Advanced HNSCC Patients Research Articles

Abstract 4169: Inhibition of EphB4-ephrin-B2 signaling results in enhanced sensitivity to cetuximab-radiation therapy in head and neck cancers

Abstract Introduction: The management of locally-advanced head and neck cancer patients relies on the combination treatment involving 7 weeks of radiation therapy (RT) with Cetuximab, an EGFR-inhibitor. Only a fraction of HNSCC patients, however, show response to cetuximab-radiation with an estimated 5-year overall survival of 46% compared to 36% with radiotherapy alone. This suggests potential involvement of redundant or compensatory tumor survival pathway that comes into action and blunts the therapeutic response. Based on our previous findings and published reports, elevated expression of EphB4, a member of Eph family of receptor tyrosine kinases and its membrane-bound ligand, ephrin-B2 might play a role in mediating this effect. In the current study, we hypothesized that EphB4-ephrin-B2 favors pro-tumorigenic survival signaling by altering the sensitivity to EGFR inhibitor and radiation. Therefore, combined inhibition of EphB4-ephrin-B2 signaling is critical to enhance sensitization of HNSCC towards anti-EGFR and radiation therapy. Materials and methods: To test the underlying hypothesis, we used a blocking protein targeting EphB4-ephrin-B2 axis in the absence and presence of EGFR inhibitor with radiation in HNSCC cell lines and patient-derived xenograft (PDX) model. Alteration in the protein levels in control and treatment groups was detected by Western blotting and immunohistochemistry. Circulating levels of cytokines was analyzed in vivo by Mesoscale U-plex assay. Tumor tissues were subjected to human PhosphoRTK array to assess changes in the activation status of downstream targets of EphB4-ephrin-B2 signaling mediating response in the triple combination groups. Results: Our data from locally advanced HNSCC patients treated with standard of care definitive Cetuximab-RT show high levels of both EphB4 and ephrin-B2 after failure of Cetuximab-RT. We observed significant tumor growth delay and increased response towards Cetuximab and radiation therapy following EphB4-ephrin-B2 inhibition resulting in better overall survival in PDX tumors and HNSCC cells. The tumor growth inhibition effects observed in vivo was accompanied by decrease in the levels of proliferation marker, angiogenic regulators, and increased apoptosis. Changes in levels of phopsho-receptor kinases and in circulating immune profile was also evident in tumors subjected to triple combination strategy. Conclusions: Overall, our data suggest that EphB4-ephrin-B2 and EGFR pathway cross-talk with each other to circumvent the therapeutic response resulting in uncontrolled tumor growth, apoptotic evasion, and immune escape. Therefore, development of combinatorial approaches targeting Eph-ephrin family of proteins might show promising outcome to enhance sensitivity towards anti-EGFR therapeutics and radiation in head and neck cancers. Citation Format: Shilpa Bhatia, Jaspreet Sharma, Sanjana Bukkapatnam, Ayman Oweida, Shelby Lennon, Parkash Gill, Antonio Jimeno, David Raben, Lynn Heasley, Sana D. Karam. Inhibition of EphB4-ephrin-B2 signaling results in enhanced sensitivity to cetuximab-radiation therapy in head and neck cancers [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 4169.

Phase II multi-site investigation of neoadjuvant pembrolizumab and adjuvant concurrent radiation and pembrolizumab with or without cisplatin in resected head and neck squamous cell carcinoma.

6017Background: 2Despite aggressive adjuvant treatment, locally advanced HNSCC patients undergoing primary surgical resection with high risk [HR] features (extracapsular extension of lymph nodes (L...

983P - An advanced tumor shape radiomic signature predicts recurrence of locally advanced (LA) HNSCC patients (pts)

983P - An advanced tumor shape radiomic signature predicts recurrence of locally advanced (LA) HNSCC patients (pts)

A radiomic signature based on advanced tumor shape parameters (Spiculated-ness) to predict the outcome of locally advanced (LA) HNSCC patients.

11571Background: Tumor invasion patterns are known to be associated with HNSCC pts outcome. Radiomics is a rapidly-emerging field involving the extraction of quantitative imaging features from stan...

Up-regulation of HB-EGF by the COX-2/PGE2 signaling associates with the cisplatin resistance and tumor recurrence of advanced HNSCC

When treating advanced HNSCC, a cisplatin-based systemic regimen benefit patient survival. However, chemoresistance will greatly reduce the effectiveness of this approach. The identification of molecules that contribute to cisplatin resistance may potentially improve the survival. Both HB-EGF and COX-2 have been reported to increase cisplatin-resistance. Here, we have focused on the regulation of HB-EGF/COX-2 and their roles in cisplatin resistance. IHC staining was used to measure the expression levels of HB-EGF and COX-2 on the tissue microarray from 43 tissue samples of patients with advanced HNSCC. siRNA, western blot and qRT-PCR were used to dissect the regulation between EGF, Akt, COX-2, PGE2, and cisplatin sensitivity. The correlation between HB-EGF, COX2 and HNSCC progression was analyzed by the receiver operating characteristic (ROC) curve and Kaplan-Meier disease free survival. Patients of advanced HNSCC patients with increased HB-EGF and COX-2 expression have higher tumor recurrent rates that was related to cisplatin resistance. The resistance was mediated via an increased expression of HB-EGF and COX-2. The activation of Akt by either EGF or areca nut extract were able to upregulate COX-2, which would increase the expression of HB-EGF in a PGE2 dependent manner. Inhibition and knockdown of COX-2 resulted in a decrease in HB-EGF. In the tissue samples from HNSCC patients, there was a significant positive correlation between the expression of COX-2 and HB-EGF. Our results suggested that COX-2 and HB-EGF are important in development of HNSCC cisplatin resistance. These findings may help the development of new strategies for overcoming cisplatin resistance.

A phase 2 randomized study to compare short course palliative radiotherapy with short course concurrent palliative chemotherapy plus radiotherapy in advanced and unresectable head and neck cancer

BackgroundTreatment of unresectable HNSCC is not well defined and has a poor outcome. This study has been designed to address the unmet needs of such groups of patients with primary end points of (a) proportion of patients eligible for radical treatment in each arm (b) loco-regional disease control at 6months between two arms. Materials and methodsLocally advanced and unresectable HNSCC patients (except Nasopharynx and Larynx) unfit for radical treatment were randomized to arm A [short course RT alone (4Gy/#/day for 5days)] or arm B [RT as arm A+concurrent cisplatin at 6mg/m2/day IV bolus for 5days]. Those with at least PR were taken for further RT to complete biological equivalent dose of 70Gy, in both the arms. In arm B, concurrent CDDP at a dose of 40mg/m2/week was administered. Results114 patients (57 in each arm) were randomized but 111 were analyzable. 15 (27.27%) patients in arm A and 28 (50%) patients in arm B had ⩾PR (p=0.01) however patients taken for FRT were 14 (25.45%) and 26 (46.42%) in arms A and B respectively (p=0.02). Locoregional control i.e. (CR+PR) at 6months was 16.36% in arm A versus 32.14% in arm B (p=0.15). Median PFS (arm A – 3.2months, arm B – 6.2months; p=0.02) and OS (arm A – 5.9months, arm B – 10.1months; p=0.03) was significantly more in arm B. There was relative improvement in quality of life for most parameters in arm B. ConclusionConcurrent low dose CTRT can be an effective treatment modality in advanced and incurable HNSCC. However, a larger phase III trial is required.

Abstract 1036: A muscle specific protein “myoferlin” modulates IL-6/STAT3 signaling by chaperoning activated STAT3 to nucleus

Abstract Head and neck squamous cell carcinoma (HNSCC) is the sixth most frequent cancer worldwide and five-year survival rates (<50%) are among the lowest of the major cancers. Although advancements in the anti-cancer treatments including surgery, radiation and chemotherapy have increased the local control of HNSCC, the overall survival rates have not improved significantly over the last three decades. This poor outcome becomes even worse (20%, 5-year survival rate) for advanced HNSCC patients that have become resistant to standard chemoradiation treatment. Therefore, there is an urgent need to identify new therapeutic targets so that novel treatment regimen(s) can be developed to improve the therapeutic efficacy while minimizing the toxicity. Recently, we made a provocative and novel discovery in our laboratory that a muscle-specific protein, myoferlin, is markedly upregulated in head and neck cancer cells. Myoferlin is a member of ferlin family of membrane proteins and most of the studies related to myoferlin's biological and molecular function(s) were carried out on muscle cells, as it was assumed that myoferlin is predominantly present only in the muscle cells. Recent studies have shown that myoferlin is also expressed in other cell types including endothelial cells and cancer cells. In our study, we observed that in addition to being expressed in tumor cells, myoferlin levels were further elevated in highly chemoresistant cells. Furthermore, myoferlin expression was also significantly higher in patients with HPV-negative tumors that show markedly poor clinical outcome and often develop resistance to chemotherapy as compared to HPV-positive tumors that normally respond better to traditional chemotherapy. Knockdown of myoferlin in head and neck cancer cells by siRNA significantly reversed chemoresistance and markedly decreased ALDH positive cancer stem cell population. These results suggest that myoferlin might be a contributing factor in the aggressive and metastatic phenotype in HNSCC. Our mechanistic studies demonstrate a novel role for myoferlin in cancer cells. Our results showed that myoferlin is bound to EHD2 protein in the resting cancer cell. Treatment of these cancer cells with IL-6 induced STAT3 phosphorylation, which led to myoferlin dissociation from EHD2 and binding to phosphorylated STAT3. Interestingly, myoferlin knock-down did not affect STAT3 phosphorylation, but completely blocked STAT3 translocation to nucleus and expression of its target proteins including Snail and Nanog. Based on these results, we have proposed a novel model for the role of myoferlin in chaperoning phosphorylated proteins to the nucleus. Taken together, our results from this study provide a novel oncogenic function of a muscle-specific protein, myoferlin, which may lead to the development of innovative therapeutic strategies to target HNSCC. Citation Format: Arti Yadav, Bhavna Kumar, Theodoros N. Teknos, Pawan Kumar. A muscle specific protein “myoferlin” modulates IL-6/STAT3 signaling by chaperoning activated STAT3 to nucleus. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1036. doi:10.1158/1538-7445.AM2015-1036

Abstract 334: PIK3CA overexpression induces epithelial-mesenchymal transition and progression of head and neck squamous cell carcinonoma.

Abstract Head and neck squamous cell carcinoma (HNSCC) patients have a poor prognosis and invasion and metastasis are the major causes of mortality. However, the molecular mechanisms of HNSCC invasion and metastasis remain obscure. The phosphatidylinositol 3-kinase (PI3K) pathway regulates a wide range of cellular processes crucial for tumorigenesis. Amplification and mutation of PIK3CA, the gene coding for the catalytic subunit of PI3K, are among the most common genetic alterations in human HNSCC. To delineate the in vivo roles of PIK3CA during head and neck tumorigenesis, we developed a PIK3CA genetically engineered mouse model (GEMM), in which PIK3CA overexpression is specifically induced in the head and neck epithelia. While overexpression of PIK3CA alone resulted in epithelial hyperplasia, and carcinoma in situ, it was not sufficient to induce visible tumor formation. In contrast, when we applied 4NQO, a DNA adduct-forming agent widely used as a tobacco surrogate together with PIK3CA overexpression, the PIK3CA-GEMM developed ∼50% poorly differentiated HNSCC compared to ∼10% of those in the control mice. PIK3CA tumors also had increased numbers of infiltrated leukocytes and angiogenesis. In addition, either regional lymph node metastases or lung metastases was observed in ∼40% of the PIK3CA-GEMM compared to no metastases in control mice. Characterizations of PIK3CA tumors revealed that PIK3CA tumors underwent epithelial and mesenchymal transition, and formed poorly differentiated metastatic tongue SCC in an orthotopical mouse model. Molecular analysis of the PIK3CA tumors suggests that rather than AKT, PDK1 facilitates progression of PI3K-driven HNSCC, and enhanced TGFbeta signaling with increased TGFbeta1 ligand and Smad3 may further contribute to this process. Lastly, overexpression of PIK3CA is associated with HNSCC progression and metastasis, and positively correlated with PDK1 expression but negatively with AKT activation, further verified the clinical relevance of our GEMM study. In summary, our results suggest that PIK3CA oncogene drives invasion and metastasis of HNSCC through PDK1 and TGFbeta signaling, and combined targeting of these pathways will reduce tumor progression in advanced HNSCC patients with PIK3CA alterations, which comprise about ∼40% of all HNSCC patients. Citation Format: Li Du, Xi Chen, Jingping Shen, Fang Zhang, Shi-Long Lu. PIK3CA overexpression induces epithelial-mesenchymal transition and progression of head and neck squamous cell carcinonoma. [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 334. doi:10.1158/1538-7445.AM2013-334