Mutations in MAP3K1 tilt the balance from SOX9/FGF9 to WNT/β-catenin signaling

In-frame missense and splicing mutations (resulting in a 2 amino acid insertion or a 34 amino acid deletion) dispersed through the MAP3K1 gene tilt the balance from the male to female sex-determining pathway, resulting in 46,XY disorder of sex development. These MAP3K1 mutations mediate this balance by enhancing WNT/β-catenin/FOXL2 expression and β-catenin activity and by reducing SOX9/FGF9/FGFR2/SRY expression. These effects are mediated at multiple levels involving MAP3K1 interaction with protein co-fact

Ectopic Otoconin 90 expression in triple negative breast cancer cell lines is associated with metastasis functions.

Triple negative breast cancer (TNBC) is an aggressive tumor with propensity to metastasize and poor treatment options. Improving treatment options would be impactful; thus, finding a tumor-specific cell surface protein with metastasis promoting functions that could be knocked out was the goal of this study. The Otoconin 90 gene (OC90), frequently amplified in tumors on chromosome 8q24.22, was identified as a potential therapeutic candidate. Normally OC90 is expressed in the cochlea with no known function in other normal tissues. In silico analysis of The Cancer Genome Atlas (TCGA) multi-tumor RNAseq cohorts revealed that OC90 is expressed in many tumor types at high prevalence and genomic amplification is associated with the elevated mRNA expression. In vitro assays in TNBC cell lines revealed OC90 expression with control over cell viability, apoptosis and invasion. RNA-seq analysis of OC90-siRNA knockdown and OC90-overexpression in BT20, BT549, HCC38 cell lines identified co-expressed transcripts, HMGA2, POLE2 and TRIB3. Altered expression of HMGA2, POLE2 and TRIB3 was predictive of survival among members of the Metabric breast cancer cohort. Thus, OC90 represents a potential therapeutic target whose knockdown could improve the treatment of TNBC

Compilation of mRNA Polyadenylation Signals in Arabidopsis Revealed a New Signal Element and Potential Secondary Structures

Using a novel program, SignalSleuth, and a database containing authenticated polyadenylation [poly(A)] sites, we analyzed the composition of mRNA poly(A) signals in Arabidopsis (Arabidopsis thaliana), and reevaluated previously described cis-elements within the 3′-untranslated (UTR) regions, including near upstream elements and far upstream elements. As predicted, there are absences of high-consensus signal patterns. The AAUAAA signal topped the near upstream elements patterns and was found within the predicted location to only approximately 10% of 3′-UTRs. More importantly, we identified a new set, named cleavage elements, of poly(A) signals flanking both sides of the cleavage site. These cis-elements were not previously revealed by conventional mutagenesis and are contemplated as a cluster of signals for cleavage site recognition. Moreover, a single-nucleotide profile scan on the 3′-UTR regions unveiled a distinct arrangement of alternate stretches of U and A nucleotides, which led to a prediction of the formation of secondary structures. Using an RNA secondary structure prediction program, mFold, we identified three main types of secondary structures on the sequences analyzed. Surprisingly, these observed secondary structures were all interrupted in previously constructed mutations in these regions. These results will enable us to revise the current model of plant poly(A) signals and to develop tools to predict 3′-ends for gene annotation

Digital electronics in fibres enable fabric-based machine-learning inference

AbstractDigital devices are the essential building blocks of any modern electronic system. Fibres containing digital devices could enable fabrics with digital system capabilities for applications in physiological monitoring, human-computer interfaces, and on-body machine-learning. Here, a scalable preform-to-fibre approach is used to produce tens of metres of flexible fibre containing hundreds of interspersed, digital temperature sensors and memory devices with a memory density of ~7.6 × 105 bits per metre. The entire ensemble of devices are individually addressable and independently operated through a single connection at the fibre edge, overcoming the perennial single-fibre single-device limitation and increasing system reliability. The digital fibre, when incorporated within a shirt, collects and stores body temperature data over multiple days, and enables real-time inference of wearer activity with an accuracy of 96% through a trained neural network with 1650 neuronal connections stored within the fibre. The ability to realise digital devices within a fibre strand which can not only measure and store physiological parameters, but also harbour the neural networks required to infer sensory data, presents intriguing opportunities for worn fabrics that sense, memorise, learn, and infer situational context.</jats:p

Characterization of TAK-264 binding, internalization, and <i>in vitro</i> cytotoxicity.

<p>(A) Unconjugated anti-GCC monoclonal antibody and TAK-264 bind to GCC-expressing cells. Flow cytometry was performed with unconjugated anti-GCC monoclonal antibody and TAK-264 on HEK293-GCC or HEK293-vector cells. Mean fluorescence intensity is plotted versus monoclonal antibody concentration ranging from 0.001–1 μg/mL. (B) TAK-264 is internalized by GCC-expressing cells. Internalization assays were performed with TAK-264 on HEK-GCC cells. Antibodies were incubated with cells at (i) 4°C for 30 min or at (ii) 37°C for 3 h, prior to fixing and staining with secondary antibody. Immunofluorescence was visualized with a confocal microscope (Zeiss Pascal) and analyzed using Axiovert software. (C) Time-dependent accumulation of MMAE in samples incubated with 1 μg TAK-264 at 37°C, 5% CO₂, and analyzed by LC/MS/MS. (D) <i>In vitro</i> cytotoxicity of TAK-264. Cell viability was assessed 4 days after incubation of HEK293-GCC cells or HEK293-vector cells with increasing concentrations of unconjugated and MMAE-conjugated anti-GCC monoclonal antibody. The average of three independent experiments is plotted with error represented as SEM. GCC, guanylyl cyclase C; LC/MS/MS, liquid chromatography–tandem mass spectrometry; MMAE, monomethyl auristatin E; SEM, standard error of the mean.</p

Phospho-histone H3 and MMAE levels in HEK293-GCC and PHTX-09c xenograft tumors following TAK-264 treatment.

<p>(A) HEK293-GCC and (B) PHTX-09 xenograft tumors were harvested at the indicated times post-treatment, with three animals per time point. Control tumors were taken from three untreated mice. The average percentage of phospho-histone H3-positive cells and average level of MMAE tumor concentration are plotted, with error represented as standard deviation. GCC, guanylyl cyclase C; MMAE, monomethyl auristatin E.</p

TAK-264 induces tumor regression and long-term tumor growth delay of GCC-expressing tumors.

<p>Female SCID mice bearing (A) GCC-expressing HEK293 xenograft tumors and (B) PHTX-09c tumors, a patient-derived xenograft model of mCRC, were treated when the tumor reached approximately 200 mm³ with either vehicle, non-targeting control antibody 209-vcMMAE, free MMAE, or TAK-264 at various doses on a weekly dosing schedule indicated by the black triangles. Average tumor volume was determined at multiple time points following start of treatment and is shown ± S.E.M.</p

A monomethyl auristatin E-conjugated antibody to guanylyl cyclase C is cytotoxic to target-expressing cells <i>in vitro</i> and <i>in vivo</i>

<div><p>Guanylyl cyclase C (GCC) is a cell-surface protein that is expressed by normal intestinal epithelial cells, more than 95% of metastatic colorectal cancers (mCRC), and the majority of gastric and pancreatic cancers. Due to strict apical localization, systemically delivered GCC-targeting agents should not reach GCC in normal intestinal tissue, while accessing antigen in tumor. We generated an investigational antibody-drug conjugate (TAK-264, formerly MLN0264) comprising a fully human anti-GCC monoclonal antibody conjugated to monomethyl auristatin E via a protease-cleavable peptide linker. TAK-264 specifically bound, was internalized by, and killed GCC-expressing cells <i>in vitro</i> in an antigen-density-dependent manner. In GCC-expressing xenograft models with similar GCC expression levels/patterns observed in human mCRC samples, TAK-264 induced cell death, leading to tumor regressions and long-term tumor growth inhibition. TAK-264 antitumor activity was generally antigen-density-dependent, although some GCC-expressing tumors were refractory to TAK-264-targeted high local concentrations of payload. These data support further evaluation of TAK-264 in the treatment of GCC-expressing tumors.</p></div