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Supplementary Fig. 6. Histone marks expression in sunitinib resistant RP-R-01 tumors

Supplementary Fig. 5. Histone marks expression associated with resistance to sunitinib

Supplementary Fig 4. Intratumoral and plasma sunitinib concentrations

Supplementary Fig. 7. EZH2 and E-cadherin mRNA expression in 786-0shRNA scramble, 786-0shEZH2 (clone a and d), and 786-0 treated with GSK126

Supplementary Figure legend

Supplementary Fig. 2. Tumor growth of RP-R-01 ccRCC xenograft treated with sunitinib and associated changes in EZH2, H3K27me3 and E-cadherin expression.

Supplementary Fig 3. Liver histology and body weights of mice on sunitinib dose escalation.

Supplementary Fig. 1. Characterization of RP-R-01 and RP-R-02 PDX models

Supplementary Figure 1. Trial schema

Supplementary Figure 3. Boxplots: PSA response versus baseline circulating androgen levels: DHEA, T and ASD

Supplementary Figure 2. Boxplot: PSA response versus time on duration of prior ketoconazole

Purpose: Patients previously treated with ketoconazole were excluded from phase III trials of abiraterone acetate due to potential overlapping mechanism of action. The purpose of this study was to determine the clinical utility of abiraterone and its impact on circulating androgens following ketoconazole.Experimental Design: Chemotherapy-naïve patients with progressive metastatic castration-resistant prostate cancer (mCRPC) and prior ketoconazole therapy ≥28 days received abiraterone acetate 1,000 mg daily and prednisone 5 mg twice daily. The primary endpoint was the proportion of patients with PSA response, defined as ≥30% PSA decline at 12 weeks. H0 = 0.30 versus H1 = 0.50 (α = 0.05, power = 0.83). Circulating androgen levels were measured using liquid chromatography tandem mass spectrometry.Results: Thirty-nine patients were included in the final analysis. Twenty (51%; 95% confidence interval, 36%–66%) patients had ≥30% PSA decline; the null hypothesis was rejected. Sixteen (41%) had ≥50% PSA decline. Median PFS (progression-free survival) was 16 weeks; median radiographic PFS (rPFS) was 36 weeks. Samples for measurement of baseline androgens were available in 37 patients. The PSA response proportion was 59% in 29 patients with DHEA ≥ limit of quantitation (LOQ), compared with 13% in 8 patients with DHEA < LOQ (P = 0.042). Median PFS was 6 and 16 weeks in DHEA < LOQ and DHEA ≥ LOQ patients, respectively (P = 0.017); median rPFS was 14 and 36 weeks in DHEA < LOQ and DHEA ≥ LOQ patients, respectively (P < 0.001).Conclusions: Abiraterone demonstrates modest clinical efficacy in mCRPC patients previously treated with ketoconazole. Patients with DHEA ≥ LOQ were more likely to demonstrate PSA responses and longer PFS. Analysis of circulating androgens merits further investigation as a biomarker for response to androgen synthesis inhibitor therapy. Clin Cancer Res; 20(24); 6269–76. ©2014 AACR.

Background Plaque psoriasis (PsO) is an inflammatory skin disease driven, in part, by the activation of Janus kinase (JAK) signalling pathways. Objectives To assess the efficacy and safety of multiple doses of topical brepocitinib, a tyrosine kinase 2/JAK1 inhibitor, in participants with mild-to-moderate PsO. Methods This phase IIb multicentre randomized double-blind study was conducted in two stages. In stage 1, participants received one of eight treatments for 12 weeks: brepocitinib 0.1% once daily, 0.3% once or twice daily, 1.0% once or twice daily, 3.0% once daily, or vehicle once or twice daily. In stage 2, participants received brepocitinib 3.0% twice daily or vehicle twice daily. The primary endpoint was the change from baseline in Psoriasis Area and Severity Index (PASI) score at week 12, analysed using analysis of covariance. The key secondary endpoint was the proportion of participants who achieved a Physician Global Assessment response [score of clear (0) or almost clear (1) and an improvement of ≥ 2 points from baseline] at week 12. Additional secondary endpoints included the difference vs. vehicle in change from baseline in PASI, using mixed-model repeated measures, and the change from baseline in Peak Pruritus Numerical Rating Scale at week 12. Safety was monitored. Results Overall, 344 participants were randomized. Topical brepocitinib did not result in statistically significant changes compared with respective vehicle controls in the primary or key secondary efficacy endpoints for any dose group. At week 12, least squares mean change from baseline in PASI score ranged from –1.4 to –2.4 for the brepocitinib once-daily groups vs. –1.6 for vehicle once daily, and from –2.5 to –3.0 for the brepocitinib twice-daily groups vs. –2.2 for vehicle twice daily. From week 8, change from baseline in PASI score separated from vehicle in all brepocitinib twice daily groups. Brepocitinib was well tolerated, with adverse events (AEs) occurring at similar rates across groups. One participant in the brepocitinib 1.0% once-daily group developed a treatment-related AE of herpes zoster in the neck area. Conclusions Topical brepocitinib was well tolerated but did not result in statistically significant changes compared with vehicle when administered at the doses evaluated to treat signs and symptoms of mild-to-moderate PsO.

Bcl-2 proteins represent a rheostat that controls cellular viability. Obatoclax, a BH3-mimetic, has been designed to specifically target and counteract anti-apoptotic Bcl-2 proteins. We evaluated the biological effects of obatoclax on the anti-tumour activity of rituximab and chemotherapy agents. Obatoclax induced cell death of rituximab/chemotherapy-sensitive (RSCL), -resistant cell lines (RRCL) and primary tumour-cells derived from patients with B-cell lymphomas (N=39). Obatoclax also enhanced the activity of rituximab and had synergistic activity when combined with chemotherapy agents. The ability of Obatoclax to induce PARP cleavage varied between patient samples and was not observed in some RRCL. Inhibition of caspase activity did not affect obatoclax activity, suggesting the existence of caspase-independent death pathways. Autophagy was detected by LC3 conversion and/or electron microscopy in RRCL and in patient-derived tumour cells. Moreover, obatoclax activity was inhibited by Beclin-1 knockdown. In summary, obatoclax is an active Bcl-2 inhibitor that potentiates the activity of chemotherapy agents and, to a lesser degree, rituximab. Defining the molecular events triggered by obatoclax is necessary to further its clinical development and identify potential biomarkers that are predictive of response.

Erlotinib is approved as treatment for metastatic non-small-cell lung cancer (NSCLC), following failure of initial therapy. Studies to define patients that derive maximal benefit from erlotinib have not dictated current practice.We retrospectively analyzed the prescription patterns and outcomes related to erlotinib use for NSCLC in a comprehensive cancer center.Of 137 consecutive patients treated with erlotinib over 2 years, 116 were evaluable. Median age was 66 years, 63% females, most common histology was adenocarcinoma (n = 58). Seventy-nine patients presented with stage IIIB-IV disease, 37 with recurrent disease. There were 109 smokers. Erlotinib was given first line in 31 (27%), second line in 52 (45%) and third line in 33 (28%) patients. Daily erlotinib dose was 100 mg in 21 (18%) and 150 mg in 91 (82%) patients. Median duration of treatment was 8 weeks (range 1-72). Median overall survival (OS) from initiation of erlotinib was 5.4 months (range 0.2-27.8). There was no significant difference in median survival by disease stage (recurrent vs. de novo IIIB-IV) (p = 0.201), whether erlotinib was used as first-, second-, third-line therapy (p = 0.971) or at different doses (100 vs. 150 mg daily dose) (p = 0.579).OS after erlotinib use was not different, whether used as first-, second- or third-line therapy, whether patients had recurrent metastatic NSCLC or de novo stage IV disease, or if erlotinib was used at a dose of 100 or 150 mg daily.

Plasma cells (PC) depend on interactions with bone marrow stroma and the resultant growth factors for their long lived survival. Very little is known about the molecular mediators of these survival signals. One such molecule that has recently come of interest is PIM2. PC malignancies such as multiple myeloma (MM) preferentially express PIM2 over its counterparts PIM1 or PIM3 downstream of the PI3K/AKT/mTOR pathway essential for differentiation and survival of PC. In this study, we explored an experimental drug JP_11646 (Jasco Pharmaceuticals LLC) that has a high potency towards PIM kinases, more specifically towards PIM2. Using MM cell lines as a model for long term PC survival, we have shown for the first time that inhibiting PIM2 deregulates survival pathways that involve IL-6, DC protection of MM and CD28 activation by deregulating key survival factors such as NFκB, decreased phosphorylation and inactivation of the PIM2 substrate and pro-apoptotic protein BAD, lower levels of the inactive phosphorylated translational repressor 4EBP1, besides affecting transcription and translation of PIM2 itself. This was recapitulated when PIM2 was silenced with siRNA. Through northerns and protein assays, we also explore the transcriptional and translational dimensions involved in regulating PIM2 in plasma cells. An understanding of the role of PIM2 in MM biology will help to understand how plasma cells survive long term and maintain humoral immunity.

Background Marrow angiogenesis plays an important role in the pathogenesis of acute myeloid leukemia (AML). Among the most prominent pro-angiogenic factors produced by AML cells are the angiopoietins (Ang-1/2), ligands of the Tie2 endothelial cell receptor. Elevated expression of Ang-2 is an independent prognostic factor for overall survival in AML. Trebananib (AMG 386) is a first-in-class neutralizing peptibody that provides potent and selective inhibition of angiopoietins by sequestering Ang1 and Ang2 and preventing their interaction with the Tie2 receptor. Purpose Here we evaluated the safety, tolerability, pharmacokinetic profile, and biologic effects of trebananib in de novo and relapsed/refractory AML patients considered to be poor candidates for standard induction chemotherapy. Methods Thirteen AML patients were enrolled in single agent arm A of this open-label, non-randomized phase Ib trial. (ClinicalTrials.gov ID: NCT01555268, NCI-2011-02979). Drug was administered at two dose levels (15 and 30 mg/kg) via weekly intravenous infusion over 1 hour. Circulating angiogenic factor levels (Ang-2, VEGF-A/C/D, HGF, IL-8, FGF-1/2, G-CSF, PlGF) in the plasma were measured by multiplex flow cytometry prior to first dose, on cycle 1 day 29 (following 4 weekly doses of trebananib), and at the end of study. Results Median age was 76 years (range 53-82). Nine patients were male (69%). Two patients had de novo, 7 had refractory, and 4 had relapsed AML. One patient had undergone prior allogeneic stem cell transplantation. Aside from one possibly drug-related incident of grade 3 mucositis on the 15 mg/kg cohort, no other dose-limiting toxicities were reported. The most common toxicities were abdominal bloating, diarrhea, edema, and weight gain. Preliminary pharmacokinetic analysis of trebananib was dose-proportional. At the 15 and 30 mg/kg dose levels, median Cmaxand AUC on Day 1 were 193 and 473 ug/mL and 11,670 and 35,217 ug-hr/mL, respectively. No neutralizing antibodies to trebananib were detected. Among seven AML patients with consecutive plasma samples, therapy was associated with marked elevations of Ang-2 levels (from 15 to 336-fold higher than baseline) after 1-4 weeks of therapy (Table 1). Induction of Ang-2 did not differ significantly between the 15 mg/kg and 30 mg/kg cohorts. Ang-2 levels measured in bone marrow or plasma samples at the same time point were comparable. In contrast, levels of other angiogenic growth factors (VEGF-A/C/D, IL-8, G-CSF, HGF, FGF-1/2, PlGF) were not consistently altered by trebananib therapy. One AML patient (on the 15 mg/kg cohort) had a partial response with >50% decrease in marrow blasts. Two patients (15 mg/kg, 30 mg/kg) had stable disease over >1 cycle. Conclusions We report here the preliminary results of phase 1b study evaluating the safety, tolerability, pharmacokinetic profile, and biologic effects of trebananib in adult AML patients. Weekly infusions of trebananib administered at 15 and 30 mg/kg were well tolerated. Drug pharmacokinetics were similar to those previously reported in solid tumor patients. Trebananib resulted in substantial elevations in measurable circulating and marrow Ang-2 levels in AML patients, presumably due to a negative compensatory feedback loop resulting from potent in vivo Ang-1/2 inhibition. Arm B of this study (trebananib plus low dose cytarabine) is currently accruing. Further in depth studies examining the biological effects of trebananib on marrow angiogenesis and clinical response in AML patients are ongoing. Disclosures: Becker: Millenium: Research Funding.