BioE PhD Thesis Presentation - Abir K. Muhuri

Advisor:

Prof. Susan N. Thomas – Georgia Institute of Technology, School of Mechanical Engineering

Committee Members:

Prof. Andres Garcia – Georgia Institute of Technology, School of Mechanical Engineering

Prof. Levi Wood - Georgia Institute of Technology, School of Mechanical Engineering

Prof. Sarwish Rafiq – Emory University School of Medicine, Department of Hematology and Medical Oncology

Prof. Yunus Alapan – University of Wisconsin-Madison, Department of Mechanical Engineering

Microfluidic-Enabled Adhesion Analysis of Therapeutic  T cells for Lymph Node Homing Potential

Adoptive cell therapies, including tumor-infiltrating lymphocytes (TILs) and chimeric antigen receptor (CAR)-T cells, have transformed cancer immunotherapy but remain limited by inefficient trafficking of transferred cells to therapeutically important tissues. In addition to direct tissue infiltration, effective T-cell therapies often depend on access to lymph nodes, which serve as specialized niches for immune surveillance, antigen presentation, T-cell maintenance and therapeutic activity across diverse disease contexts. Entry into lymph nodes is regulated by specialized vascular structures known as high endothelial venules (HEVs); however, current manufacturing approaches provide limited means to identify or enrich therapeutic T-cell populations with enhanced lymph node homing potential.

To address this gap, this dissertation develops microfluidic adhesion chromatography platforms that recreate vascular microenvironments under physiological flow to functionally assess T-cell trafficking potential and isolate cells based on adhesive behavior. Using expanded murine CD8⁺ T cells as a model of TIL products, we first demonstrate that adhesion to the HEV-associated adhesion molecule, nepmucin, identifies cell populations with enhanced early lymph node accumulation together with phenotypes associated with lymphoid cellular states. These findings were established using tumor-draining lymph nodes in a murine breast cancer model, providing a clinically relevant context for studying lymph node-directed trafficking during adoptive cell therapy. This framework is further extended to human CAR-T cells to compare adhesive phenotypes across lymph node- and tumor-mimicking vascular substrates. An integrated photoactivation strategy further distinguishes adhesive heterogeneity at both bulk-population and single-cell resolution.

Together, this work demonstrates that functional adhesion under physiological flow can distinguish T-cell populations with distinct tissue-trafficking behaviors that are not readily captured by conventional phenotypic analyses alone. By linking adhesive behavior with lymph node homing potential, this dissertation presents new tools for characterizing and enriching next-generation cellular immunotherapies with improved targeting of therapeutically relevant tissues.