J. Wesseling

Upregulation of Claudin-4, CAIX and GLUT-1 in distant breast cancer metastases

BackgroundSeveral studies have shown that the immunophenotype of distant breast cancer metastases may differ significantly from that of the primary tumor, especially with regard to differences in the level of hormone receptor protein expression, a process known as receptor conversion. This study aimed to compare expression levels of several membrane proteins between primary breast tumors and their corresponding distant metastases in view of their potential applicability for molecular imaging and drug targeting.MethodsExpression of Claudin-4, EGFR, CAIX, GLUT-1 and IGF1R was assessed by immunohistochemistry on tissue microarrays composed of 97 paired primary breast tumors and their distant (non-bone) metastases.ResultsIn both the primary cancers and the metastases, Claudin-4 was most frequently expressed, followed by GLUT-1, CAIX and EGFR.From primary breast cancers to their distant metastases there was positive to negative conversion, e.g. protein expression in the primary tumor with no expression in its paired metastasis, in 6%, 19%, 12%, 38%, and 0% for Claudin-4 (n.s), GLUT-1 (n.s), CAIX (n.s), EGFR (n.s) and IGF1R (n.s) respectively. Negative to positive conversion was seen in 65%, 47%, 43%, 9% and 0% of cases for Claudin-4 (pu2009=u20090.049), GLUT-1 (pu2009=u20090.024), CAIX (pu2009=u20090.002), EGFR (n.s.) and IGF1R (n.s.) respectively. Negative to positive conversion of Claudin-4 in the metastasis was significantly associated with tumor size (pu2009=u20090.015), negative to positive conversion of EGFR with negative PR status (pu2009=u20090.046) and high MAI (pu2009=u20090.047) and GLUT-1 negative to positive conversion with (neo)adjuvant chemotherapy (pu2009=u20090.039) and time to metastasis formation (pu2009=u20090.034). CAIX and GLUT-1 expression in the primary tumor were significantly associated with high MAI (pu2009=u20090.008 and pu2009=u20090.038 respectively).ConclusionClaudin-4 is frequently expressed in primary breast cancers but especially in their metastases and is thereby an attractive membrane bound molecular imaging and drug target. Conversion in expression of the studied proteins from the primary tumor to metastases was fairly frequent, except for IGF1R, implying that the expression status of metastases cannot always be reliably predicted from the primary tumor, thereby necessitating biopsy for reliable assessment.

Prevalence and predictors of health care use in patients with early hip or knee osteoarthritis: two-year follow-up data from the CHECK cohort

OBJECTIVEnTo describe health care utilization (HCU) and predict analgesic use and health professional (HP) contact at baseline and 2 years in individuals with early symptomatic hip and/or knee osteoarthritis (OA).nnnDESIGNnBaseline and two-year data on HCU of the 1002 participants from the multi-centre Cohort Hip & Cohort Knee study were used. Six forms of health care services were described: analgesic use, supplement use, contact with a General Practitioner (GP), contact with a HP, contact in secondary care, and alternative medicine use. Multivariable logistic regression was performed in order to identify predisposing, enabling and disease-related variables that predict analgesic use and HP contact at 2 years; treatment modalities of first choice in early OA.nnnRESULTSnFor the hip (n=170), the knee (n=414) and the hip and knee (n=418) group analgesic use (38%, 29% and 47%, respectively), contact with a GP (32%, 38% and 36%, respectively) and contact with a HP (26%, 18% and 20%, respectively), were reported most often at baseline. Contact with a GP significantly decreased, supplement use increased (to about one third), and other treatment modalities remained stable at 2 years. In all three groups, analgesic use at baseline was the strongest predictor for analgesic use at 2 years, whereas contact with a HP at baseline was the strongest predictor of contact with a HP after 2 years. Belonging to a first generation minority was a predisposing risk factor [Odds Ratio (95%-CI), 8.72 (1.55-48.97)] for analgesic use in the hip and knee group.nnnCONCLUSIONSnIn early OA, familiarity with HCU and other predisposing factors are, apart from disease-related factors strongly associated with HCU at 2 years. Further research is necessary to examine whether our findings reflect sub-optimal management of early OA in terms of efficacy and equity.

Defining hip pain trajectories in early symptomatic hip osteoarthritis – 5 year results from a nationwide prospective cohort study (CHECK)

OBJECTIVEnTo define distinct hip pain trajectories in individuals with early symptomatic hip osteoarthritis (OA) and to determine risk factors for these pain trajectories.nnnMETHODnData were obtained from the nationwide prospective Cohort Hip and Cohort Knee (CHECK) study. Participants with hip pain or stiffness and a completed 5-year follow-up were included. Baseline demographic, anamnestic, physical examination characteristics were assessed. Outcome was annually assessed by the Numeric Rating Scale (NRS) for pain. Pain trajectories were retrieved by latent class growth analysis (LCGA). Multinomial logistic regression was used to calculate risk ratios.nnnRESULTSn545 participants were included. Four distinct pain trajectories were uncovered by LCGA. We found significant differences in baseline characteristics, including body mass index (BMI); symptom severity; pain coping strategies and in criteria for clinical hip OA (American College of Rheumatology (ACR)). Lower education, higher activity limitation scores, frequent use of pain transformation as coping strategy and painful internal hip rotation were more often associated with trajectories characterized by more severe pain. No association was found for baseline radiographic features.nnnCONCLUSIONnWe defined four distinct pain trajectories over 5 years follow-up in individuals with early symptomatic hip OA, suggesting there are differences in symptomatic progression of hip OA. Baseline radiographic severity was not associated with the pain trajectories. Future research should be aimed at measuring symptomatic progression of hip OA with even more frequent symptom assessment.

OP0031 Comparison between check study (cohort hip& cohort knee) and osteoartritis initiative; development of pain and function in 3 years of follow-up

Background Pain and disability related to osteoarthritis (OA) may generally be considered to be chronic, but it is known that its course can be very different between patients. Objectives In this study, it is investigated whether there is a difference in development of pain and physical functioning during follow-up between two OA cohorts: Osteoarthritis Initiative (OAI) and Cohort Hip& Cohort knee study (CHECK). Methods For the current study, longitudinal data of three years follow-up of the CHECK study and OA Initiative were used. The CHECK study is a Dutch prospective 10-year follow-up study, initiated by the Dutch Arthritis Association, to study progression of OA in participants with early symptomatic OA of knee or hip. Individuals were eligible if they had pain of knee or hip, were aged 45-65 years, and had not yet consulted their physician for these symptoms. In the same time in the U.S. an observational 4-year follow-up study was started to create a public archive of data, biological samples and joint images to study the natural history of, and risk factors for, the onset and progression of knee OA. The WOMAC was utilized to measure pain during activities (range 0-20) and physical functioning (range 0-68). For comparison with CHECK a subgroup of the incidence OAI cohort was selected which was comparable with the CHECK cohort: participants with age 45-65 years, frequent or infrequent knee symptoms and no surgery in either knee. Generalized estimating equations were used to account for correlations within individuals (GEE) and the interaction term (OAcohort* time) was also investigated within the model. Results For this study, the data of 688 CHECK participants with knee pain at baseline were analyzed, mean age 56 years, BMI 25 kg/m2 and 79% were female. The subgroup of OAI Incidence cohort with infrequent or frequent knee pain consisted of 1417 participants, with a mean age of 56, BMI of 28 and 64% female. The CHECK cohort reported at baseline more pain and more problems in physical functioning compared to the subgroup of OAI Incidence cohort (p<0.001). A final longitudinal regression model with pain as outcome and adjusted for BMI shows that pain is 3 points lower (better health) of people in the subgroup of OAI Incidence, and improves in both cohorts after 1 year of follow-up with 0.2 points, after 2 years with 0.3 points, after 3 years with 0.3 points. In a final model with function as outcome and adjusted for BMI, this outcome is 10.2 points higher (worse health) in CHECK cohort and improves in both cohorts after 1 year of follow-up with 0.6 points, after 2 years with 0.8, after 3 years with 0.7 points. These results were not influenced by the characteristics of both cohorts (interaction term (OAcohort* time) was not statistically significant in both outcomes). Conclusions In the total group, participants of the OAI Incidence subgroup and the CHECK participants with knee pain, there is a gradually improvement over time in pain and physical functioning, despite baseline values on pain and physical functioning. In the early phase of OA there is overall a slight improvement in the complaints. Though, a difference at start between the two cohorts, completely parallel development of pain and function over 3 years. Disclosure of Interest None Declared

THU0471 Identifying Trajectories in Knee Osteophyte Formation in Early Osteoarthritis; Results of 5 Year Follow-Up in Check (Cohort HIP and Cohort Knee)

Background Osteoarthritis (OA) can be manifested in structural changes occurring at the joint level, like joint space narrowing (JSN) and osteophyte formation. Preliminary findings of the Osteoarthritis Initiative after 10 years suggest possibly different mechanisms in structural changes for JSN and osteophyte formation.1 It is proposed that formation of osteophytes should be considered as an early feature of OA.2 Objectives This study focuses on the heterogenic expression of osteophyte formation and aims to identify subgroups of participants with homogeneous patterns of osteophyte formation over time (trajectories), and to describe the prognostic factors of the different subgroups. Methods Longitudinal data of quantitative osteophyte formation, calculated as total area of mm2, over a period of five years follow-up of the early OA cohort CHECK (Cohort Hip and Cohort Knee) was used. Latent Class Growth Analysis identified homogenous subgroups with distinct trajectories of osteophyte formation. Multinomial regression analysis was used to examine the prognostic factors of the different subgroups. Results In data of 5 years follow-up of 929 participants, three subgroups were identified: an “osteophyte increase” subgroup (n=32) with an increase in osteophyte formation, “mild osteophyte” subgroup (n=181) with minimal increase, and “no osteophyte” subgroup (n=716) without increase in osteophyte formation. Being older, a BMI above 25, the existence of local clinical signs at baseline (crepitus, warmth, bony tenderness, enlargement and effusion), and radiographic characteristics (initial osteophyte formation, narrowing of JSW) suggested a higher probability of a (mild) increase in osteophyte formation. Conclusions In participants with early symptomatic OA the common course of osteophyte formation over a period of 5 years is relatively flat. However, in a small subpopulation there was a remarkable increase in osteophyte formation. Identifying which patients will develop structural changes may in the future lead to specific treatment of these patients. References Eckstein F, Kwoh CK, Link TM. Imaging research results from the Osteoarthritis Initiative (OAI): a review and lessons learned 10 years after start of enrolment. Ann Rheum Dis 2014;73:1289-300 Hart DJ, Spector TD. Kellgren & Lawrence grade 1 osteophytes in the knee – doubtful or definite? Osteoarthritis Cartilage 2003;11:149-50. Acknowledgements CHECK-cohort study is initiated by the Dutch Arthritis Foundation and performed within; Erasmus Medical Center Rotterdam; Kennemer Gasthuis Haarlem; Leiden University Medical Center; Maastricht University Medical Center; Martini Hospital Groningen /Allied Health Care Center for Rheum. and Rehabilitation Groningen; Medical Spectrum Twente Enschede /Ziekenhuisgroep Twente Almelo; Reade, formerly Jan van Breemen Institute/ VU Medical Center Amsterdam; St.Maartens-kliniek Nijmegen; University Medical Center Utrecht and Wilhelmina Hospital Assen. Disclosure of Interest None declared

AB0779 7 Years of Follow-Up of Early Symptomatic Oa: Characteristics of CHECK (Cohort HIP & Cohort Knee)

Background In the Netherlands, a prospective 10-year follow-up study was initiated and funded by the Dutch Arthritis Foundation (DAF) in order to establish the course of early symptomatic osteoarthritis (OA) in participants with early complaints of hip and/or knee: CHECK (Cohort Hip & Cohort Knee). To understand more about the disease and its course, we initiated a large independent detailed observational study starting (very) early in the stage of the disease and where subjects were characterized in 3 main domains through-out follow-up: clinical, radiographic and biochemical biomarkers. Objectives We present the results of 7 years of follow up. Methods Between October 2002 and September 2005 a cohort was formed of 1002 participants aged 45-65 years with symptomatic OA characterized by pain and/or stiffness of knee and/or hip. They were included at the first visit or within 6 months of the first visit to the general practitioner for these symptoms. Study centers comprise ten general and university hospitals located in urbanized and semi-urbanized regions in the Netherlands. A special program was developed to minimize loss to follow up, comprising newsletters, website and organizing symposia to present the results and progress of CHECK. An overview of measures that are included in the study can be found at our website www.check-research.com. Results After 7 years a total of 105 (10%) participants had dropped out. Reasons included loss of interest (21%), comorbidity that complicated participating of CHECK too much (18%), death (12%), contact lost (11%), costs incurred for the study (5%), and other reasons (33%). Dropouts did not differ significantly from other participants in baseline characteristics. Over 7 years follow-up clinical characteristics including pain, stiffness and function of disease-specific (WOMAC) and the subscales of generic self-report questionnaire (SF36) on average did not change significantly, although at least half of the patients experienced radiographic progression. Scoring of 5-year follow up is complete: at this point, 50% of the participants progressed by at least one Kellgren and Lawrence grade in the knee, and 25% in the hip. Preliminary analyses suggest the existence of several clinical and radiological phenotypes. Conclusions CHECK is a valuable source of information of early symptomatic OA with a low loss to follow-up and a multidisciplinary approach that allows the collections of high-quality data on clinical, radiological and biochemical variables. Acknowledgements CHECK-cohort study is initiated by the Dutch Arthritis Foundation and performed within; Erasmus Medical Center Rotterdam; Kennemer Gasthuis Haarlem; Leiden University Medical Center; Maastricht University Medical Center; Martini Hospital Groningen/Allied Health Care Center for Rheum. and Rehabilitation Groningen; Medical Spectrum Twente Enschede/Ziekenhuisgroep Twente Almelo; Reade, formerly Jan van Breemen Institute/VU Medical Center Amsterdam; St.Maartens-kliniek Nijmegen; University Medical Center Utrecht and Wilhelmina Hospital Assen. Disclosure of Interest None declared DOI 10.1136/annrheumdis-2014-eular.2854

OP0292 Different Pain Trajectories in Early Symptomatic Knee OA; 5 Year Follow-Up of Check (Cohort Hip & Cohort Knee) Study

Objectives Knee pain is often the first sign of knee OA and it is known that its course can be very different between patients over time. This study identifies distinct groups of patients with different trajectories of pain due to symptomatic knee OA, and describes lifestyle and coping characteristics for each trajectory of pain. Lifestyle factors might be important elements for prevention, since they are modifiable in nature. Yet de role of lifestyle factors in different pain trajectories in early OA is unclear. Methods Longitudinal data of five years follow-up of the CHECK study was used. Participants had pain of knee or hip, were aged 45-65 years, and had not yet consulted their physician for these symptoms or the consultation occurred within 6 months before inclusion. Pain severity was measured with numeric scale (0-10). Latent class growth analysis identified homogenous subgroups with distinct trajectories of pain. Multinomial regression analysis was used to examine different lifestyle and coping characteristics between the trajectories Results Longitudinal data of 5 years follow-up of 705 participants with symptomatic knee OA was analyzed. Three pain trajectories were identified based on their outcome: good, moderate and poor outcome. Participants with good outcome trajectory (n=222) had over time a slight decrease in pain severity and ended up with a low pain severity. Participants with moderate outcome trajectory (n=294) had a stable course of moderate pain over time. The poor outcome trajectory participants (n=189) had an increase of pain severity over time and ended up with severe pain. Compared to the good outcome group, participants in the moderate and poor outcome group were characterized by higher BMI (both OR’s 1.1; p=0.01), smokers (moderate outcome group OR=1.8, p=0.08; poor outcome group OR= 2.5, p=0.01) used passive coping strategies worrying (moderate outcome group OR= 2.2, p=0.01; poor outcome group OR=3.5, p<0.001) and resting (moderate outcome group OR= 1.6, p=0.09; poor outcome group OR=2.4, p=0.004). The passive coping strategy ‘retreating’ reduced the chance of belonging to the poor outcome group (moderate outcome group OR= 0.6, p=0.04; poor outcome group OR=0.5, p=0.009). Conclusions This study identified 3 trajectories of pain: good, moderate and poor outcome. Unhealthy lifestyle characteristics (high BMI and smoking) and passive coping strategies (worrying and resting) characterized the poor outcome group. Distinguishing different trajectories could have implications for the treatment. Treatment for each pain trajectory in early OA might be tailored to lifestyle and coping characteristics. Acknowledgements CHECK-cohort study is initiated by the Dutch Arthritis Association and performed within; Erasmus Medical Center Rotterdam; Kennemer Gasthuis Haarlem; Leiden University Medical Center; Maastricht University Medical Center; Martini Hospital Groningen /Allied Health Care Center for Rheum. and Rehabilitation Groningen; Medical Spectrum Twente Enschede /Ziekenhuisgroep Twente Almelo; Reade, formerly Jan van Breemen Institute/ VU Medical Center Amsterdam; St.Maartens-kliniek Nijmegen; University Medical Center Utrecht and Wilhelmina Hospital Assen Disclosure of Interest None Declared

OP0188-PC Pain trajectories in hip osteoarthritis: Results of the dutch cohort hip and cohort knee (CHECK)

Background Pain is the most important complaint reported in osteoarthritis (OA). In hip OA, a recent study showed five distinctive trajectories of pain over a two year follow-up period.[1] The next step is to validate these finding in another study population with a longer follow-up period. Objectives To identify long term trajectories of persons with hip pain due to OA over a 5-year follow-up period. Methods Data from the inception cohort CHECK[2] was used to investigate the course of pain. For this study 208 participants were included who fulfilled the clinical and/or radiographic American College of Rheumatology (ACR) criteria for hip OA. Participants filled out yearly questionnaires during the 5-year follow-up period. Pain severity was measured by an 11-point numeric rating scale (0=no pain;10=unbearable). Latent class growth analysis (LCGA) was used to determine the number of different trajectories of participants with hip pain due to OA. Multinomial regression analysis was used to predict the effect of baseline characteristics of the participants and the probability of membership to a one of the trajectories. Results Analyses yielded 4 distinctive trajectories of pain during the five year follow-up instead of five trajectories in another study sample over two years of follow-up. Participants within trajectory 1 (“mild pain”; n=78) fluctuated slightly within ranges of mild pain. Trajectory 2 (“moderate pain”; n=76) consists of participants who slowly increased within ranges of moderate pain over the 5-year period. Trajectory 3 (“decreasing pain”; n=22) comprised participants who started with severe pain at baseline, but after 5-years their pain decreased. Participants within trajectory 4 (“always pain”; n=32) had constant levels of severe pain. Compared with the “mild pain” group, participants in the “moderate pain” group have a higher body mass index (BMI), more often have morning stiffness of the hip, a decreased hip flexion, and concurrent knee complaints. Participants in the “always pain” group are more likely to be women. Conclusions LCGA identified 4 distinct trajectories in 208 participants with clinically and/or radiographic determined hip OA. Predictors for membership to the “moderate pain” group were BMI, morning stiffness of the hip, decreased hip flexion, and concurrent knee pain. For the “always pain” group this was gender. References Verkleij SPJ, Hoekstra T, Rozendaal RM, Waarsing JH, Koes BW, Luijsterburg PAJ, Bierma-Zeinstra SMA. Defining discriminative pain trajectories in hip osteoarthritis over a two-year time period. Ann Rheum Dis 2012. Wesseling J et al. CHECK (Cohort Hip and Cohort Knee): similarities and differences with the Osteoarthritis Initiative. Ann Rheum Dis 2009;68:1413-9. Disclosure of Interest None Declared

Abstract PD01-03: Phosphorylated p-70S6K predicts tamoxifen resistance in postmenopausal breast cancer patients randomized between adjuvant tamoxifen versus no systemic treatment

BACKGROUND: Activation of the PI3K/MAPK pathways results in anti-estrogen resistance in vitro , however a biomarker that can predict clinical resistance has not yet been identified. Common drivers of these pathways are PIK3CA mutations, loss of PTEN and over-expression of HER2 and IGF-1R. We aimed to test the prognostic and predictive value of PI3K/MAPK pathway drivers as well as downstream activated proteins in postmenopausal breast cancer patients. METHODS: We collected primary tumor tissue from 563 ERα positive breast cancer patients who were randomized between tamoxifen (1–3 years) versus no adjuvant systemic therapy. PIK3CA hotspot mutations were assessed by Sequenom Mass Spectrometry. Immunohistochemistry was performed for expression of PTEN, IGF-1R and the downstream markers p-AKT(Thr308), p-AKT(Ser473), p-mTOR, p-p706SK and p-ERK1/2. Cox proportional hazard models for recurrence free interval were used to assess hazard ratios and interaction between these markers and treatment. RESULTS: No significant interaction between any of the tested PI3K/MAPK pathways drivers and tamoxifen was found. [/bold]However, interactions were identified between tamoxifen and the downstream activated proteins (p-AKT(Thr308), p- mTOR, p-p70S6K and p-ERK1/2). After correcting for multiple testing, p-p70S6K remained significantly associated with tamoxifen resistance. Patients whose tumor did not express p-p70S6K did derive significant benefit from tamoxifen (HR 0.24, 95 % CI= 0.12–0.47, p CONCLUSION: We conclude that the downstream marker of PI3K/MAPK activation p-p70S6K predicts tamoxifen resistance in ERα positive postmenopausal breast cancer patients. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr PD01-03.