Cell mechanotype in cancer progression and metastasis
Cell mechanical phenotype or ‘mechanotype’ is a label-free biomarker of cell state in physiological and disease contexts ranging from stem cell differentiation to cancer progression. However, to harness deformability as a phenotype for drug screening applications requires a method that can simultaneously measure hundreds of samples in parallel. Moreover, a systems-level understanding of molecular mediators of cellular mechanotype is lacking. In this dissertation, I present a simple and scalable technique, called parallel microfiltration (PMF), to measure cell deformability of multiple samples in parallel. I also demonstrate the application of PMF to screen cancer cells based on their mechanotype across multiple cancer types. This dissertation also demonstrates how PMF can interface with existing high throughput facilities. To achieve high throughput screening using filtration, I developed high-throughput filtration (HTF), which utilizes a customized arrays of individual microfluidic filtration devices in a standard multiwell format to enable mechanotype-screening of hundreds of samples in parallel. Additionally, this dissertation presents a mechanotype-screen of cisplatin-resistant ovarian cancer cells treated with a library of small molecules to identify synergistic anti-cancer drugs. Finally, my thesis also presents an investigation of key cellular and nuclear molecular mediators of altered cell deformability using these mechanotyping methods; these findings identify novel molecular mediators of cancer cell mechanotype and also provide unique insight into potential mechanisms of a devastating neurological movement disorder, dystonia. Taken together, this dissertation presents novel high throughput cellular mechanotyping methods that enable measurements of cell deformability with unprecedented throughput, which should enable us to harness knowledge of mechanotype to identify novel treatment strategies for cancer and other diseases.