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Warthin tumor (WT; synonym: cystadenolymphoma) represents one of the most frequent salivary gland tumors with a frequency equaling or even outnumbering that of pleomorphic adenomas in some series. Histologically, the tumor displays tall columnar oncocytic cells, arranged into two cell-thick layers lining variably cystic glands within an organoid lymphoid stroma. Tumors with exuberant squamous metaplasia in response to FNA-induced or other types of tissue injury/infarction have been referred to as “metaplastic WTs.” However, the same terminology was used for tumors with variable mucinous cell and solid or stratified epidermoid proliferations (occasionally mimicking mucoepidermoid carcinoma), although the “metaplasia concept” has never been proven for the latter. We herein investigated 22 WTs showing prominent mucoepidermoid-like or solid oncocytoma-like proliferations without prior FNA or histological evidence of infarction/ trauma using the TruSight Tumor 15 gene panel and KRAS pyrosequencing. As a control, we tested 11 conventional WTs. No statistically significant differences were observed between the two subcohorts regarding patient’s age and tumor size. Six of 22 (27%) proliferating/ metaplastic WTs revealed oncogenic KRAS mutations clustering at codon 12 (exon 2), while all conventional tumors lacked these mutations. Our findings are in line with a neoplastic nature of the epidermoid/ mucoepidermoid proliferations in non-injured “metaplastic” Warthin tumors. We propose the descriptive term “de novo proliferating Warthin tumor” for this variant to distinguish it from infarcted/inflamed genuine metaplastic Warthin tumor.

With the widespread application of next-generation sequencing, the genetic landscape of uterine mesenchymal neoplasms has been evolving rapidly to include several recently identified fusion genes. Although chromosomal rearrangements involving the 10q22 and 17q21.31 loci have been reported in occasional uterine leiomyomas decades ago, the corresponding KAT6B::KANSL1 fusion has been only recently identified in 2 uterine tumors diagnosed as leiomyoma and leiomyosarcoma. We herein describe 13 uterine stromal neoplasms carrying a KAT6B::KANSL1 (n=11) and KAT6A::KANSL1 (n=2) fusion. Patient ages ranged from 33 to 81 years (median, 49 y). Tumor size was 2.6 to 23.5 cm (median, 8.2 cm). Nine tumors were myometrium-centered, and 3 had an intracavitary component. Original diagnoses were mostly low-grade endometrial stromal sarcoma (LG-ESS; 10 cases) with atypical features (limited CD10 expression, sex cord-like features, pericytic vasculature, and frequent myxoid changes). Treatment was hysterectomy±bilateral salpingo-oophorectomy (10), myomectomy (1), and curettage (2). Five patients were disease-free at 6 to 34 months, 3 (27%) died of disease at 2 to 47 months, and 3 were alive with disease at 2, 17, and 17 years. Histologically, most tumors showed variable overlap with LG-ESS, but they were generally well-circumscribed lacking the extensive permeative and angioinvasive growth typical of LG-ESS. They were composed of monotonous medium-sized oval and spindle cells arranged into diffuse sheets with prominent spiral-type arterioles and frequent pericytoma-like vascular pattern. Variable myxoid stromal changes were frequent. Mitotic activity ranged from 1 to20 in 10 HPFs. Immunohistochemistry showed variable expression of CD10 (12/13), estrogen receptor (8/11), progesterone receptor (8/11), smooth muscle actin (9/11), desmin (4/12), h-caldesmon (2/10), calretinin (3/8), inhibin (1/7), WT1 (4/7), cyclin D1 (5/11; diffuse in only 1 case), and pankeratin (5/10). This series characterizes a KAT6B/A::KANSL1 fusion-positive uterine stromal neoplasm within the morphologic spectrum of LG-ESS but with atypical features. The relationship of these neoplasms to genuine LG-ESS remains unclear. This molecular subtype of uterine endometrial stromal sarcoma has the potential for an unfavorable clinical course despite the absence of widely invasive growth; nevertheless, analysis of more cases is necessary to delineate the phenotypic spectrum and biological potential of this tumor.

Radical-based molecules are employed as functional materials in organic electronic devices.

Published online: Nov 4, 2022 Purpose: Effects related to literacy acquisition have been observed at different levels of speech processing. This study investigated the link between orthographic knowledge and children’s perception and production of specific speech sounds. Method: Sixty Spanish-speaking second graders, differing in their phonological decoding skills, completed a speech perception and a production task. In the perception task, a behavioral adaptation of the oddball paradigm was used. Children had to detect orthographically consistent /t/, which has a unique orthographic representation (hti), and inconsistent /k/, which maps onto three different graphemes (hci, hqui, and hki), both appearing infrequently within a repetitive auditory sequence. In the production task, children produced these same sounds in meaningless syllables. Results: Perception results show that all children were faster at detecting consistent than inconsistent sounds regardless of their decoding skills. In the production task, however, the same facilitation for consistent sounds was linked to better decoding skills. Conclusions: These findings demonstrate differences in speech sound processing related to literacy acquisition. Literacy acquisition may therefore affect already-formed speech sound representations. Crucially, the strength of this link in production is modulated by individual decoding skills. This project has received funding from the European Research Council under the European Union’s Horizon 2020 Research and Innovation Program (Grant Agreement No. 819093 to C.D.M.) and under the Marie Sklodowska-Curie Grant Agreement No 843533 to A.S. This work was also supported by the Spanish State Research Agency through Basque Center on Cognition, Brain and Language Severo Ochoa excellence accreditation CEX2020-001010-S, the Spanish Ministry of Economy and Competitiveness (PSI2017 82941-P and PID2020-113926GB-I00), and the Basque Government (BERC 2022-2025 and PIBA18_29). M.J. was supported by a Predoctoral fellowship (associated to the Project PSI2017 82941-P; Grant No. PRE-2018-083946) from the Spanish Ministry of Science, Innovation and Universities and the Fondo Social Europeo.

Conical deformities found at the apices of incisions after wound closure represent tissue redundancy, commonly known as "dog ears." The amount of tissue excess is related to the geometry of the incision, surrounding skin elasticity, and tension of closure. Experience with tissue handling shows that some dog ears do "settle" or resolve with time; such cases occur when local tissue has high skin elasticity, the dog ears are small in height, and tension on the line of closure is low. Dog ears that do not fall into this category can be troublesome to surgeon and patient alike as they can poorly impact aesthetic outcomes and lead to secondary revision procedures. Therefore, we present a pertinent review of the mechanisms behind dog-ear formation and the variety of surgical techniques used to minimize and correct their formation.

The mammalian Vestigial-like (VGLL) transcriptional cofactor family of proteins VGLL1-4 has recently emerged as an important player in the tumorigenesis of diverse neoplasms. The role of VGLL3 in soft tissue tumors is exemplified by its amplification in myxoinflammatory fibroblastic sarcoma and its rearrangement (fused to CHD7, CHD9, or MAMLD1) in hybrid schwannoma-perineurioma. This study characterizes a distinctive low-grade myogenic neoplasm with a striking predilection for the head and neck, characterized by VGLL3 fusions. The study includes five males and one female patient, aged 30-71 years (median, 56). Three tumors originated in the tongue, with one case each in the nasopharynx, oral cavity, and oropharynx. The VGLL3 fusion partners included TCF12 (n = 3), EP300 (n = 2), and PPARGC1A (n = 1). The tumor size range was 0.8-1.6 cm (all, but one, was1 cm). Histologically, all tumors displayed bland spindle to ovoid cells arranged into vague fascicular and diffuse patterns. Mitotic activity ranged from 1 to 7 per 10 HPFs. Five tumors were muscle-centered and infiltrative, and one was centered beneath nasopharyngeal mucosa. Immunohistochemistry revealed consistent expression of desmin (diffuse in four and patchy in two cases) associated with patchy smooth muscle actin expression (4/6), and focal reactivity for myogenin (5/6) and myoD1 (1/3). All patients were managed surgically; one patient each received adjuvant radio- or chemotherapy. Three patients with follow-up were without disease at 8, 19, and 60 months and one was alive with unknown disease status at 24 months. All VGLL3 fusions were in-frame and involved exon 2, fused with either TCF12 exon 16, EP300 exon 31, or PPARGC1A exon 5, respectively. This series characterizes a distinctive subset of spindle cell rhabdomyosarcoma (RMS) with a predilection for the head and neck in adults, defined by VGLL3 fusions, likely indolent behavior and limited rhabdomyoblastic differentiation. Further delineation of this entity and differentiation from more aggressive molecular subtypes of spindle cell RMS is mandatory to define the most appropriate therapeutic strategy and avoid overtreatment.

Purpose Treatment decisions in oncology are demanding and affect survival, general health, and quality of life. Expert systems can handle the complexity of the oncological field. We propose the application of a hybrid modeling approach for decision support models consisting of expert-based implementation of a decision model structure and machine-learning (ML) based parameter generation. We demonstrate our approach for the treatment of oropharyngeal cancer. Methods We created a clinical decision model based on Bayesian Networks and iteratively optimized its characteristics using structured knowledge engineering approaches. We combined manual adaptation of individual concepts with automatic learning of parameters and causalities. Using data from 94 patient records, we targeted the needed objectivity and clinical significance. Results In three iteration steps, we assessed the model with cross-validations. The initial aggregated accuracy of 0.529 could be increased to 0.883 in the final version. The predictive rates of the target nodes range from 0.557 to 0.960. Conclusion Combining different methodological approaches requires balancing the complexity of the clinical subject matter with the amount of information available in the dataset for ML application. Our method showed promising results because flaws of one approach can be overcome by the other approach. However, technical integrability as well as clinical acceptance must always be ensured.

Background: Making mathematics connections between home and school can have a positive impact on students’ mathematics learning. This is particularly important for students whose experiences and perspectives are underrepresented in school curricula. Although there is evidence for how crucial it is to make connections between mathematics instruction and children’s lived experiences, doing so is often a challenging task for teachers. Purpose: This study examines what teachers in two different settings—a dual language school and a Structured English Immersion school—learned from engaging in a mathematics collaboration with Latina mothers. We share the teachers’ perceptions of their students’ and families’ mathematics’ strengths and how they envisioned their future mathematics teaching. Research Design: This qualitative interpretive study explored how 15 teachers from two states made sense of their experiences working together in a two-year mathematics collaboration with mothers from their school community. We collected multiple teacher reflections and artifacts, and administered a small group interview with teachers and parents and an individual teacher interview. We used an iterative analysis by demarcating the words that pertained to the different data sources, as we looked for patterns of the teachers’ key ideas. We summarized the key ideas across the data sources that resulted in six major themes. Findings: The key findings in this study reveal that the teachers learned from the mothers more about their students’ lives outside of school, the mothers’ mathematics background, and ways to create multiple channels of communication with the mothers. The teachers also learned about the significant contributions that the mothers make to their children’s mathematics education and the importance of honoring the mothers’ ways of doing mathematics in their mathematics teaching. The teachers shared how the collaboration with the mothers helped to serve as a guide to leverage their students’ learning of mathematics in Spanish and English. Conclusions/Recommendations: This study suggests that mathematics collaborations between teachers and parents can be highly beneficial to student learning. It showcases the importance of providing teachers with opportunities to learn about the rich mathematics knowledge that parents from diverse backgrounds possess. This study also demonstrates that creating authentic relationships that enhance mutual understandings between teachers and parents can be key in supporting Latinx students’ mathematics learning. We recommend that these types of collaborations between teachers and parents be considered part of the preparation of preservice teachers as well as the professional learning of in-service teachers.

Aberrant AKT activation occurs in a number of cancers, metabolic syndrome, and immune disorders, making it an important target for the treatment of many diseases. To monitor spatial and temporal AKT activity in a live setting, we generated an Akt-FRET biosensor mouse that allows longitudinal assessment of AKT activity using intravital imaging in conjunction with image stabilization and optical window technology. We demonstrate the sensitivity of the Akt-FRET biosensor mouse using various cancer models and verify its suitability to monitor response to drug targeting in spheroid and organotypic models. We also show that the dynamics of AKT activation can be monitored in real time in diverse tissues, including in individual islets of the pancreas, in the brown and white adipose tissue, and in the skeletal muscle. Thus, the Akt-FRET biosensor mouse provides an important tool to study AKT dynamics in live tissue contexts and has broad preclinical applications. ispartof: SCIENCE ADVANCES vol:9 issue:17 ispartof: location:United States status: published

Background Clinically significant portal hypertension (CSPH) has been identified as an important prognostic factor in patients with hepatocellular carcinoma (HCC) undergoing curative treatment. This study aimed to assess PH estimates as prognostic factors in patients with HCC treated with immunotherapy. Methods All patients with HCC treated with an immunotherapeutic agent in first or subsequent lines at our tertiary care center between 2016 and 2021 were included (n = 50). CSPH was diagnosed using the established PH score for non-invasive PH estimation in pre-treatment CT data (cut-off ≥ 4). Influence of PH on overall survival (OS) and progression-free survival (PFS) was assessed in uni- and multivariable analyses. Results Based on the PH score, 26 patients (52.0%) were considered to have CSPH. After treatment initiation, patients with CSPH had a significantly impaired median OS (4.1 vs 33.3 months, p p = 0.02). In multivariable Cox regression, CSPH remained significantly associated with survival (HR 2.9, p = 0.015) when adjusted for established risk factors. Conclusions Non-invasive assessment of CSPH using routine CT data yielded an independent prognostic factor in patients with HCC and immunotherapy. Therefore, it might function as an additional imaging biomarker to detect high-risk patients with poor survival and possibly for treatment decision making.

Lipomatosis of peripheral nerves (LPN, also known as fibrolipomatous or lipofibromatous hamartoma of peripheral nerves) is a very rare, benign, intraneural, tumorous lesion that predominantly involves the median nerve but may rarely affect any peripheral nerve. Recently, PIK3CA mutations have been reported in macrodactyly, a rare condition related to LPN, and in other localized lipomatous overgrowth syndromes. In this retrospective study, we report 6 cases of FPN involving the median nerve (4 of them identified among 570 patients with carpal tunnel syndrome who underwent surgical decompression at our center from 2012 to 2022 and two seen in consultation by one of the authors). All cases were diagnosed via biopsy or resection supplemented by MRI. Patients were 4 males and 2 females aged 23 to 60 years (mean 38 years). One patient with bilateral lesions had in addition extensive angiomatosis of the paravertebral region. Histological examination showed an abnormal amount of mature fatty tissue containing disordered fibrous bands, entrapping normal-looking nerve fibers with prominent perineurial and endoneurial fibrosis. Genetic analysis using snapshot assay constructed to detect hotspots mutations in PIK3CA revealed similar PIK3CA mutations (p.H1047R; c.3140A>G) in 5/6 cases (83.3%). Our study represents a further contribution to the literature on LPN and highlights the diagnostic value of PIK3CA mutation testing as surrogate tool in equivocal cases and in those lesions without associated macrodactyly, especially as the biopsy findings of this lesion are essentially nonspecific.

Clinico-pathological features of analyzed cohorts and subgroups.

Distribution of stromal tumor infiltrating lymphocytes in analyzed cohorts and subgroups.

Figure S1. Selected images of different sTILs amounts, illustration of applied scoring method and images of IHC stainings. Figure S2. TLS in MIBC Figure S3. Subtyping of the CCC-EMN cohort and the TCGA-Cohort according to the MDACC-subtyping gene signature Figure S4. TCGA-Basal tumors (n=142). CIBERSORT and Mutational Burden. Figure S5. TCGA Luminal tumors (n=243). CIBERSORT and Mutational Burden. Figure S6. Distribution of tumor mutational burden (TMB; based on total single nucleotide variants) and neoantigens among different cut-offs of stromal TILs. Figure S7. TCGA - Antigen presenting cell signature (AGPS) and combined adjuvant platinum-containing chemotherapy cohort.

CIBERSORT - Spearman rank correlations between predicted immune cell populations in the TCGA cohort.

Inflammation gene signature TCGA: Full table of included genes.

Antigen presenting cell signature TCGA: Full table of included genes.

Multivariable Survival Models.

Reference Sequences - Nanostring analysis for subtyping (CCC-EMN cohort).

(A) Clinico-pathological cohort data and molecular genetic features of the mapping bladder (Case ID4). (B) Clinico-pathological cohort data of the tissue microarray cohort of papillary high grade carcinomas and CIS.

Suppl Figure legends

(A) Unsupervised hierarchical clustering of tumor subtyping protein markers determined by semiquantitative immunohistochemistry (CK5, CK14, CD44, CK20, GATA3, FOXA1, UPKII) reveals a basal and luminal cluster. (B) HER2/neu protein expression (left panel) and ERBB2/CEN17-ratio in the WBHM samples. HER2/neu protein expression and its genetically detected ERBB2-gene locus amplification co-occur in CIS, pT1 carcinomas and MIBCs. (C) Distribution of "Normal-like", "Transforming-like" and "Invasive-like" cluster assignments in the WBHM scheme illustrates "Invasive-like" lesions surrounded by "Transforming-like" lesions. CIS = Carcinoma in situ; DYS = Dysplasia; UPUMP/HYP = urothelial proliferation of unknown malignant potential/ hyperplasia (abbreviated as "HYP" in graphs); IRS = Immunoreactive score; MIBC = Muscle-invasive bladder cancer; pT1 = Stroma invasive bladder cancer; U = Tumor associated urothelium.

Background: PCA3 is a long noncoding RNA (lncRNA) with unknown function, upregulated in prostate cancer. LncRNAs may be processed into smaller active species. We hypothesized this for PCA3.Methods: We computed feasible RNA hairpins within the BMCC1 gene (encompassing PCA3) and searched a prostate transcriptome for these. We measured expression using qRT-PCR in three cohorts of prostate cancer tissues (n = 60), exfoliated urinary cells (n = 484 with cancer and n = 166 controls), and in cell lines (n = 22). We used in silico predictions and RNA knockup to identify potential mRNA targets of short transcribed RNAs.Results: We predicted 13 hairpins, of which PCA3-shRNA2 was most abundant within the prostate transcriptome. PCA3-shRNA2 is located within intron 1 of PCA3 and appears regulated by androgens. Expression of PCA3-shRNA2 was upregulated in malignant prostatic tissues, exfoliated urinary cells from men with prostate cancer (13–273 fold change; t test P < 0.003), and closely correlated to PCA3 expression (r = 0.84–0.93; P < 0.001). Urinary PCA3-shRNA2 (C-index, 0.75–0.81) and PCA3 (C-index, 0.78) could predict the presence of cancer in most men. PCA3-shRNA2 knockup altered the expression of predicted target mRNAs, including COPS2, SOX11, WDR48, TEAD1, and Noggin. PCA3-shRNA2 expression was negatively correlated with COPS2 in patient samples (r = −0.32; P < 0.001).Conclusion: We identified a short RNA within PCA3, whose expression is correlated to PCA3, which may target mRNAs implicated in prostate biology.Impact: This short RNA is stable ex vivo, suggesting a role as a robust biomarker. We identify cytoplasmic enrichment of this RNA and potential targeting of mRNAs implicated in prostate carcinogenesis. Cancer Epidemiol Biomarkers Prev; 24(1); 268–75. ©2014 AACR.

Suppl Figures 1-8. Supplementary Figure 1. Identification of the PCA3-shRNA2 hairpin. (a). Predicted hairpin of PCA3 RNA2. The bases in red are those identified within the prostate RNA transcriptome. (b). BLAST results of the 98bp fragment derived from PCA3-shRNA2 using 3'RACE, indicates that the sequence is found within PCA3 intron 1. Supplementary figure 2. Expression of PCA3 and PCA3-shRNA2 in cell lines representing prostate cancer (blue and labeled) and other malignancies (red). The expression of the two PCA3 RNAs is shown (normalised to PSA mRNA expression) for each of 22 cell lines. The non-prostate cancer cell lines are not labeled for clarity. In order of PCA3-shRNA2 expression these are (from HCT-116 (PCA3-shRNA2 DCt=-15.85), HEK 293, A549, NCI-H460, WM793, RT112, T47D, MRC5, AN3CA, RT4, SKOV-3, EJ, MCF-7, Jurkat and HeLa (PCA3-shRNA2 DCt=5.08)). Supplementary figure 3. Androgen regulated expression of PSA, PCA3 and PCA3 shRNA2. Expression (fold change) was determined using qrtPCR and normalized to a non-androgen regulated U1 RNA. PCA3 and PCA3 shRNA expression was directly related to testosterone concentration within growth media (0, 1nM and 10nM) Supplementary figure 4. Expression of PCA3, BMCC1, PCA3-shRNA2a and PCA3-shRNA2b in malignant and benign prostatic tissue. Significantly higher expression is seen in malignant tissues, when compared to benign tissues, for BMCC1 (DCt: -11.5{plus minus}2 vs. -14.1{plus minus}3, T Test p

Reference sequences, gene lists and gene expression distribution of utilized NanoString-assays.

(A)Overview of genes belonging to gene clusters (PanCancer Progression Panel). (B) Most differentially expressed signaling pathways and genes between lesion cluster A and C. (C) Most differentially expressed signaling pathways and genes between lesion cluster A and C.

Comparison of RNAseq and Nanostring-based subtyping in MIBCs from the CCC-EMN cohort with available RNAseq data. The 22 pairs of MIBC/CIS and the WBHM specimen were chosen from this pool of tumors. Subtyping is based on the 21 genes containing MDACC panel (Supplementary Table 2). RNAseq data were generated from FFPE tissues using QuantSeq kits (Lexogen) using 500 ng input RNA. The QuantSeq libraries were sequenced on an Illumina Hiseq platform and generated data were further processed according to the GRCh38, and further normalized. Inter-gene correlations range from 0.78-0.97 (Spearman rank correlations). Comparison of subtype assignment based on average linkage unsupervised hierarchical clustering shows a 100% overlap with no discordant classifications occurring, thus indicating that Nanostring is suitable for subtyping bladder cancer samples compared to RNAseq.

(A) Unsupervised hierarchical clustering reveals three lesion clusters (lesion cluster A-C) designated below the heat map. Histological classifications are shown above the heatmap and gene cluster families A*-D* are depicted on the left-sided cluster. Distribution of the lesion types in each cluster A-C are illustrated as pie diagrams below the heatmap. (B) Gene families from gene clusters C* and D* are mainly expressed in lesion clusters A/B in tumor associated urothelium, non-invasive urothelial lesions and CIS, while gene family A* is upregulated in CIS and pT1/MIBC samples. P-values were derived by nonparametric Kruskal-Wallis tests and are depicted above the graphs. P-values of single group comparisons derived by nonparametric Mann-Whitney tests are depicted in Suppl. Table 5. (C) Distribution of lesion cluster assignments (A-C) in the WBHM scheme. (D) Boxplots depicting differential expression of ERBB2, ERBB3, EGFR, STAT3 and CADM1 (STAT3-regulator) in luminal and basal tumors from the TCGA bladder cancer cohort. CIS = Carcinoma in situ; DYS = Dysplasia; UPUMP/HYP = urothelial proliferation of unknown malignant potential/ hyperplasia (abbreviated as "HYP" in graphs); MIBC = Muscle-invasive bladder cancer; pT1 = Stroma invasive bladder cancer; U = Tumor associated urothelium.

Suppl. Tables 1-3. Supplementary table 1. Predicted hairpin RNAs within PCA3 and BMCC1. Supplementary table 2. Predicted mRNA targets for PCA3-shRNA2. Supplementary table 3. Primers and condition used to detect target mRNAs.

Legend for Supplementary Data from Gene Expression Profiling of Progressive Papillary Noninvasive Carcinomas of the Urinary Bladder

Excel file

Purpose: The aim of the present study was to define gene expression profiles of noninvasive and invasive bladder cancer, to identify potential therapeutic or screening targets in bladder cancer, and to define genetic changes relevant for tumor progression of recurrent papillary bladder cancer (pTa).Experimental Design: Overall, 67 bladder neoplasms (46 pTa, 3 pTis, 10 pT1, and 8 pT2) and eight normal bladder specimens were investigated by a combination of laser microdissection and gene expression profiling. Eight of 16 patients with recurrent noninvasive papillary bladder tumors developed carcinoma in situ (pTis) or invasive bladder cancer (≥pT1G2) in the course of time. RNA expression results of the putative progression marker cathepsin E (CTSE) were confirmed by immunohistochemistry using high-throughput tissue microarray analysis (n = 776). Univariate analysis of factors regarding overall survival, progression-free survival, and recurrence-free survival in patients with urothelial bladder cancer was done.Results: Hierarchical cluster analyses revealed no differences between pTaG1 and pTaG2 tumors. However, distinct groups of invasive cancers with different gene expression profiles in papillary and solid tumors were found. Progression-associated gene profiles could be defined (e.g., FABP4 and CTSE) and were already present in the preceding noninvasive papillary tumors. CTSE expression (P = 0.003) and a high Ki-67 labeling index of at least 5% (P = 0.01) were the only factors that correlated significantly with progression-free survival of pTa tumors in our gene expression approach.Conclusions: Gene expression profiling revealed novel genes with potential clinical utility to select patients that are more likely to develop aggressive disease.

Basal and luminal subtypes of invasive bladder tumors have significant prognostic and predictive impacts for patients. However, it remains unclear whether tumor subtype commitment occurs in noninvasive urothelial lesions or in carcinoma in situ (CIS) and which gene pathways are important for bladder tumor progression. To understand the timing of this commitment, we used gene expression and protein analysis to create a global overview of 36 separate tissues excised from a whole bladder encompassing urothelium, noninvasive urothelial lesions, CIS, and invasive carcinomas. Additionally investigated were matched CIS, noninvasive urothelial lesions, and muscle-invasive bladder cancers (MIBC) from 22 patients. The final stage of subtype commitment to either a luminal or basal MIBC occurred at the CIS transition. For all tissues combined, hierarchical clustering of subtype gene expression revealed three subtypes: “luminal,” “basal,” and a “luminal p53-/extracellular matrix (ECM)-like” phenotype of ECM-related genes enriched in tumor-associated urothelium, noninvasive urothelial lesions, and CIS, but rarely invasive, carcinomas. A separate cohort of normal urothelium from noncancer patients showed significantly lower expression of ECM-related genes compared with tumor-associated urothelium, noninvasive urothelial lesions, and CIS. A PanCancer Progression Panel of 681 genes unveiled pathways specific for the luminal p53-/ECM-like cluster, for example, ECM remodeling, angiogenesis, epithelial-to-mesenchymal transition, cellular discohesion, cell motility involved in tumor progression, and cell proliferation and oncogenic ERBB2/ERBB3 signaling for invasive carcinomas. In conclusion, this study provides insights into bladder cancer subtype commitment and associated signaling pathways, which could help predict therapy response and enhance our understanding of therapy resistance.Significance:This study demonstrates that CIS is the stage of commitment for determining MIBC tumor subtype, which is relevant for patient prognosis and therapy response.

Pairwise group P-values of analyses with multiple group comparisons tested by non-parametric Kruskal-Wallis tests.

Genes and proteins used for calculation of gene- and protein-expression scores (luminal-ness/basal-ness/ECM/EMT/signature scores).

Description of additional methods and procedures used in the study. Also includes Supplementary References.

Sacrifice end points observed in the post-surgical model in the mock and scaffold groups with resection performed at day 10 without Gr-1 depletion.

Characterization of micro-porous PLG and PCL scaffolds (N = 10).

Micro-porous PCL scaffolds persist and maintain a space for extended times in vivo (S1); Host response following implantation of micro-porous PCL scaffolds in the dorsal subcutaneous space of an NSG mouse in vivo (S2); Dynamics of immune cell populations in the spleen of BALB/c mice with a PCL scaffold implant at day 0, 5, 10, and 15 post tumor inoculation (S3); Micro-porous PCL scaffolds enable recruitment of human MDA-MD-231BR cells in a chronic model of scaffold implantation (S4); Micro-porous PCL scaffolds do not significantly improve survival in a post-surgical model of breast cancer metastasis with day 6 post tumor inoculation resection (S5); Anti-Gr-1 depletion significantly reduces the presence of CD11b+Gr-1hiLy6C- MDSCs in the spleen primary tumor and scaffold (S6).

The onset of distant organ metastasis from primary breast cancer marks the transition to a stage IV diagnosis. Standard imaging modalities often detect distant metastasis when the burden of disease is high, underscoring the need for improved methods of detection to allow for interventions that would impede disease progression. Here, microporous poly(ϵ-caprolactone) scaffolds were developed that capture early metastatic cells and thus serve as a sentinel for early detection. These scaffolds were used to characterize the dynamic immune response to the implant spanning the acute and chronic foreign body response. The immune cell composition had stabilized at the scaffold after approximately 1 month and changed dramatically within days to weeks after tumor inoculation, with CD11b+Gr1hiLy6C− cells having the greatest increase in abundance. Implanted scaffolds recruited metastatic cancer cells that were inoculated into the mammary fat pad in vivo, which also significantly reduced tumor burden in the liver and brain. Additionally, cancer cells could be detected using a label-free imaging modality termed inverse spectroscopic optical coherence tomography, and we tested the hypothesis that subsequent removal of the primary tumor after early detection would enhance survival. Surgical removal of the primary tumor following cancer cell detection in the scaffold significantly improved disease-specific survival. The enhanced disease-specific survival was associated with a systemic reduction in the CD11b+Gr1hiLy6C− cells as a consequence of the implant, which was further supported by Gr-1 depletion studies. Implementation of the scaffold may provide diagnostic and therapeutic options for cancer patients in both the high-risk and adjuvant treatment settings. Cancer Res; 76(18); 5209–18. ©2016 AACR.

Humans use their peripheral vision during locomotion to perceive an approaching obstacle in their path, while also focusing central gaze on steps ahead of them. However, certain physiological and psychological factors may change this strategy, such as when a walker is physically fatigued. In this study, 21 healthy participants walked through a dark room while wearing eye tracking glasses before and following intense exercise. Obstacles were placed in random locations along their path and became illuminated when participants approached them. Results indicate that, when fatigued, participants had altered spatial gaze strategies, including more frequent use of central gaze to perceive obstacles and an increased gaze angular displacement. However, there were no changes in temporal gaze strategies following exercise. These findings reveal how physical fatigue alters one’s visual perception of their environment during locomotion, and may partially explain why people are at greater risk of trips and falls while fatigued.