cells in that they exhibit an ability to differentiate into many types of cells. As such, MSCs hold great potential for treating a variety of diseases. MSCs are also able to generate extracellular vesicles (EVs), which can transport bioactive molecules that prompt cellular-level responses. Though MSC-EVs have powerful immunomodulatory functions, significant challenges remain in translating these therapies into clinical practice.

There are three key obstacles this research seeks to address: 1. due to MSC heterogeneity, there are currently no reliable methods to predict how different batches of EVs will perform; 2. existing manufacturing approaches lack standardization and many studies rely on non-physiologic 2D tissue culture systems, which do not accurately mimic the body’s conditions; and 3. the scaling of EV production from 2D to 3D environments and its impact on EV quality and metabolism is not yet fully understood.

Through this research, the impact 3D hydrogel microenvironments have on MSC behavior and EV production will be investigated. This study will explore the effects of different manufacturing strategies and culture environments on the secretion of EVs, particularly focusing on the metabolic changes MSCs undergo. Prior research has shown that MSCs cultured in softer hydrogels tend to secrete more immunomodulatory factors and that cytokine priming can shift their metabolism, enhancing their function. Therefore, the goal of this project is to define metabolic pathways that influence EV production and identify how these factors can be optimized for therapeutic use.

By understanding the role of the microenvironment and metabolic processes in EV production, this research will improve the scalability, reproducibility, and effectiveness of MSC-derived EVs for clinical applications. The findings could lead to more standardized manufacturing methods, better quality control, and expanded therapeutic possibilities for MSC-derived EVs in immune disease treatment and beyond.

Collaborating institutions include Union College (Schenectady, NY), University of Georgia (Athens, GA), and University of Virginia (Charlottesville, VA). Union undergraduates will have the opportunity to gain valuable research experience and skills in cross-institutional student exchanges with the Marklein Lab at the University of Georgia and the Griffin Lab University of Virginia. Through this work each institution will empower undergraduate and/or graduate students to become members, and eventually leaders, of the STEM workforce, particularly in the area of cell manufacturing.