Prognostic role of tumour associated trypsin inhibitor in colorectal cancer patients

Colorectal cancer (CRC) is one of the most common forms of human cancer worldwide with approximately 1 million new cases detected every year. CEA is currently the only accepted CRC marker incorporated into clinical practice, where it is used for early detection of metastasis in follow-up of patients having stage II and III disease, and for monitoring response to adjuvant treatment. However there are no reliable prognostic and response predictive biomarkers for CRC. Tumour-associated trypsin inhibitor (TATI) is a form of antitrypsin, a 6kDa enzyme that inhibits trypsin with great affinity and TATI has previously been found to be of prognostic importance in mucinous ovarian cancer and other cancer forms. TATI is expressed in normal colonic mucosa, where its main function is to act as an antitrypsin, protecting the tissue from proteolysis. TATI has also been shown to promote migration and invasion in colorectal cancer cell lines in vitro. Moreover, TATI has a similar molecular structure to epidermal growth factor (EGF) and has been found to bind to EGFR and promote malignant behaviour in various cell line models. The prognostic significance of TATI in CRC patients had however not been reported and the main aim of the present investigation was therefore to examine levels of TATI in tumour tissue and serum samples from three independent CRC patient cohorts, with particular reference to their association with clinical outcome. In addition, we assessed the effect of neoadjuvant radiation therapy (RT) on TATI levels in tissue and serum of rectal cancer patients, and whether expression of epidermal growth factor receptor (EGFR) and markers of downstream signalling have a modifying effect on the prognostic value of TATI. In paper I, we investigated TATI expression by immunohistochemistry (IHC) in CRC tumours (t-TATI) in two cohorts adding up to 424 patients. High TATI expression was found to be significantly associated to short overall survival (OS), and disease free survival (DFS), as well as an increased risk of liver metastasis. In paper II, we analysed the prognostic value of TATI in serum (s-TATI) in one of the cohorts used in Paper I (n = 334). While no significant correlation was found between s-TATI and t-TATI concentrations, the prognostic value of s-TATI was more evident than for t-TATI, the former being a strong independent predictor of an impaired OS, DFS and time to recurrence (TTR), suggesting that elevated serum concentrations are not a result of TATI production by the tumour. Further results from paper II revealed that s-TATI concentrations were lower in rectal cancer patients, the majority of whom had received neoadjuvant radiotherapy (RT). In paper III, we therefore ventured to investigate whether neoadjuvant RT affects TATI concentrations in tissue and serum. To this end, we analysed serum and tissue samples collected at multiple timepoints from 53 patients with rectal cancer included in a in a case-control study. The results revealed that t-TATI and s-TATI levels remained unaffected by RT. S-TATI was significantly associated with shorter OS at all timepoints, and t-TATI assessed in surgical specimens was also a factor of poor prognosis. In paper IV we investigated the associations of s-TATI and t-TATI to EGFR expression and downstream signalling proteins pSTAT3 and pERK1/2, as well as KRAS mutation status. Apart from a significant association between t-TATI expression and KRAS mutation status, there was no intercorrelation between t-TATI or s-TATI and the investigative markers. Expression of pERK1/2 and pSTAT3, and KRAS mutation status, was significantly associated with poor prognosis. The prognostic value of EGFR expression, being non-significant per se, was modified by s-TATI concentrations in that patients with EGFR positive tumours/ high s-TATI levels had a significantly shorter survival compared to patients with EGFR negative tumours/ low s-TATI levels

Provincial coordination and inter-institutional collaboration in British Columbia's college, university college, and institute system

The purpose of this study was to better understand the historical development of the British Columbia (B.C.) community college, university college, and institute system with the focus on the changing nature of voluntary inter-institutional collaboration in relation to provincial coordination. The study also examined the related themes of centralization and decentralization within B.C.'s system and the development of a provincial system of autonomous institutions. The methodology used was qualitative, and more specifically, interpretive in nature and based on the historical method and the underlying assumptions of hermeneutics. The researcher began by analyzing pertinent primary and secondary sources of literature in relation to the study's purpose. The findings from the literature analysis formed the basis for interview questions that were asked of 10 key informants to fill gaps in understanding and confirm findings. The study found that the B.C. system began as a decentralized group of autonomous, community-oriented institutions but became more centrally coordinated by government in the late 1970s and early 1980s, largely because of increased costs and a worsening economy. The 1990s witnessed a high level of centralized decision making with stakeholder involvement, which has been replaced by a move towards decentralization and greater institutional autonomy in the early 2000s based on the market ideology of the new government. Throughout the decades, the B.C. system has had a history of voluntary collaboration but that collaboration has been gradually blended over time with provincial coordination as government built a system of autonomous institutions. The main conclusions of the study are that an appropriate balance may be achievable between centralization and decentralization in order to maintain a coherent system of accountable, autonomous institutions but would need systematic efforts by government and institutions and a policy framework for system governance. Such a balance may be achieved by learning from the lessons of B.C.'s rich history and from the experiences of other jurisdictions. To achieve system goals, the Ministry and institutions could build on the history of voluntary collaborative efforts, which seem particularly important among educators at the program level. The Ministry might reward such collaboration and hold institutions accountable for it

Cyclin D1 expression in colorectal cancer is a favorable prognostic factor in men but not in women in a prospective, population-based cohort study

<p>Abstract</p> <p>Background</p> <p>Although colorectal cancer (CRC) is generally not considered to be a hormone-dependent malignancy, several sex-related differences in incidence, molecular characteristics and survival have been reported. Epidemiological studies have consistently shown that increased exposure to female sex hormones is associated with a lower risk of CRC in women, and cyclin D1, an important downstream effector in estrogen-mediated signaling, is commonly activated in CRC. In this study, we analyzed the prognostic significance of cyclin D1 expression in CRC, with particular reference to sex-related differences, in tumors from a large, prospective, population-based cohort.</p> <p>Methods</p> <p>Using tissue microarrays and immunohistochemistry, the fraction and intensity of cyclin D1 expression was evaluated in 527 incident CRC cases from the Malmö Diet and Cancer Study. The χ²and Spearman's rho (ρ) tests were used for comparison of cyclin D1 expression and relevant clinicopathological characteristics. Kaplan-Meier analysis and Cox proportional hazards modeling were used to assess the effect of cyclin D1 expression on cancer-specific survival (CSS) in univariate and multivariate analysis, adjusted for established prognostic factors.</p> <p>Results</p> <p>Cyclin D1 intensity was significantly lower in male compared with female CRC (<it>P </it>= 0.018). In the full cohort, cyclin D1 expression was associated with a significantly prolonged CSS (hazard ratio (HR) = 0.69; 95% CI 0.49 to 0.96, <it>P </it>= 0.026) but subgroup analysis according to gender revealed a strongly accentuated prognostic effect of cyclin D1 in male CRC (HR = 0.48; 95% CI 0.31 to 0.74, <it>P </it>< 0.001), which was in contrast to female CRC, where cyclin D1 was not prognostic (HR = 1.05; 95% CI 0.62 to 1.78, <it>P </it>= 0.864) (<it>P</it>_interaction= 0.024). The prognostic value of cyclin D1 was not retained in multivariate analysis, either in the full cohort or in male CRC.</p> <p>Conclusions</p> <p>Cyclin D1 expression is strongly associated with prolonged survival in male CRC. These findings not only support an important role for cyclin D1 in colorectal carcinogenesis, but also add further weight to the accumulating evidence that CRC is indeed a hormone-dependent malignancy, for which prognostic and treatment-predictive molecular biomarkers should be evaluated differently in women and men.</p

Spatial Distribution of Macrophages During Callus Formation and Maturation Reveals Close Crosstalk Between Macrophages and Newly Forming Vessels

Macrophages are essential players in the process of fracture healing, acting by remodeling of the extracellular matrix and enabling vascularization. Whilst activated macrophages of M1-like phenotype are present in the initial pro-inflammatory phase of hours to days of fracture healing, an anti-inflammatory M2-like macrophage phenotype is supposed to be crucial for the induction of downstream cascades of healing, especially the initiation of vascularization. In a mouse-osteotomy model, we provide a comprehensive characterization of vessel (CD31+, Emcn+) and macrophage phenotypes (F4/80, CD206, CD80, Mac-2) during the process of fracture healing. To this end, we phenotype the phases of vascular regeneration-the expansion phase (d1-d7 after injury) and the remodeling phase of the endothelial network, until tissue integrity is restored (d14-d21 after injury). Vessels which appear during the bone formation process resemble type H endothelium (CD31hiEmcnhi), and are closely connected to osteoprogenitors (Runx2+, Osx+) and F4/80+ macrophages. M1-like macrophages are present in the initial phase of vascularization until day 3 post osteotomy, but they are rare during later regeneration phases. M2-like macrophages localize mainly extramedullary, and CD206+ macrophages are found to express Mac-2+ during the expansion phase. VEGFA expression is initiated by CD80+ cells, including F4/80+ macrophages, until day 3, while subsequently osteoblasts and chondrocytes are main contributors to VEGFA production at the fracture site. Using Longitudinal Intravital Microendoscopy of the Bone (LIMB) we observe changes in the motility and organization of CX3CR1+ cells, which infiltrate the injury site after an osteotomy. A transient accumulation, resulting in spatial polarization of both, endothelial cells and macrophages, in regions distal to the fracture site, is evident. Immunofluorescence histology followed by histocytometric analysis reveals that F4/80+CX3CR1+ myeloid cells precede vascularization

Validation of podocalyxin-like protein as a biomarker of poor prognosis in colorectal cancer

Background: Podocalyxin-like 1 (PODXL) is a cell-adhesion glycoprotein and stem cell marker that has been associated with an aggressive tumour phenotype and adverse outcome in several cancer types. We recently demonstrated that overexpression of PODXL is an independent factor of poor prognosis in colorectal cancer (CRC). The aim of this study was to validate these results in two additional independent patient cohorts and to examine the correlation between PODXL mRNA and protein levels in a subset of tumours. Method: PODXL protein expression was analyzed by immunohistochemistry in tissue microarrays with tumour samples from a consecutive, retrospective cohort of 270 CRC patients (cohort 1) and a prospective cohort of 337 CRC patients (cohort 2). The expression of PODXL mRNA was measured by real-time quantitative PCR in a subgroup of 62 patients from cohort 2. Spearman's Rho and Chi-Square tests were used for analysis of correlations between PODXL expression and clinicopathological parameters. Kaplan Meier analysis and Cox proportional hazards modelling were applied to assess the relationship between PODXL expression and time to recurrence (TTR), disease free survival (DFS) and overall survival (OS). Results: High PODXL protein expression was significantly associated with unfavourable clinicopathological characteristics in both cohorts. In cohort 1, high PODXL expression was associated with a significantly shorter 5-year OS in both univariable (HR = 2.28; 95% CI 1.43-3.63, p = 0.001) and multivariable analysis (HR = 2.07; 95% CI 1.25-3.43, p = 0.005). In cohort 2, high PODXL expression was associated with a shorter TTR (HR = 2.93; 95% CI 1.26-6.82, p = 0.013) and DFS (HR = 2.44; 95% CI 1.32-4.54, p = 0.005), remaining significant in multivariable analysis, HR = 2.50; 95% CI 1.05-5.96, p = 0.038 for TTR and HR = 2.11; 95% CI 1.13-3.94, p = 0.019 for DFS. No significant correlation could be found between mRNA levels and protein expression of PODXL and there was no association between mRNA levels and clinicopathological parameters or survival. Conclusions: Here, we have validated the previously demonstrated association between immunohistochemical expression of PODXL and poor prognosis in CRC in two additional independent patient cohorts. The results further underline the potential utility of PODXL as a biomarker for more precise prognostication and treatment stratification of CRC patients

Effects of radiation therapy on tissue and serum concentrations of tumour associated trypsin inhibitor and their prognostic significance in rectal cancer patients

<p>Abstract</p> <p>Background</p> <p>We have previously demonstrated that elevated concentrations of tumour-associated trypsin inhibitor (TATI) in both tumour tissue (t-TATI) and in serum (s-TATI) are associated with a poor prognosis in colorectal cancer patients. It was also found that s-TATI concentrations were lower in patients with rectal cancer compared to patients with colon cancer. In this study, we investigated the effects of neoadjuvant radiotherapy (RT) on concentrations of t-TATI and s-TATI in patients with rectal cancer.</p> <p>Methods</p> <p>TATI was analysed in serum, normal mucosa and tumour tissue collected at various time points in 53 rectal cancer patients enrolled in a case-control study where 12 patients received surgery alone, 20 patients 5 × 5 Gy (short-term) preoperative RT and 21 patients 25 × 2 Gy (long-term) preoperative RT. T-TATI was analysed by immunohistochemistry and s-TATI was determined by an immunofluorometric assay. Mann-Whitney U test and Wilcoxon Z (Z) test were used to assess t-TATI and s-TATI concentrations in relation to RT. Spearman's correlation (R) test was used to explore the associations between t-TATI, s-TATI and clinicopathological parameters. Overall survival (OS) according to high and low t-TATI and s-TATI concentrations was estimated by classification and regression tree analysis, Kaplan-Meier analysis and the log rank test.</p> <p>Results</p> <p>RT did not affect concentrations of t-TATI or s-TATI. In patients receiving short-term but not long-term RT, s-TATI concentrations were significantly higher 4 weeks post surgery than in serum drawn prior to surgery (Z = -3.366, P < 0.001). T-TATI expression correlated with male gender (R = 0.406, P = 0.008). High t-TATI expression in surgical specimens was associated with a significantly shorter OS (P = 0.045). S-TATI concentrations in serum drawn at all time points were associated with an impaired OS (P = 0.035 before RT, P = 0.001 prior to surgery, P = 0.043 post surgery). At all time points, s-TATI correlated with higher age (P < 0.001-0.021) and with increased s-creatinine concentrations assessed prior to surgery (P = 0.041).</p> <p>Conclusions</p> <p>The results presented here further validate the utility of t-TATI and s-TATI as prognostic biomarkers in patients with rectal cancer, independent of neoadjuvant RT.</p

Increased serum levels of tumour-associated trypsin inhibitor independently predict a poor prognosis in colorectal cancer patients

Peer reviewe

High RBM3 expression in prostate cancer independently predicts a reduced risk of biochemical recurrence and disease progression

<p>Abstract</p> <p>Background</p> <p>High expression of the RNA-binding protein RBM3 has previously been found to be associated with good prognosis in breast cancer, ovarian cancer, malignant melanoma and colorectal cancer. The aim of this study was to examine the prognostic impact of immunohistochemical RBM3 expression in prostate cancer.</p> <p>Findings</p> <p>Immunohistochemical RBM3 expression was examined in a tissue microarray with malignant and benign prostatic specimens from 88 patients treated with radical prostatectomy for localized disease. While rarely expressed in benign prostate gland epithelium, RBM3 was found to be up-regulated in prostate intraepithelial neoplasia and present in various fractions and intensities in invasive prostate cancer. High nuclear RBM3 expression was significantly associated with a prolonged time to biochemical recurrence (BCR) (HR 0.56, 95% CI: 0.34-0.93, <it>p </it>= 0.024) and clinical progression (HR 0.09, 95% CI: 0.01-0.71, <it>p = </it>0.021). These associations remained significant in multivariate analysis, adjusted for preoperative PSA level in blood, pathological Gleason score and presence or absence of extracapsular extension, seminal vesicle invasion and positive surgical margin (HR 0.41, 95% CI: 0.19-0.89, <it>p </it>= 0.024 for BCR and HR 0.06, 95% CI: 0.01-0.50, <it>p = </it>0.009 for clinical progression).</p> <p>Conclusion</p> <p>Our results demonstrate that high nuclear expression of RBM3 in prostate cancer is associated with a prolonged time to disease progression and, thus, a potential biomarker of favourable prognosis. The value of RBM3 for prognostication, treatment stratification and follow-up of prostate cancer patients should be further validated in larger studies.</p

Reduced expression of the polymeric immunoglobulin receptor in pancreatic and periampullary adenocarcinoma signifies tumour progression and poor prognosis

The polymeric immunoglobulin receptor (pIgR) is a key component of the mucosal immune system that mediates epithelial transcytosis of immunoglobulins. High pIgR expression has been reported to correlate with a less aggressive tumour phenotype and an improved prognosis in several human cancer types. Here, we examined the expression and prognostic significance of pIgR in pancreatic and periampullary adenocarcinoma. The study cohort encompasses a consecutive series of 175 patients surgically treated with pancreaticoduodenectomy for pancreatic and periampullary adenocarcinoma in Malmö and Lund University Hospitals, Sweden, between 2001-2011. Tissue microarrays were constructed from primary tumours (n = 175) and paired lymph node metastases (n = 105). A multiplied score was calculated from the fraction and intensity of pIgR staining. Classification and regression tree analysis was used to select the prognostic cut-off. Unadjusted and adjusted hazard ratios (HR) for death and recurrence within 5 years were calculated. pIgR expression could be evaluated in 172/175 (98.3%) primary tumours and in 96/105 (91.4%) lymph node metastases. pIgR expression was significantly down-regulated in lymph node metastases as compared with primary tumours (p = 0.018). Low pIgR expression was significantly associated with poor differentiation grade (p < 0.001), perineural growth (p = 0.027), lymphatic invasion (p = 0.016), vascular invasion (p = 0.033) and infiltration of the peripancreatic fat (p = 0.039). In the entire cohort, low pIgR expression was significantly associated with an impaired 5-year survival (HR = 2.99, 95% confidence interval (CI) 1.71-5.25) and early recurrence (HR = 2.89, 95% CI 1.67-4.98). This association remained significant for survival after adjustment for conventional clinicopathological factors, tumour origin and adjuvant treatment (HR = 1.98, 95% CI 1.10-3.57). These results demonstrate, for the first time, that high tumour-specific pIgR expression signifies a more favourable tumour phenotype and that low expression independently predicts a shorter survival in patients with pancreatic and periampullary cancer. The mechanistic basis for the putative tumour suppressing properties of pIgR in these cancers merits further study