Multi-Omics Profiling Suggesting Intratumoral Mast Cells as Predictive Index of Breast Cancer Lung Metastasis

Breast cancer lung metastasis has a high mortality rate and lacks effective treatments, for the factors that determine breast cancer lung metastasis are not yet well understood. In this study, data from 1067 primary tumors in four public datasets revealed the distinct microenvironments and immune composition among patients with or without lung metastasis. We used multi-omics data of the TCGA cohort to emphasize the following characteristics that may lead to lung metastasis: more aggressive tumor malignant behaviors, severer genomic instability, higher immunogenicity but showed generalized inhibition of effector functions of immune cells. Furthermore, we found that mast cell fraction can be used as an index for individual lung metastasis status prediction and verified in the 20 human breast cancer samples. The lower mast cell infiltrations correlated with tumors that were more malignant and prone to have lung metastasis. This study is the first comprehensive analysis of the molecular and cellular characteristics and mutation profiles of breast cancer lung metastasis, which may be applicable for prognostic prediction and aid in choosing appropriate medical examinations and therapeutic regimens.

Potential Use of CTCs as Biomarkers in Renal Cancer Patients

We demonstrated that the CellCollector is an appropriate tool for detecting CTCs in RCC patients. We examined EpCAM and MUC1 expression levels in RCC tissues and cell lines and analyzed the detection rate of CTCs in blood samples ex vivo using an anti-EpCAM antibody-covered straight or spiraled CellCollector. Eight matched samples were examined for affinity to the anti-EpCAM vs. anti-EpCAM/anti-MUC1 antibody-covered wire. The use of this combination of antibodies allowed us to classify patients with lung metastasis. Finally, four patients were analyzed in vivo. In conclusion, both straight (ex vivo, in vivo) and spiraled (ex vivo) wires detected CTCs.

Interleukin 4 controls the role of macrophages in pulmonary metastatic tumor cell seeding and growth

Metastasis is the systemic manifestation of cancer and the main cause of death from breast cancer. In mouse models of lung metastases, recruitment of classical monocytes from blood to the lung and their differentiation to metastasis-associated macrophages (MAMs) facilitate cancer cell extravasation, survival, and growth. Ablation of MAMs or their monocytic progenitors inhibits metastasis. We hypothesized that factors controlling macrophage polarization modulate tumor cell extravasation in the lung. We evaluated whether signaling by Th1 or Th2 cytokines in macrophages affected trans-endothelial migration of tumor cells in vitro. Interferon γ and LPS inhibited macrophage-dependent tumor cell extravasation while the Th2 cytokine interleukin-4 (IL4) enhanced this process. We demonstrated that IL4 receptor (IL4rα) null mice develop fewer and smaller lung metastasis. Adoptive transfer of wild type monocytes to IL4rα deficient mice rescued this phenotype. IL4 signaling in macrophages controls the expression of the chemokine receptor CXCR2, necessary for IL4-mediated tumor cell extravasation in vitro. Furthermore, IL4 signaling in macrophages transcriptionally regulates several other genes already causally associated with lung metastasis including CCL2, CSF1, CCR1, HGF and FLT1. The central role for IL4 signaling in MAMs was confirmed by high-resolution intravital imaging of the lung in mice at the time of metastatic seeding, which showed reduced physical interaction between tumor cells and IL4rα-deficient macrophages. This interaction enhances tumor cell survival. These data indicate that IL4 signaling in monocytes and macrophages is key during seeding and growth of breast metastasis in the lung as it regulates pro-tumoral paracrine signaling between cancer cells and macrophages.

JPTQ Decoction Inhibits Tumor Proliferation and Lung Metastasis in Tumor-bearing Mice with Triple Negative Breast Cancer

Background. With the increasing incidence of breast cancer and the integration of multiple methods in the treatment, traditional Chinese medicine plays an increasingly important role in the comprehensive treatment of breast cancer. we aimed to determine the anti-cancer metastasis effect of Jianpi Tiaoqi Decoction (JPTQ) on breast cancer-bearing mice by monitoring the effects of its on tumor proliferation, apoptosis, angiogenesis, epithelial to mesenchymal transition (EMT) process and regulation of immune microenvironment. Methods. The general phenotype of the Cancer-bearing mice was monitored. Bioluminescence-imaging was performed to assess the tumor status and the metastatic status of other organs. We investigated its mechanism of the effect through transcriptome analysis, Flow Cytometry(FCM) was used to analyze peripheralblood CD4+ T cells, spleen T helper 1 (Th1) cell, the proportion of MDSCs in lung. The changes of EMT process, vascular endothelial growth factor (VEGF) and Ki-67, Caspase-3 and Bcl-2 were detected by quantitative real time polymerase chain reaction (q-PCR), western blot (WB) or immunohistochemistry (IHC). Results. JPTQ inhibited the tumors proliferation and reduced lung metastasis. The transcriptome analysis of lung and tumor tissues indicated that EMT-related genes, angiogenesis, proliferation and apoptosis genes were regulated in JPTQ group, and the Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis observed enrichment of immune-related pathways. FCM suggested that JPTQ could reduced the proportion of M-MDSCs in the lung, and increased peripheral blood CD4+ T cells and Th1 cells in spleen. The q-PCR, WB or IHC assay demonstrated that E-cadherin was up-regulated in lung and tumor tissue, and Snail was down-regulated, the expression of matrix metalloprotein-9(MMP-9)was down-regulated in lung tissue. IHC showed the down-regulation of Ki67 and VEGF in lung and tumo tissues. WB found that Cleved-Caspase3 was significantly up-regulated, while Bcl-2 was down-regulated. Conclution. JPTQ can inhibit proliferation, angiogenesis, promote apoptosis and improve the immune microenvironment, and reverse the EMT process to inhibit the proliferation and metastasis of TNBC.

HIF-Dependent CKB Expression Promotes Breast Cancer Metastasis, Whereas Cyclocreatine Therapy Impairs Cellular Invasion and Improves Chemotherapy Efficacy

The oxygen-responsive hypoxia inducible factor (HIF)-1 promotes several steps of the metastatic cascade. A hypoxic gene signature is enriched in triple-negative breast cancers (TNBCs) and is correlated with poor patient survival. Inhibiting the HIF transcription factors with small molecules is challenging; therefore, we sought to identify genes downstream of HIF-1 that could be targeted to block invasion and metastasis. Creatine kinase brain isoform (CKB) was identified as a highly differentially expressed gene in a screen of HIF-1 wild type and knockout mammary tumor cells derived from a transgenic model of metastatic breast cancer. CKB is a cytosolic enzyme that reversibly catalyzes the phosphorylation of creatine, generating phosphocreatine (PCr) in the forward reaction, and regenerating ATP in the reverse reaction. Creatine kinase activity is inhibited by the creatine analog cyclocreatine (cCr). Loss- and gain-of-function genetic approaches were used in combination with cCr therapy to define the contribution of CKB expression or creatine kinase activity to cell proliferation, migration, invasion, and metastasis in ER-negative breast cancers. CKB was necessary for cell invasion in vitro and strongly promoted tumor growth and lung metastasis in vivo. Similarly, cyclocreatine therapy repressed cell migration, cell invasion, the formation of invadopodia and lung metastasis. Moreover, in common TNBC cell line models, the addition of cCr to conventional cytotoxic chemotherapy agents was either additive or synergistic to repress tumor cell growth.