23 research outputs found
Tumor Microvessel Density as a Prognostic Marker in High-Risk Renal Cell Carcinoma Patients Treated on ECOG-ACRIN E2805
Purpose—Increased vascularity is a hallmark of renal cell carcinoma (RCC). Microvessel density (MVD) is one measurement of tumor angiogenesis; however, its utility as a biomarker of outcome is unknown. ECOG-ACRIN 2805 (E2805) enrolled 1,943 resected high-risk RCC patients randomized to adjuvant sunitinib, sorafenib, or placebo. We aimed to determine the prognostic and predictive role of MVD in RCC.
Experimental Design—We obtained pretreatment primary RCC nephrectomy tissues from 822 patients on E2805 and constructed tissue microarrays. Using quantitative immunofluorescence, we measured tumor MVD as the area of CD34-expressing cells. We determined the association with disease-free survival (DFS), overall survival (OS), treatment arm, and clinicopathologic variables.
Results—High MVD (above the median) was associated with prolonged OS for the entire cohort (p = 0.021) and for patients treated with placebo (p = 0.028). The association between high MVD and OS was weaker in patients treated with sunitinib or sorafenib (p = 0.060). MVD was not associated with DFS (p = 1.00). On multivariable analysis, MVD remained independently associated with improved OS (p = 0.013). High MVD correlated with Fuhrman grade 1–2 (p \u3c 0.001), clear cell histology (p \u3c 0.001), and absence of necrosis (p \u3c 0.001) but not with gender, age, sarcomatoid features, lymphovascular invasion, or tumor size.
Conclusions—High MVD in resected high-risk RCC patients is an independent prognostic, rather than predictive, biomarker of improved OS. Further studies should assess whether incorporating MVD into clinical models will enhance our ability to predict outcome and if low MVD can be used for selection of high-risk patients for adjuvant therapy trials
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Molecular and Clinical Activity of CDX-3379, an Anti-ErbB3 Monoclonal Antibody, in Head and Neck Squamous Cell Carcinoma Patients.
PurposeErbB3 and its ligand neuregulin-1 (NRG1) are widely expressed in head and neck squamous cell carcinoma (HNSCC) and associated with tumor progression. A "window-of-opportunity" study (NCT02473731) was conducted to evaluate the pharmacodynamic effects of CDX-3379, an anti-ErbB3 mAb, in patients with HNSCC.Patients and methodsTwelve patients with newly diagnosed, operable HNSCC received two infusions of CDX-3379 (1,000 mg) at a 2-week interval prior to tumor resection. The primary study objective was to achieve ≥50% reduction in tumor ErbB3 signaling (phosphorylation of ErbB3; pErbB3) in ≥30% of patients. Other potential tumor biomarkers, pharmacokinetics, safety, and tumor measurements were also assessed.ResultspErbB3 was detectable in all tumors prior to treatment and decreased for 10 of 12 (83%) patients following CDX-3379 dosing, with ≥50% reduction in 7 of 12 (58%; P = 0.04; 95% confidence interval, 27.7%-84.8%). Target trough CDX-3379 serum levels were achieved in all patients. CDX-3379 treatment-related toxicity was grade 1-2 and included diarrhea, fatigue, and acneiform dermatitis. Five of 12 (42%) patients had shrinkage in tumor burden, including a marked clinical response in a patient with human papillomavirus-negative oral cavity HNSCC. All patients with tumor shrinkage had tumors that expressed both NRG1 and ErbB3 and demonstrated reduced pErbB3 with CDX-3379 treatment.ConclusionsThis study demonstrates that CDX-3379 can inhibit tumor ErbB3 phosphorylation in HNSCC. CDX-3379 was well tolerated and associated with measurable tumor regression. A phase II study (NCT03254927) has been initiated to evaluate CDX-3379 in combination with cetuximab for patients with advanced HNSCC
ErbB activation signatures as potential biomarkers for anti-ErbB3 treatment in HNSCC
<div><p>Head and neck squamous cell carcinoma (HNSCC) accounts for 3–5% of all tumor types and remains an unmet medical need with only two targeted therapies approved to date. ErbB3 (HER3), the kinase-impaired member of the EGFR/ErbB family, has been implicated as a disease driver in a number of solid tumors, including a subset of HNSCC. Here we show that the molecular components required for ErbB3 activation, including its ligand neuregulin-1 (NRG1), are highly prevalent in HNSCC and that HER2, but not EGFR, is the major activating ErbB3 kinase partner. We demonstrate that cetuximab treatment primarily inhibits the ERK signaling pathway and KTN3379, an anti-ErbB3 monoclonal antibody, inhibits the AKT signaling pathway, and that dual ErbB receptor inhibition results in enhanced anti-tumor activity in HNSCC models. Surprisingly, we found that while NRG1 is required for ErbB3 activation, it was not sufficient to fully predict for KTN3379 activity. An evaluation of HNSCC patient samples demonstrated that NRG1 expression was significantly associated with expression of the EGFR ligands amphiregulin (AREG) and transforming growth factor α (TGFα). Furthermore, NRG1-positive HNSCC cell lines that secreted high levels of AREG and TGFα or contained high levels of EGFR homodimers (H11D) demonstrated a better response to KTN3379. Although ErbB3 and EGFR activation are uncoupled at the receptor level, their respective signaling pathways are linked through co-expression of their respective ligands. We propose that NRG1 expression and EGFR activation signatures may enrich for improved efficacy of anti-ErbB3 therapeutic mAb approaches when combined with EGFR-targeting therapies in HNSCC.</p></div
Association of NRG1 expression with AREG and TGFα correlate with KTN3379 activity.
<p><b>(A and B)</b> A search of 303 HNSCC patient samples revealed a significant association between NRG1 expression and expression of EGFR ligands AREG (R<sup>2</sup> = 0.33) and TGFα (R<sup>2</sup> = 0.25). However, no association was found between NRG1 expression and any of the EGFR ligands in colorectal cancer patient samples (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0181356#pone.0181356.s007" target="_blank">S7 Fig</a>). <b>(C and D)</b> High levels of secreted AREG or TGFα protein were found associated with improved KTN3379 activity in HNSCC cell lines. Levels of secreted AREG and TGFα derived from a panel of 8 serum-starved HNSCC cell lines were measured after 48 hours. Supernatant-derived ligand concentrations (in pg/mL) are plotted as a function of the combined anti-proliferative activity of KTN3379 with cetuximab. A linear regression fit of the data gave R<sup>2</sup> values of 0.50 and 0.24 for AREG and TGFα, respectively. A similar result was observed when ligand concentration data were plotted against KTN3379-mediated phospho-AKT inhibition (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0181356#pone.0181356.s008" target="_blank">S8 Fig</a>). <b>(E and F)</b> High levels of EGFR homodimers (H11D) are associated with KTN33379 activity. EGFR homodimer (H11D) levels were evaluated in 8 HNSCC cell lines using a VeraTag immunoassay. H11D values were log-transformed and plotted against the combined anti-proliferative activity of KTN3379 with cetuximab (E), or KTN3379-mediated phospho-AKT inhibition in serum-grown cells (F). Linear regression analysis demonstrates a significant correlation between H11D levels and anti-proliferative activity (R<sup>2</sup> = 0.48) or phospho-AKT inhibition (R<sup>2</sup> = 0.76).</p
KTN3379 enhances cetuximab anti-proliferative activity <i>in vitro</i> and in tumor xenograft models of HNSCC.
<p><b>(A)</b> Titration of KTN3379 (red) enhanced cetuximab (blue) anti-proliferative activity in 4 of 8 HNSCC cell lines (top row) and showed modest enhancement of cetuximab-treated activity in a subset of cell lines (Detroit562 and SCC35). In contrast, neither cetuximab nor KTN3379 showed anti-proliferative activity in other HNSCC lines (UNC10 and SCC9). Combination of KTN3379 and cetuximab is shown in purple. <b>(B)</b> KTN3379 demonstrated significant single agent activity in 2 HNSCC xenograft models (FaDu and OE21), and enhanced the anti-tumor activity of cetuximab. Animals were dosed intraperitoneally twice weekly at 10 mg/kg. Asterisks denote statistical significance; * p-value <0.05; ** p-value <0.01; *** p-value <0.001; **** p-value <0.0001; ns = not significant.</p
Prevalence of EGFR, HER2, and ErbB3 expression in HNSCC.
<p><b>(A)</b> ErbB3 is widely expressed, but not overexpressed, in HNSCC. RNA overexpression was defined as > 4-fold expression over the median expression across all tumor types. TCGA data were used for this analysis <b>(B)</b> ErbB receptors were expressed in the majority of HNSCC patient tumor samples (n = 46), as determined by VeraTag proximity-based immunoassays. Total EGFR (H1T), HER2 (H2T), and ErbB3 (H3T) levels are shown <b>(C)</b> The same pattern of ErbB protein expression was observed in a panel of 8 HNSCC cell lines.</p
ErbB3 activation depends on HER2.
<p><b>(A)</b> Cal27 cells were treated with 100 nM anti-ErbB antibodies and cell lysates were analyzed for ErbB3, AKT and ERK phosphorylation. Addition of KTN3379 (KTN) or pertuzumab (Per) inhibited ErbB3 phosphorylation implicating the role of HER2 as the activating co-receptor for ErbB3. In contrast, cetuximab (Cet) did not inhibit ErbB3 phosphorylation on its own. <b>(B)</b> Cal27 cells were treated as described in (A) and phospho-ErbB levels were quantified by RPMA (Reverse-Phase Microarray Assay), yielding results consistent with (A). In addition, cetuximab treatment promoted mild stimulation of EGFR phosphorylation, consistent with previously published results.</p
Prevalence of NRG1 expression in HNSCC and other tumor types.
<p><b>(A)</b> NRG1 RNA overexpression was found to be most prevalent in HNSCC compared to other major tumor types, and overexpressed in nearly half of all head and neck tumors examined. RNA overexpression is defined as > 4-fold expression over the median expression across all samples (> 29,000). By contrast, NRG1 was overexpressed only in ~ 1% of colorectal cancer. TCGA data were used for this analysis. <b>(B and C)</b> A quantitative ISH-based assay demonstrates detectable NRG1 expression in the majority of human HNSCC tumors <b>(B)</b> and is independent of the histological site <b>(C)</b>. Values reflect the ratio of the NRG1 probe signal over that of a control probe.</p
KTN3379 demonstrates single agent <i>in vivo</i> anti-tumor activity in some, but not all NRG1-positive HNSCC models.
<p>Three patient derived xenograft (PDX) models of HNSCC and one cell line (Cal27) were evaluated for expression of NRG1 and EGFR homodimer (H11D) levels. Single agent KTN3379 dosing resulted in significant anti-tumor efficacy in two NRG1 positive models (Cal27 and CTG-0434), but had no effect in NRG1-positive model (CTG-0776) and one NRG-negative model (CTG-0840). Evaluation of EGFR homodimer (H11D) levels indicated that KTN3379 was efficacious in models that are both NRG1-positive and express high H11D levels, but not in models where H11D levels are low.</p