Geometric Mean Change Research Articles

Effect of spironolactone wash-out on albuminuria after long-term treatment in individuals with type 2 diabetes and high risk of kidney disease-An observational follow-up of the PRIORITY study.

This study aimed to explore the effect of discontinuation of long-term spironolactone treatment on markers of kidney function in individuals with type 2 diabetes (T2D) at high risk of kidney disease enrolled in the Proteomic prediction and Renin angiotensin aldosterone system Inhibition prevention Of early diabetic nephRopathy In TYpe 2 diabetic patients with normoalbuminuria (PRIORITY) study. An observational study following the nested randomised part of the PRIORITY study was conducted. A total of 115 individuals with T2D and normoalbuminuria but high risk for progression based on urinary proteomics, randomised to daily spironolactone (n = 50) or placebo (n = 65) for a median of 2.5 years, were re-examined approximately 6 weeks after the final visit in the PRIORITY study. Primary endpoint was relative change in geometric mean of urinary albumin-creatinine ratio (UACR) between the final visit in PRIORITY (baseline) and follow-up. Secondary endpoints were change in estimated glomerular filtration rate (eGFR), systolic blood pressure (SBP) and serum potassium. No change in UACR was observed in neither the spironolactone (geometric mean change: 17%; 95% CI -12, 55; p = 0.28) nor the placebo (5%; 95% CI -13, 26; p = 0.63) group at follow-up. No difference in UACR between the groups was observed at follow-up (relative difference in geometric mean: 11%, 95% CI -26, 67; p = 0.60). For eGFR and SBP, an increase after discontinuation of spironolactone was observed, as well as for SBP after placebo discontinuation. Potassium levels were lower after discontinuation of spironolactone, but higher after placebo discontinuation (all p < 0.05). UACR did not change after discontinuation of long-term treatment with spironolactone. However, an increase in eGFR was observed supporting a haemodynamic effect of spironolactone in the kidneys.

Quantitative analysis of protective T cell immunity against brucellosis

The results of study relationship between antigen reactivity of T-lymphocyte population under ex vivo conditions and the intensity of protective post-vaccination immunity to causative agent of brucellosis are presented. Тaking into account the peculiarities of immunopathogenesis brucellosis and prevailing role of adaptive T-cell immunity to protect against the causative agent of infection, possibility predictive evaluation of protective immunity against brucellosis using CAST-tests is considered as the most important aspect of brucellosis problems. There is an obvious need for an ex vivo correlation analysis of the activity of antigen stimulation of T cells and the intensity of protective immunity formed after vaccination. A close direct proportional relationship was established between the number of live microbial cells Brucella abortus 19BA vaccine strain administered and increase in ex vivo CD3-cell activation. A close correlation was revealed between ex vivo value of antigen-induced stimulation CD3-lymphocytes and level of post-vaccination immunological protection against brucellosis infection. It has been shown that in biomodels vaccinated against brucellosis with a T-lymphocyte stimulation coefficient of 50% or more (according to intensity of antigen-induced ex vivo expression CD25), 100% protection from the development of brucellosis infection after infection with Brucella melitensis at a dose of 1 × 103 live microbial cells are provided. At the same time, there was a lack of a close correlation between an increase in the dose of brucella vaccine strain administered to biomodels and a change in geometric mean antibody titer, presence of a weakly pronounced relationship between level of agglutinins and immunological protection of biomodels from development brucellosis infection and indicators bacterial contamination body.Based on results of study, it was demonstrated that it is possible to quantify the formation and protective activity of T-cell immunity to causative agent of brucellosis based on analysis of level antigen reactivity of CD3-lymphocytes ex vivo. The data obtained and described methodological approach can be used as a predictive criterion in assessing protective level of cellular immunity to causative agent of brucellosis in vaccinated or recovering patients, as well as in order to analyze effectiveness of specific prophylaxis brucellosis and study immunogenicity and protective properties candidate for brucellosis vWe present the results of studies related to antigen reactivity of T lymphocyte population under ex vivo conditions and the intensity of protective post-vaccination immunity to causative agent of brucellosis. Due to peculiarities of immunopathogenesis in brucellosis infection and prevailing role of adaptive T cell immunity for protection against the causative agent of infection, a predictive evaluation of protective immunity against brucellosis using CAST-tests is considered the most important issue in the field. There is an obvious need for ex vivo analysis of correlations between the activity of antigen stimulation of T cells, and the intensity of protective immunity raised after vaccination. A close direct relationship was established between the number of live microbial cells of Brucella abortus 19BA vaccine strain administered, and increase in ex vivo CD3 cell activation. A close correlation (r = -0.841 ÷ -0.966, R2 = 0.708 ÷ 0.969) was revealed between ex vivo values of antigeninduced stimulation of CD3 lymphocytes, and the levels of post-vaccination immunological protection against brucellosis infection. We have shown that, in biomodels vaccinated against brucellosis with a T lymphocyte stimulation coefficient of 50% or more (according to intensity of antigen-induced ex vivo CD25 expression), 100% protection against brucellosis infection was achieved after contamination with Brucella melitensis at a dose of 1×103 live microbial cells. At the same time, a lack of a close correlation was noted between an increased dose of Brucella vaccine strain administered to biomodels, and a change in geometric mean of antibody titer (R2 = 0.357÷0.404), along with a weak relationship between the levels of agglutinins and immunological protection of biomodels from developing brucellosis infection and indices of in vivo bacterial contamination.These results suggest an opportunity to quantify development and protective activity of T cell immunity to the causal agent of brucellosis based ex vivo levels of antigen reactivity of CD3 lymphocytes. A correlation analysis between the state of T cell antigen reactivity and immunological resistance to brucellosis infection indicated a high degree of closeness between these indices. The key influence on activity of protective immunity is exerted by the levels of antigen reactivity of T lymphocytes, whereas the quotient of antigenic stimulation in CD3+CD25+ population may be considered the most informative index of immune protective activity. The data obtained and the described methodology may be used as a predictive criterion in assessing protective level of cellular immunity to causative agent of brucellosis in vaccinated or recovering patients, testing the efficiency of specific prophylaxis in brucellosis and studying immunogenicity and protective properties of candidate vaccines against brucellosis.

The influence of green tea extract on nintedanib’s bioavailability in patients with pulmonary fibrosis

Nintedanib is an oral small-molecule kinase inhibitor and first-line treatment for idiopathic pulmonary fibrosis. Nintedanib is a substrate of the drug efflux transporter ABCB1. Green tea flavonoids --especially epigallocatechin gallate (EGCG)-- are potent ABCB1 modulators. We investigated if concomitant administration of green tea extract (GTE) could result in a clinically relevant herb-drug interaction.Patients were randomized between A-B and B-A, with A being nintedanib alone and B nintedanib with GTE. Both periods lasted 7 days, in which nintedanib was administered twice daily directly after a meal. In period B, patients additionally received capsules with GTE (500 mg BID, >60% EGCG). Pharmacokinetic sampling for 12 h was performed at day 7 of each period. Primary endpoint was change in geometric mean for the area under the curve (AUC0–12 h). A linear mixed model was used to analyse AUCs and maximal concentration (Cmax).In 26 included patients, the nintedanib AUC0–12 h was 21% lower (95% CI −29% to −12%; P < 0.001) in period B (with GTE) compared to period A. Cmax did not differ significantly between periods; − 14% (95% CI −29% to +4%; P = 0.12). The detrimental effect was predominant in patients with the ABCB1 3435 C>T wild type variant. No differences in toxicities were observed.Exposure to nintedanib decreased with 21% when administered 60 min after GTC for only 7 days. This is a statistically significant interaction which could potentially impair treatment efficacy. Before patients and physicians should definitely be warned to avoid this combination, prospective clinical validation of an exposure-response relationship is necessary.

Predicting Serum Ferritin Levels in Patients with Iron Overload Treated with the Film-Coated Tablet of Deferasirox during the Eclipse Study

Background: The ECLIPSE trial demonstrated that the new film-coated tablet (FCT) deferasirox (DFX) formulation and the dispersible tablet (DT) had similar safety profiles during 6 months of treatment, although fewer patients experienced severe gastrointestinal-related adverse events, and more patients experienced favorable patient-reported outcomes with FCT than with DT (Taher et al. Am J Hematol 2017). Additionally, FCT patients had a greater reduction in serum ferritin (SF) levels from baseline to end of treatment (geometric mean change −431 vs −153 ng/mL in DT patients) despite both groups experiencing similar DFX exposure, which suggests greater efficacy with FCT. In the absence of FCT efficacy data beyond 6 months, we developed a model to predict SF levels in patients from the ECLIPSE trial if they had continued DFX DT or FCT treatment for 6-12 months.Methods: ECLIPSE was a Phase II, randomized, open-label study in which patients with transfusion-dependent thalassemia (TDT) or International Prognostic Scoring System lower-risk myelodysplastic syndromes (MDS) were randomized to receive DFX DT or FCT. Planned doses in iron chelation therapy (ICT)-naïve patients were DT 20 mg/kg/day or FCT 14 mg/kg/day based on equivalent FCT doses being 30% lower than DT due to improved bioavailability. ICT pre-treated patients received either a DT dose equivalent to their pre-washout dose or a planned 30% reduction for FCT. Dose was adjusted based on SF levels and investigator's judgment every 4 weeks for ICT-naïve patients and every 3 months for ICT pre-treated patients (DT ±5-10 mg/kg/day, max 40 mg/kg/day; FCT ±3.5-7 mg/kg/day, max 28 mg/kg/day); dose adjustments for safety reasons were permitted at any time.Models for predicting ICT effect were built and tested based on 12-month data from the controlled registration study ICL670A108 (Porter et al. Eur J Haematol 2008). Most patients from ECLIPSE had received prior ICT; therefore, for the development of the model, this situation was mimicked using ICL670A108 month 3-9 data for fitting, and month 10-12 data for evaluating model prediction quality. The selected model accounted for baseline SF levels, assigned daily DFX dose per arm and patients' body weight, and predicted the ICT effect for up to 12 months if patients had continued with their therapy (DT or FCT). The model was applied to both DT and FCT arms of the ECLIPSE trial and predictions were obtained from individual estimates of each patient.Results:Trial results: 173 patients were randomized 1:1 to DT (n=86) or FCT (n=87); 70 patients in each arm had TDT and 16 patients had MDS (underlying disease missing in 1 FCT patient). Mean±SD age was 34.9±19.3 years, 51% were female and 90% had previously received ICT. Mean±SD actual daily DFX dose over 24 weeks was 27.5±7.7 mg/kg/day (DT) and 20.8±5.4 mg/kg/day (FCT). At baseline, SF geometric mean was 2381 ng/mL (DT) and 2721 ng/mL (FCT).Model prediction: Model diagnostics suggested a close approximation of the ECLIPSE data, as the predicted SF geometric mean during the first 6 months of treatment was similar to the actual SF values observed during the trial (Figure). The model predicted that SF geometric mean values would continue to decrease over the subsequent 6 months with both DT and FCT, reaching 2162 ng/mL (90% confidence interval [CI] 2029-2304) with DT and 2180 ng/mL (90% CI 2054-2311) with FCT after 12 months of treatment. This corresponds to a predicted, relative decrease of 10% with DT and 19% with FCT.Conclusions: The 6-month results of the model were similar to the actual SF values observed during the ECLIPSE trial, and reliably predicted continued ICT effect for months 7-12 if patients had continued with their assigned treatment (DT or FCT). Previous analyses have indicated that improved patient-reported outcomes, especially increased adherence, are significant mediators of the association between treatment with DT vs FCT and SF reduction from baseline over 6 months of treatment (Taher et al. Haematologica 2017;102[s2]:abst P286). According to the predicted results from this model, and at the doses given in this study, patients treated with FCT would have had a greater reduction in SF at 12 months compared with patients treated with DT; although it should be noted that FCT patients had higher baseline SF. This analysis suggests the FCT offers patients an improved DFX formulation, which could be attributed to improved treatment adherence. [Display omitted] DisclosuresTaher:Novartis Pharmaceuticals: Honoraria, Research Funding; Celgene: Research Funding. Weber:Novartis: Employment. Han:Novartis: Employment. Bruederle:Novartis: Employment. Porter:Celegene: Consultancy; Agios Pharmaceuticals: Consultancy, Honoraria; Shire: Consultancy, Honoraria; Bluebird Bio: Consultancy; Novartis: Consultancy, Honoraria, Research Funding.

Effects of an intensive lifestyle intervention on the underlying mechanisms of improved glycaemic control in individuals with type 2 diabetes: a secondary analysis of a randomised clinical trial.

The aim was to investigate whether an intensive lifestyle intervention, with high volumes of exercise, improves beta cell function and to explore the role of low-grade inflammation and body weight. This was a randomised, assessor-blinded, controlled trial. Ninety-eight individuals with type 2 diabetes (duration <10years), BMI of 25-40kg/m2, no use of insulin and taking fewer than three glucose-lowering medications were randomised (2:1) to either the standard care plus intensive lifestyle group or the standard care alone group. Standard care consisted of individual guidance on disease management, lifestyle advice and blinded regulation of medication following a pre-specified algorithm. The intensive lifestyle intervention consisted of aerobic exercise sessions that took place 5-6 times per week, combined with resistance exercise sessions 2-3 times per week, with a concomitant dietary intervention aiming for a BMI of 25kg/m2. In this secondary analysis beta cell function was assessed from the 2h OGTT-derived disposition index, which is defined as the product of the Matsuda and the insulinogenic indices. At baseline, individuals were 54.8years (SD 8.9), 47% women, type 2 diabetes duration 5years (IQR 3-8) and HbA1c was 49.3mmol/mol (SD 9.2); 6.7% (SD 0.8). The intensive lifestyle group showed 40% greater improvement in the disposition index compared with the standard care group (ratio of geometric mean change [RGM] 1.40 [95% CI 1.01, 1.94]) from baseline to 12months' follow-up. Plasma concentration of IL-1 receptor antagonist (IL-1ra) decreased 30% more in the intensive lifestyle group compared with the standard care group (RGM 0.70 [95% CI 0.58, 0.85]). Statistical single mediation analysis estimated that the intervention effect on the change in IL-1ra and the change in body weight explained to a similar extent (59%) the variance in the intervention effect on the disposition index. Our findings show that incorporating an intensive lifestyle intervention, with high volumes of exercise, in individuals with type 2 diabetes has the potential to improve beta cell function, associated with a decrease in low-grade inflammation and/or body weight. ClinicalTrials.gov NCT02417012 Graphical abstract.

A randomized masked pilot clinical trial to compare the efficacy of topical 1% voriconazole ophthalmic solution as monotherapy with combination therapy of topical 0.02% polyhexamethylene biguanide and 0.02% chlorhexidine in the treatment of Acanthamoeba keratitis.

To compare the efficacy of topical voriconazole 1% and the combination therapy of 0.02% polyhexamethylene biguanide (PHMB) and 0.02% chlorhexidine for the treatment of Acanthamoeba keratitis (AK). This is a prospective, pilot, double-masked randomized comparative study. Twenty-three eyes of 23 patients with microbiologically (smear and/or growth on culture) confirmed AK were randomized to group BG (PHMB 0.02% and chlorhexidine 0.02%) or group VZ (voriconazole 1%). Primary outcome measure was change in geometric mean (GM) of the corneal ulcer size at final visit. Secondary outcome measures were change in visual acuity. Out of 71 patients with confirmed AK seen during study period, 23 patients were recruited and 18 patients completed minimum 2 weeks of treatment and further analyzed. Ten patients received BG, whereas eight received VZ. Median ulcer size measured as GM of infiltrate decreased from 5.7 mm (IQR, 5.3-6.5 mm) (p = 0.02) to 1 mm (IQR, 0-4.3 mm) in group BG and from 4.5 mm (IQR, 1.8-5.1 mm) (p < 0.05) to 0.7 mm (IQR, 0-1.6 mm) in VZ group. Median visual acuity improved from 1.79 (IQR, 1.48-2.78) to 1.10 (IQR, 0.48-1.79) in BG group (p = 0.02) and from 1.60 (IQR, 1.00-2.78) to 0.80 (IQR, 0.48-1.30) in VZ group (p = 0.18). These outcomes suggest that topical VZ as a monotherapy in AK treatment is effective and comparable to BG combination therapy but needs trials with larger sample size and longer follow-up to provide conclusive evidence.

1540P - Influence of cow’s milk on the absorption and exposure of erlotinib in NSCLC patients

BackgroundErlotinib is an oral EGFR tyrosine kinase inhibitor used in NSCLC. Drug absorption depends largely on its solubility in the stomach and gastrointestinal tract. Potentially, erlotinib -as lipophilic drug- is ought to dissolve better in a fatty drink such as full cow’s milk compared to water. Gastric acid reducing agents like proton pump inhibitors (PPIs) decrease the solubility and thus the uptake of erlotinib. Hence, we hypothesized that administration of cow’s milk may be a feasible way to increase erlotinib uptake (both with or without PPI co-administration). We performed a two-period randomized cross-over study to investigate the influence of full cow’s milk compared to water on the exposure of erlotinib with and without the PPI esomeprazole in NSCLC patients. MethodsDuring 24hours, pharmacokinetic sampling (PK) was performed at days 7 and 14. In the 7 days prior to PK, erlotinib was taken daily with either 250mL water or full cow’s milk. Patients were assigned whether to receive erlotinib with (arm A) or without esomeprazole (40mg qd; arm B) 3hours prior to erlotinib intake starting 3 days prior to PK. Primary endpoint was change in geometric mean for the area under the curve (AUC0-24h). A linear mixed model was used to analyze AUCs and maximal concentration (Cmax). ResultsTwelve of the 20 patients used erlotinib without a PPI. Erlotinib AUC0-24h decreased non-significantly with 5% (95%CI: -14 to+5%; P=0.3) when administered with milk compared to water in the non-PPI patients. Also in the 8 patients who did use esomeprazole, erlotinib AUC0-24h did not differ between intake with water or milk (95%CI: -29 to+40%; P=1.0). Cmax did not differ in non-PPI users (P=0.6) and in PPI users (P=0.9). However, esomeprazole decreased erlotinib AUC0-24h with 48% (95%CI: -61 to -31%; P<0.001) and Cmax with 55% (95%CI: -66 to -42%; P<0.001) in 7 patients who completed both arms. No differences in toxicities were observed. ConclusionsExposure to erlotinib did not change by erlotinib intake with milk compared to water in both the PPI and non-PPI patients. Therefore, the combination with milk instead of water is safe and well tolerated. Esomeprazole strongly decreased both erlotinib AUC0-24h and Cmax, and should be avoided if possible. Clinical trial identificationNTR 6148. Legal entity responsible for the studyRon H.J. Mathijssen. FundingRoche. DisclosureR.H. Mathijssen: Research grant / Funding (institution): Roche; Research grant / Funding (institution), Travel / Accommodation / Expenses: Astellas; Research grant / Funding (institution): Boehringer; Research grant / Funding (institution): Cristal Therapeutics; Honoraria (self), Research grant / Funding (institution): Novartis; Research grant / Funding (institution): Pamgene; Research grant / Funding (institution), Travel / Accommodation / Expenses: Pfizer; Research grant / Funding (institution): Sanofi; Honoraria (self): Servier; Research grant / Funding (institution): Bayer. All other authors have declared no conflicts of interest.

Abstract PD5-06: The immunomodulatory potential of denosumab in breast cancer: results from D-BEYOND, a window of opportunity trial evaluating a RANK-ligand (RANKL) inhibitor and its biological effects in young pre-menopausal women diagnosed with early breast cancer

Abstract Background Breast cancer (BC) in young women has unique biology and poor prognosis. Previous reports suggest that they often express RANKL, which was also shown to play a role in mammary tumorigenesis and various immune processes. Here, we present the primary results of D-BEYOND, a window study investigating the biological activity of the RANKL inhibitor; denosumab in pre-menopausal BC patients. Methods D-BEYOND is a prospective, phase Iia, single-arm, multicenter study assessing the effect of denosumab on BC biology in premenopausal women with early BC (NCT01864798). Patients received two subcutaneous injections of denosumab (120mg), one week apart, followed by breast surgery. The primary endpoint was geometric mean change in tumor Ki67 assessed by immunohistochemistry (IHC). Blood, tumor and normal adjacent breast tissue were collected pre- and post-treatment. Serum levels of RANKL, OPG and CTX were assessed by ELISA. RNA was extracted from fresh-frozen tissue and RNAseq was performed. DESeq2 was used for differential expression analysis, GAGE was used for pathway analysis and CIBERSORT was used to infer immune cell subsets between pre- and post-treatment. Ki67, CD4/Foxp3 and CD4/CD8 IHC were performed on FFPE tissue to further assess the immune microenvironment. The percentage of TILs was independently evaluated by two pathologists on H&amp;E slides. Pre- and post-treatment values were compared using a paired t-test. Results A total of 27 patients were enrolled in the study between October 2013 and July 2016. The median age was 45 years (range 35-51 years). Tumors of 21 patients were hormone receptor positive (77.8%), 4 were HER2 positive (14.8%) and 2 were triple negative (7.4%). No serious adverse events were reported, the most frequent non-serious adverse event being arthralgia (14.8%). After treatment, serum levels of CTX and RANKL decreased in all patients (P &amp;lt; 0.001) whereas OPG increased in 76.9% of patients (P = 0.009, 95% CI 0.56-0.91). There was no significant reduction of Ki67 values from baseline (geometric mean [GM] change after treatment; 0.98, 95% CI 0.76-1.26; P = 0.90). Twenty-four pre- and post-treatment tumor pairs were available for RNAseq, IHC and TILs evaluation. There was a significant increase in the percentage of stromal TILs after treatment (GM change of 1.75, 95% CI 1.28–2.39; P = 0.001). 1084 differentially expressed genes were identified and pathway analysis revealed enrichment of several immune processes. CIBERSORT revealed an enrichment of CD8+ T cells (GM change 1.72, 95% CI 1.19–2.48; P = 0.006) and a decrease of Treg cells (0.71, 95% CI 0.52–0.98, P = 0.040). These results were confirmed by IHC of CD8+ and CD4+/Foxp3+ cells (GM change 1.59, 95% CI 1.14–2.21; P = 0.008 and 0.63, 95% CI 0.49–0.83, P = 0.001, respectively). Conclusion Short course of denosumab did not reduce tumor proliferation rate. However, it induced a significant increase in TILs and CD8 cytotoxic T cells, while Treg infiltration decreased. These findings suggest an immunomodulatory role for denosumab in young breast cancer and that its use in combination could boost immunotherapy efficacy. Citation Format: Nguyen B, Marion M, Salgado R, Venet D, Vuylsteke P, Polastro L, Wieldiers H, Simon P, Lindeman G, Larsimont D, Van den Eynden G, Velghe C, Rothe F, Garaud S, Michiels S, Willard-Gallo K, Azim Jr HA, Loi S, Piccart M, Sotiriou C. The immunomodulatory potential of denosumab in breast cancer: results from D-BEYOND, a window of opportunity trial evaluating a RANK-ligand (RANKL) inhibitor and its biological effects in young pre-menopausal women diagnosed with early breast cancer [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr PD5-06.

Abstract GS3-02: PALLET: A neoadjuvant study to compare the clinical and antiproliferative effects of letrozole with and without palbociclib

Abstract Background: CDK4/6 inhibitors, such as palbociclib, are used to treat ER+ metastatic breast cancer in combination with endocrine therapy with trials ongoing in patients with primary disease. No biomarkers exist to identify those who do/do not benefit from added CDK4/6 inhibition. PALLET is an investigator-initiated/led phase II randomized trial collaboration between UK and NSABP investigators evaluating the biological and clinical effects of palbociclib with letrozole combination as neoadjuvant therapy. Methods: Postmenopausal women with ER+ primary breast cancer and tumors &amp;gt;2.0cm (ultrasound) were randomized to one of 4 treatment groups (3:2:2:2 ratio): Group A: letrozole (2.5mg/d) for 14 weeks; Group B: letrozole for 2 weeks followed by letrozole + palbociclib to 14 weeks; Group C: palbociclib for 2 weeks followed by letrozole + palbociclib to 14 weeks; Group D: letrozole + palbociclib for 14 weeks. Palbociclib was given 125mg/d PO on a 21 days on, 7 days off schedule. Post-14 week treatment was at the discretion of the treating clinician including letrozole until surgery. Core-cut biopsies were taken at baseline, 2 weeks and 14 weeks. Co-primary endpoints for letrozole alone vs palbociclib groups (Group A vs Groups B+C+D) were: (i) change in Ki67 (IHC) between baseline and 14 weeks (log-fold change, Mann-Whitney test); (ii) clinical response (ultrasound) after 14 weeks (4 group, ordinal, Mann-Whitney test). Complete cell-cycle arrest (CCCA) (Ki67≤2.7%) was analyzed using a logistic regression model adjusting for recruitment region. Pre-specified exploratory biomarkers included c-PARP (apoptosis). Results: 307 patients were recruited between 27 Feb 2015 and 08 Mar 2018; 103 were randomized to letrozole alone and 204 to letrozole + palbociclib. 279 (90.9%) patients were evaluable for 14 week clinical response. Clinical response was not significantly different between letrozole vs letrozole + palbociclib groups [(p=0.20; CR+PR 49.5% (46/93) vs 54.3% (101/186) and PD 5.4% (5/93) vs 3.2% (6/186)] nor was the small proportion of patients with pathological CR (1/87, 1.1% vs 6/180, 3.3%; p=0.43). 190 (61.9%) patients were evaluable for 14 week change in Ki67. The median log-fold change in Ki67 was greater with letrozole + palbociclib vs letrozole alone (-4.1 vs -2.2; p&amp;lt;0.001) corresponding to a geometric mean change of -97.4% vs -88.5%. Similarly, a greater proportion of patients who received letrozole + palbociclib achieved CCCA (90% vs 59%, p&amp;lt;0.001). 146 (47.6%) patients were evaluable for c-PARP and the log-fold change (suppression) was greater with letrozole + palbociclib vs letrozole alone (-0.80 vs -0.42; p=0.003) corresponding to a geometric mean change of -56.8% vs -31.4%. Other biomarkers of response / resistance are being evaluated. A higher proportion of patients had a grade ≥3 toxicity on letrozole + palbociclib than letrozole alone (49.8% vs 17.0%; p&amp;lt;0.001) mainly due to asymptomatic neutropenia. Conclusion: Adding palbociclib to letrozole markedly enhanced the suppression of malignant cell proliferation as assessed by Ki67 but did not substantially increase the clinical response of primary ER+ breast cancer over a 14-week period. Concurrent reductions in cell death may have reduced the speed of tumor shrinkage. Citation Format: Dowsett M, Jacobs S, Johnston S, Bliss J, Wheatley D, Holcombe C, Stein R, McIntosh S, Barry P, Dolling D, Snowdon C, Perry S, Batten L, Dodson A, Martins V, Modi A, Cornman C, Puhalla S, Wolmark N, Julian T, Pogue-Geile K, Robidoux A, Provencher L, Boileau JF, Shalaby I, Thirlwell M, Fisher K, Huang Bartlett C, Koehler M, Osborne K, Rimawi M. PALLET: A neoadjuvant study to compare the clinical and antiproliferative effects of letrozole with and without palbociclib [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr GS3-02.

Individual and area-level determinants associated with C-reactive protein as a marker of cardiometabolic risk among adults: Results from the German National Health Interview and Examination Survey 2008-2011.

BackgroundHigh-sensitivity C-reactive protein (hsCRP) is a sensitive biomarker of systemic inflammation and is related to the development and progression of cardiometabolic diseases. Beyond individual-level determinants, characteristics of the residential physical and social environment are increasingly recognized as contextual determinants of systemic inflammation and cardiometabolic risks. Based on a large nationwide sample of adults in Germany, we analyzed the cross-sectional association of hsCRP with residential environment characteristics. We specifically asked whether these associations are observed independent of determinants at the individual level.MethodsData on serum hsCRP levels and individual sociodemographic, behavioral, and anthropometric characteristics were available from the German Health Interview and Examination Survey for Adults (2008–2011). Area-level variables included, firstly, the predefined German Index of Socioeconomic Deprivation (GISD) derived from the INKAR (indicators and maps on spatial and urban development in Germany and Europe) database and, secondly, population-weighted annual average concentration of particulate matter (PM10) in ambient air provided by the German Environment Agency. Associations with log-transformed hsCRP levels were analyzed using random-intercept multi-level linear regression models including 6,768 participants aged 18–79 years nested in 162 municipalities.ResultsNo statistically significant association of PM10 exposure with hsCRP was observed. However, adults residing in municipalities with high compared to those with low social deprivation showed significantly elevated hsCRP levels (change in geometric mean 13.5%, 95%CI 3.2%-24.7%) after adjusting for age and sex. The observed relationship was independent of individual-level educational status. Further adjustment for smoking, sports activity, and abdominal obesity appeared to markedly reduce the association between area-level social deprivation and hsCRP, whereas all individual-level variables contributed significantly to the model.ConclusionsArea-level social deprivation is associated with higher systemic inflammation and the potentially mediating role of modifiable risk factors needs further elucidation. Identifying and assessing the source-specific harmful components of ambient air pollution in population-based studies remains challenging.

Abstract CT101: D-BEYOND: A window of opportunity trial evaluating denosumab, a RANK-ligand (RANKL) inhibitor and its biological effects in young pre-menopausal women diagnosed with early breast cancer

Abstract Background Breast cancer (BC) in young women has unique biology and poor prognosis. Previous reports suggest that they often express RANKL, which was also shown to play a role in mammary tumorigenesis and the etiology of BRCA1/2 related BC. Here, we present the primary results of D-BEYOND, a window of opportunity study investigating the biological activity of the RANKL inhibitor; denosumab in pre-menopausal BC patients. Methods D-BEYOND is a prospective, phase IIa, single-arm, multicenter study assessing the effect of denosumab on BC biology in premenopausal women with early BC (NCT01864798). Patients received two subcutaneous injections of denosumab (120mg), one week apart, followed by breast surgery. Blood, tumor and normal adjacent breast tissue were collected at baseline and at surgery. The primary endpoint was geometric mean change in tumor Ki67 assessed by immunohistochemistry (IHC) from baseline to surgery. Absolute Ki67 responders were defined as having &amp;lt;2.7% IHC staining in the post-treatment tumor. Serum levels of soluble RANKL (sRANKL), OPG and C-terminal telopeptide (CTX) were assessed by ELISA. Ki67, RANK and RANKL expression were assessed by IHC. The percentage of tumor infiltrating lymphocytes (TILs) were also evaluated. Pre- and post-treatment values were compared using a paired t-test. Results A total of 27 patients were enrolled in the study between October 2013 and July 2016. The median age was 45 years (range 35-51 years). Tumors of 21 patients were hormone receptor positive (77.8%), 4 were HER2 positive (14.8%) and 2 were triple negative (7.4%). No serious adverse events were reported, the most frequent non-serious adverse event being arthralgia (14.8%). After treatment, serum levels of CTX and sRANKL decreased in all patients (P &amp;lt; 0.001) whereas serum levels of OPG increased in 76.9% of patients (P = 0.009, 95% CI 0.56-0.91). There was no significant reduction of Ki67 values from baseline (geometric mean change after treatment; 0.98, 95% CI 0.76-1.26; P = 0.90) and there were no absolute Ki67 responders. Twenty-four pre- and post-treatment tumor pairs were available for RANK/L staining and TILs assessment. There was no significant difference in RANKL and RANK H-score in tumors after treatment (P = 0.842, P = 0.142, respectively) but we observed a decrease of RANKL H-score in 3 tumors (12.5%) and an increase of RANK H-score in 5 tumors (20%). Interestingly, there was a significant increase in the percentage of stromal TILs after treatment (geometric mean change of 2.51, 95% CI 1.58-3.97; P = 0.004). There were 10/24 patients (41.7%) with a change in TILs of at least 10%, all of them having an increase in TILs presence (P = 0.002). Conclusion Short course of denosumab did not reduce tumor proliferation rate. However, it induced a significant increase in TILs. These findings suggest that denosumab may potentiate immunotherapy efficacy, at least in young BC patients. Additional results including immune cell profiling by multiplex IHC and RNA-sequencing of tumor and normal tissues will be presented at the meeting. Citation Format: Bastien Nguyen, Marion Maetens, Roberto Salgado, David Venet, Peter Vuylsteke, Laura Polastro, Hans Wieldiers, Philippe Simon, Geoff Lindeman, Denis Larsimont, Gert Van den Eynden, Chloe Velghe, Francoise Rothe, Soizic Garaud, Stefan Michiels, Karen Willard-gallo, Hatem A. Azim, Sherene Loi, Martine Piccart, Christos Sotiriou. D-BEYOND: A window of opportunity trial evaluating denosumab, a RANK-ligand (RANKL) inhibitor and its biological effects in young pre-menopausal women diagnosed with early breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr CT101.

Circulating Sphingolipids, Insulin, HOMA-IR, and HOMA-B—The Strong Heart Family Study

High levels of fasting plasma insulin, reflecting insulin resistance, are an early event in the progression to type 2 diabetes. Experimental studies implicate ceramides in insulin resistance; however, whether circulating ceramides and related sphingolipids are associated with plasma insulin in humans is uncertain. We measured 15 ceramide and sphingomyelin species with different saturated fatty acids in baseline fasting plasma samples from the Strong Heart Family Study, a prospective cohort of American Indians. We examined associations of sphingolipids with plasma insulin, HOMA-IR and HOMA-B at baseline and follow-up (median 5 years later) using linear mixed models adjusted for family structure. Among 2086 participants without diabetes, higher levels of plasma ceramides carrying the fatty acids 16:0, 18:0, 20:0 or 22:0 were associated with higher plasma insulin and higher HOMA-IR at baseline and follow-up (Table). BMI modified associations of sphingomyelin species carrying 18:0, 20:0, 22:0 or 24:0 with insulin (p&amp;lt;0.003); higher sphingomyelin levels were associated with lower fasting insulin and HOMA indices only among those with normal BMI. Our study suggests lowering circulating ceramides might be a target in prediabetes, and targeting sphingomyelins should take into account BMI. Table. Plasma Ceramides, Insulin and HOMAIR.SpeciesFold change in geometric mean with 2 fold higher ceramide (95% CI), p-valueBaseline insulinFollow-up insulinBaseline HOMAIRFollow-up HOMAIRCer-16:01.13 (1.07-1.20) 1.9x10-51.10 (1.01-1.21) 0.031.15 (1.08-1.22) 4.7x10-61.11 (1.01-1.22) 0.03Cer-18:01.11 (1.06-1.15) 1.0x10-61.12 (1.06-1.19) 1.0x10-51.12 (1.08-1.17) 6.3x10-81.13 (1.06-1.20) 2.0x10-4Cer-20:01.12 (1.07-1.17) 1.5x10-71.10 (1.03-1.18) 0.0031.13 (1.08-1.18) 1.3x10-71.11 (1.04-1.19) 0.002Cer-22:01.13 (1.08-1.19) 3.9x10-71.12 (1.04-1.20) 0.0041.15 (1.09-1.21) 1.0x10-71.13 (1.04-1.22) 0.004Cer-24:01.(0.99-1.11) 0.081.09 (1.01-1.19) 0.041.(1.00-1.13) 0.041.11 (1.01-1.21) 0.03Covariates: age, sex, site, education, smoking, log(BMI), waist, physical activity. Disclosure R.N. Lemaitre: None. C. Yu: None. A. Hoofnagle: Research Support; Self; Waters Corporation. H. Nair: None. P.N. Jensen: None. A.M. Fretts: None. J.G. Umans: None. C. Sitlani: None. D. Siscovick: None. I. King: None. N. Sotoodehnia: None. B. McKnight: None.

Abstract CT092: A phase II study of neoadjuvant abemaciclib (LY2835219) in postmenopausal women with hormone receptor positive (HR+), human epidermal growth factor receptor 2 negative (HER2-) breast cancer (neoMONARCH)

Abstract Purpose: Abemaciclib (LY2835219) is a potent, selective small molecule inhibitor of cyclin dependent kinase (CDK)4 and CDK6, which has been shown to inhibit cell cycle progression by preventing the phosphorylation and functional inactivation of the Rb tumor-suppressor protein. Cell cycle dysfunction due to abnormalities in the CDK4 and CDK6 pathway occurs in breast cancer. Abemaciclib demonstrated single-agent activity in heavily pretreated women with HR+ metastatic breast cancer (Study I3Y-MC-JPBA), and has an acceptable safety profile as a single agent and when combined with aromatase inhibitors. neoMONARCH (NCT02441946) is a randomized, open-label phase II study comparing the biologic activity of abemaciclib plus anastrozole, abemaciclib monotherapy, and anastrozole monotherapy in postmenopausal women with early-stage HR+, HER2- breast cancer by assessing percentage change from baseline in Ki67 expression after 2 weeks of therapy. Secondary objectives are to evaluate response rates, safety, quality of life, and pharmacokinetics of abemaciclib and anastrozole with up to 22 additional weeks of neoadjuvant therapy. Methods: Approximately 220 patients with HR+, HER2- breast cancer who have received no prior treatment for newly diagnosed invasive breast cancer will be randomized 1:1:1 (Arm A [abemaciclib 150 mg orally q12hrs plus anastrozole 1 mg orally q24hrs], Arm B [abemaciclib 150 mg orally q12hrs], Arm C [anastrozole 1 mg orally q24hrs]) for 2 weeks, with stratification based on progesterone receptor status and tumor size (&amp;lt;2 versus ?2 cm and &amp;lt;5 versus ?5 cm). During the subsequent 14 weeks, all patients will receive abemaciclib 150 mg orally q12hrs plus anastrozole 1 mg orally q24hrs. Patients experiencing benefit, but not a complete response, will have the option to continue combination therapy for an additional 2 cycles (up to 8 additional weeks) at the discretion of the investigators. All patients will receive prophylactic loperamide during abemaciclib therapy. Eligible patients include postmenopausal women with HR+, HER2- clinical Stage I breast tumor ?1 cm in diameter, Stage II, Stage IIIA, or IIIB breast cancer. Patients are required to have ?1 measurable lesion, adequate organ function, and an ECOG performance status of ?1. Patients’ primary breast cancer must be suitable for baseline core biopsy. Statistical methods: The study has &amp;gt;80% power to detect a superior effect of the combination therapy or of abemaciclib alone over anastrozole alone in reducing Ki67 expression at a 1-sided alpha level of 0.1, assuming a geometric mean change in Ki67 expression of -82% for anastrozole alone and -91% for combination therapy or abemaciclib alone. Present accrual: 73 patients. Citation Format: Sara A. Hurvitz, Jeanne Marie Schilder, Martin Frenzel, Miguel Martín. A phase II study of neoadjuvant abemaciclib (LY2835219) in postmenopausal women with hormone receptor positive (HR+), human epidermal growth factor receptor 2 negative (HER2-) breast cancer (neoMONARCH). [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr CT092.

THU0126 Evaluation of the Effect of Tofacitinib on Measured Glomerular Filtration Rate in Patients with Active Rheumatoid Arthritis

BackgroundIn the clinical development programme of the novel oral Janus kinase inhibitor tofacitinib for the treatment of rheumatoid arthritis (RA), small mean increases in serum creatinine (SCr) (<4 μmol/L least...

Pre-surgical study of the biological effects of the selective cyclo-oxygenase-2 inhibitor celecoxib in patients with primary breast cancer

Cyclo-oxygenase 2 (COX-2) is implicated in the regulation of aromatase transcription in malignant breast tissue and has been considered as a potential target for tissue specific aromatase suppression. We initiated a randomised controlled pre-surgical study of celecoxib versus no treatment in women with primary breast cancer to determine the effects of COX-2 inhibition on markers of biological response. Postmenopausal women (50-80 years of age) with stage I or II, primary breast cancer, were randomised 2:1 to receive 400 mg/day celecoxib or no treatment for 14 days prior to surgery. A core biopsy was obtained pre- and post-treatment. Paired baseline and endpoint biopsies were analysed for Ki67, apoptosis, COX-2, CD31, estrogen receptor (ER) and progesterone receptor (PgR). Comparisons between the treatment groups were conducted using the Mann-Whitney test with a two-sided 5% significance. Of the 25 patients treated, 23 had evaluable data and 19 (83%) were ER positive. Overall the geometric mean change in Ki67, the primary end point, relative to baseline in the celecoxib arm was -16.6% (P = 0.056). The change in the no-treatment group was -8.1% (P = 0.24). There was no statistically significant difference in the change between the two groups. Celecoxib did not significantly affect apoptosis, COX-2, ER or PgR expression. There is only modest evidence for a biological effect of celecoxib in primary breast cancer. However, the trend towards a reduction in Ki67 in ER-positive breast cancer warrants further investigations in a larger cohort of patients.

Abstract A57: High-dose resveratrol modulates drug and carcinogen metabolizing enzymes in a healthy volunteer study

Abstract Purpose: Resveratrol (RES) or 3, 4′, 5-trihydroxystilbene has been shown to inhibit carcinogenesis by affecting various molecular events in the initiation, promotion and progression stages. The cancer chemopreventive activity of RES has been demonstrated in vivo in a wide variety of tumors including skin, mammary, gastrointestinal, and liver cancer models. Modulation of Phase I and Phase II enzymes has been suggested to be one of the mechanisms responsible for the cancer preventive effect of RES. We conducted a clinical study to determine the effect of pharmacological doses of RES on drug and carcinogen metabolizing enzymes. Methods: Forty-two healthy volunteers underwent baseline assessment of Phase I and Phase II enzymes. A cocktail of cytochrome P450 (CYP) metabolic probe drugs, including caffeine, dextromethorphan, losartan, and buspirone, were administered to assess the activity of CYP1A2, 2D6, 2C9, and 3A4, respectively. Blood and urine samples were collected for 8 hours after probe drug administration to determine parent probe drug and metabolite concentrations for measurements of CYP enzyme activities. Blood lymphocyte glutathione S-transferase (GST) activity and GST-π level, and serum total and direct bilirubin, a surrogate for UDP-glucuronosyl transferase (UGT) 1A1 activity, were measured to assess Phase II enzymes. After the baseline evaluation, study participants took 1 gm of RES once daily for 4 wks. Enzyme assessment was repeated upon intervention completion. Results: RES intervention was found to suppress the activity of CYP3A4, 2D6, and 2C9. The geometric mean change of the activity index of CPY3A4, 2D6, and 2C9 was 33% (p = 0.01), 70% (p = 0.01), and 171% (p &amp;lt; 0.0001), respectively. CYP1A2 activity was induced; the geometric mean change of the activity index was 16% (p = 0.005). The overall GST and UGT1A1 activity index were minimally affected by the intervention while an induction of GST-π level (84%, p = 0.002) and UGT1A1 activity index (20%, p = 0.0089) was observed in individuals with baseline enzyme level/activity in the lowest tertile. Conclusion: We conclude that high doses of RES administration may modulate enzyme systems involved in carcinogen activation and detoxification, which may be one mechanism by which RES inhibits carcinogenesis. However, high doses of RES administration may lead to clinically relevant metabolic drug interactions. Further clinical studies are needed to determine whether lower doses of RES could be used to achieve cancer preventive activities. (Supported by N01CN35158 from the National Cancer Institute, Division of Cancer Prevention) Citation Information: Cancer Prev Res 2010;3(1 Suppl):A57.

Food Affects Zidovudine Concentration Independent of Effects on Gastrointestinal Absorption

Food can change drug concentrations by several mechanisms, including some that are independent of absorption effects. This study tests the hypothesis that a meal decreases zidovudine (ZDV) concentration in blood plasma independent of an effect on drug absorption. The study was conducted as a substudy nested in a larger protocol in which ZDV was given to 7 healthy men by continuous infusion for 5 days starting on day 1. The subjects received only a standardized breakfast meal on day 2 and were fasted on day 3 until the 8-hour sampling period each day was finished. Blood samples were collected through an indwelling cannula in the arm contralateral to the ZDV infusion at the same time of day on both days. It was found that food decreased the blood plasma ZDV concentration at 1 hour postprandial by 14% (5.0%-22%; geometric mean change with 95% confidence interval). This decrease recovered by 6 hours postprandial. Similar changes were seen with the ZDV primary metabolite, zidovudine glucuronide. The authors conclude that food decreases blood plasma ZDV concentrations independent of any absorption effects. This effect may be due to the increased hepatic metabolism because feeding increases hepatic blood flow and because ZDV has a high hepatic extraction ratio with low affinity to blood plasma proteins.

Dietary Linolenic Acid Intake Is Positively Associated with Five-Year Change in Eye Lens Nuclear Density

Objective: Dietary fat may affect lens cell membrane composition and function, which are related to age-related cataract. The present study was designed to examine the associations between dietary fat and the change in nuclear lens opacification over five years.Methods: Women aged 52 to 73 years without previously diagnosed cancer, diabetes and cataracts from the Boston, Massachusetts area were selected from the Nurses’ Health Study cohort. Four hundred forty women participated in a baseline (1993–95) and a follow-up (1998–2000) eye examination. Intakes of total fat and selected fatty acids were calculated as the average intake from five food frequency questionnaires that were collected between 1980 and baseline. Change in the degree of nuclear density (opacification) was characterized by the difference between baseline and follow-up in pixel density at the central clear zone in the Scheimpflug slit image of the lens.Results: Intake of alpha-linolenic acid (ALA) was positively associated with change in nuclear density. The geometric mean nuclear density change was 16% greater in the highest quartile category of ALA intake than in the lowest quartile category (P for trend = 0.05). For women in the high tertile category of baseline nuclear lens opacification, the geometric mean change in the highest quartile category of ALA acid intake was 70% higher than the change in the lowest quartile category (P for trend = 0.01). There were no significant associations between other dietary fats and change in nuclear density.Conclusion: Higher ALA intake was associated with a greater age-related change in lens nuclear density.

Randomized Controlled Trial of Fluticasone in Preschool Children with Intermittent Wheeze

Preschool children with intermittent wheeze are often prescribed inhaled corticosteroids, although there is no proven benefit. Measurement of airway resistance by the interrupter technique can be used to objectively assess response to treatment. If lung function improves, treatment may be justified. Children with intermittent wheeze aged 2 to <or= 5 years of age completed a 6-week randomized controlled crossover trial of fluticasone propionate (100 micro g, twice daily), followed by a 10-week parallel extension. The relationships between changes in resistance, serum immunoglobulin E and sensitization measured by skin prick testing were investigated. Sixty-one children completed the crossover trial and 44 (72%) completed the extension. After 6 weeks, geometric mean change in resistance was -16.0% (95% confidence interval, -7.0 to -25.0%, p = 0.003) in sensitized children and -3.5% (95% confidence interval, +0.7 to -7.6%, p = 0.1) in nonsensitized children. Changes in resistance were unrelated to immunoglobulin E. Sixteen weeks after stopping fluticasone, resistance returned to baseline. This is the first study of preschool children with intermittent wheeze that has related changes in lung function on treatment to aeroallergen sensitization. Lung function improved in sensitized children and deteriorated after stopping treatment. Treatment with inhaled steroids may be justified in sensitized children.

The effect of mother-infant skin-to-skin contact on plasma cortisol and β-endorphin concentrations in preterm newborns

The aim of this study was to assess the effect of mother-infant skin-to-skin contact on plasma β- endorphin and cortisol concentrations in stable, preterm infants on a newborn intensive care unit. Blood samples were obtained before and at the end of a 20-min period of skin-to-skin contact. Similarly paired samples, obtained at the same times and after the same interval, without skin-to- skin contact, were obtained on another day and served as controls. Both cortisol and β-endorphin concentrations fell significantly after the skin-to-skin session (cortisol: geometric mean change 66%, p=0.008; β-endorphin: geometric mean change 74%, p=0.002). There was also a significant fall in cortisol levels during the control session (geometric mean change 78%, p=0.02), in contrast to β-endorphin levels, in which there was no significant change. Analysis of variance showed that the fall in β-endorphin, but not the fall in cortisol, during the skin-to-skin session was significant when compared with the control session. These results emphasize the hormonal responsiveness of the preterm newborn to relatively minor interventions. We conclude that maternal skin-to-skin contact results in a significant reduction in circulating β-endorphin in the newborn; there was no evidence of any adverse effect.