Diana E. Cunningham

Extracapsular spread in head and neck carcinoma: Impact of site and human papillomavirus status

Extracapsular spread (ECS) in cervical lymph node metastases from head and neck squamous cell carcinoma (SCC) is regarded as an adverse prognostic factor and is often used to select patients who may benefit from adjuvant therapy. The prognostic value of ECS was evaluated for patients with oropharyngeal SCC (OPC; with known p16/human papillomavirus [HPV] status) and for patients with SCC of the oral cavity (OCC).

Quality of life in head and neck cancer patients: Impact of HPV and primary treatment modality

To determine posttreatment quality of life (QOL) in head and neck cancer patients, stratifying by human papillomavirus (HPV)/p16 status and primary treatment modality.

The prognostic and predictive value of KRAS oncogene substitutions in lung adenocarcinoma

The prognostic and therapeutic implications of the spectrum of v‐Ki‐ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS) oncogene substitutions in lung cancer remain poorly understood. The objective of this study was to determine whether KRAS oncogene substitutions differed with regard to prognosis or predictive value in lung adenocarcinoma.

Prevention of tobacco carcinogen-induced lung cancer in female mice using antiestrogens

Increasing evidence shows that estrogens are involved in lung cancer proliferation and progression, and most human lung tumors express estrogen receptor β (ERβ) as well as aromatase. To determine if the aromatase inhibitor anastrozole prevents development of lung tumors induced by a tobacco carcinogen, alone or in combination with the ER antagonist fulvestrant, ovariectomized female mice received treatments with the tobacco carcinogen 4-(methylnitrosoamino)-1-(3-pyridyl)-1-butanone (NNK) along with daily supplements of androstenedione, the substrate for aromatase. Placebo, anastrozole and/or fulvestrant were administered in both an initiation and a promotion protocol of lung tumorigenesis. The combination of fulvestrant and anastrozole given during NNK exposure resulted in significantly fewer NNK-induced lung tumors (mean = 0.5) compared with placebo (mean = 4.6, P < 0.001), fulvestrant alone (mean = 3.4, P < 0.001) or anastrozole alone (mean = 2.8, P = 0.002). A significantly lower Ki67 cell proliferation index was also observed compared with single agent and control treatment groups. Beginning antiestrogen treatment after NNK exposure, when preneoplastic lesions had already formed, also yielded maximum antitumor effects with the combination. Aromatase expression was found mainly in macrophages infiltrating preneoplastic and tumorous areas of the lungs, whereas ERβ was found in both macrophages and tumor cells. Antiestrogens, especially in combination, effectively inhibited tobacco carcinogen-induced murine lung tumorigenesis and may have application for lung cancer prevention. An important source of estrogen synthesis may be inflammatory cells that infiltrate the lungs in response to carcinogens, beginning early in the carcinogenesis process. ERβ expressed by inflammatory and neoplastic epithelial cells in the lung may signal in response to local estrogen production.

Oncologic and Functional Outcomes of Partial Laryngeal Surgery for Intermediate-Stage Laryngeal Cancer

Objective To evaluate the oncologic and functional outcomes of partial laryngeal surgery (PLS) using transoral laser microsurgery (TLM) and supracricoid laryngectomy (SCL) in patients with intermediate-stage laryngeal squamous cell carcinoma (LSCC). Study Design Historical cohort study. Setting Single tertiary care center. Subjects and Methods Retrospective review of oncologic and functional outcomes in intermediate-stage (T2-3/N0-1, stage II and III) LSCC patients who underwent TLM or SCL from 1998 to 2010. Results Sixty patients were included, of whom 28 (47%) underwent TLM and 32 (53%) underwent SCL. For the entire cohort, 2- and 5-year probabilities were 86.2% (95% confidence interval [CI], 73.0%-93.2%) and 72.9% (95% CI, 52.4%-85.6%), respectively, for overall survival (OS) and 79.3% (95% CI, 65.6%-88.0%) and 62.4% (95% CI, 41.9%-77.4%), respectively, for recurrence-free survival (RFS). There was no difference between the TLM and SCL cohorts in OS (P = .542) or RFS (P = .483). More than 75% of patients avoided adjuvant therapy. Communication Scale and Functional Outcome Swallowing Scale scores at median follow-up of 33 months were 2 or better in 97% and 91% of patients, respectively, reflecting functional voice and swallowing postoperatively. Eighty-eight percent of patients retained a functional larynx. Conclusion PLS provides excellent oncologic and functional outcomes for intermediate-stage LSCC and should be considered an alternative to chemoradiation or total laryngectomy in selected patients.

Co-targeting c-Met and COX-2 leads to enhanced inhibition of lung tumorigenesis in a murine model with heightened airway HGF.

Background: The hepatocyte growth factor (HGF)/c-Met pathway is often dysregulated in non–small-cell lung cancer (NSCLC). HGF activation of c-Met induces cyclooxygenase-2 (COX-2), resulting in downstream stimulation by prostaglandin E2 of additional pathways. Targeting both c-Met and COX-2 might lead to enhanced antitumor effects by blocking signaling upstream and downstream of c-Met. Methods: Effects of crizotinib or celecoxib alone or in combination were tested in NSCLC cells in vitro and in mice transgenic for airway expression of human HGF. Results: Proliferation and invasion of NSCLC cells treated with a combination of crizotinib and celecoxib were significantly lower compared with single treatments. Transgenic mice showed enhanced COX-2 expression localized to preneoplastic areas following exposure to the tobacco carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone, which was not present without carcinogen exposure. This shows that COX-2 activity is present during lung tumor development in a high HGF environment. After 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone treatment, a significant decrease in the number of lung tumors per animal was observed after 13-week treatments of crizotinib, celecoxib, or the combination compared with placebo (p < 0.001). With combination treatment, the number of tumors was also significantly lower than single agent treatment (p < 0.001). In the resulting lung tumors, P-c-Met, COX-2, prostaglandin E2, and P-MAPK were significantly downmodulated by combination treatment compared with single treatment. Expression of the epithelial-mesenchymal transition markers E-cadherin and snail was also modulated by combination treatment. Conclusions: In the presence of high HGF, dual inhibition of c-Met and COX-2 may enhance antitumor effects. This combination may have clinical potential in NSCLCs with high HGF/c-Met expression or epithelial-mesenchymal transition phenotype.

Abstract 608: Targeting the estrogen pathway in a male mouse model of lung tumor prevention

Proceedings: AACR Annual Meeting 2014; April 5-9, 2014; San Diego, CA A hormonal role in the pathology of lung cancer is well documented. We have shown that the aromatase inhibitor anastrozole and the anti-estrogen fulvestrant effectively inhibited tobacco carcinogen-induced lung tumorigenesis in a female mouse model of lung cancer prevention. Additionally, we have shown that inflammatory cells that infiltrate the lungs in response to carcinogens may be a source of estrogen synthesis, and have confirmed aromatase and estrogen receptor expression in a pure macrophage population of differentiated THP-1 cells. These results suggest that targeting the estrogen pathway may be beneficial for both treatment and prevention of lung cancer. There is no sex difference in expression of estrogen-related markers or in relation of these markers to survival in lung cancer patients, suggesting that both men and women may benefit from hormonal therapy. We hypothesized that aromatase activity is a factor in lung cancer development regardless of sex. Since testosterone can serve as an estrogen precursor, we determined whether blocking estrogen action is a feasible lung tumor prevention strategy in male mice. To standardize the amount of estrogen in each animal, orchiectomized male mice were utilized and exogenous testosterone was administered via slow release pellets or daily androstendione injections. Under these conditions, testosterone or androstendione is converted to estrogen through aromatase. The tobacco carcinogen NNK (24mg) was administered in weeks 1-4 followed by a holding period for preneoplasia development. Placebo or anastrozole (0.1mg/kg; p.o. daily) treatment was administered in weeks 9-21. Treatment group (10-11 mice per group) differences were assessed by Poisson regression for number of tumors, and by linear mixed models for tumor size. Anastrozole inhibited the mean number of NNK induced lung tumors per animal by 23% in orchiectomized male mice without hormonal supplementation (placebo treatment mean= 11; range= 8-13 vs anastrozole mean= 8; range= 6-10; p=0.11), by 62.5% in mice supplemented with testosterone (placebo treatment mean= 8; range= 6-11 vs anastrozole mean= 3; range= 2-4; p<0.001) and by 55% in mice supplemented with androstendione (placebo treatment mean= 12, range= 9-16 vs anastrozole mean= 5; range= 4-7; p<0.001). In intact male mice with no hormonal manipulation, anastrozole inhibited lung tumor formation by 33% (p=0.001). Fulvestrant also showed a significant decrease in lung tumors regardless of hormonal status. Tumor number was decreased in general by testosterone suggesting that testosterone may have an inhibitory effect on lung tumor formation. Tumor size was also decreased by anastrozole and fulvestrant under all experimental conditions. Tumor Ki67 and serum β-estradiol levels at sacrifice were significantly decreased by anastrozole. These results suggest that hormonal therapies may benefit male lung cancer patients. Supported by P50CA090440 and the Lung Cancer Research Foundation. Citation Format: Laura P. Stabile, Mary E. Rothstein, Brenda F. Kurland, Diana Cunningham, Matthew Orlowski, Jill M. Siegfried. Targeting the estrogen pathway in a male mouse model of lung tumor prevention. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 608. doi:10.1158/1538-7445.AM2014-608

Abstract 2026: Co-targeting c-Met and COX-2 leads to enhanced inhibition of lung tumorigenesis in a murine model with heightened HGF signaling in the airways.

The hepatocyte growth factor (HGF)/c-Met signaling pathway is often dysregulated in lung cancer. We previously showed that HGF activation of c-Met in non-small cell lung cancer (NSCLC) cells led to a significant induction of cyclooxygenase-2 (COX-2) followed by secretion of PGE 2 . PGE 2 caused secretion of TGFα by NSCLC cells, followed by both phosphorylation of EGFR and later, delayed phosphorylation of c-Met that does not require HGF. Secretion of PGE 2 also appears to boost c-Met pathway signaling downstream via cross-activation by EGFR, in a reinforcing loop that is independent of HGF. We hypothesized that targeting of both c-Met and COX-2 might lead to enhanced anti-tumor effects by blocking both upstream and downstream pro-tumor events. Maximum effects on cell proliferation, invasion and EMT markers were observed in vitro in NSCLC cell lines with combination celecoxib and crizotinib treatment. We next tested whether targeting c-Met with the tyrosine kinase inhibitor crizotinib combined with celecoxib to target COX-2 would result in enhanced anti-tumor effects in an animal model with heightened HGF/c-Met pathway signaling in the airways. Mice transgenic for airway expression of human HGF were treated with the tobacco carcinogen, 4-(methylnitroso-amino)-1-(3-pyridyl)-1-butanone, from weeks 1-4. Crizotinib (40mg/kg), celecoxib (50mg/kg), combined crizotinib and celecoxib or placebo control was administered by oral gavage daily from week 3 until week 15, at which time lung tumors were evaluated. A highly significant decrease in the number of lung tumors per animal was observed with crizotinib treatment (mean 3.6, range 2-6), celecoxib treatment (mean 4.3, range 3-6) and combination treatment (mean 1.5, range 0-3) compared to placebo control (mean 9.9, range 6-13; P 2 ), celecoxib (mean 0.22mm 2 ) and combination (mean 0.15mm 2 ) compared to placebo (mean 0.38mm 2 , P 2 , and P-MAPK) were optimally down-modulated by combination treatment, compared to single treatment. Additionally, E-cadherin expression was up-regulated while the E-cadherin repressor, snail, was down-regulated with dual treatment. In a pulmonary environment with heightened HGF/c-Met pathway activity, dual inhibition of the c-Met and COX-2 pathways may enhance anti-tumor effects by targeting reinforcing up- and downstream signaling. Such a combination has potential clinical benefit for lung cancer patients with dysregulated c-Met/COX-2 pathways. Supported by R01 CA79882 and P50 CA090440. Citation Format: Laura P. Stabile, Mary E. Rothstein, Christopher T. Gubish, Nathan Lee, Diana E. Cunningham, Jill M. Siegfried. Co-targeting c-Met and COX-2 leads to enhanced inhibition of lung tumorigenesis in a murine model with heightened HGF signaling in the airways. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2026. doi:10.1158/1538-7445.AM2013-2026

Partial Laryngeal Surgery for Intermediate Larynx Cancer

Objective: Chemoradiation trials over the past 2 decades have assumed that the primary surgical option for intermediate stage laryngeal cancer (T2-T3/N0-N1) was total laryngectomy. However, partial laryngeal surgery (PLS), including transoral laser microsurgery (TLM) and supracricoid partial laryngectomy (SCL), can provide sound oncologic and functional outcomes, often without adjuvant therapy. Method: We retrospectively evaluated oncologic and functional outcomes in laryngeal squamous cell carcinoma (LSCC) patients who underwent TLM or SCL from 1998-2010. Overall survival (OS) and recurrence-free survival (RFS) were calculated. The Communication Scale (CS) and Functional Outcome Swallowing Scale (FOSS) were used to evaluate speech and swallowing, respectively. Results: Eighty-one patients were analyzed with median follow-up of 31 months. Eighty-four percent were T2-T3/N0-N1. Two- and 5-year OS probabilities were 84.2% (95% CI: 73.0-91.0%) and 59.3% (95% CI: 41.4-73.3%), respectively. There was no difference between the TLM and SCL cohorts in OS (P = .781) or RFS (P = .456). Ninety-four percent of patients still had a larynx at median 31 months. Seventy-five percent of patients avoided adjuvant therapy, including 67% of TLM and 84% of SCL patients. CS and FOSS were 2-or-better in 95% and 88%, respectively, designating functional postoperative voice and swallowing. TLM patients were more likely to have a CS of 2-or-better postoperatively (P = .041). Conclusion: PLS can provide good oncologic and functional outcomes for intermediate stage LSCC and is an alternative to chemoradiation or total laryngectomy in select patients.