P53-negative Tumors Research Articles

PI3K/AKT Immunoexpression Influences the Prognostic Factors of Patients Diagnosed with Oropharyngeal Squamous Cell Carcinoma.

Squamous cell carcinoma (SCC) is the most common histological type in the oropharynx with high incidence and mortality. The Pi3K-AKT signaling pathway is important for understanding prognostic factors such as metastasis, which is involved in the aetiopathogenesis of cancer and is associated with angiogenesis, progression, and cell survival. The aim of this study was to evaluate the correlation of the Pi3K/AKT pathway with prognostic factors associated with oropharyngeal SCC, including overall survival. Patients diagnosed and treated for oropharyngeal SCC at the Haroldo Juaçaba Hospital were included in the study. Clinicopathological and socio-demographic data were collected by reviewing patient records and reports. Excisional biopsies were analyzed by immunohistochemical techniques using TMA for the detection of antibodies against ANTI-AKT, ANTI-Pi3K, ANTI-p16, ANTI-Ki67, and ANTI-p53. Immunoexpression was evaluated qualitatively and quantitatively using ImageJ software, and data were correlated with patient survival. There was a significant reduction in nuclear Pi3K immunoexpression in perilesional tissue compared to primary tumors and nodal metastases (p<0.001). Cytoplasmic Pi3K was increased in primary tumors and decreased in nodal metastases (p=0.001). pAKT expression in p16-negative tumors was associated with poorer overall survival. Further studies are needed to understand how this pathway may influence tumor progression and multimodal therapies. The PI3K/AKT pathway regulates cell survival and apoptosis and is associated with the malignant transformation of oropharyngeal tumors. AKT expression in p16-negative tumors is a strong predictor of poor prognosis.

Abstract C030: The Wip1 phosphatase activator QGC-8-52 selectively sensitizes p53-deficient cancer cells to chemotherapy

Abstract PP2C serine-threonine phosphatase Wip1 is vital for tissue homeostasis, stress signaling, and the pathogenesis of various diseases, making it a promising target for cancer therapy. Inhibiting Wip1's expression or activity is a novel strategy to prevent cancer. However, traditional Wip1 suppression may not work in cancers lacking p53. Here, we evaluated a new Wip1 activator, QGC-8-52, in preclinical breast cancer models. QGC-8-52 sensitizes p53-deficient cancer cells to chemotherapy by facilitating Wip1-FOXO3a interaction and dephosphorylation of Thr487, which enhances FOXO3a's activity on the proapoptotic TRAIL gene. This effect is specific to p53-deficient cancer cells. In normal cells, Wip1 activation reduces drug-induced apoptosis by weakening upstream signaling to p53. Thus, activating Wip1 during chemotherapy boosts apoptosis in p53-negative tumors and protects normal tissues. Our findings highlight Wip1 activation as a potentially effective and safe treatment for p53-deficient cancers. Citation Format: Ke Wu, Yong Wu, Jay Vadgama, Qiao-Hong Chen, Zhi-min Huang, Qun Li. The Wip1 phosphatase activator QGC-8-52 selectively sensitizes p53-deficient cancer cells to chemotherapy [abstract]. In: Proceedings of the 17th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2024 Sep 21-24; Los Angeles, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2024;33(9 Suppl):Abstract nr C030.

TNM 8 staging system beyond p16: Double HPV/p16 status is superior to p16 alone in predicting outcome in oropharyngeal squamous cell carcinoma

PurposeThe assessment of p16INK4a (p16) in oropharyngeal squamous cell carcinoma (OPSCC) has been incorporated into tumor classification, as p16 has been shown to impact survival probability. However, a recent study demonstrated that human papillomavirus (HPV) status in addition to p16 may have a better discriminatory effect on survival probability. This study aims to determine the impact of combined evaluation of p16 and HPV on prognosis. MethodsThis was a multicenter, multinational analysis including retrospective and prospective cohorts of patients treated for primary OPSCC with curative intent, based on the data of the HNCIG-EPIC study. The primary outcome was to determine how the combined assessment of HPV and p16 status predicts prognosis of patients with OPSCC compared to p16 assessment alone. We employed multivariable analyses models to compute hazard ratios regarding survival. Analyses were stratified by stage, smoking status, and sub-anatomical region. ResultsThe study included 7654 patients, with approximately half of the tumors being p16-negative (50.3 %, n = 3849). A total of 9.2 % of patients had discordant p16 and HPV status (n = 704). HPV status significantly impacted overall survival and disease-free survival regardless of p16 status and across both UICC 8th stage I-II and III-IVb cancers. p16-positive/HPV-positive OPSCC patients exhibited the best survival probability. ConclusionThe detection of HPV had a significant impact on survival probability for OPSCC patients with both p16-positive and p16-negative tumors. HPV testing should be integrated in cancer staging, especially in regions of low attributable fraction, alongside p16 evaluation to ensure a comprehensive assessment of prognosis.

The multicenter registry "oropharyngeal cancer" of the German ENT Study Center - First Results

The number of oropharyngeal squamous cell carcinomas (OPSCC) caused by human papillomavirus (HPV) has increased significantly in recent years. However, no robust data exist on the incidence of HPV-associated OPSCC as well as their treatment in central Europe. The aim is to establish an epidemiological register of patients with OPSCC in central Europe. To close the evidence gap on p16-positive OPSCC, a prospective multicenter registry on incidence, risk factors and treatment of OPSCC in certified Head and Neck Tumor Centers according to the German Cancer Society was established. Data collection was pseudonymously performed using the web-based Research Electronic Data Capture (REDCap). In 2022 and 2023, 1312 patients with OPSCC from 34 centers were included. 720 patients (58.2 %) were p16-positive. The most common locations of p16-positive primary tumors were the tonsil (49.9 %) and the base of the tongue (28.7 %). 63.1 % of p16-positive OPSCC received primary surgical treatment, whereas only 48.0 % of p16-negative tumors received primary surgery. Differences are also evident in the adjuvant therapy: 58 % of p16-negative OPSCC did not receive adjuvant therapy after surgery, compared with 37.1 % of p16-positive OPSCC. The start of the OPSCC registry study was technically smooth. This registry provides an insight into the current care situation of OPSCC in German-speaking countries and will gain importance through the participation of more centers.

Atox1-cyclin D1 loop activity is critical for survival of tumor cells with inactivated TP53.

The search for relevant molecular targets is one of the main tasks of modern tumor chemotherapy. To successfully achieve this, it is necessary to have the most complete understanding of the functioning of a transcriptional apparatus of the cell, particularly related to proliferation. The p53 protein plays an important role in regulating processes such as apoptosis, repair, and cell division, and the loss of its functionality often accompanies various types of tumors and contributes to the development of chemoresistance. Additionally, the proliferative activity of tumor cells is closely related to the metabolism of transition metals. For example, the metallochaperone Atox1 - a copper transporter protein - acts as a transcription activator for cyclin D1, promoting progression through the G1/S phase of the cell cycle. On the other hand, p53 suppresses cyclin D1 at the transcriptional level, thereby these proteins have divergent effects on cell cycle progression. However, the contribution of the interaction between these proteins to cell survival is poorly understood. This work demonstrates that not only exists a positive feedback loop between Atox1 and cyclin D1 but also that the activity of this loop depends on the status of the TP53 gene. Upon inactivation of TP53 in A549 and HepG2 cell lines, the expression of ATOX1 and CCND1 genes is enhanced, and their suppression in these cells leads to pronounced apoptosis. This fundamental observation may be useful in selecting more precise interventions for combined therapy of p53-negative tumors.

Association of absolute neutrophil count with outcomes in patients with head and neck cancer treated with chemoradiation.

e18055 Background: Systematic inflammation plays an integral role in tumor development and metastasis. High neutrophil counts, among various peripheral blood biomarkers, have been shown to be associated with worse survival outcomes in multiple cancer types. To evaluate the absolute neutrophil count (ANC) as a prognostic biomarker in head and neck cancer, we performed an observational cohort study of patients undergoing definitive chemoradiation. Methods: Our institutional database at the Roswell Park Comprehensive Cancer Center was queried for patients with non-metastatic head and neck cancer who underwent definitive chemoradiation between June 2007 and April 2023. ANC was analyzed as a continuous variable. Survival outcomes were assessed using Kaplan-Meier method, log-rank tests, Cox proportional hazard multivariable (MVA) analyses. Fine-Gray MVA was used to analyze failure outcomes with death as a competing event. To reduce selection bias, propensity score matching was performed using ANC as a dichotomous variable with its median value as a cutoff. Subgroup analyses were performed among p16-negative and p16-positive cohorts. Results: A total of 668 patients who met our criteria. Median follow up was 30.3 months (interquartile range 9.9-60.7). Elevated ANC as a continuous variable was associated with worse overall survival (OS; adjusted hazards ratio [aHR] 1.02, 95% confidence interval [CI] 1.01-1.04, p=0.003), progression free survival (PFS; aHR 1.03, 95% CI 1.01-1.04, p&lt;0.001), and distant failure (DF; aHR 1.03, 95% CI 1.01-1.05, p=0.007), but not locoregional failure (LRF; aHR 1.02, 95% CI 1.00-1.05, p=0.10). Similar findings were noted among 278 matched pairs (OS: hazards ratio [HR] 1.66, 95% CI 1.23-2.26, p=0.001; PFS: HR 1.74, 95% CI 1.31-2.29, p&lt;0.001; LRF: HR 1.66, 95% CI 0.98-2.80, p=0.06; DF: HR 1.64, 95% CI 1.03-2.60, p=0.04). On subgroup analysis, among 118 patients with p16-negative tumors, elevated ANC as a continuous variable was not associated with OS (aHR 1.00, 95% CI 0.90-1.11, p=0.97) and PFS (aHR 0.97, 95% CI 0.87-1.08, p=0.54). The number of LRF (n=21) and DF (n=17) were too few to analyze in this subgroup. However, among 344 patients with p16-positive tumors, high ANC was associated with worse OS (aHR 1.03, 95% CI 1.00-1.06, p=0.046), PFS (aHR 1.05, 95% CI 1.02-1.07, p&lt;0.001), LRF (aHR 1.08, 95% CI 1.05-1.12, p&lt;0.001), and DF (aHR 1.04, 95% CI 1.01-1.08, p=0.02). Conclusions: Our study suggested that elevated ANC was an independent, adverse prognostic factor for worse survival and distant metastasis outcomes. Similar findings were also observed among patients with p16-positive tumors, with high ANC also associated with worse locoregional failure. Further studies are warranted to tailor interventions based on such biomarkers.

Ten-year (2009–2019) epidemiological study of head and neck, salivary glands and upper aerodigestive tract cancers, and overall survival outcomes in the Northern Territory of Australia

BackgroundHead and neck, salivary glands and aerodigestive tract cancers (HNACs) rank sixth in cancer incidence in Australia, posing significant public health and economic challenges. However, data on HNACs in the...

P53 expression correlates with low axillary tumor burden in breast cancer.

The p53 mutation in breast cancer confers a worse prognosis and is usually associated with p53 overexpression (p53+) on immunohistochemistry. Previous studies have shown that p53+ tumors could be associated with low axillary tumor burden (ATB). We aimed to evaluate the association between p53+ and ATB in a large series of breast cancers as an aid to personalizing axillary surgical treatment. We retrieved 1762 infiltrating breast carcinomas from our database that were treated with upfront surgery in Hospital del Mar from 2004 to 2018. We compared p53+ and p53-negative (p53-) tumors in terms of the percentage of cases with high ATB and overall survival. This comparison was made overall and for each immunophenotype. Overall, 18.7% of breast tumors were p53+. High ATB was less common in p53+ tumors than in p53- tumors in the luminal B-Her2-negative immunophenotype (6.2% versus 16.9%, respectively, P =0.025), but not in the other immunophenotypes or overall. Overall survival was worse in patients with p53+ breast cancer (P =0.002). p53+ breast cancers were associated with worse overall survival. However, low ATB was more common in these tumors than in p53- tumors in the luminal B-Her2-negative subtype. Information on p53 expression could be of use to predict ATB in some breast cancer tumors.

Importance of tumor volume, overall treatment time and fractionation sensitivity for p16-positive and p16-negative oropharyngeal tumors

Background Analyses of clinical outcomes following radiotherapy (RT) have advanced our understanding of fundamental radiobiological characteristics in head and neck squamous cell carcinoma (HNSCC). Low fractionation sensitivity appears to be a common feature, as well as susceptibility to changes in overall treatment time (OTT). Large tumors should be harder to cure if a successful RT requires the sterilization of all clonogenic cells. Congruently, primary tumor volume has proven to be an important parameter. However, most findings come from an era when p16-negative HNSCC was the dominant tumor type. HPV-associated, p16-positive, oropharyngeal tumors (OPSCC) are more radiosensitive and have better outcome. The current study aims to investigate the role of primary tumor volume, OTT and estimate α-ratio for p16-positive OPSCC, and to quantify the differences in radiosensitivity depending on p16-status. Methods A cohort of 523 patients treated with RT was studied using a tumor control probability (TCP)-model that incorporates primary tumor volume (V) raised to an exponent c, OTT and α-estimation. The significance of V was also investigated in Cox-regression models. Results In the p16-positive cohort (n = 433), the volume exponent was 1.44 (95%CI 1.06–1.91), compared to 0.90 (0.54–1.32) for p16-negative tumors (n = 90). Hazard ratios per tumor volume doubling were 2.37 (1.72–3.28) and 1.83 (1.28–2.62) for p16-positive and p16-negative, respectively. The estimated α-ratio was 9.7 Gy (−2.3–21.6), and a non-significant daily loss of 0.30 Gy (−0.17–0.92) was found. An additional dose of 6.8 Gy (interquartile range 4.8-9.1) may theoretically counteract the more radioresistant behavior of p16-negative tumors. Conclusion Primary tumor volume plays a crucial role in predicting local tumor response, particularly in p16-positive OPSCC. The estimated α/β-ratio for p16-positive oropharyngeal tumors aligns with previous HNSCC studies, whereas the impact of prolonged OTT was slightly less than previously reported. The differences in radiosensitivity depending on p16-status were quantified. The findings should be validated in independent cohorts.

Role of 18F-FDG PET/CT in evaluating lymph node status in patients with head and neck squamous cell carcinoma.

The presence of cervical lymph node metastases (CLNM) at diagnosis is one of the most relevant negative prognostic factors in patients with head and neck squamous cell carcinoma (HNSCC). The aim of this study was to analyse 2-deoxy-2[18F]fluoro-D-glucose (FDG) PET/CT findings for the identification of primary tumours and CLNM in a sample of patients affected by HNSCC. Moreover, a maximum standardised uptake value (SUVmax) threshold for the detection of CLNM was estimated. Clinical variables (i.e. smoking and alcohol habits), and tumour features (i.e. EBV and HPV positivity) were also evaluated in relation to FDG PET/CT findings. We retrospectively analysed patients who underwent FDG PET/CT for HNSCC staging between 2015-2020 at the University Hospital of Ferrara. All patients had cytological or histological confirmation of suspected cervical lymph nodes. In total, 65 patients were enrolled (53 males, 12 females, median age 65.7 years). CLNM of patients with smoking habit had significantly higher SUVmax values than those of patients with previous smoking habit and non-smokers (p = 0.04). p16 positive HNSCC demonstrated a trend for higher SUVmax values on CLNM, in comparison to p16 negative tumours (p = 0.089). ROC curve analysis identified 5.8 as the best cut-off value of SUVmax for the detection of CLNM (AUC = 0.62, sensitivity 71.4% and specificity 72.7%). FDG PET/CT is a useful tool to evaluate CLNM in patients with HNSCC, particularly in those with smoking habit and p16 positive disease. A SUVmax cut-off of 5.8, combined with the use of conventional radiological investigations, may represent a useful tool in the identification of CLNM.

P16 expression in urothelial carcinoma: Experience from a tertiary care center in coastal South India.

Urothelial carcinomas (UC) account for 6 and 2% of all cancers in men and women, respectively. Human papillomavirus (HPV) is one of the causative agents in cancers of the uterine cervix and head and neck. The role of HPV is also being studied in cancers of the urinary bladder, penis, and prostate. As p16-INK4a is a surrogate marker for high-risk HPVE7 oncoprotein, this study aims to highlight the utility of p16 immunohistochemistry (IHC) in the evaluation of HPV-associated UC. A retrospective study was conducted on UC of the bladder received in the Pathology department between January 2013 and December 2018. Bladder biopsies from non-neoplastic lesions served as controls. IHC was done for the detection of the p16 antigen. The p16 staining was recorded as positive, when there was strong staining in >50% of tumor nuclei. The p16 positive and negative tumors were compared based on age, gender, tumor size, grade, and muscle invasion. P value <0.05 was considered statistically significant. The expression of p16 was analyzed in 72 UC and compared with 20 non-neoplastic cases, of which 26.4% of the cases showed p16 expression. The p16 expression was absent in the non-neoplastic lesions. While the majority (87.5%) of the low-grade tumors were negative for p16 expression, 43.8% high-grade tumors were positive. Similarly, a larger proportion of invasive carcinomas (38.8%) expressed p16 as compared to non-invasive carcinomas (13.8%). Thus, p16 expression showed a significant association with grade and stage in these malignancies (P < 0.05). The p16 expression was associated with high-grade and muscle-invasive UC. The p16 was absent in all non-neoplastic and precursor lesions. Thus, it can provide essential information not only about HPV association but also on the prognostic implications for the patients.

Sinonasal Adenosquamous Carcinoma - Morphology and Genetic Drivers Including Low- and High-Risk Human Papillomavirus mRNA, DEK::AFF2 Fusion, and MAML2 Rearrangement.

Sinonasal adenosquamous carcinoma is rare, and there are almost no studies detailing morphology or characterizing their genetic driver events. Further, many authors have termed sinonasal tumors with combined squamous carcinoma and glands as mucoepidermoid carcinoma but none have analyzed for the presence of MAML2 rearrangement. Cases from 2014 to 2020 were collected and diagnosed using World Health Organization criteria. They were tested for p16 expression by immunohistochemistry (70% cut-off), DEK::AFF2 fusion by fluorescence in situ hybridization (FISH) and AFF2 immunohistochemistry, MAML2 rearrangement by FISH, and low- and high-risk HPV by RNA ISH and reverse transcription PCR, respectively. Detailed morphology and clinical features were reviewed. There were 7 male (64%) and 4 female (36%) patients with a median age of 69 years, most Caucasian (10 of 11 or 91%). Most had tobacco exposure (8/11, 73%) and most presented with epistaxis, a visible nasal mass, and/or facial pain. Several had a precursor papillomas (3 of 11, 27%). The squamous component had variable keratinization, 5 of 11 (46%) of which would be described as keratinizing, 3 non-keratinizing, and 2 with mixed features. All had gland formation, by definition, and 2 of 11 (18%) had ciliated tumor cells. None of the 11 cases had MAML2 rearrangement and one had DEK::AFF2 fusion with associated positive nuclear AFF2 protein immunostaining. Most were p16 positive (7 of 11, 64%) and all 7 of these were hrHPV positive either by RNA ISH or RT-PCR. Two of the p16-negative tumors were positive for lrHPV by RNA ISH. Treatment included surgery alone (4 of 11, 36%), surgery with adjuvant radiation (5 of 11, 45%), and surgery with radiation and chemotherapy (2 of 11, 18%). Four of 11 patients (36%) suffered disease recurrence, two requiring re-operation and who were disease free at last follow-up, one receiving additional chemotherapy and who was alive with disease. The other elected to undergo palliative therapy and died of disease. Sinonasal adenosquamous carcinoma is a somewhat heterogeneous tumor not infrequently arising ex papilloma and having various drivers including high- and low-risk HPV and rarely DEK::AFF2 fusion. The prognosis appears favorable when proper treatment is possible.

Gastrostomy Tube Use in Head and Neck Cancer Survivors, Pre-Treatment Factors Supersede Treatment Intensification: A Second Analysis of RTOG 0129, RTOG 0522, and NRG/RTOG 1016

<h3>Purpose/Objective(s)</h3> Clinical and treatment factors that engender long-term gastrostomy tube (g-tube) dependence are poorly defined. We attempted to identify potential predictors. <h3>Materials/Methods</h3> All eligible patients treated on RTOG 0129, RTOG 0522, and NRG/RTOG 1016 were grouped according to treatment received: standard fractionation RT (3DSFX) + cisplatin (C), accelerated fractionation RT (3DAFX) + C, accelerated IMRT (AFX) + C, AFX + C + cetuximab (cetux) and AFX + cetux. G-tube rates were compared at baseline (pre-treatment), 6-months after radiation completion, and annually from years 1 to 9 post treatment. Time to g- tube placement was analyzed using Cox proportional hazards models for variables of interest. Additionally, a separate review was performed for patients with no g-tube at 3 years' post-treatment (survivors with acceptable post-treatment swallowing) to evaluate late g-tube placement. <h3>Results</h3> 2389 patients were identified, 1839 of which had oropharynx cancer (OPC). AFX+C was the most common management (n=843, 35%). The G-tube rate at treatment initiation was 14%. At one-year post-treatment the rate was 19% and at 9 years the number was 8%. The RTOG 0129 g-tube rate was significantly higher than that of later studies (RTOG 0522 HR 0.7, 0.6-0.8; NRG/RTOG1016 HR 0.85 (0.72-0.99). Patients treated with 3D regimens had significantly higher feeding tube rate at baseline, 6 months, 1 year, and 2 years post-treatment (3DSFX + C: 25%/45%/29%/13% and 3DAFX + C: 22%/41%/26%/17%, respectively) when compared to accelerated IMRT regimens (all time points p < 0.001). There was no difference in feeding tube rate between treatment groups from years 3 – 9. Among OPC patients the predictors of feeding tube placement were older age (HR 1.01, 1.00-1.02), T4 primary (1.3, 1.1-1.6) and >10 pack year smoking history (HR 1.2, 1.0-1.4). Treatment regimen, tumor location, and p16-status did not affect g-tube placement. Among all patients, predictors of g-tube placement were older age (HR 1.01, 1.01-1.02); T4 primary (HR 1.3, 1.2-1.5), AJCC 7^th ed N2c/N3 (1.3, 1.1-1.6); and >10 pack year smoking history (HR 1.2, 1.0-1.3). At 3 years there were 989 patients alive with follow-up who never required a gastrostomy tube. Of the 249 of these patients with 9 years' follow-up the g-tube rate was 6%. From univariable model predictors of feeding tubes placed more than 5 years after treatment completion were T3 category, T4 category, >10 pack years smoking, non-tonsil or tongue base primary, and p16-negative tumors. <h3>Conclusion</h3> G-tube rates were highest in patients managed with 3DCRT on RTOG 0129; utilization has significantly decreased over time. Radiation acceleration treatment intensification does not increase g-tube use during treatment or survivorship. Predictors of feeding tube use are increasing age, T4 primary tumors and >10 pack year smoking history.

Effect of p16 Status on Survival Outcomes in Sinonasal Squamous Cell Carcinoma.

Evaluate the effect of p16 status on disease-free survival (DFS) and overall survival (OS) in patients with sinonasal squamous cell carcinoma (SCC) undergoing treatment with curative intent; and to assess how p16 status may affect patterns of recurrence. Retrospective cohort study. Tertiary medical center. Patients with sinonasal SCC treated with curative intent from 2012 to 2018 were identified. Independent variable of interest was p16 status, which was assessed using immunohistochemistry (IHC) with a 70% staining cutoff for positivity. Kaplan Meier survival curve was plotted to assess correlation between p16 status and DFS and OS. Association between recurrence patterns and p16 status was conducted using chi square and fisher's exact tests. Multivariable Cox proportional hazard analysis was conducted to assess association between independent variables and DFS. Fifty patients with sinonasal SCC met inclusion criteria. Patients were p16 positive in 28/50 (56%) of cases. Kaplan Meier survival curve revealed no statistically significant association between p16 status and DFS or OS survival (P = .780, P = .474). There was no difference in recurrence patterns in patients with p16 positive versus negative tumors. p16 status did not have prognostic value on DFS and OS in our cohort of patients with sinonasal SCC undergoing treatment with curative intent. There was no difference in recurrence patterns between the 2 populations. Based on the results of this study, p16 status should not impact counseling of patients as it relates to their prognosis from SNM.

Primary tumor volume and prognosis for patients with p16-positive and p16-negative oropharyngeal squamous cell carcinoma treated with radiation therapy

BackgroundThe prescribed radiation dose to patients with oropharyngeal squamous cell carcinoma (OPSCC) is standardized, even if the prognosis for individual patients may differ. Easy-at-hand pre-treatment risk stratification methods are valuable to individualize therapy. In the current study we assessed the prognostic impact of primary tumor volume for p16-positive and p16-negative tumors and in relationship to other prognostic factors for outcome in patients with OPSCC treated with primary radiation therapy (RT).MethodsFive hundred twenty-three OPSCC patients with p16-status treated with primary RT (68.0 Gy to 73.1 Gy in 7 weeks, or 68.0 Gy in 4.5 weeks), with or without concurrent chemotherapy, within three prospective trials were included in the study. Local failure (LF), progression free survival (PFS) and overall survival (OS) in relationship to the size of the primary gross tumor volume (GTV-T) and other prognostic factors were investigated. Efficiency of intensified RT (RT with total dose 73.1 Gy or given within 4.5 weeks) was analyzed in relationship to tumor volume.ResultsThe volume of GTV-T and p16-status were found to be the strongest prognostic markers for LF, PFS and OS. For p16-positive tumors, an increase in tumor volume had a significantly higher negative prognostic impact compared with p16-negative tumors. Within a T-classification, patients with a smaller tumor, compared with a larger tumor, had a better prognosis. The importance of tumor volume remained after adjusting for nodal status, age, performance status, smoking status, sex, and hemoglobin-level. The adjusted hazard ratio for OS per cm3 increase in tumor volume was 2.3% (95% CI 0–4.9) for p16-positive and 1.3% (95% 0.3–2.2) for p16-negative. Exploratory analyses suggested that intensified RT could mitigate the negative impact of a large tumor volume.ConclusionsOutcome for patients with OPSCC treated with RT is largely determined by tumor volume, even when adjusting for other established prognostic factors. Tumor volume is significantly more influential for patients with p16-positive tumors. Patients with large tumor volumes might benefit by intensified RT to improve survival.

Laser tongue base mucosectomy is a useful diagnostic tool in the management of unknown primary cancers of the head and neck region.

Tongue base mucosectomy identified cancer in 78 per cent of cancers of unknown primary in a recent meta-analysis. The carbon dioxide laser is an alternative technique if there is no access to a robot. This study aimed to describe the steps for undertaking tongue base mucosectomy using the carbon dioxide laser and its diagnostic utility in cancers of unknown primary. This was a prospective feasibility study utilising carbon dioxide laser for tongue base mucosectomy in cancers of unknown primary. Data collected included demographic data and p16 status. There were 14 cancers of unknown primary with 86 per cent p16 positivity on immunohistochemistry. Laser tongue base mucosectomy alone identified the cancer primary in 7 of 12 (58 per cent) cancers of unknown primary among p16 positive tumours and 0 of 2 (0 per cent) among p16 negative tumours. Combining bilateral tonsillectomy with laser tongue base mucosectomy resulted in identification of the primary cancer in 8 of 12 (67 per cent) p16 positive tumours. In centres without a robot, tongue base mucosectomy using the carbon dioxide laser is a viable alternative, especially in combination with bilateral tonsillectomy in p16 positive cases.

Pathologic Comparison of Locally Advanced p16-Negative and p16-Positive Larynx Squamous Cell Carcinoma

<h3>Purpose/Objective(s)</h3> The significance of p16 as a biomarker in the management of squamous cell carcinoma of the larynx is controversial. Large database analyses demonstrate that patients with p16-positive disease exhibit more favorable outcomes compared to those with tumors negative for p16. The reasons underlying this association are unknown. Here, we examined the relationship between pathologic findings and p16-status. <h3>Materials/Methods</h3> We performed an IRB-approved retrospective study of patients with locally advanced, treatment-naïve, larynx cancer who underwent total laryngectomy. IHC was performed. Patients that exhibited 70% or greater staining were considered positive. The chi-squared test was used to analyze disease and patient characteristics. <h3>Results</h3> We identified 46 patients that met our selection criteria. The patient population was predominantly Caucasian (78%) with a median age of 62 years at treatment. Seven percent had pathologic stage I disease, 13% stage II, 21% stage III, 41% stage IVA and 17% stage IVB. Twenty percent (9/46) were positive for p16 over-expression. Pathologic nodal disease was present in 33% (3/9) and 27% (10/37) of patients with p16-negative and p16-positive disease respectively (p=0.7) Extranodal extension was identified in 33% (1/3) of patients with p16-positive tumors who had nodal disease and 90% (9/10) of patients with p16-negative disease (p=0.04). Sixty-eight percent (n=15) of p16-negative tumors exhibited lymphovascular space invasion compared to 11% (n=1) of p16-positive tumors (p=0.09). Grade 2/3 tumors were identified in 84% (n=31) of p16-negative tumors compared to 55% (n=5) of p16-positive tumors (p=0.06). <h3>Conclusion</h3> We identified a statistically significant association of extranodal extension with p16-negative larynx cancers that may contribute worse overall survival of this cohort. Additional studies are warranted to confirm these findings.

The Vulvar Immunohistochemical Panel (VIP) Project: Molecular Profiles of Vulvar Squamous Cell Carcinoma.

Simple SummaryThis study investigated the immunohistochemical expression of 14 biological markers as potential prognostic/therapeutic factors in vulvar squamous cell carcinoma, comparing 53 node-negative (Group A) and 48 node-positive (Group B) patients. Our results show a significantly higher p16 expression (surrogate of HPV-related tumors) in the vulvar samples of non-metastatic patients. In Group B, PD-L1 positivity and high EGFR expression were found in the vast majority of vulvar and/or nodal specimens. VEGF showed strong/moderate-diffuse expression in almost 14% of all vulvar samples. A mutated p53 and over-expressed PD-L1 showed a significant association with nodal metastasis. Our results support a potential role of immune checkpoint inhibitors and anti-VEGF and anti-EGFR drugs, especially in patients with worse prognosis (metastatic, HPV-independent). A panel including EGFR, VEGF, PDL1, p16, and p53 might be performed routinely in primary tumor and repeated in case of lymph node metastases to identify changes in marker expression.Introduction: The study’s aim was to investigate the immunohistochemical (IHC) expression of biological markers as potential prognostic/therapeutic factors in vulvar squamous cell carcinoma (VSCC). Methodology: A series of 101 patients surgically treated at our center from 2016 to 2020 were retrospectively enrolled: 53 node-negative (Group A) and 48 node-positive (Group B). A total of 146 samples, 101 from primary tumor (T) and 45 from nodal metastases (N), were investigated. The IHC panel included: p16, p53, MLH1, MSH2, MSH6, PMS2, PD-L1, CD3, HER2/neu, ER, PR, EGFR, VEGF, and CD31. The reactions were evaluated on qualitative and semi-quantitative scales. Generalized Linear Model (GLM) and cluster analysis were performed in R statistical environment. A distance plot compared the IHC panel of T with the correspondent N. Results: In Group A: p16-positive expression (surrogate of HPV-dependent pathway) was significantly higher (20.8% vs. 6.2%, p = 0.04). In Group B: PD-L1 positivity and high EGFR expression were found, respectively, in 77.1% and 97.9% patients (T and/or N). Overall, p16-negative tumors showed a higher PD-L1 expression (60.9% vs. 50.0%). In both groups: tumoral immune infiltration (CD3 expression) was mainly moderate/intense (80% vs. 95%); VEGF showed strong/moderate-diffuse expression in 13.9% of T samples; CD31, related to tumoral microvessel density (MVD), showed no difference between groups; a mutated p53 and over-expressed PD-L1 showed significant association with nodal metastasis, with Odds Ratios (OR) of 4.26 (CI 95% = 1.14–15.87, p = 0.03) and 2.68 (CI 95% = 1.0–7.19, p < 0.05), respectively; since all mismatch repair proteins (MMR) showed a retained expression and ER, PR, and HER2/neu were negative, they were excluded from further analysis. The cluster analysis identified three and four sub-groups of molecular profiles, respectively, in Group A and B, with no difference in prognosis. The molecular signature of each N and corresponding T diverged significantly in 18/41 (43.9%) cases. Conclusions: Our results support a potential role of immune checkpoint inhibitors and anti-VEGF and anti-EGFR drugs especially in patients with worse prognosis (metastatic, HPV-independent). A panel including EGFR, VEGF, PDL1, p16, and p53 might be performed routinely in primary tumor and repeated in case of lymph node metastases to identify changes in marker expression.

Abstract 2045: Aneuploidy, immunohistochemical overexpression of MSH-2, P53, P21^WAF1/CIP1, P16^INK4A, and correlation to epithelial ovarian carcinoma patient prognosis

Abstract Aneuploidy, immunohistochemical overexpression of MSH-2, P53, P21WAF1/CIP1, P16INK4A, and correlation to epithelial ovarian carcinoma patient prognosis Overexpression of MSH-2, P53, P21WAF1/CIP1, P16INK4A and DNA status in epithelial ovarian carcinoma (EOC) was determined in paraffin-embedded tissue (PET) to predict prognosis. Immunoreactivity, DNA and % S content were evaluated for their potential role in survival and relapse-free survival (RFS) in 112 patients diagnosed with EOC at Saskatoon Cancer Center, Canada. Immunostaining studies were with the following mouse monoclonal antibodies: MSH-2, FE11; P53, D07; P21WAF1/CIP1, EA 10; except for P16INK4A, rabbit, G175-405, 13251A. DNA and % S content (median, &amp;lt; % 5 = low-risk or &amp;gt; % 5 = high-risk) were assessed by flow-cytometric analysis1. Survival curves were determined by Kaplan-Meier method, mean survival time, and log rank test. P-values &amp;lt; 0.05 were considered significant; X2-test examined association between variables. Patients with aneuploid DNA had 59 months mean survival time, compared to cases with diploid DNA which had 89 months, (p = 0.03). RFS in those with diploid DNA lasted longer (84 months) while aneuploid cases had shorter time to progression (47 months, p = 0.01). % S content in the high-risk group, 47 cases, had worse survival (57 months, p = 0.0008) and RFS (52 months, p = 0.004). Aneuploidy was not related to overall survival within the subsets of early and advanced stage patients. Grade 2 patients with abnormal DNA and P53 expression were adversely associated with survival and RFS. Diploid DNA was observed in all mucinous tumors except in one case with aneuploid DNA. P53 immunoreactivity was related to shorter survival and RFS in 44 cases. P21WAF1/CIP1 overexpression was associated with P53 overexpression in 11 cases (p = 0.04) and shorter median survival (29 months) compared to negative P21WAF1/CIP1 staining (median survival time 62 months, p = 0.03). 11 grade 2 patients with P53 positivity had worse survival (59 months) and RFS (47 months) than the P53 negative tumors (p = 0.05, and p = 0.01 respectively). P16INK4A overexpression (46 cases) had no adverse effect on survival and RFS. No significant prognostic difference was found between MSH-2 positive and negative tumors. We conclude that % S content was a more beneficial prognostic marker than ploidy status. In grade 2 EOC, the effects of DNA content and P53 overexpression were significant variables related to outcome. For pathologists, these variables in grade 2 tumors can be adjunctive aid for reaching an agreement to diagnose the grade of EOC patients.Footnotes1 A. Geissel and J. L. Griffin, “Preparation of nuclei for flow cytometry,” in AFIP Advances in Laboratory Methods in histology and Pathology, ed. Mikel UV (Washington DC: American Registry of Pathology, 1944), 111-121. Citation Format: Kavitha Advikolanu Rao, Antony M. Magliocco, Andrew W. Maksymiuk. Aneuploidy, immunohistochemical overexpression of MSH-2, P53, P21^WAF1/CIP1, P16^INK4A, and correlation to epithelial ovarian carcinoma patient prognosis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2045.

Co-expression of fibroblast growth factor receptor 3 with mutant p53, and its association with worse outcome in oropharyngeal squamous cell carcinoma.

Fibroblast growth factor receptor 3 (FGFR3) is expressed in squamous cell carcinoma of the head and neck (SCCHN) including oropharyngeal squamous cell carcinoma (OPSCC) and is a potential therapeutic target. However, information on its correlation with other relevant cancer related proteins stratified by p16 status and its prognostic significance in OPSCC is limited. We examined FGFR3 expression and its correlation with clinical characteristics, p16 status, and mutant p53 (mp53) among 220 retrospectively collected OPSCC cases and 40 prospectively collected SCCHN cases, including a majority of OPSCC. Correlations of FGFR3 Weighted Index (WI) with p16 status and mp53 WI as well as its association with disease-free survival (DFS) and overall survival (OS) were evaluated. FGFR3 expression was detected in 61% and 70% of cases in cohorts 1 and 2, respectively. FGFR3 level was significantly higher in p16-negative tumors in both cohorts (p<0.001 and 0.006). FGFR3 expression was highly correlated with mp53 expression in both p16 + and p16– OPSCC (p<0.0001 and p = 0.0006, respectively). In cohort 1, univariate analysis showed that FGFR3 was associated with DFS but not OS. Kaplan-Meier analysis showed that higher FGFR3 and mp53 level correlated with worse DFS (p = 0.025) and OS (p = 0.009). As expected, p16 positive status was associated with improved OS and DFS (p<0.001 for both). Our results suggest that high FGFR3 expression is associated with p16 negative status and mp53 expression in OPSCC and correlates with a worse clinical outcome. The biological relationship between FGFR3 and mp53 in OPSCC deserves further investigation.