Tissue microarrays enabling high-throughput molecular pathology

Takikita, Mikiko; Chung, Joon‐Yong; Hewitt, Stephen M.

doi:10.1016/j.copbio.2007.05.007

Cited by 36 publications

(23 citation statements)

References 43 publications

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“…Although immunohistochemistry is considered a gold standard for evaluation of protein expression in histopathology, several difficult issues remain in evaluation of immunohistochemistry, including a subjective of determination of intensity (29). Image analysis overcomes some of these issues, but it lacks normalization to account for differences in tissue handling and processing (30). In order to overcome this limitation of immunohistochemistry, we examined protein expression with MTI, which could measure protein expression, incorporating quantitation and normalization.…”

Section: Discussionmentioning

confidence: 99%

“…In order to overcome this limitation of immunohistochemistry, we examined protein expression with MTI, which could measure protein expression, incorporating quantitation and normalization. Normalization methods for immunohistochemistry have been suggested (31), but remain technically challenging (30). The MTI method permits rigorous comparisons of protein expression across histogeographic zones of histology as well as interrogation of multiple antigens from a single section of tissue (16 -19).…”

Section: Discussionmentioning

confidence: 99%

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The Expression of Phospho-AKT, Phospho-mTOR, and PTEN in Extrahepatic Cholangiocarcinoma

Chung

Hong

Choi

et al. 2009

Clinical Cancer Research

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107

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Purpose: The protein kinase B (AKT) pathway plays a key role in the regulation of cellular survival, apoptosis, and protein translation, and has been shown to have prognostic significance in a number of cancers.We sought to define its role in extrahepatic cholangiocarcinoma. Experimental Design: Two hundred twenty-one extrahepatic cholangiocarcinoma patients with clinicopathologic data, including survival, were arrayed into tissue microarrays. Phosphorylated AKT (p-AKT), phosphorylated mammalian target of rapamycin (p-mTOR), and total phosphatase and tensin homolog deleted on chromosome 10 (PTEN) protein expressions were studied with multiplex tissue immunoblotting assay. Results: Expressions of p-AKT and p-mTOR were significantly increased in extrahepatic cholangiocarcinoma cases compared with normal and dysplastic bile duct epithelium (P < 0.05 both).Decreased PTEN expression was observed in patients with increasing depth of invasion (P < 0.05),Tclassification (P < 0.05), and stage grouping (P < 0.05), and the presence of invasion of the pancreas (P < 0.05) and duodenum (P < 0.05). Decreased PTEN expression (P = 0.004) as well as decreased PTEN/p-AKT (P = 0.003) and PTEN/p-mTOR (P = 0.009) expression showed shorter survival by univariate but not by multivariate analysis. Conclusions: The AKT pathway is activated in a subset of extrahepatic cholangiocarcinoma. Elevated PTEN expression correlates with longer survival. Quantitative data obtained by multiplex tissue immunoblotting may provide additional information than assessment of immunohistochemistry alone. Quantitative analysis of PTEN, PTEN/p-AKT and PTEN/p-mTOR shows differences in survival by univariate analysis.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

The Expression of Phospho-AKT, Phospho-mTOR, and PTEN in Extrahepatic Cholangiocarcinoma

Chung

Hong

Choi

et al. 2009

Clinical Cancer Research

Self Cite

107

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“…TMAs are collections of multiple tissue cores placed in parallel in a single acceptor paraffin block, thereby enabling simultaneous and standardized analyses of many samples concurrently. 16,17 The application of TMA technology in this type of vascular disease analyses places our study among the very few that have attempted the same. 18 By this approach, we demonstrated upregulation of genes involved in inflammation, lipid metabolism, cell proliferation, and coagulation in symptomatic compared with asymptomatic carotid plaques, as well as differential expression of several proteins.…”

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confidence: 99%

Profiling of Atherosclerotic Lesions by Gene and Tissue Microarrays Reveals PCSK6 as a Novel Protease in Unstable Carotid Atherosclerosis

Perisic

Hedin

Razuvaev

et al. 2013

ATVB

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Objective— Carotid plaque instability is a major cause of ischemic stroke, but detailed knowledge about underlying molecular pathways is still lacking. Here, we evaluated large-scale transcriptomic and protein expression profiling in a biobank of carotid endarterectomies followed by characterization of identified candidates, as a platform for discovery of novel proteins differentially regulated in unstable carotid lesions. Approach and Results— Genes highly upregulated in symptomatic versus asymptomatic plaques were selected from Affymetrix microarray analyses (n=127 plaques), and tissue microarrays constructed from 34 lesions were assayed for 21 corresponding proteins by immunohistochemistry. Quantification of stainings demonstrated differential expression of CD36, CD137, and DOCK7 ( P <0.05) in unstable versus stable lesions and the most significant upregulation of a proprotein convertase, PCSK6 ( P <0.0001). Increased expression of PCSK6 in symptomatic lesions was verified by quantitative real-time polymerase chain reaction (n=233), and the protein was localized to smooth muscle α-actin positive cells and extracellular matrix of the fibrous cap by immunohistochemistry. PCSK6 expression positively correlated to genes associated with inflammation, matrix degradation, and mitogens in microarrays. Stimulation of human carotid smooth muscle cells in vitro with cytokines caused rapid induction of PCSK6 mRNA. Conclusions— Using a combination of transcriptomic and tissue microarray profiling, we demonstrate a novel approach to identify proteins differentially expressed in unstable carotid atherosclerosis. The proprotein convertase PCSK6 was detected at increased levels in the fibrous cap of symptomatic carotid plaques, possibly associated with key processes in plaque rupture such as inflammation and extracellular matrix remodeling. Further studies are needed to clarify the role of PCSK6 in atherosclerosis.

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“…Our method can classify the images with approximately 90% specificity and above 92% sensitivity. Automation has been tried at different stages of the TMA process by various groups [22][23][24][25][26][27][28]. Automation of construction of TMA has been quite satisfactory.…”

Section: Discussionmentioning

confidence: 99%

Datamining Approach for Automation of Diagnosis of Breast Cancer in Immunohistochemically Stained Tissue Microarray Images

Prasad¹

2010

TOMINFOJ

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Abstract:Cancer of the breast is the second most common human neoplasm, accounting for approximately one quarter of all cancers in females after cervical carcinoma. Estrogen receptor (ER), Progesteron receptor and human epidermal growth factor receptor (HER-2/neu) expressions play an important role in diagnosis and prognosis of breast carcinoma. Tissue microarray (TMA) technique is a high throughput technique which provides a standardized set of images which are uniformly stained, facilitating effective automation of the evaluation of the specimen images. TMA technique is widely used to evaluate hormone expression for diagnosis of breast cancer. If one considers the time taken for each of the steps in the tissue microarray process workflow, it can be observed that the maximum amount of time is taken by the analysis step. Hence, automated analysis will significantly reduce the overall time required to complete the study. Many tools are available for automated digital acquisition of images of the spots from the microarray slide. Each of these images needs to be evaluated by a pathologist to assign a score based on the staining intensity to represent the hormone expression, to classify them into negative or positive cases. Our work aims to develop a system for automated evaluation of sets of images generated through tissue microarray technique, representing the ER expression images and HER-2/neu expression images. Our study is based on the Tissue Microarray Database portal of Stanford university at http://tma.stanford.edu/cgibin/cx?n=her1, which has made huge number of images available to researchers. We used 171 images corresponding to ER expression and 214 images corresponding to HER-2/neu expression of breast carcinoma. Out of the 171 images corresponding to ER expression, 104 were negative and 67 were representing positive cases. Out of the 214 images corresponding to HER-2/neu expression, 112 were negative and 102 were representing positive cases. Our method has 92.31% sensitivity and 93.18% specificity for ER expression image classification and 96.67% sensitivity and 88.24% specificity for HER-2/neu expression image classification.

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Tissue microarrays enabling high-throughput molecular pathology

Cited by 36 publications

References 43 publications

The Expression of Phospho-AKT, Phospho-mTOR, and PTEN in Extrahepatic Cholangiocarcinoma

The Expression of Phospho-AKT, Phospho-mTOR, and PTEN in Extrahepatic Cholangiocarcinoma

Profiling of Atherosclerotic Lesions by Gene and Tissue Microarrays Reveals PCSK6 as a Novel Protease in Unstable Carotid Atherosclerosis

Datamining Approach for Automation of Diagnosis of Breast Cancer in Immunohistochemically Stained Tissue Microarray Images

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