It is well established that transforming growth factor-β (TGFβ) switches its function from being a tumor suppressor to a tumor promoter during the course of tumorigenesis, which involves both cell-intrinsic and environment-mediated mechanisms. We are interested in breast cancer cells, in which SMAD mutations are rare and interactions between SMAD and other transcription factors define pro-oncogenic events. Here, we have performed chromatin immunoprecipitation (ChIP)-sequencing analyses which indicate that the genome-wide landscape of SMAD2/3 binding is altered after prolonged TGFβ stimulation. De novo motif analyses of the SMAD2/3 binding regions predict enrichment of binding motifs for activator protein (AP)1 in addition to SMAD motifs. TGFβ-induced expression of the AP1 component JUNB was required for expression of many late invasion-mediating genes, creating a feed-forward regulatory network. Moreover, we found that several components in the WNT pathway were enriched among the late TGFβ-target genes, including the invasion-inducing WNT7 proteins. Consistently, overexpression of WNT7A or WNT7B enhanced and potentiated TGFβ-induced breast cancer cell invasion, while inhibition of the WNT pathway reduced this process. Our study thereby helps to explain how accumulation of pro-oncogenic stimuli switches and stabilizes TGFβ-induced cellular phenotypes of epithelial cells.

Dysregulated bone morphogenetic protein (BMP) signaling in endothelial cells (ECs) is implicated in vascular diseases such as pulmonary arterial hypertension (PAH). Here, we showed that the transcription factor ATOH8 was a direct target of SMAD1/5 and was induced in a manner dependent on BMP but independent of Notch, another critical signaling pathway in ECs. In zebrafish and mice, inactivation of Atoh8 did not cause an arteriovenous malformation–like phenotype, which may arise because of dysregulated Notch signaling. In contrast, Atoh8-deficient mice exhibited a phenotype mimicking PAH, which included increased pulmonary arterial pressure and right ventricular hypertrophy. Moreover, ATOH8 expression was decreased in PAH patient lungs. We showed that in cells, ATOH8 interacted with hypoxia-inducible factor 2α (HIF-2α) and decreased its abundance, leading to reduced induction of HIF-2α target genes in response to hypoxia. Together, these findings suggest that the BMP receptor type II/ALK-1/SMAD/ATOH8 axis may attenuate hypoxic responses in ECs in the pulmonary circulation and may help prevent the development of PAH.

Over 350 million people across the world suffer from major depressive disorder (MDD). More than 10% of MDD patients have suicide intent, while it has been reported that more than 40% patients did not consult their doctors for MDD. In order to increase consultation rate of potential MDD patients, we developed a novel MDD screening system which can be used at home without help of health-care professionals. Using a fingertip photoplethysmograph (PPG) sensor as a substitute of electrocardiograph (ECG), the system discriminates MDD patients from healthy subjects using autonomic nerve transient responses induced by a mental task (random number generation) via logistic regression analysis. The nine logistic regression variables are averages of heart rate (HR), high frequency (HF) component of heart rate variability (HRV), and the low frequency (LF)/HF ratio of HRV before, during, and after the mental task. We conducted a clinical test of the proposed system. Participants were 6 MDD patients (4 females and 2 males, aged 23–60 years) from Shizuoka Saiseikai General Hospital psychiatry outpatient unit and 14 healthy volunteers from University of Electro-Communications (6 females and 8 males, aged 21–63 years). The average PPG- and ECG (as a reference)-derived HR, HF and LF/HF were significantly correlated with each other (HR; r = 1.00, p < 0.0001, HF; r = 0.98, p < 0.0001, LF/HF; r = 0.98, p < 0.0001). Leave-one-out cross validation (LOOCV) revealed 83% sensitivity and 93% specificity. The proposed system appears promising for future MDD self-screening at home and are expected to encourage psychiatric visits for potential MDD patients.

Cyclin‐dependent kinase (CDK) 4 and CDK6 inhibitors are effective therapeutic options for hormone receptor (HR)‐positive, human epidermal growth factor receptor 2 (HER2)‐negative advanced breast cancer. Although CDK4/6 inhibitors mainly target the cyclin D‐CDK4/6‐retinoblastoma tumor suppressor protein (RB) axis, little is known about the clinical impact of inhibiting phosphorylation of other CDK4/6 target proteins. Here, we focused on other CDK4/6 targets, SMAD proteins. We showed that a CDK4/6 inhibitor palbociclib and activin‐SMAD2 signaling cooperatively inhibited cell cycle progression of a luminal‐type breast cancer cell line T47D. Palbociclib enhanced SMAD2 binding to the genome by inhibiting CDK4/6‐mediated linker phosphorylation of the SMAD2 protein. We also showed that cyclin G2 plays essential roles in SMAD2‐dependent cytostatic response. Moreover, comparison of the SMAD2 ChIP‐seq data of T47D cells with those of Hs578T (triple‐negative breast cancer cells) indicated that palbociclib augmented different SMAD2‐mediated functions based on cell type, and enhanced SMAD2 binding to the target regions on the genome without affecting its binding pattern. In summary, palbociclib enhances the cytostatic effects of the activin‐SMAD2 signaling pathway, whereas it possibly strengthens the tumor‐promoting aspect in aggressive breast cancer.

Since objective biomarkers for major depressive disorder (MDD) are not readily available, clinical psychiatrists diagnose patients with MDD subjectively based on clinical interviews and diagnostic criteria. It often raises various concerns, including false responses by patients, subjective factors, and inexperience of the attendants leading to incorrect diagnosis. Here, we developed a self-monitoring system for simple and objective screening of MDD using a photoplethysmography (PPG) sensor and a 24-GHz microwave radar, which was based on the analysis of heart rate variability (HRV) during paced respiration and mental task conditions. In our previous study, we assessed the reactivity of HRV measurements during a mental task (random number generation) condition in patients with MDD and healthy control subjects. The HRV indices are less reactive in patients with MDD compared to healthy subjects during the mental task, which enabled us to identify the patients at risk for depression. In this study, the reactivity of HRV was measured not only in the mental task but also during paced respiration (i.e., 5-s inhalation and 5-s exhalation) conditions, thereby assessing more detailed autonomic nervous system (ANS) activity via HRV indices. To investigate the effect of paced respiration on MDD screening, we compared the ANS activity via HRV indices in with/without paced respiration conditions in 28 drug-naïve patients with MDD and 27 healthy control subjects. The result showed that ANS significantly activated during the paced respiration condition (p<;0.05). The sensitivity in detecting patients with MDD was 86% under paced respiration and mental task conditions, which was higher than the sensitivity (68%) under mental task condition alone.