Characterization of transcriptome diversity and in vitro behavior of primary human high-risk breast cells

Abstract

Biology and transcriptomes of non-cancerous human mammary epithelial cells at risk for breast cancer development were explored following primary isolation utilizing conditional reprogramming cell technology from mastectomy tissue ipsilateral to invasive breast cancer. Cultures demonstrated consistent categorizable behaviors. Relative viability and mammosphere formation differed between samples but were stable across three different mammary-specific media. E2F cell cycle target genes expression levels were positively correlated with viability and advancing age was inversely associated. Estrogen growth response was associated with Tissue necrosis factor signaling and Interferon alpha response gene enrichment. Neoadjuvant chemotherapy exposure significantly altered transcriptomes, shifting them towards expression of genes linked to mammary stem cell formation. Breast cancer prognostic signature sets include genes that in normal development are limited to specific stages of pregnancy or the menstrual cycle. Sample transcriptomes were queried for stage specific gene expression patterns. All cancer samples and a portion of high-risk samples showed overlapping stages reflective of abnormal gene expression patterns, while other high-risk samples exhibited more stage specific patterns. In conclusion, at-risk cells preserve behavioral and transcriptome diversity that could reflect different risk profiles. It is possible that prognostic platforms analogous to those used for breast cancer could be developed for high-risk mammary cells.

Preclinical Models of Neuroblastoma—Current Status and Perspectives

“…Conditional reprogramming is the epitome of personalized medicine; the cells can be directly obtained from a patient, cultured, and used for potential therapeutic approaches [ 23 ]. They can be also used as models for certain cancer types, including rare diseases, e.g., neuroendocrine cervical carcinoma [ 24 , 27 , 132 ], as well as biobanked for future basic and preclinical applications, for example genetic and chemosensitivity testing [ 133 , 134 , 135 , 136 ]. Conditional reprogramming is an inexpensive, easy to use, and robust method [ 25 ].…”

Unlocking Translational Potential: Conditionally Reprogrammed Cells in Advancing Breast Cancer Research

“…It is interesting to note that the phenotype is completely reversible if these factors are eliminated [66]. Therefore, CR technology can be an ideal model for performing gene profiling analysis [65,67] and drug sensitivity testing for use in breast and other cancer research [68], regenerative medicine, and xenograft studies. An important feature is that these cultures can be used for establishing xenografts [69], patient-derived xenograft cell lines [70], cell cultures from PDXs, and organoid cultures.…”

“…One of the limitations is that these cells have a short lifespan when cultured under conventional tissue culture conditions [95]. CR system demonstrated that feeder cell-conditioned medium, together with a Rho-associated kinase (ROCK) inhibitor, can induce inexhaustible and rapid in vitro proliferation of primary epithelial cells from normal and malignant breast tissues [63,65,67,69,80,96].…”

“…Both the construction of different 3D organoid structures and the development of milk-producing cells are supported by this culture technique. Alothman et al established PHMEC to study the behavior and transcriptomes of non-cancerous human mammary epithelial cells at risk for breast cancer development [67] (Figure 2). 2), and deidentified tissue acquisition following pathological review (3), with initial isolation of mammary epithelial cells using CRC technology with assignment of a unique GUMC identifier to viable cell pellets (4), sample validation with secondary pathology review (5), and acquisition of deidentified medical records (6).…”

“…Then, exploiting the ability of utilizing CRC-isolated cells for parallel studies of behavior and gene expression, showed that estrogen growth response was associated with tissue necrosis factor signaling and interferon alpha response gene enrichment while neoadjuvant chemotherapy exposure significantly altered transcriptomes, shifting them towards expression of genes linked to mammary stem cell formation. Finally, they showed gene expression patterns in mammary cells normally linked to pregnancy can be found as an abnormal finding in non-pregnant at-risk and breast cancer cells [67]. Alamri et al used genetically engineered mice carrying Brca1 and Trp53 mutations to explore how CRC methodology compared to his-torical methods of mammary epithelial cell isolation for initial primary cell isolation, allograft generation, and impact of CRC and passage on the transcriptome [65].…”