A comprehensive cellular anatomy of normal human prostate is essential for solving the cellular origins of benign prostatic hyperplasia and prostate cancer. The tools used to analyze the contribution of individual cell types are not robust. We provide a cellular atlas of the young adult human prostate and prostatic urethra using an iterative process of single-cell RNA sequencing (scRNA-seq) and flow cytometry on ∼98,000 cells taken from different anatomical regions. Immunohistochemistry with newly derived cell type-specific markers revealed the distribution of each epithelial and stromal cell type on whole mounts, revising our understanding of zonal anatomy. Based on discovered cell surface markers, flow cytometry antibody panels were designed to improve the purification of each cell type, with each gate confirmed by scRNA-seq. The molecular classification, anatomical distribution, and purification tools for each cell type in the human prostate create a powerful resource for experimental design in human prostate disease.

Objective: ARID1A is commonly mutated in pancreatic ductal adenocarcinoma (PDAC), but the functional effects of ARID1A mutations in the pancreas are unclear. Understanding the molecular mechanisms that drive PDAC formation may lead to novel therapies. Design: Concurrent conditional Arid1a deletion and Kras activation mutations were modelled in mice. Small-interfering RNA (siRNA) and CRISPR/Cas9 were used to abrogate ARID1A in human pancreatic ductal epithelial cells. Results: We found that pancreas-specific Arid1a loss in mice was sufficient to induce inflammation, pancreatic intraepithelial neoplasia (PanIN) and mucinous cysts. Concurrent Kras activation accelerated the development of cysts that resembled intraductal papillary mucinous neoplasm. Lineage-specific Arid1a deletion confirmed compartment-specific tumour-suppressive effects. Duct-specific Arid1a loss promoted dilated ducts with occasional cyst and PDAC formation. Heterozygous acinar-specific Arid1a loss resulted in accelerated PanIN and PDAC formation with worse survival. RNA-seq showed that Arid1a loss induced gene networks associated with Myc activity and protein translation. ARID1A knockdown in human pancreatic ductal epithelial cells induced increased MYC expression and protein synthesis that was abrogated with MYC knockdown. ChIP-seq against H3K27ac demonstrated an increase in activated enhancers/promoters. Conclusions: Arid1a suppresses pancreatic neoplasia in a compartment-specific manner. In duct cells, this process appears to be associated with MYC-facilitated protein synthesis.

The MYC oncogene is frequently amplified in triple-negative breast cancer (TNBC). Here, we show that MYC suppression induces immune-related hallmark gene set expression and tumor-infiltrating T cells in MYC-hyperactivated TNBCs. Mechanistically, MYC repressed stimulator of interferon genes (STING) expression via direct binding to the STING1 enhancer region, resulting in downregulation of the T-cell chemokines CCL5, CXCL10, and CXCL11. In primary and metastatic TNBC cohorts, tumors with high MYC expression or activity exhibited low STING expression. Using a CRISPR-mediated enhancer perturbation approach, we demonstrated that MYC-driven immune evasion is mediated by STING repression. STING repression induced resistance to PD-L1 blockade in mouse models of TNBC. Finally, a small-molecule inhibitor of MYC combined with PD-L1 blockade elicited a durable response in immune-cold TNBC with high MYC expression, suggesting a strategy to restore PD-L1 inhibitor sensitivity in MYC-overexpressing TNBC.

Significance: Clathrin-mediated endocytosis (CME), the major pathway for uptake into cells, is a multistep process involving a myriad of endocytic accessory proteins (EAPs). Although the biochemical activities of many EAPs have been extensively studied, their stage-specific role(s) during clathrin-coated pit (CCP) initiation, stabilization, and/or maturation are poorly defined. Here, using quantitative total internal reflection fluorescence microscopy and a rigorous experimental and multiparametric analytical pipeline, we study the effect of CRISPR interference (CRISPRi)- or small interfering RNA (siRNA)-mediated knockdown of 67 individual EAPs on CCP dynamics. Our comprehensive analyses combined with unsupervised phenotypic clustering reveal the complex and overlapping roles of EAPs during early, critical stages of CME, providing a valuable resource to spur further research into their function.

Autism spectrum disorder (ASD) is a neurodevelopmental disorder characterized by deficits in social communication and social interaction and restricted, repetitive patterns of behavior, interests, or activities. Given the lack of specific pharmacological therapy for ASD and the clinical heterogeneity of the disorder, current biomarker research efforts are geared mainly toward identifying markers for determining ASD risk or for assisting with a diagnosis. A wide range of putative biological markers for ASD is currently being investigated. Proteomic analyses indicate that the levels of many proteins in plasma/serum are altered in ASD, suggesting that a panel of proteins may provide a blood biomarker for ASD. Serum samples from 76 boys with ASD and 78 typically developing (TD) boys, 18 months-8 years of age, were analyzed to identify possible early biological markers for ASD. Proteomic analysis of serum was performed using SomaLogic’s SOMAScan^TMassay 1.3K platform. A total of 1,125 proteins were analyzed. There were 86 downregulated proteins and 52 upregulated proteins in ASD (FDR < 0.05). Combining three different algorithms, we found a panel of 9 proteins that identified ASD with an area under the curve (AUC) = 0.8599±0.0640, with specificity and sensitivity of 0.8217±0.1178 and 0.835±0.1176, respectively. All 9 proteins were significantly different in ASD compared with TD boys, and were significantly correlated with ASD severity as measured by ADOS total scores. Using machine learning methods, a panel of serum proteins was identified that may be useful as a blood biomarker for ASD in boys. Further verification of the protein biomarker panel with independent test sets is warranted.

In this work, we analyze the baseline, signal strength, aortic augmentation index (AIx), radial AIx, time to reflection and P_T2 at Chon, Gwan, and Cheok, which are the three pulse diagnosis positions in Oriental medicine. For the pulse measurement, we used the SphygmoCor apparatus, which has been widely used for the evaluation of the arterial stiffness at the aorta. By two‐way repeated measures analysis of variance, we tested two independent measurements for repeatability and investigated their mean differences among Chon, Gwan and Cheok. To characterize further the parameters that were shown to be different between each palpation position, we carried out Duncan′s test for the multiple comparisons. The baseline and signal strength were statistically different (P< .05) among Chon, Gwan and Cheok, respectively, which supports the major hypothesis of Oriental medicine that all of the three palpation positions contain different clinical information. On the other hand, aortic AIx and time to reflection were found to be statistically different between Chon and the others, and radial AIx and P_T2 did not show any difference between pulse positions. In the clinical sense, however, the aortic AIx at each palpation position was found to fall within the 90% confidence interval of normal arterial compliance. The results of the multiple comparisons indicate that the parameters of arterial stiffness were independent of the palpation positions. This work is the first attempt to characterize quantitatively the pulse signals at Chon, Gwan and Cheok with some relevant parameters extracted from the SphygmoCor apparatus.