RESUMO
Cell-cell communication and physical interactions play a vital role in cancer initiation, homeostasis, progression, and immune response. Here, we report a system that combines live capture of different cell types, co-incubation, time-lapse imaging, and gene expression profiling of doublets using a microfluidic integrated fluidic circuit that enables measurement of physical distances between cells and the associated transcriptional profiles due to cell-cell interactions. We track the temporal variations in natural killer-triple-negative breast cancer cell distances and compare them with terminal cellular transcriptome profiles. The results show the time-bound activities of regulatory modules and allude to the existence of transcriptional memory. Our experimental and bioinformatic approaches serve as a proof of concept for interrogating live-cell interactions at doublet resolution. Together, our findings highlight the use of our approach across different cancers and cell types.
Assuntos
Transcriptoma , Neoplasias de Mama Triplo Negativas , Humanos , Microfluídica , Perfilação da Expressão Gênica/métodos , Regulação da Expressão GênicaRESUMO
Background: The emergency department (ED) help desk is an undergraduate-run service learning program that screens ED patients for social needs, connects them to community resources, and follows-up to promote connections with resources. Students accepted to the program participate in a didactic course on the fundamentals of social emergency medicine as well as available community resources. Students also receive training around interviewing patients and use of screening software. Students commit to at least three quarters of service, during which they attend weekly team meetings. Methods: This qualitative study explores the impact of this service learning experience for students. Current and former students were identified by the director of the program. Purposive and snowball sampling was used to select a sample of participants that participated in a semistructured interview. Our codebook was developed inductively using thematic analysis. Themes were presented and discussed with the entire research team for further analysis and refinement. Data collection and analysis used a constant comparative approach, and data collection ceased when saturation was achieved. Results: Study participants consisted of current and former ED help desk student volunteers (n = 21). All participants believed that the ED help desk service learning experience prepared them for future careers by providing an experience that filled a gap in their education. We identified four main themes: (1) participants' perceived impact on patients, (2) learning from patients' experiences and differences, (3) appreciating patients' vulnerability and collaboratively addressing patients' needs, and (4) learning to navigate patients' social needs within the broader health care system. Conclusions: Our ED help desk service learning program offers a unique experience for students to learn about patients' social needs, participate in meaningfully interactions with patients, and empower themselves and patients to work together as coproducers of patients' care.
RESUMO
Patient-derived orthotopic xenograft (PDOX) models have been verified as a useful method for studying human cancers in mice. Previous studies on the extent of metastases in these models have been limited by the necessity of welfare euthanasia (primary tumors reaching threshold size), at which point metastases may only be micrometers in diameter, few in number, and solely identified by step-sectioning of formalin-fixed paraffin-embedded tissue. These small micro-metastases are less suitable for many downstream molecular analyses than macro-metastases. Resection of the primary tumor by survival surgery has been proven to allow further time for metastases to grow. Although PDOX models of triple-negative breast cancer (TNBC) shed circulating tumor cells (CTCs) into the bloodstream and metastasize, similar to human TNBC, little data has been collected in these TNBC PDOX models regarding the association between CTC characteristics and distant metastasis following excision of the primary tumor xenograft. This study assembles a timeline of PDOX tumor shedding and metastatic tumor progression before and after tumor excision surgery. We report the ability to use tumorectomies to increase the lifespan of TNBC PDOX models with the potential to obtain larger metastases. CTC clusters and CTCs expressing a mesenchymal marker (vimentin) were associated with metastatic burden in lung and liver. The data collected through these experiments will guide the further use of PDOX models in studying metastatic TNBC.
