High-Throughput Prostate Cancer Gland Detection, Segmentation, and Classification from Digitized Needle Core Biopsies

BACKGROUND Biomarker validation remains one of the most challenging constraints to the development of new diagnostic assays. To facilitate biomarker validation, we previously developed a chromatography-free stable isotope standards and capture by antipeptide antibodies (SISCAPA)-MALDI assay allowing rapid, high-throughput quantification of protein analytes in large sample sets. Here we applied this assay to the measurement of a surrogate proteotypic peptide from protein C inhibitor (PCI) in sera from patients with prostate cancer. METHODS A 2-plex SISCAPA-MALDI assay for quantification of proteotypic peptides from PCI and soluble transferrin receptor (sTfR) was used to measure these peptides in 159 trypsin-digested sera collected from 51 patients with prostate cancer. These patients had been treated with radiation with or without neoadjuvant androgen deprivation. RESULTS Patients who experienced biochemical recurrence of prostate cancer showed decreased serum concentrations of the PCI peptide analyte within 18 months of treatment. The PCI peptide concentrations remained increased in the sera of patients who did not experience cancer recurrence. Prostate-specific antigen concentrations had no predictive value during the same time period. CONCLUSIONS The high-throughput, liquid chromatography–free SISCAPA-MALDI assay is capable of rapid quantification of proteotypic PCI and sTfR peptide analytes in complex serum samples. Decreased serum concentrations of the PCI peptide were found to be related to recurrence of prostate cancer in patients treated with radiation with or without hormone therapy. However, a larger cohort of patients will be required for unequivocal validation of the PCI peptide as a biomarker for clinical use.

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Abstract LB-332: High-throughput proteomic analysis identifies protein S as a potential modulator of androgen-independent prostate cancer

10.1158/1538-7445.am2011-lb-332 ◽

2011 ◽

Author(s):

Punit Saraon ◽

Keith Jarvi ◽

Eleftherios P. Diamandis

Keyword(s):

Prostate Cancer ◽

High Throughput ◽

Proteomic Analysis ◽

Protein S ◽

Androgen Independent

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Analytical Currents: Defining the molecular forces that shape protein nanofibrils | Understanding noise in solid-state nanopores | DataChip tackles high-throughput drug screening | Dielectrophoresis with reconfigurable oil barriers | A multiwell, multicantilever, protein detection system | Proteomic profiling method not suitable for detecting prostate cancer | Whole-organism imaging MS | MALDI MS of transition-metal catalysts | Bard and Moerner win the 2008 Wolf Prize in Chemistry

Analytical Chemistry ◽

10.1021/ac0860572 ◽

2008 ◽

Vol 80 (5) ◽

pp. 1349-1352

Keyword(s):

Prostate Cancer ◽

Solid State ◽

High Throughput ◽

Detection System ◽

Protein Detection ◽

Transition Metal Catalysts ◽

Proteomic Profiling ◽

High Throughput Drug Screening ◽

Molecular Forces ◽

Imaging Ms

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Detection of Intracellular Granularity Induction in Prostate Cancer Cell Lines by Small Molecules Using the HyperCyt® High-Throughput Flow Cytometry System

CrossRef Listing of Deleted DOIs ◽

10.1177/1087057109335671 ◽

2009 ◽

Vol 14 (6) ◽

pp. 596-609 ◽

Cited By ~ 21

Author(s):

Mark K. Haynes ◽

J. Jacob Strouse ◽

Anna Waller ◽

Andrei Leitao ◽

Ramona F. Curpan ◽

...

Keyword(s):

Prostate Cancer ◽

Flow Cytometry ◽

Small Molecules ◽

Cell Lines ◽

Cancer Cell ◽

High Throughput ◽

Prostate Cancer Cell ◽

Cancer Cell Lines ◽

Phenotypic Change ◽

Prostate Cancer Cell Lines

Prostate cancer is a leading cause of death among men due to the limited number of treatment strategies available for advanced disease. Discovery of effective chemotherapeutics involves the identification of agents that inhibit cancer cell growth. Increases in intracellular granularity have been observed during physiological processes that include senescence, apoptosis, and autophagy, making this phenotypic change a useful marker for identifying small molecules that induce cellular growth arrest or death. In this regard, epithelial-derived cancer cell lines appear uniquely susceptible to increased intracellular granularity following exposure to chemotherapeutics. We have established a novel flow cytometry approach that detects increases in side light scatter in response to morphological changes associated with intracellular granularity in the androgen-sensitive LNCaP and androgen-independent PC3 human prostate cancer cell lines. A cell-based assay was developed to screen for small molecule inducers of intracellular granularity using the HyperCyt® high-throughput flow cytometry platform. Validation was performed using the Prestwick Chemical Library, where known modulators of LNCaP intracellular granularity, such as testosterone, were identified. Nonandrogenic inducers of granularity were also detected. A further screen of ~25,000 small molecules led to the identification of a class of aryl-oxazoles that increased intracellular granularity in both cell lines, often leading to cell death. The most potent agents exhibited submicromolar efficacy in LNCaP and PC3 cells. ( Journal of Biomolecular Screening. 2009:596-609)

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miR-26a regulates extracellular vesicle secretion from prostate cancer cells via targeting SHC4, PFDN4, and CHORDC1

Science Advances ◽

10.1126/sciadv.aay3051 ◽

2020 ◽

Vol 6 (18) ◽

pp. eaay3051 ◽

Cited By ~ 4

Author(s):

Fumihiko Urabe ◽

Nobuyoshi Kosaka ◽

Yurika Sawa ◽

Yusuke Yamamoto ◽

Kagenori Ito ◽

...

Keyword(s):

Prostate Cancer ◽

Cancer Cells ◽

Cancer Progression ◽

High Throughput ◽

Intercellular Communication ◽

Extracellular Vesicle ◽

Prostate Cancer Cells ◽

Biological Phenomena ◽

Vesicle Secretion

Extracellular vesicles (EVs) are involved in intercellular communication during cancer progression; thus, elucidating the mechanism of EV secretion in cancer cells will contribute to the development of an EV-targeted cancer treatment. However, the biogenesis of EVs in cancer cells is not fully understood. MicroRNAs (miRNAs) regulate a variety of biological phenomena; thus, miRNAs could regulate EV secretion. Here, we performed high-throughput miRNA-based screening to identify the regulators of EV secretion using an ExoScreen assay. By using this method, we identified miR-26a involved in EV secretion from prostate cancer (PCa) cells. In addition, we found that SHC4, PFDN4, and CHORDC1 genes regulate EV secretion in PCa cells. Furthermore, the progression of the PCa cells suppressing these genes was inhibited in an in vivo study. Together, our findings suggest that miR-26a regulates EV secretion via targeting SHC4, PFDN4, and CHORDC1 in PCa cells, resulting in the suppression of PCa progression.

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