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

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

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

2009 Status Report of AD-4 Experimen

Breast induration and irradiated volume in the DBCG HYPO trial: The impact of age, smoking, and boost

PurposeTo investigate the association between irradiated breast volume and grade 2–3 breast induration three years after radiotherapy in the phase III Danish Breast Cancer Group HYPO trial randomizing patients ≥ 41 years to whole breast irradiation (WBI) with 40 Gy/15fr versus 50 Gy/25fr.MethodsTreatment plans were available for all Danish patients. Associations between frequency of induration and irradiated volume, age, smoking status, and boost were assessed by logistic regression. A sequential boost was given to patients &lt; 50 years or in case of a narrow (&lt;2 mm) resection margin.ResultsRT plans from 1,333 patients were analyzed with 178 (13 %) having grade 2–3 induration. 1135 patients had only WBI. For this group, induration was correlated with irradiated breast volume for patients ≥ 65 years (n = 343, 10 %/22 % for small/large irradiated volumes, p = 0.005) but not for patients aged 50–64 years (n = 792, 11 % for both small and large volumes, p = 0.82). Smoking doubled the frequency irrespective of irradiated volume and age. All patients &lt; 50 years (n = 156) had a boost. A volume effect was found for this group (5 %/21 % induration for small/large volume, p = 0.002). 42 patients ≥ 50 years had a boost and 14 (33 %) had grade 2–3 induration, however, with a p-value &gt; 0.05 due to the few numbers of patients.ConclusionA relationship between irradiated breast volume and 3-year frequency of breast induration was found for patients ≥ 65 years, whilst not for patients aged 50–64 years. Smoking doubled the risk of induration irrespective of volume and age. A dose-induration relationship was seen for boost patients &lt; 50 years

Management of head and neck cancer of unknown primary: A phase IV study by DAHANCA

BACKGROUND: Diagnostic and therapeutic management of patients with head and neck squamous cell carcinoma of unknown primary (HNSCCUP) remains a challenge. The aim of the present phase IV study was to assess adherence to the current Danish guidelines and evaluate the treatment outcome in HNSCCUP patients.MATERIALS AND METHODS: Prospectively collected data in the DAHANCA database from patients treated between 2014 and 2020 was evaluated. The median follow-up was 6.7 years. Treatment included definitive neck dissection (dND), primary (chemo-)radiotherapy ((C-)RT), neck dissection (ND) followed by postoperative (C-)RT (ND + (C-)PORT). Outcome were reported as five-year estimates of loco-regional failure (LRF), ultimate LRF (ULRF), disease specific mortality (DSM), overall survival (OS), and toxicity scores ≥ 3.RESULTS: A total of 288 patients were treated, of which 254 (88 %) received treatment with curative intent and were eligible for adherence assessment. These were allocated to dND (n = 60), (C-)RT (n = 81) and ND + (C-)PORT (n = 113). The HPV/p16 status was known in 95 % of patients with 109 (43 %) positive cases. The 5-year LRF, DSM, and OS for patients treated with curative intent was 22 %, 15 % and 73 %, and in patients with p16 positive disease 16 %, 5 %, and 85 %. The overall guideline adherence was 76 % (192/254). In the adherent group the LRF, ULRF, DSM, and OS were 22 %, 11 %, 16 %, and 73 %, respectively.CONCLUSION: The study revealed good treatment outcome measures in HNSCCUP patients subject to the Danish guidelines, comparable to other head and neck cancer patients. The observed guideline-deviations did not affect outcome.</p

A genome-wide association study of radiotherapy induced toxicity in head and neck cancer patients identifies a susceptibility locus associated with mucositis

PURPOSE: A two-stage genome-wide association study was carried out in head and neck cancer (HNC) patients aiming to identify genetic variants associated with either specific radiotherapy-induced (RT) toxicity endpoints or a general proneness to develop toxicity after RT.MATERIALS AND METHODS: The analysis included 1780 HNC patients treated with primary RT for laryngeal or oro/hypopharyngeal cancers. In a non-hypothesis-driven explorative discovery study, associations were tested in 1183 patients treated within The Danish Head and Neck Cancer Group. Significant associations were later tested in an independent Dutch cohort of 597 HNC patients and if replicated, summary data obtained from discovery and replication studies were meta-analysed. Further validation of significantly replicated findings was pursued in an Asian cohort of 235 HNC patients with nasopharynx as the primary tumour site.RESULTS: We found and replicated a significant association between a locus on chromosome 5 and mucositis with a pooled OR for rs1131769*C in meta-analysis = 1.95 (95% CI 1.48-2.41; ppooled = 4.34 × 10-16).CONCLUSION: This first exploratory GWAS in European cohorts of HNC patients identified and replicated a risk locus for mucositis. A larger Meta-GWAS to identify further risk variants for RT-induced toxicity in HNC patients is warranted.</p