Characterization of organoid cultured human breast cancer

Abstract

Organoid cultures are increasingly used to model human cancers experimentally with a view to tailoring personalized medicine and predicting drug responses. Breast cancer is no exception, but in particular, primary breast cancer poses some inherent difficulties due to the frequent presence of residual non-malignant cells in the biopsies. We originally developed an assay for the distinction between malignant and non-malignant structures in primary breast cancer organoid cultures (Petersen et al., Proc Natl Acad Sci (USA) 89(19):9064–8, 1992). Here, we apply this assay to assess the frequency of normal-like organoids in primary breast carcinoma cultures and the cellular composition as a consequence of passaging. We find that in consecutively collected samples of primary human breast cancers, residual non-malignant tissues were observed histologically in five out of ten biopsies. Based on relevant morphogenesis and correct polarization as recorded by expression in luminal epithelial cells of mucin 1 (Muc1), occludin, and keratin 19 (K19) and expression in basal cells of integrin β4, p63, and K14, non-malignant organoids were present in all primary human breast cancer-derived cultures. Furthermore, passaging in a contemporary culture medium was in favor of the selective expansion of basal-like cells. We conclude that organoid cultures of human breast cancers are most representative of the tissue origin in primary culture.

Three-Dimensional Spheroids as In Vitro Preclinical Models for Cancer Research

“…Cells grown in culture environment of low binding or absence of adhesive surface can assemble into 3D spheroids, as these conditions favor cell-cell and cell-ECM interactions over cell-substrate Therefore, 3D culture systems recapitulate many characteristics of in vivo tumors, such as cell-cell and cell-ECM interactions, nutrient and oxygen gradients, and distinct layers of cell populations. Besides, the morphology and polarity of the cells, as well as gene expression and activation of cell signaling pathways, are also close to those of real tumors [8,9,60,61]. These features make spheroids a promising model for the study of cancer biology, cancer initiation, invasion and metastatic processes, as well as drug testing.…”

Contralateral prophylactic mastectomy: A narrative review of the evidence and acceptability

“…Three different contralateral breast cancer risk prediction models have been described in the literature, including the Manchester formula (part of the Manchester guidelines for contralateral risk-reducing mastectomy), CBCrisk, and PredictCBC [ 13 , [34] , [35] , [36] ]. These models calculate an individual’s risk of contralateral breast cancer in different ways using patient and tumor characteristics such as age at first primary breast cancer diagnosis, family history, ER status, breast density, first breast cancer type, and adjuvant treatments.…”

Breast Cancer Organoids Model Patient-Specific Response to Drug Treatment

“…Candidate pathogenic mutations in untranslated regions (UTRs) and BC-relevant genes regions [18,19], including oncogenes (i.e., KRAS, ERBB3, PIK3C2G) and tumor suppressor genes (i.e., FANCC, ATR, SMAD2), were found in organoids, and the majority of them were conserved within the tumor-organoid pair ( Figure 3b, Table S5). In a few cases, mutations in driver genes comprising ATRX, BRCA2, and NF1 were found in primary tumors, but not in the derived organoids (Table S5), likely because of the selection in culture of less aberrant cell subpopulations and/or the presence of cell subpopulations at low frequencies in the originating tumors [12]. Next, we interrogated WES data for the presence of BC distinctive mutational signatures [20,21].…”

“…Recently, in a seminal work, the generation of a biobank of patient-derived breast cancer organoids has been described for its application as a patient-specific model for preclinical studies [ 10 ]. In another recent study [ 12 ], breast tumor organoids were generated by similar procedures, though these 3D models were described to also contain a significant amount of normal-like cells.…”