Summary Introduction : Head and neck squamous cell carcinomas (HNSSCs) are one of the leading causes of cancer-associated death worldwide. Although certain behavioral risk factors are well recognized ...as tumor promoting, there is very little known about the presence of predisposing germline mutations in HNSCC patients. Methods : In this study, we analyzed 121 individuals with HNSCCs collected at our institution for germline alterations in the newly identified cancer susceptibility gene RAD51C. Results : Sequencing of all exons and the adjacent introns revealed five distinct heterozygous sequence deviations in RAD51C in seven patients (5.8%). A female patient without any other risk factors carried a germline mutation that disrupted the canonical splice acceptor site of exon 5 (c.706-2A>G). Conclusions : As there are only a few publications in the literature identifying germline mutations in head and neck cancer patients, our results provide the first indication that paralogs of RAD51, recently described as mutated in breast and ovarian cancer patients, might also be candidates for genetic risk factors in sporadic squamous cell carcinomas of the head and neck.
(1) Background: Evaluation of impact of adjuvant radiation therapy (RT) in patients with oral squamous cell carcinoma of the oral cavity/oropharynx (OSCC) of up to 4 cm (pT1/pT2) and solitary ...ipsilateral lymph node metastasis (pN1). A non-irradiated group with clinical follow-up was chosen for control, and survival and quality of life (QL) were compared; (2) Methods: This prospective multicentric comprehensive cohort study included patients with resected OSCC (pT1/pT2, pN1, and cM0) who were allocated into adjuvant radiation therapy (RT) or observation. The primary endpoint was overall survival. Secondary endpoints were progression-free survival and QL after surgery; (3) Results: Out of 27 centers, 209 patients were enrolled with a median follow-up of 3.4 years. An amount of 137 patients were in the observation arm, and 72 received adjuvant irradiation. Overall survival did not differ between groups (hazard ratio (HR) 0.98 0.55-1.73,
= 0.94). There were fewer neck metastases (HR 0.34 0.15-0.77;
= 0.01), as well as fewer local recurrences (HR 0.41 0.19-0.89;
= 0.02) under adjuvant RT. For QL, irradiated patients showed higher values for the symptom scale pain after 0.5, two, and three years (all
< 0.05). After six months and three years, irradiated patients reported higher symptom burdens (impaired swallowing, speech, as well as teeth-related problems (all
< 0.05)). Patients in the RT group had significantly more problems with mouth opening after six months, one, and two years (
< 0.05); (4) Conclusions: Adjuvant RT in patients with early SCC of the oral cavity and oropharynx does not seem to influence overall survival, but it positively affects progression-free survival. However, irradiated patients report a significantly decreased QL up to three years after therapy compared to the observation group.
Therapeutic strategies attacking oral squamous cell carcinoma have not essentially succeeded to improve long-term prognosis and overall survival over the last decades. Therefore, in this study, we ...aimed to illuminate the molecular regulation of angiogenesis in this tumour entity in order to demask novel markers of prognosis or therapeutic approach.
A panel of significant transcriptional alterations in angiogenic genes of 83 cancer samples was established by comparison to 30 samples of healthy oral mucosa with microarray technique. Immunohistochemistry (IHC) was performed to trace the signalling cascade from gene to protein level.
A distinctive expression profile of VEGFA, EFNB2, PECAM1/CD31, ANGPT1 and ANGPT2 was revealed: VEGFA, EFNB2, and ANGPT2 were found overexpressed in 84 % to 95 % of tumour samples. In contrast, the expression of CD31 and ANGPT1 was downregulated in 80 % to 95 % of tumour samples. IHC confirmed results of the microarray analysis. Tumours with lymphatic spread showed higher gene expression rates of VEGFA, EFNB2 and ANGPT2 in moderately differentiated tumours and of VEGFA and EFNB2 in small tumours, respectively. The ANGPT1/ ANGPT2 transcription ratio was found decreased in larger tumours and especially in tumours without lymphatic spread.
A characteristic expression profile of angiogenic markers was established. The specific overexpression of EFNB2 in small tumours with lymphatic spread and the typical decrease of the ANGPT1/ ANGPT2 ratio in larger tumours give weight to EFNB2 and angiopoietins as prognostic factors and potential therapeutic targets.
BACKGROUND: Circulating tumor cells (CTCs) are promising surrogate markers for systemic disease, and their molecular characterization might be relevant to guide more individualized cancer therapies. ...To enable fast and efficient purification of individual CTCs, we developed a work flow from CellSearchTM cartridges enabling high-resolution genomic profiling on the singlecell level. METHODS: Single CTCs were sorted from 40 CellSearch samples from patients with metastatic breast cancer using a MoFlo XDP cell sorter. Genomes of sorted single cells were amplified using an adapter-linker PCR. Amplification products were analyzed by array-based comparative genomic hybridization, a gene-specific quantitative PCR (qPCR) assay for cyclin D1 (CCND1) locus amplification, and genomic sequencing to screen for mutations in exons 1, 9, and 20 of the phosphatidylinositol-4,5bisphosphate 3-kinase, catalytic subunit alpha (PIK3CA) gene and exons 5, 7, and 8 of the tumor protein p53 (TP53) gene. RESULTS: One common flow-sorting protocol was appropriate for 90% of the analyzed CellSearch cartridges, and the detected CTC numbers correlated positively with those originally detected with the CellSearch system (R.sup.2 = 0.9257). Whole genome amplification was successful in 72.9% of the sorted single CTCs. Over 95% of the cells displayed chromosomal aberrations typical for metastatic breast cancers, and amplifications at the CCND1 locus were validated by qPCR. Aberrant CTCs from 2 patients harbored mutations in exon 20 of the PIK3CA gene. Conclusions: This work flow enabled effective CTC isolation and provided insights into genomic alterations of CTCs in metastatic breast cancer. This approach might facilitate further molecular characterization of rare CTCs to increase understanding of their biology and as a basis for their molecular screening in the clinical setting.
Circulating tumor cells (CTCs) are promising surrogate markers for systemic disease, and their molecular characterization might be relevant to guide more individualized cancer therapies. To enable ...fast and efficient purification of individual CTCs, we developed a work flow from CellSearch(TM) cartridges enabling high-resolution genomic profiling on the single-cell level. Single CTCs were sorted from 40 CellSearch samples from patients with metastatic breast cancer using a MoFlo XDP cell sorter. Genomes of sorted single cells were amplified using an adapter-linker PCR. Amplification products were analyzed by array-based comparative genomic hybridization, a gene-specific quantitative PCR (qPCR) assay for cyclin D1 (CCND1) locus amplification, and genomic sequencing to screen for mutations in exons 1, 9, and 20 of the phosphatidylinositol-4,5-bisphosphate 3-kinase, catalytic subunit alpha (PIK3CA) gene and exons 5, 7, and 8 of the tumor protein p53 (TP53) gene. One common flow-sorting protocol was appropriate for 90% of the analyzed CellSearch cartridges, and the detected CTC numbers correlated positively with those originally detected with the CellSearch system (R^sup 2^ = 0.9257). Whole genome amplification was successful in 72.9% of the sorted single CTCs. Over 95% of the cells displayed chromosomal aberrations typical for metastatic breast cancers, and amplifications at the CCND1 locus were validated by qPCR. Aberrant CTCs from 2 patients harbored mutations in exon 20 of the PIK3CA gene. This work flow enabled effective CTC isolation and provided insights into genomic alterations of CTCs in metastatic breast cancer. This approach might facilitate further molecular characterization of rare CTCs to increase understanding of their biology and as a basis for their molecular screening in the clinical setting.
Objectives
The repair of skin defects in the head and neck region still poses a significant problem for many clinicians. Tissue expansion is described as a treatment option providing good color, ...texture, and thickness match of the expanded skin. Unfortunately, the complication rates for tissue expansion range from 0 to 48 %. Therefore, the purpose of this study was to investigate risk factors for the use of tissue expanders in head and neck reconstructions.
Materials and methods
Forty-nine patients with skin deficits in the head and neck area underwent tissue expansion. Sixty-two implanted expanders were analyzed regarding the various complications and the success rate.
Results
The success rate of treated patients was 37 (75.5 %) of all 49 included patients. The most frequent cause for the skin deficit was a tumor resection near the tip of the nose followed by skin deficits resulting after craniectomy. Interestingly, a higher number of expanders and a larger volume were significantly associated with a worse outcome. There was a trend of association between larger defect size and failure, too.
Conclusions
The internal tissue expansion is a suitable technique for skin reconstruction in the head and face area. Compared to distant or free flaps, it often offers a better cosmetic outcome. In very large defects (>100 cm
2
) or when more than two expanders are needed, the failure rate increases. In these cases, other treatment options are recommended.
Clinical relevance
The internal tissue expansion is a suitable technique for skin reconstruction in the head and face area.
Circulating tumor cells (CTCs) are promising surrogate markers for systemic disease, and their molecular characterization might be relevant to guide more individualized cancer therapies. To enable ...fast and efficient purification of individual CTCs, we developed a work flow from CellSearch(TM) cartridges enabling high-resolution genomic profiling on the single-cell level.
Single CTCs were sorted from 40 CellSearch samples from patients with metastatic breast cancer using a MoFlo XDP cell sorter. Genomes of sorted single cells were amplified using an adapter-linker PCR. Amplification products were analyzed by array-based comparative genomic hybridization, a gene-specific quantitative PCR (qPCR) assay for cyclin D1 (CCND1) locus amplification, and genomic sequencing to screen for mutations in exons 1, 9, and 20 of the phosphatidylinositol-4,5-bisphosphate 3-kinase, catalytic subunit alpha (PIK3CA) gene and exons 5, 7, and 8 of the tumor protein p53 (TP53) gene.
One common flow-sorting protocol was appropriate for 90% of the analyzed CellSearch cartridges, and the detected CTC numbers correlated positively with those originally detected with the CellSearch system (R(2) = 0.9257). Whole genome amplification was successful in 72.9% of the sorted single CTCs. Over 95% of the cells displayed chromosomal aberrations typical for metastatic breast cancers, and amplifications at the CCND1 locus were validated by qPCR. Aberrant CTCs from 2 patients harbored mutations in exon 20 of the PIK3CA gene.
This work flow enabled effective CTC isolation and provided insights into genomic alterations of CTCs in metastatic breast cancer. This approach might facilitate further molecular characterization of rare CTCs to increase understanding of their biology and as a basis for their molecular screening in the clinical setting.
Purpose Integrins are adhesion molecules and play a role in many cellular processes, e. g. angiogenesis, tumor development and metastasis. Monoclonal antibodies directed to α v β 3-integrins are ...tested in clinical studies for the treatment of advances malignant melanoma and could be an option for the treatment of oral cancer. Material and methods Biopsies were taken from 83 histological proven oral squamous cell cancers, the mRNA was isolated and the gene expression was measured with “whole genome microarrays“. Expression patterns were compared to biopsies from the oral mucosa of healthy volunteers. The expression of the α v- and β 3-subunit was analyzed. In an orthotopic xenograft mouse model of oral cancer tumors were induced by sonographic controlled injection of 1 × 106 SCC-9 cells, a human squamous cell cancer cell line, in the floor of the mouth in Fox-n1 nude mice. Tumors or recurrent tumors ( n = 10) were visualized in a fluorescence-chamber (Berthold NightOWL) after the application of “Integrisense” (visen medical) via the tail vein. Integrisense binds to α v β 3-integrins and is labeled with a fluorochrome. Results The microarray-analysis showed an upregulation of the α v- and β 3-subunit in 40% of the tumor samples. Even though the cell line SCC-9 showed in vitro no upregulation of α v β 3-integrins, it could be shown that Integrisense binds to the tumors that were induced in vivo in the mouse model. The best signal-to-noise-ratio was measured one day after the application. The intensity of the fluorescence-signal was between 245 nm and 510 nm counts per second. Conclusion α v β 3-Integrins are a possible target for the visualisation of the tumor with fluorochrome-labeled probes and are a possible therapeutic target for selected oral squamous cell carcinomas.
Integrins are adhesion molecules and play a role in many cellular processes, e. g. angiogenesis, tumor development and metastasis. Monoclonal antibodies directed to αvβ3-integrins are tested in ...clinical studies for the treatment of advances malignant melanoma and could be an option for the treatment of oral cancer.
Biopsies were taken from 83 histological proven oral squamous cell cancers, the mRNA was isolated and the gene expression was measured with “whole genome microarrays“. Expression patterns were compared to biopsies from the oral mucosa of healthy volunteers. The expression of the αv- and β3-subunit was analyzed.
In an orthotopic xenograft mouse model of oral cancer tumors were induced by sonographic controlled injection of 1×106 SCC-9 cells, a human squamous cell cancer cell line, in the floor of the mouth in Fox-n1 nude mice. Tumors or recurrent tumors (n=10) were visualized in a fluorescence-chamber (Berthold NightOWL) after the application of “Integrisense” (visen medical) via the tail vein. Integrisense binds to αvβ3-integrins and is labeled with a fluorochrome.
The microarray-analysis showed an upregulation of the αv- and β3-subunit in 40% of the tumor samples. Even though the cell line SCC-9 showed in vitro no upregulation of αvβ3-integrins, it could be shown that Integrisense binds to the tumors that were induced in vivo in the mouse model. The best signal-to-noise-ratio was measured one day after the application. The intensity of the fluorescence-signal was between 245nm and 510nm counts per second.
αvβ3-Integrins are a possible target for the visualisation of the tumor with fluorochrome-labeled probes and are a possible therapeutic target for selected oral squamous cell carcinomas.
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