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It is postulated that the complexity and heterogeneity in cancer may hinder most efforts that target a single pathway. Thus, discovery of novel therapeutic agents targeting multiple pathways, such as miRNAs, holds promise for future cancer therapy. One such miRNA, miR-489, is downregulated in a majority of breast cancer cells and several drug-resistant breast cancer cell lines, but its role and underlying mechanism for tumor suppression and drug resistance needs further investigation. The current study identifies autophagy as a novel pathway targeted by miR-489 and reports Unc-51 like autophagy activating kinase 1 (ULK1) and lysosomal protein transmembrane 4 beta (LAPTM4B) to be direct targets of miR-489. Furthermore, the data demonstrate autophagy inhibition and LAPTM4B downregulation as a major mechanism responsible for miR-489–mediated doxorubicin sensitization. Finally, miR-489 and LAPTM4B levels were inversely correlated in human tumor clinical specimens, and more importantly, miR-489 expression levels predict overall survival in patients with 8q22 amplification (the region in which LAPTM4B resides).Implications: These findings expand the understanding of miR-489–mediated tumor suppression and chemosensitization in and suggest a strategy for using miR-489 as a therapeutic sensitizer in a defined subgroup of resistant breast cancer patients. Mol Cancer Res; 16(9); 1348–60. ©2018 AACR.

Figure S1. miR-489 modulates autophagy. Figure S2. Western blot analysis of autophagy flux in breast cancers. Figure S3. Validation of microarray using real-time RT-PCR analysis. Figure S4. miR-489 restoration in HCC1954 and T47D cells under starvation. Figure S5. Endogenous expression level of miR-489 in different classes of breast cancer cell lines. Figure S6. Drug sensitivity assay with cisplatin and doxorubicin. Figure S7. ULK1 restoration rescues MDA-MB-231 cells from cytotoxic effect of miR-489. Figure S8. Characterization and validation of nanoparticles in vitro and in vivo. Figure S9. Correlation of miR-489 and ULK1 mRNA expression in breast cancers. Supplemental Experimental Procedures Supplementary Table 1: Primer list and sequences. Supplementary Table 2: Demographic and histopathology data of the patient samples.

Supplementary Figures with legends- Supplementary Figures S1 to S14. S1. Shows gene ontology enrichment analysis between MCF10A-HER2 and MCF10A-vector cells. S2 shows western blot analysis of HER2 signaling after IL-1β treatment. S3, S4 and S9 show knockout of IL1A and IL6 using CRISPR/Cas9 system. S5 shows western blot analysis of HER2-signalign pathways in cells treated with IRAK1 and JAK2 inhibitors. S6 shows that activation of NF-κB and STAT3 is not mediated by HER2-immediate downstream molecules. S7 and S10 shows western blot analysis of total HER2, phosphorylated p65, STAT3 and AKT in mammary gland tissues and primary tumors. S8 shows the effects of blockades of HER2-downstream signaling pathways on cell viability. S11 shows representative images of Immunohistochemical staining of HER2, IL-1α and IL6 in primary breast cancer tissues. S12 shows Kaplan-Meier plots of distant metastasis-free survival of patients. S13 shows combination drug effects of IRAK inhibitor (IRAKi) plus paclitaxel (PCL) or cisplatin (CPT). S14 shows tumor volume growth curves for individual mice after last treatment.

Supplementary Table S2 shows the clinicopathological characteristics of clinical samples.

Supplementary methods and References - Supplementary methods describing key reagents, plasmids, cell culture, CRISP/Cas9 system, HER2 agonist preparation, RT-PCR, western blot, flow-cytometry, tumorsphere culture, ChIP assay, animal experiments, Immunohistochemically staining and microarray analysis.

Systemic inflammation in breast cancer correlates with poor prognosis, but the molecular underpinnings of this connection are not well understood. In this study, we explored the relationship between HER2 overexpression, inflammation, and expansion of the mammary stem/progenitor and cancer stem–like cell (CSC) population in breast cancer. HER2-positive epithelial cells initiated and sustained an inflammatory milieu needed to promote tumorigenesis. HER2 induced a feedforward activation loop of IL1α and IL6 that stimulated NFκB and STAT3 pathways for generation and maintenance of breast CSC. In mice, Il1a genetic deficiency delayed MMTV-Her2–induced tumorigenesis and reduced inflammatory cytokine expression as well as CSC in primary tumors. In clinical specimens of human breast tumor tissues, tissue microarray analysis revealed a strong positive correlation between IL1α/IL6 expression and CSC-positive phenotype. Pharmacologic blockade of IL1α signaling reduced the CSC population and improved chemotherapeutic efficacy. Our findings suggest new therapeutic or prevention strategies for HER2-positive breast cancers.Significance: IL1α signaling driven by HER2 promotes chronic inflammation needed to support cancer stem-like cell maintenance in HER2-positive breast cancers. Cancer Res; 78(8); 2040–51. ©2018 AACR.

Supplementary Tables - Supplementary Tables S1, S3 and S4. S1 shows frequency of CSCs after transplantation of the indicated cells. S3 shows multivariate Cox regression data. S4 Positive correlation between IL-1α and HER2 or IL6 expression in breast cancer tissues.

Supplementary Data from Oncogenic Wip1 Phosphatase Is Inhibited by miR-16 in the DNA Damage Signaling Pathway

Supplementary Figures 1-7 from Hoxb7 Inhibits Transgenic HER-2/neu–Induced Mouse Mammary Tumor Onset but Promotes Progression and Lung Metastasis

Supplementary Figure Legends 1-7 from Hoxb7 Inhibits Transgenic HER-2/neu–Induced Mouse Mammary Tumor Onset but Promotes Progression and Lung Metastasis

Table S1. The clinicopathological characteristics of clinical samples. Table S2. Primer sequences used in the cloning and quantitative PCR analysis. Table S3. Primer sequences used for the CHIP assay.

Wild-type p53-induced phosphatase 1 (Wip1) was identified as an oncogene amplified and overexpressed in several human cancers. Recent evidence suggested that Wip1 is a critical inhibitor in the ATM/ATR-p53 DNA damage signaling pathway. Wip1 dephosphorylates several key DNA damage–responsive proteins and reverses DNA damage–induced cell cycle checkpoints. Previous reports showed that Wip1 was transcriptionally induced by p53 at the early stage of the DNA damage response. To investigate the temporal and functional regulation of Wip1, we identified a microRNA, miR-16, that specifically targets the mRNA of Wip1 and thus negatively regulates the expression level of Wip1. miR-16 itself is induced immediately after DNA damage. Therefore, the increase in Wip1 protein level is significantly postponed compared with that of its mRNA level, preventing a premature inactivation of ATM/ATR signaling and allowing a functional completion of the early DNA damage response. To better understand miR-16 biological functions in the context of cancer cells, we examined its expression in mammary tumor stem cells and found it to be markedly downregulated in mammary tumor stem cells. Overexpression of miR-16 or inhibition of Wip1 suppresses the self-renewal and growth of mouse mammary tumor stem cells and sensitizes MCF-7 human breast cancer cells to the chemotherapeutic drug doxorubicin. Together, our results suggest an important role of miR-16 in the regulation of Wip1 phosphatase in the DNA damage response and mammary tumorigenesis. Cancer Res; 70(18); 7176–86. ©2010 AACR.

Fig. S1. TGF-β/SMAD3 signaling mediates the promotion of migration and invasion by HOXB7. Fig. S2 Effects of HOXB7 overexpression and TGF-β2 knockdown on cellular morphology and proliferation rate. Fig. S3 Effects of HOXB7 overexpression on cellular morphology and proliferation rate of Her2 mouse mammary tumor cell line, H605

The trillions of microorganisms inhabiting the human gut are intricately linked to human health. At the species abundance level, correlational studies have connected specific bacterial taxa to various diseases. While the abundances of these bacteria in the gut serve as good indicators for disease progression, understanding the functional metabolites they produce is critical to decipher how these microbes influence human health. Here, we report a unique biosynthetic enzyme-guided disease correlation approach to uncover microbial functional metabolites as potential molecular mechanisms in human health. We directly connect the expression of gut microbial sulfonolipid (SoL) biosynthetic enzymes to inflammatory bowel disease (IBD) in patients, revealing a negative correlation. This correlation is then corroborated by targeted metabolomics, identifying that SoLs abundance is significantly decreased in IBD patient samples. We experimentally validate our analysis in a mouse model of IBD, showing that SoLs production is indeed decreased while inflammatory markers are increased in diseased mice. In support of this connection, we apply bioactive molecular networking to show that SoLs consistently contribute to the immunoregulatory activity of SoL-producing human microbes. We further reveal that sulfobacins A and B, two representative SoLs, primarily target Toll-like receptor 4 (TLR4) to mediate immunomodulatory activity through blocking TLR4’s natural ligand lipopolysaccharide (LPS) binding to myeloid differentiation factor 2, leading to significant suppression of LPS-induced inflammation and macrophage M1 polarization. Together, these results suggest that SoLs mediate a protective effect against IBD through TLR4 signaling and showcase a widely applicable biosynthetic enzyme-guided disease correlation approach to directly link the biosynthesis of gut microbial functional metabolites to human health.

Immunotherapy is an emerging form of cancer therapy that is associated with promising outcomes. However, most cancer patients either do not respond to immunotherapy or develop resistance to treatment. The resistance to immunotherapy is poorly understood compared to chemotherapy and radiotherapy. Since immunotherapy targets cells within the tumor microenvironment, understanding the behavior and interactions of different cells within that environment is essential to adequately understand both therapy options and therapy resistance. This review focuses on reviewing and analyzing the special features of cancer stem cells (CSCs), which we believe may contribute to cancer resistance to immunotherapy. The mechanisms are classified into three main categories: mechanisms related to surface markers which are differentially expressed on CSCs and help CSCs escape from immune surveillance and immune cells killing; mechanisms related to CSC-released cytokines which can recruit immune cells and tame hostile immune responses; and mechanisms related to CSC metabolites which modulate the activities of infiltrated immune cells in the tumor microenvironment. This review also discusses progress made in targeting CSCs with immunotherapy and the prospect of developing novel cancer therapies.

PurposeThis study examines the stakeholder perception of the sustainability risks, challenges and benefits arising from managing these risks in the Singapore construction industry.Design/methodology/approachA questionnaire consisting of 89 risk factors, challenges and benefits, was administered, with 216 responses received from various stakeholders. Regression analyses were used to estimate the relationships between sustainability and business risk factors, challenges and benefits associated with business sustainability practices.FindingsStakeholders recognise the importance of the emerging sustainability risk factors, and indeed rank these almost on a par with conventional business risk factors. The inherent business risks determine the nature of sustainability risk factors for construction firms, which in turn can affect their business risks and the performance and value creation of firms. However, most stakeholders, while acknowledging that business sustainability practices can provide benefits as well as posing challenges, do not believe that they can derive net benefits from such practices.Research limitations/implicationsThrough this perception study, there is an urgent need to turn the existing awareness of the importance of business sustainability (BS) practices into more consistent and solid actions among construction firms in Singapore.Practical implicationsThis study’s results imply construction firms to incorporate BS practices more systematically into their business strategies and operations, and to include sustainability risk factors alongside conventional business risks in their risk registers and risk management frameworks.Originality/valueThis study consolidates various variables and constructs of BS matters in the literature and practice into a meaningful framework for the management of BS in the construction industry.