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Development of novel diagnostic system for pancreatic cancer, including early stages, measuring mRNA of whole blood cells.

ABSTRACT: Pancreatic ductal adenocarcinoma (PDAC) is the most life-threating disease among all digestive system malignancies. We developed a blood mRNA PDAC screening system using real-time detection PCR to detect the expression of 56 genes, to discriminate PDAC from noncancer subjects. We undertook a clinical study to assess the performance of the developed system. We collected whole blood RNA from 53 PDAC patients, 102 noncancer subjects, 22 patients with chronic pancreatitis, and 23 patients with intraductal papillary mucinous neoplasms in a per protocol analysis. The sensitivity of the system for PDAC diagnosis was 73.6% (95% confidence interval, 59.7%-84.7%). The specificity for noncancer volunteers, chronic pancreatitis, and patients with intraductal papillary mucinous neoplasms was 64.7% (54.6%-73.9%), 63.6% (40.7%-82.8%), and 47.8% (26.8%-69.4%), respectively. Importantly, the sensitivity of this system for both stage I and stage II PDAC was 78.6% (57.1%-100%), suggesting that detection of PDAC by the system is not dependent on the stage of PDAC. These results indicated that the screening system, relying on assessment of changes in mRNA expression in blood cells, is a viable alternative screening strategy for PDAC.

SUBMITTER: Sakai Y

PROVIDER: S-EPMC6447845 | biostudies-literature | 2019 Apr

REPOSITORIES: biostudies-literature

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Development of novel diagnostic system for pancreatic cancer, including early stages, measuring mRNA of whole blood cells.

Sakai Yoshio Y Honda Masao M Matsui Shigeyuki S Komori Osamu O Murayama Toshinori T Fujiwara Tadami T Mizuno Masaaki M Imai Yasuhito Y Yoshimura Kenichi K Nasti Alessandro A Wada Takashi T Iida Noriho N Kitahara Masaaki M Horii Rika R Toshikatsu Tamai T Nishikawa Masashi M Okafuji Hirofumi H Mizukoshi Eishiro E Yamashita Tatsuya T Yamashita Taro T Arai Kuniaki K Kitamura Kazuya K Kawaguchi Kazunori K Takatori Hajime H Shimakami Tetsuro T Terashima Takeshi T Hayashi Tomoyuki T Nio Kouki K Kaneko Shuichi S

Cancer science 20190327 4

Pancreatic ductal adenocarcinoma (PDAC) is the most life-threating disease among all digestive system malignancies. We developed a blood mRNA PDAC screening system using real-time detection PCR to detect the expression of 56 genes, to discriminate PDAC from noncancer subjects. We undertook a clinical study to assess the performance of the developed system. We collected whole blood RNA from 53 PDAC patients, 102 noncancer subjects, 22 patients with chronic pancreatitis, and 23 patients with intra ...[more]

PMID: 30742728

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Project description:ESSENTIALS: How platelets organize their ?-granule cargo and use their canalicular system remains controversial. Past structural studies were limited due to small sampling volumes or decreased resolution. Our analyses revealed homogeneous granules and a closed canalicular system that opened on activation. Understanding how platelets alter their membranes during activation and secretion elucidates hemostasis.Platelets survey the vasculature for damage and, in response, activate and release a wide range of proteins from their ?-granules. Alpha-granules may be biochemically and structurally heterogeneous; however, other studies suggest that they may be more homogeneous with the observed variation reflecting granule dynamics rather than fundamental differences.Our aim was to address how the structural organization of ?-granules supports their dynamics.To preserve the native state, we prepared platelets by high-pressure freezing and freeze-substitution; and to image nearly entire cells, we recorded tomographic data in the scanning transmission electron microscope (STEM).In resting platelets, we observed a morphologically homogeneous ?-granule population that displayed little variation in overall matrix electron density in freeze-substituted preparations (i.e., macro-homogeneity). In resting platelets, the incidence of tubular granule extensions was low, ~4%, but this increased by > 10-fold during early steps in platelet secretion. Using STEM, we observed that the initially decondensing ?-granules and the canalicular system remained as separate membrane domains. Decondensing ?-granules were found to fuse heterotypically with the plasma membrane via long, tubular connections or homotypically with each other. The frequency of canalicular system fusion with the plasma membrane also increased by about three-fold. Our results validate the utility of freeze-substitution and STEM tomography for characterizing platelet granule secretion and suggest a model in which fusion of platelet ?-granules with the plasma membrane occurs via long tubular connections that may provide a spatially limited access route for the timed release of ?-granule proteins.

Dataset Information

Development of novel diagnostic system for pancreatic cancer, including early stages, measuring mRNA of whole blood cells.

Publications

Development of novel diagnostic system for pancreatic cancer, including early stages, measuring mRNA of whole blood cells.

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