MAPK inhibitors induce serine peptidase inhibitor Kazal type 1 (SPINK1) secretion in BRAF V600E-mutant colorectal adenocarcinoma

The mitogen-activated protein kinase (MAPK) pathway plays a central role in colorectal cancers (CRC). In particular, BRAF V600E-mutant tumors, which represent around 10% of CRCs, are refractory to current therapies. Overexpression and secretion of serine peptidase inhibitor Kazal type 1 (SPINK1) are observed in around 50% of CRCs, and its serum level can be used as a biomarker for poor prognosis. Utilizing a recently developed extendable blocking probe assay, we analyzed the BRAF mutation status in a CRC patient cohort (N=571) using tissue-derived RNA as the starting material. From the same RNA samples, we measured the relative SPINK1 expression levels using a quantitative real-time PCR method. Expression of mutant BRAF V600E correlated with poor prognosis, as did low expression of SPINK1 mRNA. Further, BRAF V600E correlated negatively with SPINK1 levels. In order to investigate the effect of MAPK pathway-targeted therapies on SPINK1 secretion, we conducted invitro studies using both wild-type and V600E CRC cell lines. BRAF inhibitor vemurafenib, and subsequent MAPK pathway inhibitors trametinib and SCH772984, significantly increased SPINK1 secretion in V600E CRC cell lines Colo205 and HT-29 with a concomitant decrease in trypsin-1 and -2 secretion. Notably, no SPINK1 increase or trypsin-1 decrease was observed in BRAF wild-type CRC cell line Caco-2 in response to MAPK pathway inhibitors. In further mechanistic studies, we observed that only trametinib was able to diminish completely both MEK and ERK phosphorylation in the V600E CRC cells. Furthermore, the key regulator of integrated stress response, activating transcription factor 4 (ATF-4), was downregulated both at mRNA and at protein level in response to trametinib treatment. In conclusion, these data suggest that sustained inhibition of not only MAPK pathway activation, but also ATF-4 and trypsin, might be beneficial in the therapy of BRAF V600E-mutant CRC and that SPINK1 levels may serve as an indicator of therapy response.Peer reviewe

Extendable blocking probe in reverse transcription for analysis of RNA variants with superior selectivity

Peer reviewe

Classification of Inhibitors of Hepatic Organic Anion Transporting Polypeptides (OATPs): Influence of Protein Expression on Drug–Drug Interactions

ABSTRACT: The hepatic organic anion transporting poly-peptides (OATPs) influence the pharmacokinetics of several drug classes and are involved in many clinical drug−drug interactions. Predicting potential interactions with OATPs is, therefore, of value. Here, we developed in vitro and in silico models for identification and prediction of specific and general inhibitors of OATP1B1, OATP1B3, and OATP2B1. The maximal transport activity (MTA) of each OATP in human liver was predicted from transport kinetics and protein quantification. We then used MTA to predict the effects of a subset of inhibitors on atorvastatin uptake in vivo. Using a data set of 225 drug-like compounds, 91 OATP inhibitors were identified. In silico models indicated that lipophilicity and polar surface area are key molecular features of OATP inhibition. MTA predictions identified OATP1B1 and OATP1B3 as major determinants of atorvastatin uptake in vivo. The relative contributions to overall hepatic uptake varied with isoform specificities of the inhibitors

Contrast medium-induced nephropathy. Aspects on incidence, consequences, risk factors and prevention

Contrast media-induced nephropathy (CIN) is a well-known complication of radiological examinations employing iodine contrast media (I-CM). The rapid development and frequent use of coronary interventions and multi-channel detector computed tomography with concomitant administration of relatively large doses of I-CM has contributed to an increasing number of CIN cases during the last few years. Reduced renal function, especially when caused by diabetic nephropathy or renal arteriosclerosis, in combination with dehydration, congestive heart failure, hypotension, and administration of nephrotoxic drugs are risk factors for the development of CIN. When CM-based examinations cannot be replaced by other techniques in patients at risk of CIN, focus should be directed towards analysis of number and type of risk factors, adequate estimation of GFR, institution of proper preventive measures including hydration and post-procedural observation combined with surveillance of serum creatinine for 1-3 days. For the radiologist, there are several steps to consider in order to minimise the risk for CIN: use of “low-“ or “iso-osmolar” I-CM and dosing the I-CM in relation to GFR and body weight being the most important as well as utilizing radiographic techniques to keep the I-CM dose in gram iodine as low as possible below the numerical value of estimated GFR. There is as yet no pharmacological prevention that has been proven to be effective

Combining CRP and CA19-9 in a novel prognostic score in pancreatic ductal adenocarcinoma

Inflammation promotes tumor progression, induces invasion and metastatic spread. This retrospective study explored CRP, CA19-9, and routine laboratory values as preoperative prognostic factors in pancreatic cancer patients. Between 2000 and 2016, there were 212 surgically treated pancreatic cancer patients at Helsinki University Hospital, Finland. Out of these, 76 borderline resectable patients were treated with neoadjuvant therapy (NAT); 136 upfront resected patients were matched for age and sex at a 1:2 ratio. We analyzed preoperative CRP, CA19-9, CEA, leukocytes, albumin, bilirubin and platelets. CRP and CA19-9 were combined into a prognostic score: both CRP and CA19-9 below the cut-off values (3 mg/l and 37 kU/l, respectively), either CRP or CA19-9 above the cut-off value, and finally, both CRP and CA19-9 above the cut-off values. Among all patients, median disease-specific survival times were 54, 27 and 16 months, respectively (pPeer reviewe

Back pain and health status in patients with clinically diagnosed ankylosing spondylitis, psoriatic arthritis and other spondyloarthritis : a cross-sectional population-based study

BACKGROUND: In the broader spectrum of back pain, inflammatory back pain (IBP) is a symptom that may indicate axial spondyloarthritis (SpA). The objectives of this study were to determine the frequency of current IBP, as a hallmark sign of possible axial SpA, in patients with ankylosing spondylitis (AS), psoriatic arthritis (PsA) and other SpA and to compare self-reported health between the groups with current IBP. METHODS: Five-thousand seven hundred seventy one patients identified in the regional healthcare register of the most southern county of Sweden, diagnosed at least once by a physician (based on ICD-codes) with any type of SpA in 2003-2007, were sent a postal survey in 2009. Patients with current IBP were identified, based on self-reported back pain ≥3 months in the preceding year and fulfilling the Berlin criteria for IBP. The frequencies of IBP in AS, PsA and other SpA (including the remaining subgroups of SpA) were determined, and the groups were compared with regard to patient reported outcome measures (PROMs). RESULTS: The frequency and proportion of patients with current IBP in AS, PsA and other SpA were 319 (43 %), 409 (31 %) and 282 (39 %) respectively, within the responders to the survey (N = 2785). The proportion was statistically higher in AS, compared to PsA (p &lt; 0.001), but not for AS compared to other SpA (p = 0.112). PsA and other SpA, with current IBP, had similar (BASFI, EQ-5D, patients global assessment, fatigue, spinal pain) or worse (BASDAI) PROMs, compared to AS with current IBP. PsA with current IBP received pharmacological, anti-rheumatic, treatment more frequently than AS with current IBP, while AS and other SpA received treatment to a similar degree. CONCLUSION: The proportion of patients with current IBP was substantial in all three groups and health reports in the non-AS groups were similar or worse compared to the AS group supporting the severity of IBP in these non-AS SpA groups. These findings may indicate a room for improvement concerning detection of axial disease within different subtypes of non-AS SpA, and possibly also for treatment.The work was supported by funds from Lund University, Gothenburg University, the Swedish Rheumatism Association, Region Skåne, Region Halland, and Göteborgsregionens stiftelse för reumatologisk forskning (the Gothenburg region’s fund for rheumatology research).</p