Gene fusions involving ETS (erythroblastosis virus E26 transformation-specific) family transcription factors are found in ~50% of prostate cancers and as such can be used as a basis for the molecular ...subclassification of prostate cancer. Previously, we showed that marked overexpression of SPINK1 (serine peptidase inhibitor, Kazal type 1), which encodes a secreted serine protease inhibitor, defines an aggressive molecular subtype of ETS fusion-negative prostate cancers (SPINK1+/ETS⁻, ~10% of all prostate cancers). Here, we examined the potential of SPINK1 as an extracellular therapeutic target in prostate cancer. Recombinant SPINK1 protein (rSPINK1) stimulated cell proliferation in benign RWPE as well as cancerous prostate cells. Indeed, RWPE cells treated with either rSPINK1 or conditioned medium from 22RV1 prostate cancer cells (SPINK1+/ETS⁻) significantly increased cell invasion and intravasation when compared with untreated cells. In contrast, knockdown of SPINK1 in 22RV1 cells inhibited cell proliferation, cell invasion, and tumor growth in xenograft assays. 22RV1 cell proliferation, invasion, and intravasation were attenuated by a monoclonal antibody (mAb) to SPINK1 as well. We also demonstrated that SPINK1 partially mediated its neoplastic effects through interaction with the epidermal growth factor receptor (EGFR). Administration of antibodies to SPINK1 or EGFR (cetuximab) in mice bearing 22RV1 xenografts attenuated tumor growth by more than 60 and 40%, respectively, or ~75% when combined, without affecting PC3 xenograft (SPINK1⁻/ETS⁻) growth. Thus, this study suggests that SPINK1 may be a therapeutic target in a subset of patients with SPINK1+/ETS⁻ prostate cancer. Our results provide a rationale for both the development of humanized mAbs to SPINK1 and evaluation of EGFR inhibition in SPINK1+/ETS⁻ prostate cancers.
Previous studies have suggested that markers of TH2-associated inflammation in asthma predict clinical responsiveness or resistance to corticosteroid therapy.8,9 We therefore used gene expression ...data in a subset of the 56 patients with asthma with RNA-seq data to determine TH2-high and TH2-low status defined by ±2 SD of mean values in control subjects and confirmed by hierarchical clustering (Fig 1, C). ...in our study, sHLA-G levels were highest in patients with asthma with a low TH2 gene score. ...rather than influencing asthma susceptibility per se, sHLA-G may be a modifier of disease severity and a marker of a distinct clinical endotype characterized by less airway inflammation. Supplementary data 1 T. Fujii, A. Ishitani, D.E. Geraghty, A soluble form of the HLA-G antigen is encoded by a messenger ribonucleic acid containing intron 4, J Immunol, Vol. 153, 1994, 5516-5524 2 A. Ishitani, D.E. Geraghty, Alternative splicing of HLA-G transcripts yields proteins with primary structures resembling both class I and class II antigens, Proc Natl Acad Sci U S A, Vol. 89, 1992, 3947-3951 3 D. Nicolae, N.J. Cox, L.A. Lester, D. Schneider, Z. Tan, C. Billstrand, Fine mapping and positional candidate studies identify HLA-G as an asthma susceptibility gene on chromosome 6p21, Am J Hum Genet, Vol. 76, 2005, 349-357 4 F. Tahan, T. Patiroglu, Plasma soluble human leukocyte antigen G levels in asthmatic children, Int Arch Allergy Immunol, Vol. 141, 2006, 213-216 5 S.R. White, D.A. Loisel, J.F. McConville, R. Stern, Y. Tu, B.A. Marroquin, Levels of soluble human leukocyte antigen-G are increased in asthmatic airways, Eur Respir J, Vol. 35, 2010, 925-927 6 Z. Tan, G. Randall, J. Fan, B. Camoretti-Mercado, R. Brockman-Schneider, L. Pan, Allele-specific targeting of microRNAs to HLA-G and risk of asthma, Am J Hum Genet, Vol. 81, 2007, 829-834 7 N. Rouas-Freiss, A. Naji, A. Durrbach, E.D. Carosella, Tolerogenic functions of human leukocyte antigen G: from pregnancy to organ and cell transplantation, Transplantation, Vol. 84, Iss. 1 Suppl, 2007, S21-5 8 P.G. Woodruff, B. Modrek, D.F. Choy, G. Jia, A.R. Abbas, A. Ellwanger, T-helper type 2-driven inflammation defines major subphenotypes of asthma, Am J Respir Crit Care Med, Vol. 180, 2009, 388-395 9 M.C. Peters, Z.K. Mekonnen, S. Yuan, N.R. Bhakta, P.G. Woodruff, J.V. Fahy, Measures of gene expression in sputum cells can identify TH2-high and TH2-low subtypes of asthma, J Allergy Clin Immunol, Vol. 133, 2014, 388-394 10 J. Nicodemus-Johnson, B. Laxman, R.K. Stern, J. Sudi, C.N. Tierney, L. Norwick, Maternal asthma and microRNA regulation of soluble HLA-G in the airway, J Allergy Clin Immunol, Vol. 131, 2013, 1496-1503
Abstract Objectives We sought to develop a clinical algorithm combining serum PSA with detection of TMPRSS2:ERG fusion and PCA3 in urine collected after digital rectal exam (post-DRE urine) to ...predict prostate cancer on subsequent biopsy. Materials and methods Post-DRE urine was collected in 48 consecutive patients before prostate biopsy at 2 centers; quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to detect PCA3 and TMPRSS2:ERG fusion transcript expression. Serum PSA was measured by clinical assay. The performance of TMPRSS2:ERG fusion, PCA3, and serum PSA as biomarkers predicting prostate cancer at biopsy was measured; a clinically practical algorithm combining serum PSA with TMPRSS2:ERG and PCA3 in post-DRE urine to predict prostate cancer was developed. Results Post-DRE urine sediment provided informative RNA in 45 patients; prostate cancer was present on subsequent biopsy in 15. TMPRSS2:ERG in post-DRE urine was associated with prostate cancer (OR = 12.02; P < 0.001). PCA3 had the highest sensitivity in predicting prostate cancer diagnosis (93%), whereas TMPRSS2:ERG had the highest specificity (87%). TMPRSS2:ERG had the greatest discriminatory value in predicting prostate cancer (AUC = 0.77 compared with 0.65 for PCA3 and 0.72 for serum PSA alone). Combining serum PSA, PCA3, and TMPRSS2:ERG in a multivariable algorithm optimized for clinical utility improved cancer prediction (AUC = 0.88; specificity = 90% at 80% sensitivity). Conclusions A clinical algorithm specifying biopsy for all patients with PSA ≥ 10 ng/ml, while restricting biopsy among those with PSA <10 ng/ml to only those with detectable PCA3 or TMPRSS2:ERG in post-DRE urine, performed better than the individual biomarkers alone in predicting prostate cancer.
Basal airway epithelial cells (AEC) constitute stem/progenitor cells within the central airways and respond to mucosal injury in an ordered sequence of spreading, migration, proliferation, and ...differentiation to needed cell types. However, dynamic gene transcription in the early events after mucosal injury has not been studied in AEC. We examined gene expression using microarrays following mechanical injury (MI) in primary human AEC grown in submersion culture to generate basal cells and in the air-liquid interface to generate differentiated AEC (dAEC) that include goblet and ciliated cells. A select group of ~150 genes was in differential expression (DE) within 2-24 hr after MI, and enrichment analysis of these genes showed over-representation of functional categories related to inflammatory cytokines and chemokines. Network-based gene prioritization and network reconstruction using the PINTA heat kernel diffusion algorithm demonstrated highly connected networks that were richer in differentiated AEC compared to basal cells. Similar experiments done in basal AEC collected from asthmatic donor lungs demonstrated substantial changes in DE genes and functional categories related to inflammation compared to basal AEC from normal donors. In dAEC, similar but more modest differences were observed. We demonstrate that the AEC transcription signature after MI identifies genes and pathways that are important to the initiation and perpetuation of airway mucosal inflammation. Gene expression occurs quickly after injury and is more profound in differentiated AEC, and is altered in AEC from asthmatic airways. Our data suggest that the early response to injury is substantially different in asthmatic airways, particularly in basal airway epithelial cells.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
ETS gene fusions have been characterized in a majority of prostate cancers; however, the key molecular alterations in
ETS-negative cancers are unclear. Here we used an outlier meta-analysis ...(meta-COPA) to identify
SPINK1 outlier expression exclusively in a subset of
ETS rearrangement-negative cancers (∼10% of total cases). We validated the mutual exclusivity of
SPINK1 expression and
ETS fusion status, demonstrated that
SPINK1 outlier expression can be detected noninvasively in urine, and observed that
SPINK1 outlier expression is an independent predictor of biochemical recurrence after resection. We identified the aggressive 22RV1 cell line as a
SPINK1 outlier expression model and demonstrate that
SPINK1 knockdown in 22RV1 attenuates invasion, suggesting a functional role in
ETS rearrangement-negative prostate cancers.
Breast cancer patients have benefited from the use of targeted therapies directed at specific molecular alterations. To identify additional opportunities for targeted therapy, we searched for genes ...with marked overexpression in subsets of tumors across a panel of breast cancer profiling studies comprising 3,200 microarray experiments. In addition to prioritizing ERBB2, we found AGTR1, the angiotensin II receptor type I, to be markedly overexpressed in 10-20% of breast cancer cases across multiple independent patient cohorts. Validation experiments confirmed that AGTR1 is highly overexpressed, in several cases more than 100-fold. AGTR1 overexpression was restricted to estrogen receptor-positive tumors and was mutually exclusive with ERBB2 overexpression across all samples. Ectopic overexpression of AGTR1 in primary mammary epithelial cells, combined with angiotensin II stimulation, led to a highly invasive phenotype that was attenuated by the AGTR1 antagonist losartan. Similarly, losartan reduced tumor growth by 30% in AGTR1-positive breast cancer xenografts. Taken together, these observations indicate that marked AGTR1 overexpression defines a subpopulation of ER-positive, ERBB2-negative breast cancer that may benefit from targeted therapy with AGTR1 antagonists, such as losartan.
Soluble human leukocyte antigen-G (sHLA-G), an immunomodulatory molecule associated with suppression of inflammation, is elevated in the airways of asthmatic patients with a low inflammatory endotype ...as evidenced by low airway eosinophils and low exhaled nitric oxide.
Recurrent gene fusions involving E26 transformation-specific (ETS) transcription factors ERG, ETV1, ETV4, or ETV5 have been identified in 40% to 70% of prostate cancers. Here, we used a comprehensive ...fluorescence in situ hybridization (FISH) split probe strategy interrogating all 27 ETS family members and their five known 5' fusion partners in a cohort of 110 clinically localized prostate cancer patients. Gene rearrangements were only identified in ETS genes that were previously implicated in prostate cancer gene fusions including ERG, ETV1, and ETV4 (43%, 5%, and 5%, respectively), suggesting that a substantial fraction of prostate cancers (estimated at 30-60%) cannot be attributed to an ETS gene fusion. Among the known 5' gene fusion partners, TMPRSS2 was rearranged in 47% of cases followed by SLC45A3, HNRPA2B1, and C15ORF21 in 2%, 1%, and 1% of cases, respectively. Based on this comprehensive FISH screen, we have made four noteworthy observations. First, by screening the entire ETS transcription factor family for rearrangements, we found that a large fraction of prostate cancers (44%) cannot be ascribed to an ETS gene fusion, an observation which will stimulate research into identifying recurrent non-ETS aberrations in prostate cancers. Second, we identified SLC45A3 as a novel 5' fusion partner of ERG; previously, TMPRSS2 was the only described 5' partner of ERG. Third, we identified two prostate-specific, androgen-induced genes, FLJ35294 and CANT1, as 5' partners to ETV1 and ETV4. Fourth, we identified a ubiquitously expressed, androgen-insensitive gene, DDX5, fused in frame with ETV4, leading to the expression of a DDX5-ETV4 fusion protein.
We recently reported the identification of recurrent gene fusions in the majority of prostate cancers involving the 5V untranslated region of the androgenregulated gene TMPRSS2, the ETS family ...members ERG, ETV1, ETV4. Here we report the noninvasive detection of these gene fusions in the urine of patients with clinically localized prostate cancer. By quantitative polymerase chain reaction, we assessed the expression of ERG, TMPRSS2:ERG transcripts in urine samples obtained after prostatic massage from 19 patients (11 prebiopsy, 8 pre-radical prostatectomy) with prostate cancer. We observed a strong concordance between ERG overexpression, TMPRSS2:ERG expression, with 8 of 19 (42%) patients having detectable TMPRSS2:ERG transcripts in their urine. Importantly, by fluorescence in situ hybridization, we confirmed the presence or the absence of TMPRSS2:ERG gene fusions in matched prostate cancer tissue samples from three of three patients with fusion transcripts in their urine, from two of two patients without fusion transcripts in their urine. These results demonstrate that TMPRSS2:ERG gene fusions can be detected in the urine of patients with prostate cancer, support larger studies on prospective cohorts for noninvasive detection of prostate cancer.
Background We previously reported an interaction between maternal asthma and the child's HLA-G genotype on the child's subsequent risk for asthma. The implicated single nucleotide polymorphism ...at +3142 disrupted a target site for the microRNA (miR)-152 family. We hypothesized that the interaction effect might be mediated by these miRs. Objective The objective of this study was to test this hypothesis in adults with asthma who are a subset of the same subjects who participated in our earlier family-based studies. Methods We measured soluble HLA-G (sHLA-G) concentrations in bronchoalveolar lavage fluid (n = 36) and plasma (n = 57) from adult asthmatic subjects with and without a mother with asthma, and HLA-G and miR-152 family ( miR-148a , miR-148b , and miR-152 ) transcript levels in airway epithelial cells from the same subjects. Results miR-148b levels were significantly increased in airway epithelial cells from asthmatic subjects with an asthmatic mother compared with those seen in asthmatic subjects without an asthmatic mother, and +3142 genotypes were associated with sHLA-G concentrations in bronchoalveolar lavage fluid among asthmatic subjects with an asthmatic mother but not among those with a nonasthmatic mother. Neither effect was observed in the plasma (sHLA-G) or white blood cells (miRNA). Conclusion These combined results are consistent with +3142 allele–specific targeting of HLA-G by the miR-152 family and support our hypothesis that miRNA regulation of sHLA-G in the airway is influenced by both the asthma status of the subject's mother and the subject's genotype. Moreover, we demonstrate that the effects of maternal asthma on the gene regulatory landscape in the airways of the mother's children persist into adulthood.
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