Estrogen-related receptor alpha (ERRα) promotes the migration, invasion and angiogenesis of breast cancer stem cell-like cells.

Breast cancer progression, metastasis and recurrence are largely driven by breast cancer stem cells (BCSCs), which constitute a subset of tumor cells exhibiting stem cell characteristics. In this study, we evaluated the role of estrogen-related receptor alpha (ERRα) in the migration, invasion and angiogenesis of BCSCs. The inhibition of ERRα using XCT790 or knockdown of ERRα using shRNA inhibited the mammosphere formation efficiency, as well as the migration and invasion of BCSCs derived from the mammospheres of MCF7 and MDA-MB-231 (MB231) cells. Conversely, the overexpression of ERRα significantly increased the migration and invasion of BCSCs derived from the mammosphere. In addition, the XCT790 treatment or shERRα significantly downregulated the epithelial-mesenchymal transition (EMT), as evidenced by the downregulation in the expression of vimentin, Snail, Slug and N-cadherin in the mammospheres of MCF7 and MB231 cells. The chorioallantoic membrane assay showed that the conditioned media from XCT790-treated and shERRα cells significantly inhibited blood vessel formation and vessel length. Furthermore, XCT790 treatment or shERRα also downregulated the expression of molecular markers of angiogenesis, such as VEGF-A and Ang-2 in the mammospheres. Conversely, the overexpression of ERRα in MCF7 cells significantly increased both EMT and angiogenesis. These findings suggest that ERRα inhibits the migration, invasion and angiogenesis of BCSCs, suggesting as a potential target for breast cancer therapy.

Use of inferior vena cava guided fluid therapy in the treatment of septic shock: A randomised controlled trial.

INTRODUCTION: By administering inferior vena cava (IVC) directed fluid, it is possible to avoid the use of additional fluid and fluid overload in patients with septic shock (SS) and sepsis-induced hypoperfusion (SIH). METHODOLOGY: In patients with SIH and SS, we conducted prospective observational research on fluid therapy. A time-motion trace of the IVC diameter was created using M-mode imaging. The ability to predict fluid responsiveness was based on the IVC collapsibility index (cIVC) > 40%. Participants were randomised into 2 groups using a permuted block-of-four randomization list, with the investigators being blinded prior to patient allocation. They were split equally between the usual-care (UC) group, which received sepsis-guided fluid treatment, and the interventional ultrasound-guided fluid therapy (UGFT) group. RESULTS: The average age of the participants was 63.2 years (62.8 years for the UGFT group and 63.7 years for the UC group). Co-morbid health conditions were practically the same in both arms at baseline. Prior to enrolment, both groups received the same quantity of fluid as part of resuscitation (UGFT arm received 2.4 0.6 L, UC group received 2.2 0.7 L). The UGFT group outperformed the UC group with a P value of 0.02 due to a significantly lower positive fluid balance after 72 hours of ICU discharge (-1.37 L), which rendered the UGFT group superior to the UC group. Even after accounting for the fluids consumed before enrolment, there was still a sizable difference in the fluids infused. When the pre-enrolment fluids were counted at 72 hours, UGFT participants still displayed a decreased positive fluid balance. However, there was no discernible difference in the 30-day mortality rate overall (6.3% difference, UGFT: 15.7%, and UC: 22.0%). CONCLUSIONS: In contrast to the UC group, the UGFT arm of our study demonstrated a statistically significant benefit of Point of Care USG (POCUS) guided fluid therapy during resuscitation in sepsis in reducing the positive fluid balance in 72 hours, preventing fluid overload, and reducing the need for dialysis and invasive ventilation. However, there was no statistically significant variation in the 30-day mortality rate.

Estrogen-Related Receptor Alpha (ERRα) Promotes Cancer Stem Cell-Like Characteristics in Breast Cancer.

Cancer stem cells drive tumor initiation, invasion, metastasis and recurrence. In the present study, we have evaluated the role of ERRα in the maintenance of breast cancer stem cells (BCSCs) using breast cancer cell lines. The inhibition of ERRα with the inverse agonist, XCT-790, or the knockdown of ERRα in breast cancer cells significantly reduced the mammosphere formation efficiency and mammosphere size along with a significant reduction in the CD44+/CD24- BCSCs. Treatment with XCT-790 significantly downregulated expression of the transcription factors involved in stem cell maintenance such as Oct4, Klf4, Sox2, Nanog and c-Myc in the mammosphere forming stem cells of MCF7 and MDA-MB-231. In addition, XCT-790 induced cell cycle arrest and apoptosis in the mammosphere-forming cells. The knockdown or inhibition of ERRα downregulated the expression of Notch1 and ß-catenin, whereas the overexpression of ERRα in MCF7 cells upregulated the expression of these proteins. Moreover, the inhibition of ERRα synergistically enhanced the efficacy of paclitaxel in inhibiting the BCSCs. These results show that ERRα is crucial for the maintenance of BCSCs and suggest that ERRα could be a potential target for breast cancer treatment.

In Vitro Characterization of Histone Chaperones using Analytical, Pull-Down and Chaperoning Assays.

Histone proteins associate with DNA to form the eukaryotic chromatin. The basic unit of chromatin is a nucleosome, made up of a histone octamer consisting of two copies of the core histones H2A, H2B, H3, and H4, wrapped around by the DNA. The octamer is composed of two copies of an H2A/H2B dimer and a single copy of an H3/H4 tetramer. The highly charged core histones are prone to non-specific interactions with several proteins in the cellular cytoplasm and the nucleus. Histone chaperones form a diverse class of proteins that shuttle histones from the cytoplasm into the nucleus and aid their deposition onto the DNA, thus assisting the nucleosome assembly process. Some histone chaperones are specific for either H2A/H2B or H3/H4, and some function as chaperones for both. This protocol describes how in vitro laboratory techniques such as pull-down assays, analytical size-exclusion chromatography, analytical ultra-centrifugation, and histone chaperoning assay could be used in tandem to confirm whether a given protein is functional as a histone chaperone.