Abstract
Simple SummaryCells communicate mainly through the secretion of proteins. Impaired protein secretion can indicate the development of disease. Cancer cell heterogeneity and acquired resistance to therapy are, however, reducing the effectiveness of cancer treatments. As cancer cells change during the course of the disease, sampling of cancer cells at the time of treatment is needed in order to determine which drugs will be effective. This paper describes a method for measuring secreted prostate specific antigen (PSA) protein from thousands of prostate cancer (PCa) cells. Furthermore, we show that the PSA secretion of individual cells in microwells can be stimulated or inhibited with drugs. To this end, we believe that this method could accelerate the development of new drugs, improve our understanding of resistance to therapy, and, ultimately, improve personalized cancer therapy.The treatment of cancer faces a serious challenge as cancer cells within patients are heterogeneous and frequently resistant to therapeutic drugs. Here, we introduce a technology enabling the assessment of single cancer cells exposed to different drugs. PCa cells were individually sorted in self-seeding microwells, cultured for 24 h, and then exposed to several drugs to induce (R1881) or inhibit (Enzalutamide/Abiraterone) the secretion of a protein (PSA). Cell viability and PSA secretion of each individual prostate cell were monitored over a 3-day period. The PSA protein secreted by each cell was captured on a PVDF membrane through a pore in the bottom of each well. The basal PSA secretion was found to be 6.1 ± 4.5 and 3.7 ± 1.9 pg/cell/day for LNCaP and VCaP, respectively. After exposure to R1881, the PSA secretion increased by ~90% on average and was not altered for ~10% of the cells. PSA production decreased in the majority of cells after exposure to enzalutamide and abiraterone.
Highlights
Cell-to-cell heterogeneity plays an important role in normal tissue homeostasis and is the driving factor in many diseases [1]
To measure secreted prostate-specific antigen (PSA), cells from the prostate cancer cell lines LNCaP and VCaP were loaded as single cells into the individual microwells
The secreted PSA was captured at the bottom of the microwell array using a poly-vinylidenefluoride (PVDF) membrane that was coated with commercially available anti-PSA antibodies (Figure 1d)
Summary
Cell-to-cell heterogeneity plays an important role in normal tissue homeostasis and is the driving factor in many diseases [1]. The heterogeneity of cancer cells and acquired resistance to therapy are, reducing the effectiveness of cancer treatments. As cancer cells change during the course of the disease, sampling of cancer cells at the time of treatment is needed in order to determine which drugs will be effective. In patients with metastatic disease, circulating tumor cells can be obtained from blood [8,9]. In cases where the number of these circulating tumor cells (CTCs) is too low, diagnostic leukapheresis (DLA) can be used to obtain a larger number of cancer cells [10,11,12]. New tools are needed to assess the cancer cell heterogeneity, determine the effectiveness of therapeutic drugs to be administered, and investigate the mechanisms leading to drug resistance
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