Cushing's disease, also known as adrenocorticotropic hormone (ACTH)-secreting pituitary adenomas (PAs) that cause excess cortisol production, accounts for up to 85% of corticotrophin-dependent Cushing's syndrome cases. However, the genetic alterations in this disease are unclear. Here, we performed whole-exome sequencing of DNA derived from 12 ACTH-secreting PAs and matched blood samples, which revealed three types of somatic mutations in a candidate gene, USP8 (encoding ubiquitin-specific protease 8), exclusively in exon 14 in 8 of 12 ACTH-secreting PAs. We further evaluated somatic USP8 mutations in additional 258 PAs by Sanger sequencing. Targeted sequencing further identified a total of 17 types of USP8 variants in 67 of 108 ACTH-secreting PAs (62.04%). However, none of these mutations was detected in other types of PAs (n = 150). These mutations aggregate within the 14-3-3 binding motif of USP8 and disrupt the interaction between USP8 and 14-3-3 protein, resulting in an elevated capacity to protect EGFR from lysosomal degradation. Accordingly, PAs with mutated USP8 display a higher incidence of EGFR expression, elevated EGFR protein abundance and mRNA expression levels of POMC, which encodes the precursor of ACTH. PAs with mutated USP8 are significantly smaller in size and have higher ACTH production than wild-type PAs. In surgically resected primary USP8-mutated tumor cells, USP8 knockdown or blocking EGFR effectively attenuates ACTH secretion. Taken together, somatic gain-of-function USP8 mutations are common and contribute to ACTH overproduction in Cushing's disease. Inhibition of USP8 or EGFR is promising for treating USP8-mutated corticotrophin adenoma. Our study highlights the potentially functional mutated gene in Cushing's disease and provides insights into the therapeutics of this disease.

MicroRNAs (miRNAs) have emerged as important regulators in cancer that are implicated in regulation of various cellular processes. miR-382 has been proposed as a tumor suppressor by several recent studies. However, the function of miR-382 in prostate cancer remains unknown. In this study, we aimed to investigate the potential function of miR-382 in prostate cancer. We found that miR-382 was significantly decreased in prostate cancer specimens and cancer cell lines. The overexpression of miR-382 in prostate cancer cells markedly inhibited cell proliferation, migration, and invasion. In contrast, miR-382 suppression exhibited an opposite effect. Target analysis predicted that chicken ovalbumin upstream promoter transcription factor II (COUP‑TFII) was a direct target of miR-382. This prediction was experimentally confirmed by dual-luciferase reporter assay, real-time quantitative polymerase chain reaction, and western blot analysis. Our results further demonstrated that miR-382 inhibited the downstream genes of COUP‑TFII, including Snail and matrix metalloproteinase 2 (MMP2). Moreover, the restoration of COUP‑TFII expression significantly blocked the inhibitory effect of miR-382 on cell proliferation, migration, and invasion, and Snail expression. Taken together, this study suggests that miR-382 inhibits prostate cancer cell proliferation and metastasis through inhibiting COUP‑TFII, representing an important new mechanism for understanding prostate cancer pathogenesis and providing a novel therapeutic candidate target for prostate cancer therapy.

To maintain brain homeostasis, a unique interface known as the blood-brain barrier (BBB) is formed between the blood circulation and the central nervous system (CNS). Major facilitator superfamily domain-containing 2a (Mfsd2a) is a specific marker of the BBB. However, the mechanism by which Mfsd2a influences the BBB is poorly understood. In this study, we demonstrated that Mfsd2a is essential for sphingosine-1-phosphate (S1P) export from endothelial cells in the brain. We found that Mfsd2a and Spinster homolog 2 (Spns2) form a protein complex to ensure the efficient transport of S1P. Furthermore, the S1P-rich microenvironment in the extracellular matrix (ECM) in the vascular endothelium dominates the formation and maintenance of the BBB. We demonstrated that different concentrations of S1P have different effects on BBB integrity. These findings help to unravel the mechanism by which S1P regulates BBB and also provide previously unidentified insights into the delivery of neurological drugs in the CNS.

A stable GFP-expressing (GFP + LJES1) cell strain was developed from the LJES1 cells obtained from sea perch (Lateolabrax japonicus,) embryos. GFP + LJES1 cells were induced in vitro by RA to differentiate into a variety of cell types and also had the ability to form embryoid body-like structures in suspension culture. To determine the differentiation potential of LJES1 cells in vivo, GFP + LJES1 cells were transplanted into sea perch and zebrafish embryos at midblastula stage. Twenty out of 478 transplanted sea perch embryos contained GFP-expressing LJES1 cells 24 h after microinjection. Fifteen chimera embryos developed into fry. In these chimeras, the GFP + LJES1 cells contributed to a variety of tissues including the head and trunk. In zebrafish, 221 embryos were microinjected with GFP + LJES1 cells and 22 chimera embryos and fries expressing GFP were obtained. Donor GFP + LJES1 cells contributed to various tissues in head and trunk of zebrafish embryos and hatched fry.

No abstract

No abstract

No abstract

The morphology and surroundings of cells have been routinely used by pathologists to diagnose the pathological subtypes of gliomas and to assess the malignancy of tumors. Thanks to the advent and development of digital pathology technology, it is possible to automatically analyze whole slides of tissue and focus on the nucleus in order to derive a quantitative assessment that can be used for grading, classification, and diagnosis. During the process of computer-assisted diagnosis, the accurate location and segmentation of nuclei from hematoxylin and eosin (H&E)-stained histopathological images is an important step. In this paper, we proposed a U-Net-based multi-task learning network in which the boundary and region information is utilized to improve the segmentation accuracy of glioma nuclei, especially overlapping ones. To refine the segmentation, a classification model is used to predict the boundary, a regression model is used to predict the distance map, and the final segmentation is obtained by using the fusion layers. The proposed approach was compared with other specially designed boundary-aware methods by using a pathological section dataset that consists of 320 glioma cases from the Huashan Hospital at Fudan University. Both the pixel-level and object-level evaluations showed that the structural modification is effective in segmentation with an F1-score of 0.82, a Hausdorff distance (HD) of 3.95, and an aggregated Jaccard index (AJI) of 0.66 (+0.46%, −3.75%, and +4.09% compared with the unimproved methods, respectively). In addition, comparative experiments on multi-organ nuclei segmentation (MoNuSeg) open dataset proved the advanced nature of the proposed method in the field of nuclei segmentation, especially separating touching objects. The proposed method obtains an AJI of 0.59 and an F1-score of 0.79.INDEX TERMS Cancer research, deep learning, digital pathology, nuclei segmentation.