Reliable PCR quantitation of estrogen, progesterone and ERBB2 receptor mRNA from formalin-fixed, paraffin-embedded tissue is independent of prior macro-dissection

Gene expression analysis on messenger RNA (mRNA) purified from formalin-fixed, paraffin-embedded tissue is increasingly used for research purposes. Tissue heterogeneity may question specificity and interpretation of results from mRNA isolated from a whole slide section, and thresholds for minimal tumor content in the paraffin block or macrodissection are used to avoid contamination from non-neoplastic tissue. The aim was to test if mRNA from tissue surrounding breast cancer affected quantification of estrogen receptor α (ESR1), progesterone receptor (PGR) and human epidermal growth factor receptor 2 (ERBB2), by comparing gene expression from whole slide and tumor-enriched sections, and correlating gene expression from whole slide sections with corresponding immunohistochemistry. Gene expression, based on mRNA extracted from a training set (36 paraffin blocks) and two validation sets (133 + 1,083 blocks), were determined by quantitative reverse transcription polymerase chain reaction for all samples, as well as by microarray for 133 validation samples. In the training set, agreement between high vs. low mRNA expression from whole slide and tumor-enriched sections was absolute for ESR1 and ERBB2, and 83 % for PGR. Overall agreements, when comparing mRNA expression to immunohistochemistry, were 100 % (ERBB2), 89 % (ESR1) and 83 % (PGR), which was confirmed in the validation sets. Percentage of tumor in the sections did not influence the results. In conclusion, reliable quantification of ESR1, PGR and ERBB2 mRNA expression can be obtained from a whole slide section, and correlates well with immunohistochemistry. Prior removal of surrounding tissue was found to be unnecessary even with minimal tumor content in the section

Status Report and Beam Time Request for Experiment AD-4

Summary of current status and plans for October 200

Biological Effects of Antiprotons Are Antiprotons a Candidate for Cancer Therapy?

2009 Status Report of AD-4 Experimen

Hypoxia in human soft tissue sarcomas: Adverse impact on survival and no association with p53 mutations

Clinical and experimental studies have suggested that tumour hypoxia is associated with poor treatment outcome and that loss of apoptotic potential may play a role in malignant progression of neoplastic cells. The tumour suppressor gene p53 induces apoptosis under certain conditions and microenvironmental tumour hypoxia may select for mutant tumour cells with diminished apoptotic potential due to lack of p53 function. The aim of this study was to evaluate the prognostic relevance of oxygenation status for treatment outcome and to compare pre-treatment tumour oxygenation measurements were done in 31 of those by PCR using DNA extracted from paraffin-embaedded sections (n = 2) or frozen biopsies (n = 29). The overall median of the tumour median pO 2 was 19 mmHg (range 1–58 mmHg). Only 6 tumours had functional p53 mutations and no association was found between mutant p53 and tumour hypoxia. Five out of 6 STS with lower histopathological grade were well-oxygenated whereas high-grade STS were both hypoxic and well-oxygenated. At a median follow-up of 74 months, 16 patients were still alive among 28 available for survival analysis. When stratifying into hypoxic and well-oxygenated tumours patients with the most hypoxic tumours has a statistically poorer disease-specific and overall survival at 5 years. In conclusion hypoxia was an indicator for both a poorer disease specific and overall survival in human STS but hypoxic tumours were not characterized by mutations in the p53 gene. © 2001 Cancer Research Campaign http://www.bjcancer.co

Subsite variation of HPV-related p16-expression in oropharynx cancer:Incidence and prognostic impact in a population-based DAHANCA cohort 1986–2020

Background and purpose: Separate staging criteria based on human papillomavirus (HPV)-related p16-expression are implemented in the TNM8 classification of oropharyngeal cancer (OPSCC). Based on a nationwide cohort, we provide a detailed description of subsite variation in the age-standardised inci-dence-rates of OPSCC alongside an evaluation of the prognostic impact of p16-expression according to subsite after primary radiotherapy (RT). Patient/material and methods: A total of 8,462 Danish OPSCC patients from 1986 to 2020 were identified in the DAHANCA-database, and tumours were grouped into ‘tonsil/base of tongue (BOT)’, ‘neighbouring subsites’ and ‘distant subsites’. Subsite-specific age-standardised incidence-rates were calculated, and out-come-analysis (loco-regional control, disease-free survival and overall-survival 5 years after the completion of RT) stratified by p16-status/subsite and restricted to curatively treated patients only (N = 3,387) was performed. Results: A 5-fold increase in the age-standardised incidence of OPSCC was observed and could be ascribed to the rise in p16-positive tumours of tonsil/BOT and neighbouring subsites only as neither the incidence rates nor the proportion of p16-positivity in distant subsites tumours changed over time. The prognostic impact of p16-status for all endpoints differed significantly across tumour subsites with the strongest association found in tonsil/BOT tumours, a diminishing but still significant impact in neighbouring subsite tumours and no significant impact in tumours arising in distant subsites. Interpretation: Our findings suggest that grouping all p16-positive OPSCC as one entity for staging and prognostication, as currently done in TNM8, is too simple as it does not accurately depict the differences in tumour biology and the consequent treatment response.</p

DAHANCA19:A randomized phase III study of primary curative (chemo)-radiotherapy and the EGFR-inhibitor zalutumumab for squamous cell carcinoma of the head and neck

Background and purpose: Antibodies against the Epidermal Growth Factor receptor is suggested to decrease tumour failure and increase survival rates of patients (pts) with Head and Neck Squamous Cell Carcinomas (HNSCC) when combined with radiotherapy. This study aimed to evaluate if concurrent treatment with the EGFR inhibitor zalutumumab during (chemo-)radiotherapy improved outcome in pts with HNSCC. Materials and methods: Overall, 608 eligible pts with biopsy-verified HNSCC of the oral cavity, pharynx and larynx were accrued November 2007 to June 2012. Pts were randomized to a control-arm of primary accelerated radiotherapy predominantly 66–68 Gy, 2 Gy/fraction, 6fx/week and concomitant daily hypoxic radiosensitisation with nimorazole. St. III-IV carcinomas received weekly cisplatin 40 mg/m2 in addition to nimorazole. The zalutumumab-arm was identical to the control-arm plus zalutumumab 8 mg/kg. First dose was given the week before start of treatment and continued weekly during radiotherapy. Analyses were performed as intention-to-treat. Primary endpoint was loco-regional failure. Secondary endpoints were disease-specific survival and overall survival. Results: In total, 307 pts were in the control-arm and 301 in the zalutumumab-arm. Median follow-up was 59 months. Patient and tumour parameters were well balanced. The 5-year loco-regional failure rate was 24 % in the zalutumumab-arm and 18 % in the control-arm; Hazard Ratio (HR) 1.16 (95 % CI 0.84–1.59); disease-specific survival; HR 1.04 (95 % CI 0.73–1.50) and overall survival; HR 1.21, (0.91–1.61). Effect of zalutumumab was not influenced by HPV/p16 status. Conclusion: Addition of concomitant zalutumumab to primary (chemo-)radiotherapy and concomitant nimorazole for HNSCC did not increase loco-regional control nor disease-specific or overall survival.</p

A systematic review and proportional meta-analysis of image-based pattern of loco-regional failure analyses outcomes in head and neck squamous cell carcinoma

Background and purpose: The prognosis following loco-regional failure after primary radiotherapy (RT) for head and neck squamous cell carcinoma (HNSCC) is poor. The hypothesis that most failures occur as a consequence of tumor radioresistance, can be evaluated by proxy as the proportion of failures that occur in the high-dose region. Several studies have investigated possible reasons for treatment failure by an image-based pattern of failure analyses (POF), comparing the initial planning CT scan with a scan conducted upon failure. The aim of the present systematic review and meta-analysis was to evaluate the proportion of failures that occurred in the high-dose region of all analyzed failures. Materials and Methods: A systematic database search from 2000 to 2023, was performed for studies including results from image-based loco-regional POF, regardless of the method, after primary RT for HNSCC. Proportions of volumetrically in-field (opposed to marginal or outfield) failures, point of origin-based inside high-dose targets, or covered by curative doses for both the number of patients and the number of failure sites were analyzed in proportional meta-analyses. The review was registered at Prospero (CRD42023412545). Results: Out of 56 included studies, accumulated image-based POF results were available from 1,161 patients and 658 individual failure sites. The majority of patients had in-field failures in volumetric-based studies (84 % (95 % CI: 77;90)), inside failures in point of origin-based studies (82 % (95% CI:61;85)) or failures covered by 95 % of dose prescribed to CTV1 (84 % (95% CI:69;95)). A trend toward increasing proportions of non-high-dose failures in more recently treated patients was observed. Conclusion: Most loco-regional failures for patients treated with primary RT for HNSCC are related to the high-dose volume. Therefore, a focus on biomarkers predicting individual tumor radiosensitivity is warranted to enable individualized treatment intensification to increase loco-regional control.</p

A gene signature for selecting benefit from hypoxia modification of radiotherapy for high risk bladder cancer patients

PURPOSE: Hypoxia modification improves overall survival in muscle invasive bladder cancer patients who undergo radiotherapy. There is evidence that hypoxic tumors benefit most from hypoxia modification. The study aimed to identify or derive a hypoxia gene signature that predicts benefit from hypoxia-modifying treatment in bladder cancer.Experimental Design: Published hypoxia signatures were tested and a new one derived by analyzing bladder cancer transcriptomic data from public databases. Tumor samples were available from the BCON phase III randomized trial of radiotherapy alone or with carbogen and nicotinamide (CON). Gene expression data were generated for 151 tumors using Affymetrix Human 1.0 Exon ST arrays and used for independent validation. Results: A 24-gene signature was derived, which was prognostic in four out of six independent surgical cohorts (n=679, meta HR 2·32, 95% CI 1·73-3·12, P<0·0001). The signature was also prognostic in BCON patients receiving radiotherapy alone (n=75, HR for local relapse free survival 2·37, 95% CI 1·26-4·47, P=0·0076). The signature predicted benefit from CON (n=76, HR 0·47, 95% CI 0·26-0·86, P=0·015). Prognostic (P=0·017) and predictive (P=0·058) significance remained after adjusting for clinicopathological variables. A test for interaction between hypoxia status and treatment arms was significant (P=0·0094).Conclusions: A 24-gene hypoxia signature has strong and independent prognostic and predictive value for muscle invasive bladder cancer patients.  The signature can aid identification of patients likely to benefit from the addition of carbogen and nicotinamide to radiotherapy