Triple Tracer Heterogeneity in Dual Neoplasia​​​​: The "Three Musketeers" in Discerning Squamous Cell Carcinoma and Paraganglioma in the Same Patient.

CHK1 plays a critical role in DNA damage repair (DDR) pathways as well as in coordinating DNA replication. Selective CHK1/CHK2 compounds are being tested in clinical trials but predictive biomarkers of patient response are lacking. A phase 1b expansion cohort study (I4D-MC-JTJA, NCT01115790) with the CHK 1 inhibitor, prexasertib, included patients with advanced, metastatic head and neck squamous cell carcinoma (HNSCC) or squamous cell carcinoma of the anus (SCCA). To identify genomic biomarkers associated with single-agent drug response, pre-treatment tissues (archived or biopsy) from 71 consented patients (HNSCC=47, SCCA=24) were subjected to next-generation sequencing (NGS) using the FoundationOne gene panel. In this subset of patients, the disease control rate (DCR) (Complete Response (CR) + Partial Response (PR) + Stable Disease (SD) based on RECIST Criteria (v 1.1)) was 60% (28/47) and 75% (18/24), respectively. We present here the observed genetic alterations corresponding to three pathways, Cell Cycle, DNA Damage Repair (DDR) and PI3K. In addition, patients’ human papillomavirus (HPV) carrier status was inferred from DNA sequencing using HPV-specific capture probes. HPV+ was 47% for HNSCC and 87% for SCCA. HPV+ and TP53 mutations were mutually exclusive across the two patient cohorts. In HNSCC patients with evaluable progression-free survival (PFS) data, greater clinical efficacy was observed in the HPV+ cohort (median PFS: 4.5 vs 1.4 months, log-rank p = 0.0008). Known or likely loss-of-function (LOF) mutations in FBXW7 and PARK2, two genes implicated in Cyclin E1 proteolysis, were noted in patients with favorable response in both tumor types. Across both HNSCC and SCCA cohorts, mutations and/or germline variants in the DDR genes BRCA1, BRCA2, MRE11A and ATR but not in Fanconi (FANC) pathway genes, were found in patients with treatment benefit. Whereas PIK3CA mutations were infrequent in the HNSCC cohort, in SCCA, mutations occurred in 5/8 (63%) patients with disease control vs 1/6 (17%) with PD. All 7 PI3KCA mutations observed in HPV+ HNSCC and SCCA patients mapped to the helical domain suggestive of Apobec-induced mutagenesis as their source of origin. The enhanced clinical benefit to prexasertib associated with HPV+ in HNSCC may reflect a prognostic effect. Alternatively, the clinical biomarker findings may support the hypothesis that oncogene-induced replication stress (RS) (i.e. arising from HPV E6/E7 and/or FBXW7 loss-dependent Cyclin E1 dysregulation) in the context of attenuated DDR (i.e. BRCA1/2, MRE11A mutations) may sensitize patients to prexasertib monotherapy. Citation Format: Ricardo Martinez, Sameera R. Wijayawardana, David Hong, Johanna Bendell, Anna Maria Russell, Richard P. Beckmann, Aimee Bence Lin. A clinical genomic biomarker study of the CHK1 inhibitor prexasertib in advanced head and neck squamous cancer and squamous cell carcinoma of the anus [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1778. doi:10.1158/1538-7445.AM2017-1778

Oral squamous cell carcinoma positive for p16/human papilloma virus in post allogeneic stem cell transplantation: 2 cases and review of the literature

51. Role of biomarkers in prognosis and treatment of head and neck squamous cell carcinomas

R421 – Ubc9: A Novel Marker of Lymph Nodes Metastases in SCCHN

Role of Hyaluronan-Mediated CD44 Signaling in Head and Neck Squamous Cell Carcinoma Progression and Chemoresistance

The functional GRHL3-filaggrin axis maintains a tumor differentiation potential and influences drug sensitivity

Purpose: Numerous studies have indicated that Sex-determining region Y-box 2 (SOX2) is involved in many squamous cell carcinomas. However, the role of SOX2 in head and neck squamous cell carcinoma (HNSCC) remains unclear. Therefore, we investigated whether activation of SOX2 is significantly associated with prognosis in HNSCC. Methods: Gene expression signature reflecting SOX2 activation was identified in HNSCC cohort, and patients were stratified into two groups according to this signature: SOX2-high group or SOX2-low group. Validation of the signature was sought in two independent patient groups. The association between the signature and prognosis of patients was assessed. Results: The SOX2-low group was associated with poor prognosis for HNSCC in three independent patient cohorts. In a multivariate analysis, the impact of the SOX2 signature on overall survival (OS) was independent of other clinical variables [hazard ratio (HR), 1.45; 95% confidence interval (CI), 1.09 - 1.92; P=0.01]. In patients who received radiotherapy (RT), SOX2-low group had significantly poor OS than those in SOX2-high group. Conclusions: SOX2-low signature is associated with poor prognosis in patients with HNSCC and could be used to predict patients who would benefit from RT. Note: This abstract was not presented at the meeting. Citation Format: Young-Gyu Eun, Young Chan Lee, Jun Eul Hwang. SOX2-related gene signature predicting the prognosis in head and neck squamous cell cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4373. doi:10.1158/1538-7445.AM2017-4373

The increasing incidence of oropharyngeal squamous cell cancer (OPSCC) is well established. However, up-to-date incidence estimates and trends for head and neck squamous cell cancers (HNSCCs) overall, including major anatomic sites, and nonoropharyngeal (non-OP) HNSCCs by sex, race, and age in the United States are not well described. A retrospective analysis of incident HNSCCs during 1992 through 2014 using the Surveillance, Epidemiology, and End Results database was performed to evaluate the incidence of HNSCCs overall, OPSCC, and non-OP HNSCC (those of the larynx, oral cavity, hypopharynx, nasopharynx, and nasal cavity). Incidence rates were calculated overall and by subgroups of interest, and incidence rate ratios were used to compare rates between groups. The incidence rates presented were per 100,000 population and were age adjusted to the 2000 US standard population (19 age groups; Census P25-1130). The annual percent change (APC) was modeled with and without joinpoints. The incidence of HNSCC overall declined (average APC [aAPC], -0.8; P<.001) despite significant increases in the incidence of OPSCCs, most notably between 2000 and 2014 (APC, 2.1; P<.001). Significant declines in incidence were observed for all non-OP HNSCC sites for both women and men (P<.001 each). Among women, the risk of OPSCC also significantly decreased (aAPC, -0.8; P = .002), whereas the risk among men was stable during 1992 through 2001 (APC, 0.4; P = .42) and then significantly increased from 2001 to 2014 (APC, 2.7; P<.001). Decreases in the risk of non-OP HNSCC were especially large for black women (aAPC, -2.6; P<.001) and men (aAPC, -3.0; P<.001). Although the incidence of HNSCC previously was highest among black individuals, since 2009 its incidence has been higher among white compared with black individuals. The incidence of HNSCC is declining, especially for non-OP HNSCC and among black individuals. Cancer 2018;124:2125-33. © 2018 American Cancer Society.

Activating mutations in the HRAS oncogene are present in 5-11% of head and neck squamous cell cancers (HNSCC). We set out to determine if HRAS is a driver of HNSCC growth and whether pharmacologic inhibitors of HRAS membrane association and/or RAS effector signaling may have therapeutic value in these cancers. To determine the importance of HRAS, we depleted it genetically from HRAS-mutant HNSCC cell lines by shRNA and evaluated the effects on cell growth and survival. To determine the potential therapeutic value of blocking HRAS membrane association and/or signaling through the key RAS effector, ERK MAPK, we treated HNSCC cells with inhibitors of farnesyltransferase (FTIs, tipifarnib and lonafarnib) or ERK1/2 (ERKi, SCH772984), respectively. We monitored target inhibition by aqueous/detergent fractionation, gel shift and immunoblotting; quantitated cell viability by Alamar blue assay; and assessed cell cycle distribution and apoptosis by flow cytometry. We found that genetic depletion of mutant HRAS inhibited the growth of HRAS-mutant HNSCC cell lines in vitro, caused a modest G1 cell cycle arrest and robustly induced apoptosis. Since genetic depletion is not clinically feasible, we wished to identify targeted therapies suitable for this patient population. Several cancer types driven by mutant RAS are sensitive to inhibitors of the ERK pathway downstream of RAS, and HRAS is the sole RAS isoform whose activity is blocked by FTIs. We found that FTIs and ERKi each inhibited their respective targets at nanomolar range but only FTIs inhibited the growth of HRAS-mutant HNSCC in a target-dependent manner. In cell lines where FTI treatment upregulated ERK signaling, ERKi enhanced sensitivity to FTI. We established that genetic suppression of mutant HRAS blocks HNSCC growth, validating HRAS as a potential therapeutic target. HRAS-mutant HNSCC cell lines are sensitive to FTI but, surprisingly, less sensitive to ERKi. The combination of FTI and ERKi may further sensitize a subset of HRAS-mutant HNSCC. Other targeted combinations will also be discussed. Citation Format: Sehrish Javaid, Andrew M. Waters, Victoria V. Nguyen, Channing J. Der, Adrienne D. Cox. Role of mutant HRAS in growth and drug sensitivity of head and neck squamous cell cancers (HNSCC) [abstract]. In: Proceedings of the AACR Special Conference on Targeting RAS-Driven Cancers; 2018 Dec 9-12; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(5_Suppl):Abstract nr B40.

Sphingosine kinase 1 (SphK1) is an important regulator of apoptosis, survival, and proliferation in cancer cells. SphK1 expression in head and neck squamous cell cancer (HNSCC) cell lines and tumor tissue was assessed, and the efficacy of SphK1 knockdown in increasing tumor radiosensitivity was evaluated in vitro and in vivo. Expression of SphK1 was determined by immunohistochemistry, Western blot, and real-time polymerase chain reaction (RT-PCR) in 34 prospectively collected HNSCC tumor samples. HNSCC cell lines squamous cell carcinoma (SCC)-15 and SCC-25 were treated with SphK1 inhibitor SKI-II and siRNA targeting SphK1 with and without radiation, and the cell viability was assessed. SCC-15 cells with and without transfection of SphK1 siRNA were then injected into athymic nude mice to develop tumor xenografts, and these 2 groups were further divided into 1 group that received radiation and 1 group that did not. Tumor size was measured over 18 days, when the animals were killed and the tumors were evaluated by immunohistochemistry. SphK1 is found in both HNSCC cell lines and human tumor samples, with higher expression correlated with advanced tumor stage, nodal involvement, and recurrence. In vitro, both SCC-15 and SCC-25 were found to be radioresistant; however, they were sensitized by administration of SKI-II and transfection with siRNA targeting SphK1. In vivo, SphK1-siRNA transfected xenografts were decreased in size compared with both nonradiated control and radiated control mice, whereas mice with both SphK1-siRNA and radiation treatment showed a synergistic reduction in tumor volume. Histopathologic analysis demonstrated a decreased proliferative state in SphK1-siRNA transfected tumors. SphK1 is upregulated in HNSCC, and inhibition of SphK1 sensitizes HNSCC to radiation-induced cytotoxicity.

Low-Level Expression of miR-375 Correlates with Poor Outcome and Metastasis While Altering the Invasive Properties of Head and Neck Squamous Cell Carcinomas

e17547 Background: Lymph node metastasis is a major prognostic determinant for patients with head and neck squamous cell cancer (HNSCC). This study aimed to construct a predictive model using CT characteristics of lymph node and tumor for patients with HNSCC to stratify the risk of lymph node metastasis. Methods: The study population was obtained from historical cohort of 472 cervical lymph nodes from 191 patients with HNSCC in a tertiary referral hospital. We analyzed preoperative CT images of lymph nodes according to diameter, ratio of long-to-short axis diameter, necrosis or cystic change, conglomeration, infiltration to adjacent soft tissue, and laterality (ipsilateral vs contralateral) and analyzed T stage. Reference standard was surgical pathologic results. Multivariate logistic regression analysis was performed to predict whether nodules were diagnosed as metastasis or benign. Results: CT features of lymph nodes, including minimal axial diameter, ratio of long-to-short axis diameter, necrosis or cystic change, and T stage were selected as predictors for lymph nodes metastasis. A 10-point risk scoring system was developed, and the risk of malignancy ranged from 7.3% to 99.8%, which was positively associated with increases in risk scores. The areas under the receiver operating characteristic curve of the development and validation sets were 0.886 and 0.879, respectively. Conclusions: We have devised a simple predictive model using the CT characteristics of lymph nodes and tumor for HNSCC to stratify the risk of cervical lymph node metastasis. [Table: see text]

Squamous cell carcinomas (SCCs) comes from different parts, but there may be similar tumorigenic signaling pathways and metabolism, and different squamous cell carcinoma has a similar mutation landscape and squamous differentiation expression. Studying the expression profile of common SCCs is helpful to find biomarkers with diagnostic and prognostic significance for a variety of squamous cell carcinoma. Lung squamous cell carcinoma (LUSC), head and neck squamous cell carcinoma (HNSC), and ‘squamous cell cancer’ in esophageal carcinoma (ESCA) and cervical squamous cell carcinoma and endocervical adenocarcinoma (CESC) in The Cancer Genome Atlas (TCGA) database were used as training sets. The relevant data sets in the Gene Expression Omnibus (GEO) database were selected as validation sets. Machine learning algorithms were used to screen out factors with high accuracy in the diagnosis of SCCs as core genes, and explore their effects on patient prognosis and immunotherapy. COL7A1 (Collagen Type VII Alpha 1 Chain) has high accuracy in the diagnosis of LUSC and HCSC, whether in the training set (LUSC _ AUC: 0.987; HNSC _ AUC: 0.933) or validation set (LUSC _ AUC: 1.000; HNSC _ AUC: 0.967). Moreover, the expression of COL7A1 was significantly correlated with shorter OS and DSS in HNSC and LUSC patients, and was also significantly negatively correlated with IPS in LUSC patients treated with CTLA4 (-) PD1 (+), CTLA4 (+) PD1 (-) and CTLA4 (+) PD1 (+). COL7A1 has the potential to be used as a diagnostic and prognostic marker for LUSC and HNSC and to predict the efficacy of LUSC immunotherapy.

Despite aggressive therapies, head and neck squamous cell carcinoma (HNSCC), which affects 50,000 new patients annually in the United States, is associated with less than 50% 5-year survival. HNSCC tumors display increased glycolysis, even in the presence of oxygen. Consequently, there is an increase in lactic acid (LA) production. However, the effect of lactic acid in the tumor microenvironment and the mechanisms whereby HNSCC tumors survive in highly acidic conditions remain unknown. HNSCC consist of up to 80% tumor-associated fibroblasts (TAFs). We previously reported that activation of receptor tyrosine kinase, c-Met, by TAF-secreted hepatocyte growth factor (HGF) contributes to HNSCC progression (1,2). The mechanism associated with tumor-stroma metabolic symbiosis is not well understood. On this basis, we hypothesized that TAF-secreted HGF regulates HNSCC glycolysis. We demonstrate that HNSCC-secreted basic fibroblast growth factor (bFGF) induces oxidative phosphorylation (OXPHOS) in TAFs. In addition, bFGF regulates secretion of HGF from TAFs. TAF-secreted HGF increases HNSCC glycolysis and induces bFGF secretion from HNSCC. Thus, HNSCC and TAFs engage in reciprocal signaling through paracrine effects of HGF and bFGF. Inhibition of c-Met with small molecule inhibitor PF-02341066 or specific knockdown with c-Met siRNA decreased TAF-facilitated HNSCC glycolysis, lactic acid production and bFGF expression. In addition, inhibition of FGFR with small molecule inhibitor AZD-4547 decreased OXPHOS and HGF expression in TAFs. Inhibition of FGFR also reduced HNSCC-stimulated phosphorylation of p42/44 mitogen activated protein kinase, proliferation and migration in TAFs. Further, we tested the efficacy of AZD-4547 in combination with c-Met inhibitor PF-02341066 in mitigating TAF-induced HNSCC growth in vitro and in vivo. Combined treatment with AZD-4547 and PF-02341066 significantly inhibited TAF-induced proliferation of HNSCC in vitro compared to the vehicle control treated cells (p&amp;lt;0.001). Further, combined treatment significantly reduced growth of admixed HNSCC and TAF xenograft tumors (p&amp;lt;0.001). Our cumulative findings underscore the therapeutic potential of combinatorial treatment with PF-02341066 and AZD-4547 in HNSCC. The therapeutic approach developed in this study may be a feasible in HNSCC. 1. Wheeler SE, Shi H, Lin F, Dasari S, Bednash J, Thorne S, et al. Enhancement of head and neck squamous cell carcinoma proliferation, invasion, and metastasis by tumor-associated fibroblasts in preclinical models. Head &amp; neck 2014;36(3):385-92. 2. Knowles LM, Stabile LP, Egloff AM, Rothstein ME, Thomas SM, Gubish CT, et al. HGF and c-Met participate in paracrine tumorigenic pathways in head and neck squamous cell cancer. Clinical cancer research : an official journal of the American Association for Cancer Research 2009;15(11):3740-50. Citation Format: Dhruv Kumar, Vikalp Vishwakarma, Jacob New, Wade Gutierrez, Hemant Chavan, Partha Kasturi, Ossama Tawfik, Douglas Girod, Bennett Van Houten, George Leef, Radhika Joshi, Shary Shelton, Jeffrey Straub, Yelizaveta Shnayder, Kiran Kakarala, Terance Tsue, Fangchen Lin, Sumana Dasari, Sufi Thomas. Targeting tumor-stroma metabolic symbiosis for head and neck cancer therapy. [abstract]. In: Proceedings of the AACR Special Conference: Function of Tumor Microenvironment in Cancer Progression; 2016 Jan 7–10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2016;76(15 Suppl):Abstract nr B37.

Functional RNAi Screens Define Distinct Protein Kinase Vulnerabilities in EGFR-Dependent HNSCC Cell Lines