Biology subjects

Rochefort, W. E.

Publications and source records attributed to Rochefort, W. E..

3 recordsLinked to original sources

bioRxiv2024

Engineering a three-dimensional multilayer multicellular model of endometrial cancer for high throughput drug screening and novel treatment methods

Endometrial cancer is one of the most common gynecological cancers in the world, with an estimated 382,000 new cases and 90,000 deaths each year. There is no specific treatment, as the underlying causes of endometrial cancer neoplasia are poorly understood. This study focuses on the development and validation of a three-dimensional (3D) in vitro multilayer, multicellularhydrogel that facilitates drug screening analysis. We hypothesized that a specific combination of natural (collagen type I and IV, fibrinogen, fibronectin, Laminin) and synthetic (GELMA, PEGDA) polymers would maximize microvessel formation and cell invasion. The 3D model incorporates human microvascular endothelial cells (hMVEC) and endometrial cancer cells (HEC-1A) atop hydrogel formulations mimicking cell-specific extracellular matrix components. Using a D-optimal experimental design, 45 hydrogel combinations were generated. The predicted hydrogel formulation to maximize all cell responses enhanced higher microvessel formation and cancer invasion compared to the gold standard Matrigel. Subsequent validation emphasizes the importance of a disease-specific model and cell crosstalk in maximizing microvessel formation and cancer invasion. The optimized 3D model adeptly captures variances in cell responses among endometrial cancer cell lines from distinct stages. Finally, the platform is employed to compare cell viability, microvessel formation, and cancer invasion across Ishikawa, KLE, and HEC-1A cells after Paclitaxel exposure, delivered both as a free drug and loaded in poly(caprolactone) (PCL) nanoparticles. Overall, this study provides a valuable tool for exploring intricate interactions within the tumor microenvironment, offering a holistic understanding of cell responses and fostering the development of targeted therapeutic strategies for endometrial cancer.

bioengineering↗

bioRxiv2024

Development of a 3D in vitro human-sized model of cervical dysplasia to evaluate the delivery of ethyl cellulose-ethanol injection for the treatment of cervical dysplasia ablation

Cervical cancer, the second leading cause of cancer-related death for women worldwide, remains a preventable yet persistent disease that disproportionately affects women in low and middle-income countries (LMICs). While existing therapies for treating cervical dysplasia are effective, they are often inaccessible in LMICs. Ethanol ablation is an alternative low-cost, accessible therapy that we previously enhanced into an ethyl cellulose (EC)-ethanol gel formulation to improve efficacy. When seeking to evaluate EC-ethanol for cervical dysplasia, we found a paucity of relevant animal models. Thus, in this study, we developed a 3D in vitro model of cervical dysplasia featuring a central lesion of cervical cancer cells surrounded by fibroblasts and keratinocytes to enable the evaluation of EC-ethanol and other novel therapeutics. Our GelMA-based 3D model successfully captured the architectural complexity of cervical dysplasia, showcasing cell response and high viability. The GelMA hydrogel formulation (8.7% w/v) exhibited viscoelastic properties akin to human cervical tissue. Using micro-CT imaging, we assessed EC-ethanol injection deposition in the hydrogel, revealing retention of virtually the entire injected volume near the injection site. Finally, we evaluated the EC-ethanol injections efficacy in eliminating cervical cancer cells. The EC-ethanol injection led to a significant decrease in cancer cell viability while preserving healthy cells in the 3D in vitro model. Taken together, our in vitro model mirrored the architecture of cervical dysplasia and demonstrated the potential of EC-ethanol for localized treatment of cervical dysplasia.

bioengineering↗

bioRxiv2022

Engineering High Throughput Screening Platforms of Cervical Cancer

There is a critical need for complex multicellular three-dimensional physiomimetic models of cancer that can interface with high throughput drug screening methods to assess anti-metastatic and anti-angiogenic drug efficacy in a rapid yet high content manner. We report a multilayer multicellular platform of human cervical cancer cell lines and primary human microvascular endothelial cells that incorporates critical biophysical and extracellular matrix cues, interfaces with standard high throughput drug screening methods, and can evaluate cervical cancer invasion and endothelial microvessel formation over time. Through the use of Design of Experiments statistical optimization, we identified the specific concentrations of collagen I, fibrinogen, fibronectin, GelMA, and PEGDA in each hydrogel layer that maximized cervical cancer invasion and endothelial microvessel length simultaneously. We then validated the optimized platform and assessed the viscoelastic properties of the composite hydrogels as well as their individual constituents. Finally, using this optimized platform, we conducted a targeted drug screen of four clinically relevant drugs on two cervical cancer cell lines. From these data we identified each of the cervical cancer cell lines (SiHa and Ca Ski) as either responsive or refractive to Paclitaxel, Dasitinib, Dovitinib, or Pazopanib. Overall, we developed a phenotypic drug screening platform of cervical cancer that captures cell behavior present in the cervical cancer tumor microenvironment, captures patient to patient variability, and integrates with standard high throughput high content drug screening methods. This work provides a valuable platform that can be used to screen large compound libraries for mechanistic studies, drug discovery, and precision oncology for cervical cancer patients.

bioengineering↗