Journal: Breast Cancer Research : BCR

Article Title: High levels of circulating miR-19a-3p in patients with metastatic HER2 + breast cancer are associated with a favorable prognosis and anti-tumor immune responses

doi: 10.1186/s13058-025-02174-8

Figure Lengend Snippet: Expression and secretion levels of miR-19a-3p in breast cancer cells. a) Breast cancer cells express different levels of miR-19a-3p depending on the subtypes, with TNBC (basal) expressing the highest levels compared to luminal B, PAM50 Normal, HER2+, luminal A, and normal cells (TCGA data). The data are shown as box and whisker plots, where the box encompasses the 25th percentile, median, and 75th percentile, and the whiskers extend to 1.5 times the interquartile range; b) the two HER2+ breast cancer cell lines KPL-4 and SKBR3 express higher level of miR-19a-3p than the luminal A MCF-7, consistent with TCGA data; c) the three breast cancer cell lines secrete varying amounts of miR-19a-3p, following an expression pattern similar to that in their parental cells. The results are shown as the mean ± SD of three technical replicates from 3 experimental replicates

Article Snippet: MCF-7 (ER +) and SKBR3 (HER2 +) cells were obtained from the American Type Culture Collection (Manassas, VA, USA), and KPL-4 (HER2 +) cells were provided courtesy of Dr. Junichi Kurebayashi (Kawasaki Medical School, Kurashiki, Japan) [ ].

Techniques: Expressing, Whisker Assay

Journal: Breast Cancer Research : BCR

Article Title: High levels of circulating miR-19a-3p in patients with metastatic HER2 + breast cancer are associated with a favorable prognosis and anti-tumor immune responses

doi: 10.1186/s13058-025-02174-8

Figure Lengend Snippet: Central memory T (T CM ) cell phenotype of CD4 + Th1 cells and immune cell populations in the peripheral blood of patients with metastatic HER2 + breast cancer. a-b) CD4 + Th1 cells expressing high levels of miR-19a-3p developed a central memory T (T CM ) cell phenotype (CD45RO + CCR7 + CD62L +) by the end of the cell culture; c-e) in a small cohort of patients with metastatic HER2 + breast cancer, those with a good prognosis and high serum levels of miR-19a-3p showed a trend of higher percentage of CD4 + Th1 than CD4 + Th2 cells (ratio: IFN-γ + /IL-4 +), although not statistically significant (p = 0.08130) and activated CD4 + T and NK cells circulating in their blood. The results of panel a are shown as the mean ± SD of 3 technical replicates. The results in panel b are presented as the most representative of 3 technical replicates and two experimental replicates. The results in panels c-e are shown as the median with a 95% confidence interval (CI)(Available data from n = 15)

Article Snippet: MCF-7 (ER +) and SKBR3 (HER2 +) cells were obtained from the American Type Culture Collection (Manassas, VA, USA), and KPL-4 (HER2 +) cells were provided courtesy of Dr. Junichi Kurebayashi (Kawasaki Medical School, Kurashiki, Japan) [ ].

Techniques: Expressing, Cell Culture

Journal: Breast Cancer Research : BCR

Article Title: High levels of circulating miR-19a-3p in patients with metastatic HER2 + breast cancer are associated with a favorable prognosis and anti-tumor immune responses

doi: 10.1186/s13058-025-02174-8

Figure Lengend Snippet: Proposed model of TME of HER2 + breast cancer modulated by NK cell-mediated ADCC. The proposed model is based on findings from our current and previous work. Further studies are needed to validate and confirm our hypothetical model. Trastuzumab binds to HER2 + cells and engages CD16 on NK cells, activating them and inducing ADCC. This leads to perforin/granzyme B-mediated apoptosis and death of HER2 + breast cancer cells, as well as the release of miR-19a-3p into the TME and eventually into the bloodstream. ADCCalso stimulates the production and secretion of IFN-γ and T cell-recruiting chemokines from NK cells, which activate DCs, promote their migration to draining lymph nodes, and induce migration of lymphocytes into the TME. This process enables TAA cross-presentation to T cells (both CD4 + and CD8 +) and triggers the production and secretion of IL-12 by DCs. IL-12 and IFN-γ promote the differentiation of CD4 + and CD8 + T cells into CD4 + Th1 cells and CD8 + CTLs and enhance the cytotoxic activity of CD8 + CTLs and NK cells. IL-2 promotes the proliferation of CD4 + Th1 cells, CD8 + CTLs, and NK cells, while IFN-γ from CD4 + Th1 and NK cells further activates DCs (enhancing TAA cross-presentation and cytokine release). The activation and differentiation of CD4 + Th1 cells increase the expression and secretion of miR-19a-3p into the TME and bloodstream. Activated CD8 + CTLs infiltrate tumor tissue and work with NK cells to kill tumor cells, further increasing the release of miR-19a-3p. Additionally, CD4 + Th1 cells differentiate into CD4 + Th1 T CM cells (CD45RO + CCR7 + CD62L +). TAAs from the TME and presented by DCs can maintain the expression and secretion of miR-19a-3p by stimulating CD4 + Th1 T CM cells in draining lymph nodes and contributing to higher serum levels of miR-19a-3p. (This figure was created in BioRender )

Article Snippet: MCF-7 (ER +) and SKBR3 (HER2 +) cells were obtained from the American Type Culture Collection (Manassas, VA, USA), and KPL-4 (HER2 +) cells were provided courtesy of Dr. Junichi Kurebayashi (Kawasaki Medical School, Kurashiki, Japan) [ ].

Techniques: Migration, Activity Assay, Activation Assay, Expressing

Journal: Human Genetics and Genomics Advances

Article Title: Understanding exceptional response: The role of MINDY1 SNP in CDK4/6 inhibitor therapy for ER + , HER2 − advanced breast cancer

doi: 10.1016/j.xhgg.2025.100560

Figure Lengend Snippet: CRISPR-spCas9-mediated engineering of breast cancer cell models (A) Schematic illustration of the CRISPR-spCas9-mediated homology-directed repair strategy used to introduce the rs771205 SNP into the MINDY1 locus of MCF-7 cells. (B) Sanger sequencing of RT-PCR amplicon from parental MCF-7 cells showing the SNP at position c.130 of MINDY1 transcript. (C) Sanger sequencing of RT-PCR amplicon from CRISPR-edited MCF-7 cells confirming successful SNP edit at position c.130 of the MINDY1 transcript. (D) Western blot shows MINDY1 protein levels in ExR (c.130G) and RefG (c.130A) cell lines. (E) Real-time qPCR indicates MINDY1 mRNA expression levels in ExR and RefG cell lines. Error bars indicate the standard deviation (SD). Statistical significance was assessed using a two-tailed Student’s t test.

Article Snippet: The ER + /HER2 − breast cancer cell line MCF-7 (ATCC) was selected ( ).

Techniques: CRISPR, Introduce, Sequencing, Reverse Transcription Polymerase Chain Reaction, Amplification, Western Blot, Expressing, Standard Deviation, Two Tailed Test

Journal: Cancer Discovery

Article Title: Sevabertinib, a Reversible HER2 Inhibitor with Activity in Lung Cancer

doi: 10.1158/2159-8290.CD-25-0605

Figure Lengend Snippet: Sevabertinib shows potent antiproliferative activity in isogenic Ba/F3 cells ectopically expressing HER2 exon 20 insertions or HER2 missense mutations. A, HER2 exon 20 insertions and single-nucleotide HER2 missense mutations in the extracellular domain, transmembrane domain (TM), and kinase domain of HER2 protein. B, Dose–response curves of sevabertinib in isogenic Ba/F3 cell lines ectopically expressing HER2 exon 20 insertions. C, IC 50 values of sevabertinib and the EGFR/HER2 TKIs zongertinib, tucatinib, lapatinib, and neratinib in isogenic Ba/F3 cell lines ectopically expressing HER2 missense mutations. D, Dose–response curves of sevabertinib in isogenic Ba/F3 cell lines ectopically expressing HER2 missense mutations. E, IC 50 values of sevabertinib and the EGFR/HER2 TKIs zongertinib, tucatinib, lapatinib, and neratinib in isogenic Ba/F3 cell lines ectopically expressing HER2 missense mutations. Dose–response curves of sevabertinib in isogenic Ba/F3 cell lines ectopically expressing HER2 exon 20 insertions (A775insYVMA, P780insGSP) or point mutation (S310F) coupled with ( F ) HER2 C805S mutation or ( G ) HER2 T798I or T798M mutations, predicted to cause TKI resistance. Ba/F3 cells transduced with an empty vector and maintained in the presence of IL3 were included as a control.

Article Snippet: EGFR/HER2 TKIs tucatinib, lapatinib, neratinib, and zongertinib were purchased from Selleck Chemicals, and STX-721 from MedChemExpress.

Techniques: Activity Assay, Expressing, Mutagenesis, Transduction, Plasmid Preparation, Control