Abstract
BackgroundMammary Paget’s disease (MPD) is an uncommon cutaneous intraepithelial malignancy with ulceration of the nipple or areola. Its pathogenesis and genomic mutation remain largely unknown and no cell lines are established from primary tumors.MethodsWe collected surgical tumor specimens from a 65-year-old Chinese woman diagnosed with MPD and established patient-derived breast cancer (BC) organoids from MPD using organoid culture technology.ResultsWe successfully propagated BC organoids from a patient with MPD for more than 6 months. The organoids were cultured for long-term expansion without any change in spherical organoid morphology. Besides, the spherical organoid morphology did not change when they underwent cryopreservation after resuscitation. The H&E staining and immunohistochemistry analyses showed the similar morphological and histological features of the organoids compared with their paired original BC tissues. The organoids retained positive expression of breast cancer biomarkers: estrogen receptor, progesterone receptor, antigen Ki-67 and negative expression of human epidermal growth factor receptor 2. We also showed that MPD organoids recapitulated the unique genomic landscape including copy number alterations, mutational load, mutational signatures and cancer gene mutations by whole genome sequencing. In situ senescence-associated acid beta galactosidase assay confirmed senescence phenomenon existed in the process of organoids culture and there was no significant difference in the proportion of senescent organoids after organoid passage and resuscitation.ConclusionsOur results suggested that an effective platform for ex vivo BC organoids from MPD patients could be used to explore clinicopathological and genomic characteristics of these patients.
Highlights
Mammary Paget’s disease (MPD) is an uncommon cutaneous intraepithelial malignancy with ulceration of the nipple or areola
They found that MPD and extramammary Paget’s disease (EMPD) had the similar genomic aberrations, especially in genes involved in chromatin remodeling processes, such as KMT2C and ARID2
Establishing MPD patient‐derived organoids In order to evaluate the feasibility of primary culture of MPD-derived tumor cells, we used organoid technology to culture these tumor cells from our patient
Summary
Establishing MPD patient‐derived organoids In order to evaluate the feasibility of primary culture of MPD-derived tumor cells, we used organoid technology to culture these tumor cells from our patient. The culture state of nipple tissue-derived tumor cells under 3D condition was poor in the process of culture and died gradually even after passage (Fig. 2). The tumor cells grew rapidly under 2D condition and the cultivation process at 1 day, 4 days and 7 days was recorded When they were cultured for 4 days, they began to grow fast and fill up the entire cultivation space quickly after 4 days of cultivation (Additional file 2: Figure S1). MPD patient‐derived organoids match the original histological characteristics We performed histopathological analysis of H&E stained tissues and MPD patient-derived organoids sections, and confirmed that the phenotypes of tumor cells matched the histological characteristics of BC. Senescence phenomenon existed in the process of organoids culture and there was no significant difference in the proportion of senescent organoids after organoid passage and resuscitation
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