Intratumor heterogeneity index of breast carcinomas based on DNA methylation profiles

Chen

et al. 2023

Environ. Sci. Technol.

While exogenous metal/metalloid (metal) exposure has been associated with reduced human semen quality, no study has assessed the associations of exogenous metals in human spermatozoa with semen quality. Here, we developed a strategy to explore the associations between exogenous metals in spermatozoa at single-cell resolution and human semen quality among 84 men screened as sperm donors, who provided 266 semen samples within 90 days. A cellular atlas of exogenous metals at the single-cell level was created with mass cytometry (CyTOF) technology, which concurrently displayed 18 metals in more than 50 000 single sperm. Exogenous metals in spermatozoa at single-cell resolution were extremely heterogeneous and diverse. Further analysis using multivariable linear regression and linear mixed-effects models revealed that the heterogeneity and prevalence of the exogenous metals at single-cell resolution were associated with semen quality. The heterogeneity of lead (Pb), tin (Sn), yttrium (Y), and zirconium (Zr) was negatively associated with sperm concentration and count, while their prevalence showed positive associations. These findings revealed that the heterogeneous properties of exogenous metals in spermatozoa were associated with human semen quality, highlighting the importance of assessing exogenous metals in spermatozoa at single-cell resolution to evaluate male reproductive health risk precisely.

Section: Discussionmentioning

confidence: 99%

Exogenous Metals Atlas in Spermatozoa at Single-Cell Resolution in Relation to Human Semen Quality

Chen

et al. 2023

Environ. Sci. Technol.

“…In our experiments, we found that many compounds significantly inhibited the average cell motility but not the fast-moving cells, especially for the MDA-MB-231 cell line, which is known to be highly aggressive and heterogeneous. [80][81][82] The inability to stop those fast-moving cells might be explained by cancer cellular heterogeneity and the existence of redundant pathways. In our prior work, we have performed scRNA-Seq for highly migratory breast cancer cells.…”

Section: Discussionmentioning

confidence: 99%

High‐Throughput Cellular Heterogeneity Analysis in Cell Migration at the Single‐Cell Level

Zhou

Chianga

et al. 2022

Small

cancer metastasis is a complicated multistep process, migration and intravasation of tumor cells from the tumor stroma to a capillary bed or lymphatic vessel represent critical early steps. [3] In addition, considerable evidence suggests that cancer metastasis can be driven by epithelialto-mesenchymal transition (EMT), [4][5][6] a developmental program in which epithelial cells acquire migratory and invasive phenotypes. [7][8][9][10] Inhibition or activation of EMT significantly limits or increases metastasis, respectively. [11][12][13] To elucidate the correlation between cancer migration and metastasis, we previously developed a high-throughput microfluidic platform that can measure the individual migratory capability of thousands of cancer cells and selectively isolate fast-moving subpopulations. [14,15] As expected, we found that highly migratory breast cancer cells metastasized more than non-migratory cells. [15] Furthermore, through whole transcriptome sequencing of migratory and control cancer cells, differentially expressed genes that correlate with clinical outcomes in breast cancer were identified. [16] The distinct gene expression profile of migratory cancer cells highlights the possibility to selectively inhibit this metastatic sub-population. As migratory breast Cancer cell migration represents an essential step toward metastasis and cancer deaths. However, conventional drug discovery focuses on cytotoxic and growthinhibiting compounds rather than inhibitors of migration. Drug screening assays generally measure the average response of many cells, masking distinct cell populations that drive metastasis and resist treatments. Here, this work presents a high-throughput microfluidic cell migration platform that coordinates robotic liquid handling and computer vision for rapidly quantifying individual cellular motility. Using this innovative technology, 172 compounds were tested and a surprisingly low correlation between migration and growth inhibition was found. Notably, many compounds were found to inhibit migration of most cells while leaving fast-moving subpopulations unaffected. This work further pinpoints synergistic drug combinations, including Bortezomib and Danirixin, to stop fast-moving cells. To explain the observed cell behaviors, single-cell morphological and molecular analysis were performed. These studies establish a novel technology to identify promising migration inhibitors for cancer treatment and relevant applications.

Diagnostic Cytopathology

“…Therefore, e‐ITH, due to their dynamism and reversibility, evolve during a tumorigenic process in parallel to the acquisition of genomic alterations 99 . Specifically, in breast cancer it was reported that tumor subtype's behavior differ in terms of epigenetic heterogeneity, which could contribute to understand the different prognosis of these groups 100 …”

Section: Discussionmentioning

confidence: 99%

DNA methylation patterns of RAR‐β2 and RASSF1A gene promoters in FNAB samples from Greek population with benign or malignant breast lesions

Kougioumtsidou

Vavoulidis

Nasioutziki

et al. 2020

Background Promoter hypermethylation is common in Breast Cancer (BC) with studies mainly in histological specimens showing frequent methylation of tumor suppressor genes (TSGs) compared with normal tissues. The aim of this study was to estimate the frequency of promoter methylation of RAR‐β2 and RASSF1A genes in breast FNAB material aiming to evaluate the methylation status of these two genes as biomarker for detecting BC in Greek population. Methods FNAB material from 104 patients was collected for cytological evaluation and epigenetic analysis. DNA was extracted and subjected to bisulfite conversion. A methylation‐specific PCR was carried out and the final products were separated with electrophoresis in 2% agarose gels. Results From 104 samples, RASSF1A hypermethylation was observed in 78 (75%) and RAR‐β2 hypermethylation in 64 (61.6%). 84% and 78% of the cases diagnosed with breast malignancy (n = 50) were methylated for RASSF1A and RAR‐β2, respectively. Methylated RASSF1A and RAR‐β2 were also detected in 88.3% and 76.5% in samples diagnosed as suspicious for malignancy (n = 17) and in 57.2% of samples diagnosed with atypia (n = 14). The Odds Ratio for breast malignancy was 4.545 in patients with RASSF1A hypermethylation and 9.167 in patients with RAR‐β2 hypermethylation underlying their promoter's methylation positive correlation with breast malignancy. Conclusion To optimize the sensitivity and specificity of this epigenetic setting, more TSGs related to BC should be gradually imported in our evaluated methylation panel and be validated in a larger study sample with the aim that the obtained epigenetic profiles will provide clinicians with valuable tools for management of BC patients in Greece.