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Bochdalek hernias are the most common congenital diaphragmatic hernias and are usually diagnosed during childhood1. They can present in adulthood and, in uncommon circumstances, result in gastric herniation with strangulation. We present a case of an adult Bochdalek hernia resulting in total gastric necrosis necessitating Roux-en-Y esophagojejunostomy in an otherwise healthy 39-year-old male. This case highlights the potential morbidity associated with unrepaired congenital diaphragmatic hernias and the need for appropriate referral.

Microscopic vascular invasion (VI) is predictive of recurrence in stage I lung adenocarcinoma (LUAD) but is difficult to assess in resection specimens and cannot be accurately predicted prior to surgery. Thus, new biomarkers are needed to identify this aggressive subset of stage I LUAD tumors. To assess molecular and tumor microenvironment (TME) features associated with angioinvasive LUAD we profiled 171 resected stage I tumors with and without VI by RNA-seq, including 16 tumors by high-resolution spatial transcriptomics (10x Genomics Visium). Visium capture areas were selected by an experienced thoracic pathologist to include invasive foci, tumor regions distal to foci, and tumors without invasive foci. We identified a robust molecular signature of VI from the RNA-seq data containing subclusters of genes involved in hallmark programs of tumor suppression, EMT, angiogenesis, growth and metabolism. This VI-associated signature increases across the spectrum of indolent to aggressive stage I LUAD histopathology and is predictive of recurrence-free survival. Analysis of 43,421 Visium spots across 16 tumors revealed high inter-tumor patient heterogeneity, with most spots clustering by tumor identity, suggesting tumor-intrinsic properties. Scoring the Visium data for VI signature expression revealed spatial variability around VI foci, with distinct spatial distributions of different signature subclusters among tumor and stromal compartments within and adjacent to regions of tumor intravasation. We found increased expression of the bulk VI signature in invasive tumors, regardless of whether the capture area contained the invasive focus. We independently verified this finding by performing laser-capture microdissection and RNA-seq from invaded vessel regions and uninvaded parenchyma of 8 additional tumors with VI and tumor regions of non-VI tumors. Given increased signature expression in regions of invasive tumors at a distance from VI foci, we leveraged our bulk RNA-seq dataset to develop a transcriptomic predictor of VI. We applied a nested cross-validation approach within a training cohort to select a machine learning model. We then evaluated the performance of our model in an independent test set. Finally, we generated over 200 pseudo-bulked in silico biopsies similar in size to standard transthoracic needle biopsies using the spatial data. The scores from these in silico biopsies were similar to predictions from matched bulk RNA-seq data, suggesting robustness to intra-tumor heterogeneity. Our combined bulk and spatial transcriptomics analysis suggests that VI-associated gene expression extends far from the site of intravasation and can be used to predict the presence of VI. This may enable the prediction of angioinvasive LUAD from small biopsy specimens, allowing for more tailored treatment prior to surgery. Citation Format: Dylan Steiner, Lila Sultan, Travis Sullivan, Emily Green, Hanqiao Liu, Xiaohui Xiao, Gang Liu, Avrum Spira, Sarah Mazzilli, Kimberly Rieger-Christ, Eric Burks, Jennifer Beane, Marc Lenburg. Evaluating a novel molecular biomarker of angioinvasive lung adenocarcinoma with spatial transcriptomics. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5632.

Background Early cancer detection could identify tumors at a time when outcomes are superior and treatment is less morbid. This prospective case-control sub-study (from NCT02889978 and NCT03085888) assessed the performance of targeted methylation analysis of circulating cell-free DNA (cfDNA) to detect and localize multiple cancer types across all stages at high specificity. Participants and methods The 6689 participants [2482 cancer (>50 cancer types), 4207 non-cancer] were divided into training and validation sets. Plasma cfDNA underwent bisulfite sequencing targeting a panel of >100 000 informative methylation regions. A classifier was developed and validated for cancer detection and tissue of origin (TOO) localization. Results Performance was consistent in training and validation sets. In validation, specificity was 99.3% [95% confidence interval (CI): 98.3% to 99.8%; 0.7% false-positive rate (FPR)]. Stage I–III sensitivity was 67.3% (CI: 60.7% to 73.3%) in a pre-specified set of 12 cancer types (anus, bladder, colon/rectum, esophagus, head and neck, liver/bile-duct, lung, lymphoma, ovary, pancreas, plasma cell neoplasm, stomach), which account for ∼63% of US cancer deaths annually, and was 43.9% (CI: 39.4% to 48.5%) in all cancer types. Detection increased with increasing stage: in the pre-specified cancer types sensitivity was 39% (CI: 27% to 52%) in stage I, 69% (CI: 56% to 80%) in stage II, 83% (CI: 75% to 90%) in stage III, and 92% (CI: 86% to 96%) in stage IV. In all cancer types sensitivity was 18% (CI: 13% to 25%) in stage I, 43% (CI: 35% to 51%) in stage II, 81% (CI: 73% to 87%) in stage III, and 93% (CI: 87% to 96%) in stage IV. TOO was predicted in 96% of samples with cancer-like signal; of those, the TOO localization was accurate in 93%. Conclusions cfDNA sequencing leveraging informative methylation patterns detected more than 50 cancer types across stages. Considering the potential value of early detection in deadly malignancies, further evaluation of this test is justified in prospective population-level studies.

Purpose To better characterize the urinary microbiome in males and contribute to overall understanding of the urinary microbiota specifically in patients undergoing evaluation for possible bladder cancer, stratified by risk exposure to smoking. Patients and methods Recruitment of 43 male patients in a sequential manner presenting for hematuria evaluation to a single institution was undertaken. Mid-stream urine specimen pellets were processed through a DNA isolation protocol before undergoing PCR amplification, purification, and 16S rRNA gene sequencing. Gene sequences were clustered into operational taxonomic units and statistical analysis was performed to determine specimen diversity and phylogenetic trends. Results No significant difference in microbial diversity was found between the specimens. On subgroup analysis, no significant difference was observed when stratified by either tobacco smoking history or by newly diagnosed urothelial bladder cancer. Variation in microbial diversity was seen amongst all analyzed specimens. Conclusion The results of our analysis of carefully selected subjects help to better characterize the urinary microbiome in males and supplements the limited available information on the interrelationship between the urinary microbiome and development of genitourinary malignancy. No significant difference was observed in our small sample size when stratified by tobacco exposure or newly diagnosed bladder cancer.

Background: SARS-CoV-2 infection and disease severity are influenced by viral entry (VE) gene expression patterns in airway epithelium. The similarities and differences of VE gene expression (ACE2, TMPRSS2, and CTSL) across nasal and bronchial compartments has not been fully characterized using matched samples from large cohorts. Results: Gene expression data from 793 nasal and 1,673 bronchial brushes obtained from individuals participating in lung cancer screening or diagnostic workup revealed that smoking was the only clinical factor significantly and reproducibly associated with VE gene expression. ACE2 and TMPRSS2 expression were higher in smokers in the bronchus but not in the nose. scRNA-seq of nasal brushings indicated that ACE2 co-expressed genes were highly expressed in club and C15orf48+ secretory cells while TMPRSS2 co-expressed genes were highly expressed in keratinizing epithelial cells. In contrast, these ACE2 and TMPRSS2 modules were highly expressed in goblet cells in scRNA-seq from bronchial brushings. Cell-type deconvolution of the RNA-seq confirmed that smoking increased the abundance of several secretory cell populations in the bronchus, but only goblet cells in the nose. Conclusions: The association of ACE2 and TMPRSS2 with smoking in the bronchus is due to their high expression in goblet cells which increase in abundance in current smoker airways. In contrast, in the nose these genes are not predominantly expressed in cell populations modulated by smoking. Smoking-induced VE gene expression changes in the nose likely has minimal impact on SARS-CoV-2 infection, but in the bronchus, smoking may lead to higher viral loads and more severe disease.

INTRODUCTION AND OBJECTIVE:Identifying non-invasive biomarkers that distinguish Lichen Sclerosus (LS) induced from non-LS induced urethral strictures would have broad implications for the diagnosis...

INTRODUCTION AND OBJECTIVE:Lichen sclerosus (LS) is a chronic inflammatory condition that can affect both the penile skin and urethral epithelium in men. Over time, severe impairment of sexual and ...

We evaluated the pathophysiology of lichen sclerosus and nonlichen sclerosus urethral stricture disease by comparing protein expression related to inflammation, cell cycle disruption, oxidative stress, hormone receptor status and infection.Tissue samples were collected from the urethral strictures of 81 patients undergoing urethroplasty. Clinical and demographic data were obtained by chart review. After identifying areas pathognomonic for lichen sclerosus a tissue microarray was created with cores from each sample and immunohistochemistry was performed.Patients had similar baseline demographics and comorbidities. Of the 81 strictures 58 were and 23 were not due to lichen sclerosus. Lichen sclerosus strictures were significantly longer and showed higher levels of inflammation. The proportion of T cells which stained positive for CD8 was significantly higher in strictures due to lichen sclerosus (50% vs 13%, p = 0.004). CCL-4 was expressed significantly more in strictures due to lichen sclerosus (76% vs 42%, p = 0.01). Several other inflammatory markers were only found in strictures due to lichen sclerosus. Block-like p16, a surrogate for high risk human papillomavirus infection, and varicella zoster virus were found only in lichen sclerosus urethral stricture disease samples, although both were rare. Epstein-Barr virus RNA was found in significantly more lichen sclerosus samples (37% vs 10%, p = 0.024).To our knowledge this is the first study to evaluate protein expression in lichen sclerosus urethral stricture disease. These strictures demonstrate increased inflammation compared to nonlichen sclerosus urethral strictures. Markers of oxidative stress, cell cycle dysregulation and the androgen receptor do not appear to be uniquely associated with lichen sclerosus urethral stricture disease. Positive staining for several viruses in samples of lichen sclerosus urethral stricture disease suggests a possible infectious etiology.

Coronavirus Disease 2019 (COVID-19) is caused by severe acute respiratory syndrome coronavirus 2 SARS-CoV-2), which infects host cells with help from the Viral Entry (VE) proteins ACE2, TMPRSS2, and CTSL1–4. Proposed risk factors for viral infection, as well as the rate of disease progression, include age5,6, sex7, chronic obstructive pulmonary disease7,8, cancer9, and cigarette smoking10–13. To investigate whether the proposed risk factors increase viral infection by modulation of the VE genes, we examined gene expression profiles of 796 nasal and 1,673 bronchial samples across four lung cancer screening cohorts containing individuals without COVID-19. Smoking was the only clinical factor reproducibly associated with the expression of any VE gene across cohorts. ACE2 expression was significantly up-regulated with smoking in the bronchus but significantly down-regulated with smoking in the nose. Furthermore, expression of individual VE genes were not correlated between paired nasal and bronchial samples from the same patients. Single-cell RNA-seq of nasal brushings revealed that an ACE2 gene module was detected in a variety of nasal secretory cells with the highest expression in the C15orf48+ secretory cells, while a TMPRSS2 gene module was most highly expressed in nasal keratinizing epithelial cells. In contrast, single-cell RNA-seq of bronchial brushings revealed that ACE2 andTMPRSS2 gene modules were most enriched in MUC5AC+ bronchial goblet cells. The CTSL gene module was highly expressed in immune populations of both nasal and bronchial brushings. Deconvolution of bulk RNA-seq showed that the proportion of MUC5AC+ goblet cells was increased in current smokers in both the nose and bronchus but proportions of nasal keratinizing epithelial cells, C15orf48+ secretory cells, and immune cells were not associated with smoking status. The complex association between VE gene expression and smoking in the nasal and bronchial epithelium revealed by our results may partially explain conflicting reports on the association between smoking and SARS-CoV-2 infection.

Introduction: Although three randomized control trials have proven mortality benefit of CT lung cancer screening (CTLS)

BACKGROUND: MicroRNAs (miRNAs), a group of non-coding post-transcriptional regulators of gene expression, are dysregulated in clear cell renal cell carcinoma (ccRCC) and play an important role in carcinogenesis. Our prior work identified a subset of miRNAs in pT1 ccRCC tumors associated with progression to metastatic disease. OBJECTIVE: To investigate the impact of two of these dysregulated miRNA, miR-15a-5p and -26a-5p, in an effort to elucidate the mechanisms underpinning aggressive forms of stage I ccRCC. METHODS: The ccRCC cell line 786-O was transfected with pre-miRs-15a-5p and -26a-5p to rescue expression. Cell proliferation was measured via MT Cell Viability Assay. O-GlcNAc-transferase (OGT), a known protein in ccRCC proliferation, was identified by bioinformatics analysis as a target of both miRNA and validated via luciferase reporter assay to confirm binding of each miR to the 3′ untranslated region (UTR). OGT protein expression was evaluated via western blotting. RESULTS: Luciferase assay confirmed specificity of miR-15a-5p and -26a-5p for the OGT UTR. Western blot analysis for OGT showed reduced expression following co-transfection of both miRNAs compared to negative control or individual transfection. Co-transfection of these miRNAs greatly reduced proliferation when compared to negative control or the individual transfections. CONCLUSION: Our results indicate that the dysregulation of miR-15a-5p and -26a-5p contribute cooperatively to the proliferation of ccRCC through their regulation of OGT. These results give insight into the pathogenesis of aggressive early stage ccRCC and suggest potential therapeutic targets for future research.

Lung cancer screening has improved mortality among high-risk smokers but has coincidentally detected a fraction of nonprogressive adenocarcinoma historically classified as bronchoalveolar carcinoma (BAC). In the National Lung Screening Trial (NLST) the majority of BAC-comprising 29% of computed tomography-detected stage I lung adenocarcinoma-were considered overdiagnosis after extended follow-up comparison with the control arm. In the current classification, adenocarcinoma in situ and minimally invasive adenocarcinoma have replaced BAC but together comprise only ∼5% of stage I lung adenocarcinoma. Lepidic and subsets of papillary and acinar adenocarcinoma also infrequently recur. We, therefore, propose criteria for low malignant potential (LMP) adenocarcinoma among nonmucinous adenocarcinoma measuring ≤3 cm in total, exhibiting ≥15% lepidic growth, and lacking nonpredominant high-grade patterns (≥10% cribriform, ≥5% micropapillary, ≥5% solid), >1 mitosis per 2 mm2, angiolymphatic or visceral pleural invasion, spread through air spaces or necrosis. We tested these criteria in a multi-institutional cohort of 328 invasive stage I (eighth edition) and in situ adenocarcinomas and observed 16% LMP and 7% adenocarcinoma in situ/minimally invasive adenocarcinoma which together (23%) approximated the frequency of overdiagnosed stage I BAC in the NLST. The LMP group had 100% disease-specific survival. The proposed LMP criteria, incorporating multiple histologic parameters, may be a clinically useful "low-grade" prognostic group. Validation of these criteria in additional retrospective cohorts and prospective screen-detected cohorts should be considered.

Palliative care is a medical discipline that emphasizes quality of life and can be provided in parallel with recovery-directed treatments in colon and rectal surgery. Palliative care is receiving increasing attention and investigation for its potential to improve quality and outcomes for a wide spectrum of patients by benefiting symptom management, supporting complex health care decision making and facilitating care transitions. Primary palliative care refers to the application of palliative care principles by clinicians of all disciplines whereas specialty palliative care is a multidisciplinary approach and includes a clinician with advanced training and experience.

This study sought to identify microRNA (miRNA) profiles of small, pathologically confirmed stage 1 clear cell renal cell carcinoma (ccRCC) tumors that are associated with progression to metachronous metastatic disease.Fifty-five pathologic stage 1 ccRCC tumors ≤5cm, from 2 institutions, were examined in a miRNA screening, followed by a validation study. For the screening phase 752 miRNA were evaluated on each sample to identify those with differential expression between tumors that subsequently did (n = 10) or did not (n = 10) progress to metastatic disease. For the validation, 35 additional samples (20 nonprogressors and 15 with distant progression) were utilized to investigate 20 miRNA to determine if a miRNA panel could differentiate aggressive tumors: associations of miRNA expression with cancer specific survival was also investigated.In the screening analysis, 35 miRNA were differentially expressed (P0.05, FDR0.1) between the groups. In the validation, 11 miRNA were confirmed to have differential expression. The miRNA -10a-5p, -23b-3p, and -26a-5p differentiated nonprogressive and distant progressive disease with a sensitivity of 73.3% and a specificity of 85% (AUC=0.893). In addition, levels of miR-30a-3p and -145-5p were identified as independent prognostic factors of cancer specific survival.This investigation identified miRNA biomarkers that may differentiate between non-progressive ccRCC tumors and those that progress to metastatic disease in this group of stage I tumors. The miRNA profiles determined in this study have the potential to identify patients with small renal masses who are likely to have progressive ccRCC. Such information may be valuable to incorporate into predictive models.

Background There is significant variability in weight loss after bariatric surgery. We hypothesize that part of this variability may be predetermined by genetic differences associated with metabolic homeostasis. MicroRNA (miRNA) are short pieces of RNA that regulate gene expression and are readily detectable in serum. They are implicated in numerous metabolic processes, including weight homeostasis. In this pilot study, we briefly review the role of miRNA, and assess the feasibility of using them in the clinical setting of obesity treatment. Objectives To evaluate the feasibility of using miRNA to predict weight loss after bariatric surgery. Setting Academic medical center. Methods Serum was collected from patients at the initial bariatric surgery consultation. Weight loss data were collected 6 to 12 months postoperatively. Individuals experiencing the least and the greatest amount of percentage of excess weight lost at 6 months were analyzed to assess for genetic differences in miRNA expression. Results The median percentage of excess weight lost was 51% (range, 34%–63%) for those who lost the least and 87% (range, 82%–111%) for those who lost the most weight. Groups were similar in age, sex, diabetic status, and type of surgery. In total, of the 119 miRNA detected in the serum of the patients, 6 demonstrated potential for discriminating between the high and low weight loss groups. These miRNA have previously been implicated in regulation of fatty acid biosynthesis, adipocyte proliferation, type 2 diabetes, and obesity. Conclusions In this pilot study, we demonstrated the feasibility of identifying genetic differences between high and low weight loss groups by identifying distinct serum miRNA. In the near future, these biomarkers could facilitate informed decisions about surgery. In addition, these miRNA could open new genetic pathways that describe the pathophysiology of obesity, and provide targets for future treatment.

Objective To identify miRNA characteristic of metastatic clear cell renal cell carcinoma (ccRCC) and those indicative of cancer specific survival in nephrectomy and biopsy specimens. We also sought to determine if a miRNA panel could differentiate benign from ccRCC tissue. Materials and Methods RNA was isolated from nephrectomy and kidney biopsy specimens (n=156; n=46 respectively). Samples were grouped: benign, non-progressive and progressive ccRCC. MiRNA were profiled by microarray and validated by qRT-PCR. Biomarker signatures were developed to predict cancer status in nephrectomy and biopsy specimens. Cancer specific survival was examined using Kaplan-Meier and Cox proportional hazards analyses. Results Microarray analysis revealed 20 differentially expressed miRNA comparing non-progressive with progressive tumors. A biomarker signature validated in nephrectomy specimens had a sensitivity of 86.7% and a specificity of 92.9% for differentiating benign and ccRCC. A second signature differentiated non-progressive versus progressive ccRCC with a sensitivity of 93.8% and a specificity of 83.3%. These biomarkers also discriminated cancer status in biopsy specimens. Levels of miR-10a-5p, -10b-5p, and -223-3p were associated with cancer specific survival. Conclusion This study identified miRNA differentially expressed in ccRCC samples; as well as those correlating with cancer specific survival. Biomarkers identified in this study have the potential to identify patients who are likely to have progressive ccRCC, and although preliminary, these results may aid in differentiating aggressive and indolent ccRCC based on biopsy specimens. This article is protected by copyright. All rights reserved.

Urethroplasty of lichen sclerosus strictures has a significantly higher failure rate than strictures due to other causes. We sought to determine predictors of urethroplasty failure in men with lichen sclerosus urethral stricture disease by evaluating protein expression profiles.Urethral tissue was excised from patients with lichen sclerosus who were undergoing urethroplasty of urethral stricture disease at a single institution. A tissue microarray was created with cores from each sample. Immunohistochemistry was performed to compare protein expression related to inflammation, cell cycle disruption, oxidative stress, hormone receptor status and infection. Stricture recurrence was defined by the need for a subsequent unanticipated procedure for urethral stricture disease.We evaluated 50 men with lichen sclerosus urethral stricture disease, including 31 with successful reconstruction and 19 with recurrent stricture. Recurrent strictures expressed lower levels of several inflammatory markers and had a lower Ki-67 mitotic index and significantly higher vascular endothelial growth factor levels than nonrecurrent strictures.To our knowledge this is the first study to use tissue protein expression to identify risk factors for urethroplasty failure among men with lichen sclerosus urethral stricture disease. Our findings suggest that recurrent lichen sclerosus strictures demonstrate a suppressed inflammatory response, a decreased cell turnover rate, and poor oxygenation and nutrient delivery. Prospective studies are needed to clarify the role of these pathways in the pathophysiology of lichen sclerosus urethral stricture disease, determine whether preoperative biopsy can predict urethroplasty success, help counsel patients and develop future treatments.

Introduction: Bariatric surgery has been shown to lead to resolution of type 2 diabetes mellitus (T2DM) in 78.1% of patients (Buchwald, Am J Med 2009), however, it also confers significant risk for potential complications. It is unclear in the preoperative setting which patients will have resolution of T2DM following bariatric surgery. The objective of this study is to identify if patients who have resolution of T2DM following bariatric surgery express different biomarkers preoperatively from their counterparts who do not have resolution of T2DM. Methods: Blood samples were obtained from patients who presented to the Center for Surgical Weight Loss with T2DM and either BMI ≥ 40 or BMI ≥ 35 with weight-related comorbidities. All 20 patients studied then underwent gastric bypass or gastric sleeve surgery. At 6 to 12 months postoperatively the patients were evaluated for resolution of T2DM; defined as a hemoglobin A1c less than 6.5 and off all diabetic medications. Extracellular vesicles were isolated from patient serum and analyzed to identify and quantify microRNA and protein biomarkers. The biomarkers in patients who had resolution of T2DM were compared to the biomarkers in patients who had persistent T2DM. Results: Out of the 395 microRNA detected, there were 10 that were significantly differentially expressed between patients who had resolution of T2DM after bariatric surgery and those who did not. Out of the 71 proteins detected, there were 5 identified that differed significantly between the two groups. Conclusion: Preoperatively, patients with T2DM who undergo bariatric surgery with subsequent resolution of T2DM may have a different biomarker profile than patients whose T2DM does not resolve following bariatric surgery. If these biomarkers are validated in a larger population of patients, they could be used to help clinicians select patients in the preoperative setting who would benefit most from bariatric surgery. Disclosure R. Hillyer: None. T. Sullivan: None. K. Christ: None. A. Hodarkar: None. M. Hodge: None. Funding Ellison Foundation; Cancer Prevention and Research Institute of Texas

Intrahepatic cholangiocarcinoma (ICC) is an aggressive liver bile duct malignancy exhibiting frequent isocitrate dehydrogenase (IDH1/IDH2) mutations. Through a high-throughput drug screen of a large panel of cancer cell lines, including 17 biliary tract cancers, we found that IDH mutant (IDHm) ICC cells demonstrate a striking response to the multikinase inhibitor dasatinib, with the highest sensitivity among 682 solid tumor cell lines. Using unbiased proteomics to capture the activated kinome and CRISPR/Cas9-based genome editing to introduce dasatinib-resistant “gatekeeper” mutant kinases, we identified SRC as a critical dasatinib target in IDHm ICC. Importantly, dasatinib-treated IDHm xenografts exhibited pronounced apoptosis and tumor regression. Our results show that IDHm ICC cells have a unique dependency on SRC and suggest that dasatinib may have therapeutic benefit against IDHm ICC. Moreover, these proteomic and genome-editing strategies provide a systematic and broadly applicable approach to define targets of kinase inhibitors underlying drug responsiveness.Significance: IDH mutations define a distinct subtype of ICC, a malignancy that is largely refractory to current therapies. Our work demonstrates that IDHm ICC cells are hypersensitive to dasatinib and critically dependent on SRC activity for survival and proliferation, pointing to new therapeutic strategies against these cancers. Cancer Discov; 6(7); 727–39. ©2016 AACR.This article is highlighted in the In This Issue feature, p. 681

INTRODUCTION AND OBJECTIVE:The pathophysiology of Lichen Sclerosus (LS) urethral stricture disease (USD) is poorly understood. MicroRNA (miRNA) are non-coding genetic material involved in the regul...

Objective To evaluate microRNA (miRNA) biomarkers for upper tract urothelial carcinoma (UTUC) to improve risk stratification. Methods miRNA was isolated from 157 radical nephroureterectomy specimens from 2 institutions. The relative expression of miRNA was examined for high grade vs low grade tumors as well as muscle invasive vs nonmuscle invasive tumors. Recurrence free survival (RFS) and overall survival (OS) were also stratified using relative expression of specific miRNA. Results The optimized model to identify high grade UTUC included miR-29b-2-5p, miR-18a-5p, miR-223-3p, and miR-199a-5p, generating a sensitivity of 83%, specificity of 85%, and generated a receiver operating characteristic (ROC) curve with area-under-the-curve of 0.86. Similarly, the model classifier for predicting ≥pT2 disease incorporated miR-10b-5p, miR-26a-5p-5p, miR-31-5p, and miR-146b-5p, producing a sensitivity of 64%, specificity of 96%, and area-under-the-curve of 0.90. RFS was best reflected by a combination of miR-10a-5p, miR-30c-5p, and miR-10b-5p, while OS was best predicted by miR-10a-5p, miR-199a-5p, miR-30c-5p, and miR-10b-5p. Conclusion High-grade vs low-grade as well as muscle invasive vs nonmuscle invasive UTUC can be reliable distinguished with unique miRNA signatures. Furthermore, differential expression of UTUC miRNA produces robust classifiers for predicting RFS and OS that may help identify patients who would most benefit from adjuvant therapies.

We report a comprehensive analysis of 412 muscle-invasive bladder cancers characterized by multiple TCGA analytical platforms. Fifty-eight genes were significantly mutated, and the overall mutational load was associated with APOBEC-signature mutagenesis. Clustering by mutation signature identified a high-mutation subset with 75% 5-year survival. mRNA expression clustering refined prior clustering analyses and identified a poor-survival "neuronal" subtype in which the majority of tumors lacked small cell or neuroendocrine histology. Clustering by mRNA, long non-coding RNA (lncRNA), and miRNA expression converged to identify subsets with differential epithelial-mesenchymal transition status, carcinoma in situ scores, histologic features, and survival. Our analyses identified 5 expression subtypes that may stratify response to different treatments.

The objective of this study was to identify a panel of microRNAs (miRNAs) differentially expressed in high-grade non-muscle invasive (NMI; TaG3–T1G3) urothelial carcinoma that progress to muscle-invasive disease compared to those that remain non-muscle invasive, whether recurrence happens or not. Eighty-nine high-grade NMI urothelial carcinoma lesions were identified and total RNA was extracted from paraffin-embedded tissue. Patients were categorized as either having a non-muscle invasive lesion with no evidence of progression over a 3-year period or as having a similar lesion showing progression to muscle invasion over the same period. In addition, comparison of miRNA expression levels between patients with and without prior intravesical therapy was performed. Total RNA was pooled for microarray analysis in each group (non-progressors and progressors), and qRT-PCR of individual samples validated differential expression between non-progressive and progressive lesions. MiR-32-5p, -224-5p, and -412-3p were associated with cancer-specific survival. Downregulation of miR-203a-3p and miR-205-5p were significantly linked to progression in non-muscle invasive bladder tumors. These miRNAs include those implicated in epithelial mesenchymal transition, previously identified as members of a panel characterizing transition from the non-invasive to invasive phenotype in bladder tumors. Furthermore, we were able to identify specific miRNAs that are linked to postoperative outcome in patients with high grade NMI urothelial carcinoma of the bladder (UCB) that progressed to muscle-invasive (MI) disease.

RATIONALE: The National Lung Screening and Nelson Trials demonstrated a 20% and 26% (for men) reduction, respectively in lung cancer mortality for patients screened using low-dose CT. However, lung cancer screening has the potential for over-diagnosis of indolent tumors. Therefore, we sought to identify molecular features that could distinguish indolent from aggressive early stage lung tumors based on histologic aggressiveness profiling, with the ultimate goal being to translate these findings into biomarkers that could inform post-biopsy/surgery management. METHODS: 63 FFPE samples of stage I lung adenocarcinomas were included for pathologic annotation and Whole Exome sequencing. An average of 48.2±15.6 million total reads per sample were generated with 78.4%±4.8% reads uniquely mapped. Tumors were categorized into 3 groups based on histologic features previously associated with tumor aggressiveness: Minimally-Aggressive (n=18): containing zero aggressive components (e.g. zero solid/cribriform); Medium-Aggressive (n=19): not yet dominated by aggressive components; Highly-Aggressive (n=26): solid/cribriform predominant. Tumor histology was characterized for percentage of lepidic, acinar, papillary, micro-papillary, solid and cribriform components. Negative binomial models were used to identify genes whose expression was associated with tumor histology. Estimate Algorithm was used to estimate immune cell type infiltration of these tumors. RESULTS: 430 genes (FDR q CONCLUSION: We identified gene expression alterations among stage I adenocarcinomas associated with histologic features of tumor aggressiveness. This gene signature may help identify more indolent tumors and potentially impact their post biopsy/surgery clinical management. Citation Format: Jiarui Zhang, Eric Burks, Jennifer Beane, Travis Sullivan, Gang Liu, Steven Dubinett, Marc Lenburg, Avrum Spira. Gene expression alterations associated with histologic aggressiveness in stage I lung adenocarcinomas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3530.