Circulating metastasis associated in colon cancer 1 transcripts in gastric cancer patient plasma as diagnostic and prognostic biomarker

METHODS: We provide for the first time a blood-based assay for transcript quantification of the metastasis inducer MACC1 in a prospective study of gastric cancer patient plasma. MACC1 is a strong prognostic biomarker for tumor progression and metastasis in a variety of solid cancers. We conducted a study to define the diagnostic and prognostic power of MACC1 transcripts using 76 plasma samples from gastric cancer patients, either newly diagnosed with gastric cancer, newly diagnosed with metachronous metastasis of gastric cancer, as well as follow-up patients. Findings were controlled by using plasma samples from 54 tumor-free volunteers. Plasma was separated, RNA was isolated, and levels of MACC1 as well as S100A4 transcripts were determined by quantitative RT-PCR. RESULTS: Based on the levels of circulating MACC1 transcripts in plasma we significantly discriminated tumor-free volunteers and gastric cancer patients (P < 0.001). Levels of circulating MACC1 transcripts were increased in gastric cancer patients of each disease stage, compared to tumor-free volunteers: patients with tumors without metastasis (P = 0.005), with synchronous metastasis (P = 0.002), with metachronous metastasis (P = 0.005), and patients during follow-up (P = 0.021). Sensitivity was 0.68 (95%CI: 0.45-0.85) and specificity was 0.89 (95%CI: 0.77-0.95), respectively. Importantly, gastric cancer patients with high circulating MACC1 transcript levels in plasma demonstrated significantly shorter survival when compared with patients demonstrating low MACC1 levels (P = 0.0015). Furthermore, gastric cancer patients with high circulating transcript levels of MACC1 as well as of S100A4 in plasma demonstrated significantly shorter survival when compared with patients demonstrating low levels of both biomarkers or with only one biomarker elevated (P = 0.001). CONCLUSION: Levels of circulating MACC1 transcripts in plasma of gastric cancer patients are of diagnostic value and are prognostic for patient survival in a prospective study

The Value of an Automated Ultrasound System in the Detection of Synovitis

Background The detection of joint swelling caused by synovitis is important for the diagnosis of inflammatory arthritis. Ultrasound (US) and MRI have proven to be more sensitive and reliable than physical examination, but they are time-consuming and expensive. The automated breast volume scanner was developed to acquire serial B-mode pictures of the female breast and these can be analyzed in all three dimensions. Objectives To analyze the value of automated B-mode ultrasound employing the ABVS system in detecting synovitis of the finger joints compared to manual ultrasound (mUS) and physical examination, using MRI as the gold standard. Methods 19 consecutive patients suffering from active rheumatoid (n=15) or psoriatic (n=4) arthritis were included. Automated and mUS were conducted with a linear array (ACUSON S2000, 11MHz). Multiplanar reconstruction enabled examination of the images for the presence of synovitis. Results 90% of the hand joints were assessable by automated ultrasound. Automated US detected 12.0, mUS 14.2, MRI 13.4, and clinical examination 4.1 positive joints - i.e. joints with synovitis - on average per patient. The inter-observer reliability of both assessors for automated and mUS, MRI, and physical examination, was 66.9%, 72.7%, 95.1%, and 88.9%, respectively. 84.3% of the joints classified as positive on MRI were confirmed by automated ultrasound, 85.5% on mUS, and 36.0 on physical examination. This translated into a sensitivity of 83.5%, 85.5%, and 36.0% for the three methods, respectively. Conclusion: Automated ultrasound is a promising ultrasound method for assessing small joints in patients with inflammatory arthritis

AP-1 recruitment to VAMP4 is modulated by phosphorylation-dependent binding of PACS-1

The R-SNARE VAMP4, which contains a dileucine motif, binds to the AP-1 (adaptor protein-1) subunit μ1a, but not μ1b, or the GGAs (Golgi-associated gamma ear containing ARF binding proteins). Serine 20 and leucines 25,26 are essential for this binding. AP-1 association with VAMP4 is enhanced when serine 30, in an acidic cluster, is phosphorylated by casein kinase 2. This phosphorylation-dependent modulation of AP-1 binding is mediated by PACS-1 (phosphofurin acidic cluster sorting protein). Ablation of both the dileucine motif and serine 30 results in a dramatic mislocalization of VAMP4 in the regulated secretory pathway in AtT20 cells. A dominant-negative PACS-1, which binds acidic clusters but not AP-1, also causes mislocalization of VAMP4. Our data support a model whereby phosphorylation-dependent recruitment of PACS-1 enhances AP-1 association to cargo, and suggest that efficient retrieval depends on the formation of a complex between cargo, such as VAMP4, AP-1 and PACS-1

Circulating MACC1 transcript levels in plasma discriminate cancer patients from tumor-free volunteers.

<p>A. Plasma separation conditions for determination of circulating MACC1 transcripts. Plasma separation of samples from tumor-free volunteers (n = 12) was done immediately, 7, 24, 48, and 72 hours after taking blood, either from blood samples kept at 4°C or at room temperature. Subsequently, plasma was generated within the first 7 hours from 4°C-cooled blood. B. MACC1 transcripts in plasma of tumor-free volunteers (n = 54). No difference in MACC1 transcript levels were found in two independently analyzed cohorts of tumor-free volunteers (n = 34 and n = 20, respectively). C. MACC1 transcripts in plasma of all colorectal cancer patients (n = 312). All patient cohorts bearing colorectal, colon (n = 151), or rectal (n = 161) cancer expressed significantly higher MACC1 transcript levels than tumor-free volunteers (<i>P</i><.001 for all comparisons).</p

Colon cancer patients characteristics and circulating MACC1 transcript levels in plasma.

<p>Colon cancer patients characteristics and circulating MACC1 transcript levels in plasma.</p

Colorectal cancer patients characteristics and circulating MACC1 transcript levels in plasma.

<p>Colorectal cancer patients characteristics and circulating MACC1 transcript levels in plasma.</p

Survival of CRC patients based on circulating transcript levels of MACC1 or a combination of MACC1 and S100A4.

<p>A, B. Kaplan-Meier analysis for newly diagnosed (A, n = 49) and all (B, n = 294) CRC patients, based on MACC1 alone (A, low MACC1 n = 24, high MACC1 n = 25; B, low MACC1 n = 147, high MACC1 n = 147). Newly diagnosed CRC patients (A) as well as all CRC patients (B) with high circulating MACC1 transcript levels demonstrated shorter survival when compared with patients demonstrating low MACC1 levels (<i>P</i> = .003 and <i>P</i><.0001, respectively). C, D. Kaplan-Meier analysis for newly diagnosed (C, n = 49) and all (D, n = 294) CRC patients, based on a combination of MACC1 and S100A4. Newly diagnosed CRC patients (C) and all CRC patients (D) were classified into groups of low expressors of both genes (n = 13 and n = 85, respectively), of patients with high S100A4 levels (n = 25 and n = 147, respectively), of patients with high MACC1 levels (n = 25 and n = 147, respectively), or with high expression of both biomarkers (n = 14 and n = 85, respectively). For newly diagnosed patients (C), survival was reduced with increasing levels of circulating transcripts of S100A4, of MACC1, or of both. Expression induction of either MACC1 or S100A4 or of both biomarkers correlated to reduced patients survival.</p

Diagnostic value of circulating MACC1 transcript levels in plasma of newly diagnosed colorectal, colon, and rectal cancer patients with a primary tumor with or without synchronous metastasis.

<p>Optimal cut offs were calculated with a fourfold table.</p

Rectal cancer patients characteristics and circulating MACC1 transcript levels in plasma.

<p>Rectal cancer patients characteristics and circulating MACC1 transcript levels in plasma.</p

Circulating MACC1 transcripts in colorectal, colon, and rectal cancer patient plasma.

<p>A, B, C. All colorectal (A, n = 312), colon (B, n = 151), and rectal (C, n = 161) cancer patient sub-cohorts showed significantly higher circulating MACC1 transcript levels than tumor-free volunteers (n = 54). Higher MACC1 levels were also found for colorectal (<i>P</i> = .006; A) and rectal (<i>P</i> = .041; C) cancer patients with synchronous metastases (n = 20 and n = 10, respectively) compared to patients without distant metastases (n = 51 and n = 39, respectively). Box plot analysis, based on quantitative real-time RT-PCR.</p