The Somatic Genomic Landscape of Chromophobe Renal Cell Carcinoma

We describe the landscape of somatic genomic alterations of 66 chromophobe renal cell carcinomas (ChRCCs) based on multidimensional and comprehensive characterization, including mitochondrial DNA (mtDNA) and whole genome sequencing. The result is consistent that ChRCC originates from the distal nephron compared to other kidney cancers with more proximal origins. Combined mtDNA and gene expression analysis implicates changes in mitochondrial function as a component of the disease biology, while suggesting alternative roles for mtDNA mutations in cancers relying on oxidative phosphorylation. Genomic rearrangements lead to recurrent structural breakpoints within TERT promoter region, which correlates with highly elevated TERT expression and manifestation of kataegis, representing a mechanism of TERT up-regulation in cancer distinct from previously-observed amplifications and point mutations

Engaging African Americans in Smoking Cessation Programs

African Americans are disproportionately exposed to and targeted by prosmoking advertisements, particularly menthol cigarette ads. Though African Americans begin smoking later than whites, they are less likely to quit smoking than whites. This study was designed to explore African American smoking cessation attitudes, behaviors, and resources to gain insights on how to enhance the appeal of smoking cessation interventions in African American communities. Ten focus groups were conducted with urban, suburban, and rural African American adult smokers and ex-smokers in Maryland who also completed brief questionnaires. Although knowledge of negative health effects of smoking and motivation for smoking cessation were high, participants lacked confidence in their ability to quit successfully and were poorly informed about resources and programs for smoking cessation. Findings from this study suggest that cultural tailoring, neighborhood focus, and strong marketing may enhance the appeal of smoking cessation programs to African American smokers. Programs also may be more attractive if they use respected nonsmoking role models and peer support

Expression of Ror2 mediates invasive phenotypes in renal cell carcinoma.

Ror2 is a Wnt ligand receptor that is overexpressed in a variety of tumors including clear cell renal cell carcinoma (ccRCC). Here we demonstrate that expression of wild type Ror2 results in increased tumorigenic properties in in vitro cell culture and in vivo xenograft models. In addition, Ror2 expression produced positive changes in both cell migration and invasion, which were dependent on matrix metalloprotease 2 (MMP2) activity. Mutations in key regions of the kinase domain of Ror2 resulted in the abrogation of increased tumor growth, cell migration, and cell invasion observed with expression of wild-type Ror2. Finally, we examined Ror2 expression as a prognostic biomarker for ccRCC utilizing the TCGA ccRCC dataset. High expression of Ror2 showed a significant correlation with higher clinical stage, nuclear grade, and tumor stage. Furthermore, high expression of Ror2 in ccRCC patients correlated with significant lower overall survival, cancer specific survival, and recurrence free survival. Together, these findings suggest that Ror2 plays a central role in influencing the ccRCC phenotype, and can be considered as a negative prognostic biomarker and potential therapeutic target in this cancer

Ror2 regulation of MMP2 expression in RCC cells is dependent upon an intact kinase domain.

<p>Quantitative RT-PCR for 786-0 cells shows a significant correlated decrease in <b>A</b>) Ror2 and <b>B</b>) MMP2 mRNA levels in both shRNA knockdown cell lines relative to control. <b>C</b>) Additionally, induction of wildtype Ror2 and Ror2-DM is evident following treatment with doxycycline (500 ng/ml). <b>D</b>) But a matching increase in MMP2 is only seen with overexpression of wildtype Ror2. For all quantitative RT-PCR assays, transcript values were normalized to β-actin RNA internal standard with fold change calculated in reference to either 786-0 control or unstimulated GFP expressing cells. Error bars represent SEM across triplicates of a representative duplicated experiment. Gelatin Zymography shows increased levels of both the precursor pro-MMP2 and its cleaved active form with overexpression of wildtype Ror2 relative to GFP vector control in both <b>E</b>) 786-0 and <b>F</b>) HEK293T cells. Quantification of the levels of active MMP2 normalized to the media only control (Con) is shown below each gel. <b>G</b>) Invasion of 786-0 cells seeded in Boyden matrigel coated chambers under normal culture conditions show a significant decrease upon treatment with MMP2 inhibitor (OA-Hy - 60 uM) in both independent shRNA knockdowns and Ror2 overexpression in comparison with vehicle control. The percentage of area covered was quantitated from random fields using ImageJ. Error bars represent stdev across duplicates of a representative duplicated experiment. <i>P</i>-values were calculated using an Anova one-way analysis (*<.05, **<.001).</p

Ror2 expression regulates cell invasion in RCC cells.

<p>Analysis of invasive potential using Boyden matrigel coated chamber assays under normal culture conditions with <b>A</b>) 786-0 cells show a significant reduction in invasion in both independent shRNA Ror2 knockdowns in comparison with parental 786-0 and vector control cells (representative images shown below). Additionally, <b>B</b>) 786-0 & <b>C</b>) HEK293T cells under normal culture conditions show a significant increase in cell invasion with overexpression of wild-type Ror2 that is reduced with expression of Ror2-DM (representative images shown below). The percentage of area covered was quantitated from random fields using ImageJ. <i>P</i>-values were calculated using an Anova one-way analysis with error bars representing stdev shown (*<.05, **<.001).</p

Summary of clinical characteristics of patients in the TCGA ccRCC dataset.

<p>Summary of clinical characteristics of patients in the TCGA ccRCC dataset.</p

Expression of Ror2 promotes <i>in vivo</i> tumor growth and invasion.

<p><b>A&C</b>) Tabulated rates of successful injection and growth of all 786-0 xenograft tumors are shown. <b>B</b>) Average tumor size (cm) with error bars showing stdev for resulting xenograft tumors following paired orthotropic injections of 786-0 cells containing either the control empty vector, pcDNA6 or hRor2 exhibited a trend to increased <i>in vivo</i> tumor growth with Ror2 overexpression. <b>D</b>) Average tumor size (mm) with error bars showing stdev for resulting xenograft tumors following subcutaneous injection of 786-0 cells containing either the control empty vector, GFP, wild-type Ror2, or Ror2-DM showed a significant increase in <i>in vivo</i> tumor growth with wild-type Ror2 overexpression. <i>P</i>-values were calculated using an Anova one-way analysis (*<.05 **<.01). <b>E</b>) Representative image of an orthotopic xenograft consisting of 786-0 vector control cells. <b>F</b>) Increased invasion of local tissues was evident with Ror2 expression, with arrows highlighting areas of continued invasion into the spleen.</p

High Ror2 expression correlates with increased tumor growth and stage in primary human RCC tumors.

<p>Tumor percentage breakdowns of TCGA ccRCC cases (468) stratified by Ror2-High and Ror2-Low expression show Ror2-High tumors exhibiting significant increases in <b>A</b>) tumor size, <b>B</b>) higher nuclear grade, <b>C</b>) clinical stage, and <b>D</b>) tumor stage. <i>P</i>-values were calculated using either an Anova one-way analysis or Fisher's exact test (*<.05, **<.001).</p

Expression of Ror2 Mediates Invasive Phenotypes in Renal Cell Carcinoma

Ror2 is a Wnt ligand receptor that is overexpressed in a variety of tumors including clear cell renal cell carcinoma (ccRCC). Here we demonstrate that expression of wild type Ror2 results in increased tumorigenic properties in in vitro cell culture and in vivo xenograft models. In addition, Ror2 expression produced positive changes in both cell migration and invasion, which were dependent on matrix metalloprotease 2 (MMP2) activity. Mutations in key regions of the kinase domain of Ror2 resulted in the abrogation of increased tumor growth, cell migration, and cell invasion observed with expression of wild-type Ror2. Finally, we examined Ror2 expression as a prognostic biomarker for ccRCC utilizing the TCGA ccRCC dataset. High expression of Ror2 showed a significant correlation with higher clinical stage, nuclear grade, and tumor stage. Furthermore, high expression of Ror2 in ccRCC patients correlated with significant lower overall survival, cancer specific survival, and recurrence free survival. Together, these findings suggest that Ror2 plays a central role in influencing the ccRCC phenotype, and can be considered as a negative prognostic biomarker and potential therapeutic target in this cancer