Collective Invasion In Heterogeneous Bladder Cancer Tumors: A Biomanufacturing Approach
Open Access
- Author:
- Torab, Peter
- Graduate Program:
- Mechanical Engineering
- Degree:
- Doctor of Philosophy
- Document Type:
- Dissertation
- Date of Defense:
- October 07, 2020
- Committee Members:
- Pak Kin Wong, Dissertation Advisor/Co-Advisor
Pak Kin Wong, Committee Chair/Co-Chair
Jing Du, Committee Member
Tak Sing Wong, Committee Member
William O Hancock, Outside Member
Daniel Connell Haworth, Program Head/Chair - Keywords:
- bladder
cancer
heterogeneity
invasion
heterogeneous
biomanufacturing
biosensor
biomanufactured
biomanufacture
3D
extracellular matrix
leader cells
collective invasion
invasion assay
in vitro - Abstract:
- Bladder cancer is an increasingly common malignancy, and muscle invasive bladder cancer is associated with particularly high rates of morbidity and mortality. The morphologic and molecular diversity of bladder cancer poses significant challenges in elucidating the invasion mechanisms responsible for disease progression. Furthermore, conventional invasion assays do not provide a physiological context for studying bladder cancer invasion within 3D microenvironments and have limited ability to capture the contribution of cellular phenotypic and molecular heterogeneity to disease progression. Here, we describe the development of biomanufactured three-dimensional (3D) microtumor invasion models suitable for the analysis of collective invasion in cancer cell lines and primary tumor cells while accounting for the heterogeneity of bladder cancer. These models incorporate a self-assembly approach for recapitulating features of bladder cancer tumors in 3D microenvironments and probing the invasive cell subpopulations. The gene expression profiles of invading microtumors were analyzed by incorporating gold-nanorod-locked-nucleic-acid (GNR-LNA) biosensors. Incorporation of single cell biosensors and transient gene knockdown into the system revealed the formation of invasive leader cells with upregulated Delta-like ligand 4 (DLL4) expression, as well as the role of NOTCH1-DLL4 signaling in collective bladder cancer invasion. The involvement of DLL4 expressing cells in bladder cancer invasion was also observed in patient samples obtained from transurethral resection. Collectively, this study demonstrates a biomanufactured 3D invasion assay approach for investigating collective invasion in bladder cancer microtumors with cellular and molecular heterogeneity, including those generated from patient-derived samples, toward personalized medicine applications.