Asporin binds to TGF-β1 and inhibits its downstream signaling and function.

<p>(A) Western blot analysis of phospho-SMAD2 (p-SMAD2) and SMAD2 total protein extracts from MDA-MB-468 breast cancer cells treated for 15 min with TGF-β1 and/or human recombinant asporin (Rec. ASPN). (B) Western blot analysis of p-SMAD2 in total protein extracts from MDA-MB-468 breast cancer cells treated with TGF-β1 and/or asporin peptide corresponding to the 159–205 amino acid region (ASPNpep.). (C) Western blot analysis of p-SMAD2 and SMAD2 in total protein extracts from EpRAS cells treated for 15 min with TGF-β1 (5 ng/ml) and/or asporin peptide. (D) EMT induction in EpRAS cells in the presence of TGF-β1 and/or asporin peptide. EMT was monitored both at the phenotype level (upper panel) and using Western blot evaluation of VIM expression in total protein extracts from EpRAS cells (lower panel). (A–D): HSC70 was used as loading control. (E) Transwell migration assay of EpRAS cells pretreated with TGF-β1 (5 ng/ml) and/or asporin peptide (10 μg/ml). (F) Quantification of the CSC population in EpRAS cells following TGF-β1 and/or asporin peptide treatment. (E and F): The data are presented as mean ± SD. All panels: statistical significance was calculated using the Student’s <i>t</i>-test (as described in the <a href="http://www.plosmedicine.org/article/info:doi/10.1371/journal.pmed.1001871#sec010" target="_blank">Methods</a> section). Western blots show representative data of three independent experiments.</p

Asporin is overexpressed in breast cancer tissues.

<p>(A) Tissue-specific pattern of mRNA expression of asporin (ASPN), biglycan (BGN), and decorin (DCN). Source: BioGPS (<a href="http://biogps.org/" target="_blank">http://biogps.org</a>). The data are presented as mean ± standard deviation (SD). (B) Representative IHC staining of asporin expression in ductal carcinoma and adjacent non-tumoral breast tissue (left panel) and normal breast tissue obtained from patients undergoing mammary reduction surgery (right panel). Asporin is almost exclusively expressed in breast cancer lesions, while a very low signal is detectable in the adjacent non-tumoral regions. Normal breast tissues are negative. Images of representative fields were taken at 100× and 400× magnification. (C) Western blot analysis of asporin expression in tumoral breast cancer tissues (T) and the adjacent normal counterpart (AdN) of six ductal adenocarcinoma patients. Ponceau red staining was used as loading control.</p

Co-injection of cancer cells and fibroblasts overexpressing asporin reduces primary breast cancer tumor growth and lung metastasis formation in vivo.

<p>(A) Western blot analysis of asporin expression in CM of MDA-MB-468 cells and in NBF stable clones used for subcutaneous injection in mice. Ponceau red is shown as loading control. (B) Bioluminescence imaging of control and asporin-expressing xenografts at day 28 after tumor engraftment. The color scale indicates the fluorescent intensity. (C) The volume (in cubic millimeters) of primary tumors measured weekly (from day 7 onwards). The data are presented as mean ± standard error of the mean (SEM) (<i>n</i> = 10 for each group). Statistical significance was calculated using Student’s <i>t</i>-test (**0.01 < <i>p</i> < 0.001; ***0.001 < <i>p</i> < 0.0001). (D) Human-specific Alu-PCR performed on genomic DNA isolated from dissected lungs was used to detect human cancer cells. The data are presented as mean ± SD. (E) Western blot analysis of asporin expression in mice primary tumors monitored for several weeks. HSC70 was used as loading control.</p

Asporin is produced by breast fibroblasts in response to conditioned medium from breast cancer cells.

<p>(A) Western blot of total cell extracts (upper panel) and qRT-PCR analysis for asporin expression (lower panel) in breast cancer cell lines and NBFs incubated for 48 h with CM collected from a panel of breast cancer cells. (B) Western blot of total cell extracts (upper panel) and qRT-PCR analysis of asporin expression (lower panel) in non-cancerous epithelial breast cell line MCF-10A cells and NBFs incubated for 48 h with CM collected from MCF-10A. Fibroblasts treated with MCF-7 CM were used as the positive control for asporin expression induction. (C) Validation of NBFs and CAFs isolated from patient material. MCF-7 and MDA-MB-231 cells were used as epithelial controls. (D) Western blot analysis of asporin expression in total cell extracts of CAFs obtained from three different patients and treated with the CM of breast cancer cell lines. (A and B): The data are presented as mean ± SD. All panels: HSC70 was used as loading control; Western blots show representative data of three independent experiments.</p

COVID-19 Host Genetics Initiative. A first update on mapping the human genetic architecture of COVID-19

The COVID-19 pandemic continues to pose a major public health threat, especially in countries with low vaccination rates. To better understand the biological underpinnings of SARS-CoV-2 infection and COVID-19 severity, we formed the COVID-19 Host Genetics Initiative1. Here we present a genome-wide association study meta-analysis of up to 125,584 cases and over 2.5 million control individuals across 60 studies from 25 countries, adding 11 genome-wide significant loci compared with those previously identified2. Genes at new loci, including SFTPD, MUC5B and ACE2, reveal compelling insights regarding disease susceptibility and severity.</p