Diagnostic criteria for oncocytic renal neoplasms:a survey of urologic pathologists

Renal oncocytoma and chromophobe renal cell carcinoma (RCC) have been long recognized as distinct tumors; however, it remains unknown if uniform diagnostic criteria are used to distinguish these tumor types in practice. A survey was distributed to urologic pathologists regarding oncocytic tumors. Responses were received from 17/26 invitees. Histologically, >1 mitotic figure was regarded as most worrisome (n=10) or incompatible (n=6) with oncocytoma diagnosis. Interpretation of focal nuclear wrinkling, focal perinuclear clearing, and multinucleation depended on extent and did not necessarily exclude oncocytoma if minor. Staining techniques most commonly used included: CK7 (94%), KIT (71%), vimentin (65%), colloidal iron (59%), CD10 (53%), and AMACR (41%). Rare CK7-positive cells (≤5%) was regarded as most supportive of oncocytoma, although an extent excluding oncocytoma was not universal. Multiple chromosomal losses were most strongly supportive for chromophobe RCC diagnosis (65%). Less certainty was reported for chromosomal gain or a single loss. For tumors with mixed or inconclusive features, many participants use an intermediate diagnostic category (82%) that does not label the tumor as unequivocally benign or malignant, typically "oncocytic neoplasm" or "tumor" with comment. The term "hybrid tumor" was used variably in several scenarios. A slight majority (65%) report outright diagnosis of oncocytoma in needle biopsies. The morphologic, immunohistochemical, and genetic characteristics that define oncocytic renal tumors remain incompletely understood. Further studies correlating genetics, behavior, and histology are needed to define which tumors truly warrant classification as carcinomas for patient counseling and follow-up strategies

ADAM15 Is Functionally Associated with the Metastatic Progression of Human Bladder Cancer.

ADAM15 is a member of a family of catalytically active disintegrin membrane metalloproteinases that function as molecular signaling switches, shed membrane bound growth factors and/or cleave and inactivate cell adhesion molecules. Aberrant metalloproteinase function of ADAM15 may contribute to tumor progression through the release of growth factors or disruption of cell adhesion. In this study, we utilized human bladder cancer tissues and cell lines to evaluate the expression and function of ADAM15 in the progression of human bladder cancer. Examination of genome and transcriptome databases revealed that ADAM15 ranked in the top 5% of amplified genes and its mRNA was significantly overexpressed in invasive and metastatic bladder cancer compared to noninvasive disease. Immunostaining of a bladder tumor tissue array designed to evaluate disease progression revealed increased ADAM15 immunoreactivity associated with increasing cancer stage and exhibited significantly stronger staining in metastatic samples. About half of the invasive tumors and the majority of the metastatic cases exhibited high ADAM15 staining index, while all low grade and noninvasive cases exhibited negative or low staining. The knockdown of ADAM15 mRNA expression significantly inhibited bladder tumor cell migration and reduced the invasive capacity of bladder tumor cells through MatrigelTM and monolayers of vascular endothelium. The knockdown of ADAM15 in a human xenograft model of bladder cancer inhibited tumor growth by 45% compared to controls. Structural modeling of the catalytic domain led to the design of a novel ADAM15-specific sulfonamide inhibitor that demonstrated bioactivity and significantly reduced the viability of bladder cancer cells in vitro and in human bladder cancer xenografts. Taken together, the results revealed an undescribed role of ADAM15 in the invasion of human bladder cancer and suggested that the ADAM15 catalytic domain may represent a viable therapeutic target in patients with advanced disease

Novel ADAM15-specific sulfonamide inhibitor reduces viability of bladder cancer cells.

<p><i>A)</i> Surface model of adamastat binding to the ADAM15 catalytic. <i>B)</i> A mesh representation of adamastat binding in the S1’ pocket of the catalybtic domain of ADAM15. <i>C)</i> Fluorescence Resonance Energy Transfer assay indicating that inhibition of the recombinant ADAM15 catalytic domain (rA15cat) by adamastat is as effective as two other previously tested metalloproteinase inhibitors over the same concentration range. FI, Fluorescence Intensity. <i>D)</i> Viability of adamastat treated SV-HUC-1, UM-UC-9 and UM-UC-6 cells. Vehicle represents 0.1% DMSO control. Bars show mean cell viability % ± SD in 1 out of three independent experiments.</p

Integration of Architectural and Cytologic Driven Image Algorithms for Prostate Adenocarcinoma Identification

Introduction:: The advent of digital slides offers new opportunities within the practice of pathology such as the use of image analysis techniques to facilitate computer aided diagnosis (CAD) solutions. Use of CAD holds promise to enable new levels of decision support and allow for additional layers of quality assurance and consistency in rendered diagnoses. However, the development and testing of prostate cancer CAD solutions requires a ground truth map of the cancer to enable the generation of receiver operator characteristic (ROC) curves. This requires a pathologist to annotate, or paint, each of the malignant glands in prostate cancer with an image editor software - a time consuming and exhaustive process. Recently, two CAD algorithms have been described: probabilistic pairwise Markov models (PPMM) and spatially-invariant vector quantization (SIVQ). Briefly, SIVQ operates as a highly sensitive and specific pattern matching algorithm, making it optimal for the identification of any epithelial morphology, whereas PPMM operates as a highly sensitive detector of malignant perturbations in glandular lumenal architecture. Methods:: By recapitulating algorithmically how a pathologist reviews prostate tissue sections, we created an algorithmic cascade of PPMM and SIVQ algorithms as previously described by Doyle el al. [1] where PPMM identifies the glands with abnormal lumenal architecture, and this area is then screened by SIVQ to identify the epithelium. Results:: The performance of this algorithm cascade was assessed qualitatively (with the use of heatmaps) and quantitatively (with the use of ROC curves) and demonstrates greater performance in the identification of malignant prostatic epithelium. Conclusion:: This ability to semi-autonomously paint nearly all the malignant epithelium of prostate cancer has immediate applications to future prostate cancer CAD development as a validated ground truth generator. In addition, such an approach has potential applications as a pre-screening/quality assurance tool

ADAM15 staining index in bladder cancer microarrays.

<p>ADAM15 staining index in bladder cancer microarrays.</p

Knockdown of ADAM15 inhibits bladder tumor cell motility.

<p><i>A)</i> ADAM15 immunoblot of UM-UC-9 bladder cancer cells expressing empty vector and scrambled ADAM15 sequences (ctl) and short hairpin RNA (shA15) and UM-UC-6 expressing scrambled ADAM15 sequences (ctl) and short hairpin RNA (shA15). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as loading control. <i>B)</i> Wound healing assay comparing extent of wound closure in shA15 UM-UC-9 and shA15 UM-UC-6 cells compared to controls (ctl). Bars represent mean ± SD (n = 4) of one of two independent experiments (UM-UC-9, <i>top</i>) and mean ± SEM (N = 3) (UM-UC-6, <i>bottom</i>). <i>C)</i> Proliferation assays of both knockdown (shA15) and control (ctl) UM-UC-9 and UM-UC-6 cells over a 72 hour time course. Results represent mean ± SD of one of three independent experiments.</p

<i>In vivo</i> inhibition of ADAM15 activity reduces tumor growth.

<p><i>A)</i> ADAM15 immunoblot of UM-UC-6 bladder cancer cells expressing empty vector (Vec), control scrambled ADAM15 sequences (ctlA15UC6) and short hairpin RNA (shRNA) sequence (shA15UC6). GAPDH was utilized as loading control (<i>top</i>). Subcutaneous inoculation of shA15UC6 cells (n = 10) in SCID mice led to decreased tumor weight compared to ctlA15UC6 control cells (n = 10) (<i>bottom</i>). <i>B)</i> UM-UC-6 cells were subcutaneously implanted in SCID mice. Specific inhibition of the ADAM15 catalytic activity by daily treatment with adamastat 100 mg/kg (n = 5) during 3 weeks led to decrease in tumor growth compared to vehicle treated mice (n = 5). Bars represent mean tumor weight (mg) ± SD (<i>5A</i>,<i>B</i>).</p

Integration of Architectural and Cytologic Driven Image Algorithms for Prostate Adenocarcinoma Identification

Introduction: The advent of digital slides offers new opportunities within the practice of pathology such as the use of image analysis techniques to facilitate computer aided diagnosis (CAD) solutions. Use of CAD holds promise to enable new levels of decision support and allow for additional layers of quality assurance and consistency in rendered diagnoses. However, the development and testing of prostate cancer CAD solutions requires a ground truth map of the cancer to enable the generation of receiver operator characteristic (ROC) curves. This requires a pathologist to annotate, or paint, each of the malignant glands in prostate cancer with an image editor software - a time consuming and exhaustive process

Knockdown of ADAM15 decreases bladder cancer cell invasion and transmigration through vascular endothelium.

<p><i>A)</i> Extent of Matrigel^TM invasion of UM-UC-9 and UM-UC-6 bladder cancer cells expressing empty vector and scrambled ADAM15 sequences (ctl) and short hairpin RNA (shA15). The bars indicate mean invading cells ± SD (n = 3). The data are representative of 1 of 5 UM-UC-9 or 1 of 3 UM-UC-6 independent experiments. A representative 20X image of each condition is shown. <i>B)</i> Transendothelial migration assay revealed reduced transmigration through an endothelial HUV-EC-C monolayer of shA15UM-UC-9 and shA15UM-UC-6 cells when compared with control cells (ctl). Bars represent mean ± SEM (N = 3). A representative 20X image of each condition is shown.</p