Luc Bron

Pattern and clinical significance of cancer-testis gene expression in head and neck squamous cell carcinoma

Cancer‐testis (CT) antigens comprise families of tumor‐associated antigens that are immunogenic in patients with various cancers. Their restricted expression makes them attractive targets for immunotherapy. The aim of this study was to determine the expression of several CT genes and evaluate their prognostic value in head and neck squamous cell carcinoma (HNSCC). The pattern and level of expression of 12 CT genes (MAGE‐A1, MAGE‐A3, MAGE‐A4, MAGE‐A10, MAGE‐C2, NY‐ESO‐1, LAGE‐1, SSX‐2, SSX‐4, BAGE, GAGE‐1/2, GAGE‐3/4) and the tumor‐associated antigen encoding genes PRAME, HERV‐K‐MEL, and NA‐17A were evaluated by RT‐PCR in a panel of 57 primary HNSCC. Over 80% of the tumors expressed at least 1 CT gene. Coexpression of three or more genes was detected in 59% of the patients. MAGE‐A4 (60%), MAGE‐A3 (51%), PRAME (49%) and HERV‐K‐MEL (42%) were the most frequently expressed genes. Overall, the pattern of expression of CT genes indicated a coordinate regulation; however there was no correlation between expression of MAGE‐A3/A4 and BORIS, a gene whose product has been implicated in CT gene activation. The presence of MAGE‐A and NY‐ESO‐1 proteins was verified by immunohistochemistry. Analysis of the correlation between mRNA expression of CT genes with clinico‐pathological characteristics and clinical outcome revealed that patients with tumors positive for MAGE‐A4 or multiple CT gene expression had a poorer overall survival. Furthermore, MAGE‐A4 mRNA positivity was prognostic of poor outcome independent of clinical parameters. These findings indicate that expression of CT genes is associated with a more malignant phenotype and suggest their usefulness as prognostic markers in HNSCC.

Prognostic value of arginase‐II expression and regulatory T‐cell infiltration in head and neck squamous cell carcinoma

Tumor‐infiltrating lymphocytes are present in a variety of tumors and play a central role in antitumor immune responses. Nevertheless, most cancers progress probably because tumors are only weakly immunogenic and develop multiple immunosuppressive mechanisms. In the present study, on head and neck squamous cell carcinoma, we found high intraepithelial infiltration of regulatory FOXP3+ T cells, and relatively high levels of BDCA2+ and FOXP3+ cells in stromal (peripheral) regions of the tumors. Tumor‐infiltrating (intraepithelial) FOXP3+ T cells were significantly more frequent in patients with oropharynx and oral cavity squamous cell carcinoma and in patients without lymph node metastasis. Furthermore, arginase‐II (ARG2) was expressed by 60%, inducible nitric oxide synthetase by 9%, cyclooxygenase‐2 by 43%, and B‐cell lymphoma 2 (BCL2) by 26% of tumors. Interestingly, the absence of ARG2 expression, enhanced stromal infiltration of CD11c+ myeloid dendritic cells, and high numbers of FOXP3+ T cells were each significantly associated with prolonged overall survival, and the latter two parameters were also confirmed by multivariate analysis. For disease‐free survival, multivariate analysis revealed significant negative correlations with BCL2 and ARG2 expression by tumor cells. These findings shed new light on mechanisms of cancer progression, and provide rationales for therapeutic inhibition of immunosuppressive mechanisms in head and neck squamous cell carcinoma.

Decreased Local Control Following Radiation Therapy Alone in Early-Stage Glottic Carcinoma with Anterior Commissure Extension*

Purpose:To assess the patterns of failure in the treatment ofnearly-stage squamous cell carcinoma of the glotticnlarynx.Patients and Methods:Between 1983–2000, 122 consecutive patients treated fornearly laryngeal cancer (UICC T1N0 and T2N0) by radical radiationntherapy (RT) were retrospectively studied. Male-to-female rationwas 106 : 16, and median age 62 years (35–92 years). There weren68 patients with T1a, 18 with T1b, and 36 with T2 tumors.nDiagnosis was made by biopsy in 104 patients, and by lasernvaporization or stripping in 18. Treatment planning consisted ofnthree-dimensional (3-D) conformal RT in 49 (40%) patientsnincluding nine patients irradiated using arytenoid protection. Anmedian dose of 70 Gy (60–74 Gy) was given (2 Gy/fraction) over anmedian period of 46 days (21–79 days). Median follow-up periodnwas 85 months.Results:The 5-year overall, cancer-specific, and disease-freensurvival amounted to 80%, 94%, and 70%, respectively. 5-yearnlocal control was 83%. Median time to local recurrence in 19npatients was 13 months (5–58 months). Salvage treatmentnconsisted of surgery in 17 patients (one patient refused salvagenand one was inoperable; total laryngectomy in eleven, andnpartial laryngectomy or cordectomy in six patients). Sixnpatients died because of laryngeal cancer. Univariate analysesnrevealed that prognostic factors negatively influencing localncontrol were anterior commissure extension, arytenoidnprotection, and total RT dose < 66 Gy. Among the factorsnanalyzed, multivariate analysis (Cox model) demonstrated thatnanterior commissure extension, arytenoid protection, and malengender were the worst independent prognostic factors in terms ofnlocal control.Conclusion:For early-stage laryngeal cancer, outcome after RT isnexcellent. In case of anterior commissure extension, surgery ornhigher RT doses are warranted. Because of a high relapse risk,narytenoid protection should not be attempted.Ziel:Ergründung der Versagensmechanismen bei der Therapie desnLarynxkarzinoms im Frühstadium.Patienten und Methodik:Zwischen 1983 und 2000 wurden 122 konsekutive Patienten,ndie wegen eines Larynxkarzinoms (UICC T1N0 und T2N0) einenStrahlentherapie erhielten, retrospektiv untersucht. DasnVerhältnis von Frauen zu Männern betrug 106 : 16, das mittlerenAlter lag bei 62 Jahren. Es handelte sich um 68 Patienten mitnT1a-, 18 mit T1b- und 36 mit T2-Tumoren. Die Diagnose wurde bein104 Patienten mit Hilfe einer Biopsie und bei 18 Patienten mitnLaservaporisation oder Stripping gestellt. Bei 49 Patientenn(40%) bestand die Behandlungsplanung aus einer dreidimensionalennkonformalen Strahlentherapie, einschließlich neun Patienten, dienunter Arytänoidprotektion bestrahlt wurden. Die mittlere Dosisnvon 70 Gy (60–74 GT) wurde über einen mittleren Zeitraum von 46nTagen verabreicht. Die mittlere Nachbehandlungszeit erstrecktensich über 85 Monate.Ergebnisse:Das 5-Jahres-Überleben betrug 80%. Das tumorspezifischen5-Jahres-Überleben lag bei 94%, und 70% der Patienten bliebennwährend dieses Zeitraums erkrankungsfrei. 83% wiesen nach 5nJahren kein Lokalrezidiv auf. Der mittlere Zeitraum bis zumnAuftreten eines lokalen Rückfalls belief sich bei 19 Patientennauf 13 Monate (5–58 Monate). Die Rezidivbehandlung bestand bein17 Patienten aus einem chirurgischen Eingriff (ein Patientnlehnte die Rezidivbehandlung ab, ein anderer war inoperabel;ntotale Laryngektomie bei elf und partielle Laryngektomie bzw.nKordektomie bei sechs Patienten). Sechs Patienten starben annihrem Larynxkarzinom. Eine einseitige Varianzanalyse zeigte,ndass die Ausbreitung auf die vordere Kommissur, dienArytänoidprotektion oder eine Strahlendosis < 66 Gy dienPrognose der Lokalrezidive verschlechterte. EinenMultivarianzanalyse (Cox-Modell) belegte, dass unter dennberücksichtigten Faktoren die Ausbreitung auf die vorderenKommissur, die Protektion des Aryknorpels und männlichesnGeschlecht die schlechtesten unabhängigen Prognosefaktoren imnHinblick auf Lokalrezidive sind.Schlussfolgerung:Beim Larynxkarzinom im Frühstadium erbringt dienStrahlentherapie hervorragende Ergebnisse. Im Fall einernAusbreitung auf die vordere Kommissur ist ein chirurgischernEingriff oder eine höhere Strahlendosis erforderlich. Wegen desnhohen Rezidivrisikos sollte keine Protektion des Aryknorpelsnvorgenommen werden.

Decreased local control following radiation therapy alone in early-stage glottic carcinoma with anterior commissure extension.

Purpose:To assess the patterns of failure in the treatment ofnearly-stage squamous cell carcinoma of the glotticnlarynx.Patients and Methods:Between 1983–2000, 122 consecutive patients treated fornearly laryngeal cancer (UICC T1N0 and T2N0) by radical radiationntherapy (RT) were retrospectively studied. Male-to-female rationwas 106 : 16, and median age 62 years (35–92 years). There weren68 patients with T1a, 18 with T1b, and 36 with T2 tumors.nDiagnosis was made by biopsy in 104 patients, and by lasernvaporization or stripping in 18. Treatment planning consisted ofnthree-dimensional (3-D) conformal RT in 49 (40%) patientsnincluding nine patients irradiated using arytenoid protection. Anmedian dose of 70 Gy (60–74 Gy) was given (2 Gy/fraction) over anmedian period of 46 days (21–79 days). Median follow-up periodnwas 85 months.Results:The 5-year overall, cancer-specific, and disease-freensurvival amounted to 80%, 94%, and 70%, respectively. 5-yearnlocal control was 83%. Median time to local recurrence in 19npatients was 13 months (5–58 months). Salvage treatmentnconsisted of surgery in 17 patients (one patient refused salvagenand one was inoperable; total laryngectomy in eleven, andnpartial laryngectomy or cordectomy in six patients). Sixnpatients died because of laryngeal cancer. Univariate analysesnrevealed that prognostic factors negatively influencing localncontrol were anterior commissure extension, arytenoidnprotection, and total RT dose < 66 Gy. Among the factorsnanalyzed, multivariate analysis (Cox model) demonstrated thatnanterior commissure extension, arytenoid protection, and malengender were the worst independent prognostic factors in terms ofnlocal control.Conclusion:For early-stage laryngeal cancer, outcome after RT isnexcellent. In case of anterior commissure extension, surgery ornhigher RT doses are warranted. Because of a high relapse risk,narytenoid protection should not be attempted.Ziel:Ergründung der Versagensmechanismen bei der Therapie desnLarynxkarzinoms im Frühstadium.Patienten und Methodik:Zwischen 1983 und 2000 wurden 122 konsekutive Patienten,ndie wegen eines Larynxkarzinoms (UICC T1N0 und T2N0) einenStrahlentherapie erhielten, retrospektiv untersucht. DasnVerhältnis von Frauen zu Männern betrug 106 : 16, das mittlerenAlter lag bei 62 Jahren. Es handelte sich um 68 Patienten mitnT1a-, 18 mit T1b- und 36 mit T2-Tumoren. Die Diagnose wurde bein104 Patienten mit Hilfe einer Biopsie und bei 18 Patienten mitnLaservaporisation oder Stripping gestellt. Bei 49 Patientenn(40%) bestand die Behandlungsplanung aus einer dreidimensionalennkonformalen Strahlentherapie, einschließlich neun Patienten, dienunter Arytänoidprotektion bestrahlt wurden. Die mittlere Dosisnvon 70 Gy (60–74 GT) wurde über einen mittleren Zeitraum von 46nTagen verabreicht. Die mittlere Nachbehandlungszeit erstrecktensich über 85 Monate.Ergebnisse:Das 5-Jahres-Überleben betrug 80%. Das tumorspezifischen5-Jahres-Überleben lag bei 94%, und 70% der Patienten bliebennwährend dieses Zeitraums erkrankungsfrei. 83% wiesen nach 5nJahren kein Lokalrezidiv auf. Der mittlere Zeitraum bis zumnAuftreten eines lokalen Rückfalls belief sich bei 19 Patientennauf 13 Monate (5–58 Monate). Die Rezidivbehandlung bestand bein17 Patienten aus einem chirurgischen Eingriff (ein Patientnlehnte die Rezidivbehandlung ab, ein anderer war inoperabel;ntotale Laryngektomie bei elf und partielle Laryngektomie bzw.nKordektomie bei sechs Patienten). Sechs Patienten starben annihrem Larynxkarzinom. Eine einseitige Varianzanalyse zeigte,ndass die Ausbreitung auf die vordere Kommissur, dienArytänoidprotektion oder eine Strahlendosis < 66 Gy dienPrognose der Lokalrezidive verschlechterte. EinenMultivarianzanalyse (Cox-Modell) belegte, dass unter dennberücksichtigten Faktoren die Ausbreitung auf die vorderenKommissur, die Protektion des Aryknorpels und männlichesnGeschlecht die schlechtesten unabhängigen Prognosefaktoren imnHinblick auf Lokalrezidive sind.Schlussfolgerung:Beim Larynxkarzinom im Frühstadium erbringt dienStrahlentherapie hervorragende Ergebnisse. Im Fall einernAusbreitung auf die vordere Kommissur ist ein chirurgischernEingriff oder eine höhere Strahlendosis erforderlich. Wegen desnhohen Rezidivrisikos sollte keine Protektion des Aryknorpelsnvorgenommen werden.

Mixed medullary-papillary carcinoma of the thyroid: Report of a case and review of the literature

Mixed medullary‐follicular thyroid carcinoma denotes a rare and heterogeneous group of tumors displaying morphological and immunophenotypical features of both origins within the same lesion.

MAGE-A3 and MAGE-A4 specific CD4+ T cells in head and neck cancer patients: detection of naturally acquired responses and identification of new epitopes

Frequent expression of cancer testis antigens (CTA) has been consistently observed in head and neck squamous cell carcinomas (HNSCC). For instance, in 52 HNSCC patients, MAGE-A3 and -A4 CTA were expressed in over 75% of tumors, regardless of the sites of primary tumors such as oral cavity or hypopharynx. Yet, T-cell responses against these CTA in tumor-bearing patients have not been investigated in detail. In this study, we assessed the naturally acquired T-cell response against MAGE-A3 and -A4 in nonvaccinated HNSCC patients. Autologous antigen-presenting cells pulsed with overlapping peptide pools were used to detect and isolate MAGE-A3 and MAGE-A4 specific CD4+ T cells from healthy donors and seven head and neck cancer patients. CD4+ T-cell clones were characterized by cytokine secretion. We could detect and isolate MAGE-A3 and MAGE-A4 specific CD4+ T cells from 7/7 cancer patients analyzed. Moreover, we identified six previously described and three new epitopes for MAGE-A3. Among them, the MAGE-A3111–125 and MAGE-A3161–175 epitopes were shown to be naturally processed and presented by DC in association with HLA-DP and DR, respectively. All of the detected MAGE-A4 responses were specific for new helper epitopes. These data suggest that naturally acquired CD4+ T-cell responses against CT antigens often occur in vivo in HNSCC cancer patients and provide a rationale for the development of active immunotherapeutic approaches in this type of tumor.

Feasibility and Efficacy of Accelerated Weekly Concomitant Boost Postoperative Radiation Therapy Combined with Concomitant Chemotherapy in Patients with Locally Advanced Head and Neck Cancer

BackgroundThe aim of this study was to assess feasibility and efficacy of weekly concomitant boost accelerated postoperative radiation therapy (PORT) with concomitant chemotherapy (CT) in patients with locally advanced head and neck cancer (LAHNC).Methods and MaterialsConformal or intensity-modulated 66-Gy RT was performed in 5.5xa0weeks in 40 patients. Cisplatin was given at days 1, 22, and 43. Median follow-up was 36xa0months.Results and DiscussionGrade 3 mucositis, dysphagia, and erythema was observed in ten (25%), nine (23%), and six (13%) patients, respectively. Grade 3 or more anemia was observed in two (6%) patients, and leukopenia in five (13%) patients. No grade 3 or 4 thrombocytopenia was observed. Grade 3 nephrotoxicity was observed in one patient (3%). No treatment-related mortality was observed. Grade 2 or more xerostomia and edema were observed in ten (25%) and one (3%) patient, respectively. Locoregional relapse occurred in eight patients, and seven patients developed distant metastases. Median time to locoregional relapse was 6xa0months. Three-year overall, disease-free survival, and locoregional control rates were 63%, 62%, and 81%, respectively. Multivariate analysis revealed that the only prognostic factor was nodal status.ConclusionReducing overall treatment time using accelerated PORT/CT by weekly concomitant boost (six fractions per week) combined with concomitant cisplatin CT is easily feasible with acceptable morbidity.

Decrease in Hemoglobin Levels Following Surgery Influences the Outcome in Head and Neck Cancer Patients Treated with Accelerated Postoperative Radiotherapy

AimTo assess the influence of hemoglobin (Hb) levels in locally advanced head and neck cancer (LAHNC) patients treated with surgery and postoperative radiotherapy (PORT).Material and MethodsPre- and postoperative Hb levels were collected in 79 patients treated with surgery followed by accelerated PORT for LAHNC. Median follow-up was 52xa0months (range 12–95xa0months).Results and DiscussionFour-year overall survival (OS) rate was 51%. Neither pre- nor postoperative Hb level (<120 or 130xa0g/l in women or men, respectively) influenced the outcome. However, when Hb decrease between pre- and postoperative Hb values was taken into account, 4-year OS was significantly higher in patients with Hb difference less than 38xa0g/l (quartile value) compared with those with Hb decrease 38xa0g/l or more (61% versus 16%, Pxa0=xa00.008).ConclusionDecrease in Hb level by more than 38xa0g/l after surgery secondary to blood loss influences the outcome when postoperative RT is indicated.